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NIESA Training Manual
NEMATOLOGY TRAINING
MANUAL
FUNDED BY NIESA and UNIVERSITY OF NAIROBI, CROP PROTECTION

DEPARTMENT
CONTRIBUTORS: J. Kimenju, Z. Sibanda, H. Talwana and W. Wanjohi
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CHAPTER 1
TECHNIQUES FOR NEMATODE DIAGNOSIS AND
HANDLING
Herbert A. L. Talwana
Department of Crop Science, Makerere University
P. O. Box 7062, Kampala Uganda
Section Objectives
Going through this section will enrich you with skill to be able to:
 diagnose nematode problems in the field considering all aspects involved in sampling, extraction and
counting of nematodes from soil and plant parts,
 make permanent mounts,
 set up and maintain nematode cultures,
 design experimental set-ups for tests with nematodes
Section Content
 sampling and quantification of nematodes
 extraction methods for plant-parasitic nematodes, free-living nematodes from soil and plant parts
 mounting of nematodes,
 drawing and measuring of nematodes,
 preparation of nematode inoculum and culturing nematodes,
 set-up of tests for research with plant-parasitic nematodes,
A. Nematode sampling
Unlike some pests and diseases, nematodes cannot be monitored by observation in the field.
Nematodes must be extracted for microscopic examination in the laboratory. Nematodes
can be collected by sampling soil and plant materials. There is no problem in finding
nematodes, but getting the species and numbers you want may be trickier. In general,
natural and undisturbed habitats will yield greater diversity and more slow-growing
nematode species, while temporary and/or disturbed habitats will yield fewer and fast-
multiplying species.
Sampling considerations
Getting nematodes in a sample that truly represent the underlying population at a given
time requires due attention to sample size and depth, time and pattern of sampling, and
handling and storage of samples. Since plant parasitic nematodes feed on plant tissue, their
distribution is influenced by the distribution of their hosts, soil types, nematode species
involved and season. In most situations, nematodes are clustered or aggregated. When the
host crop is present, depth and lateral distribution of nematodes in soil generally mirrors the
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degree of proliferation of the root system (particularly the fine roots on which nematodes
feed). When the field is fallowed, the existing distribution will remain except that the bulk
of the population may be slightly deeper in soil (largely due to desiccation of nematodes
near the surface). Nematodes are rarely distributed evenly across a field. Some species are
favoured by certain soils and their distribution may change with subtle changes in soil
texture.
Besides, nematode populations fluctuate over time; numbers increase in the presence of a
host crop, the rate of increase being greatest when the environmental conditions are
favourable. When the field is under fallow or a non-host crop is present, nematodes die of
starvation and populations decline. The number of nematodes in a sample will therefore be
affected by sampling time.
Additionally, many plant parasitic nematodes have the capacity to survive periods of
dryness through a behavioural adaptation in which their surface area is reduced by a
process known as coiling. Since nematodes are very susceptible to mechanical damage in
this desiccated state, the process of collecting samples from dry soils may damage
nematodes. Nematode population densities are therefore likely to be underestimated in dry
soil and it is preferable to wait until soil moisture is adequate before collecting samples.
This uneven distribution makes collection of truly representative samples difficult as it

introduces sampling errors. Other sources of sampling error are low population densities,
latent infections, sub-sampling and counting of nematodes. Therefore, absence of
nematodes in a sample may indicate their absence in the sampled field but may also
indicate that the populations are too low to be detected by sampling.
In order to increase the probability of detecting nematode infestations, roots of weeds and
volunteer crops should be collected when soil is sampled.
Sampling equipment
For collecting soil samples to a depth of 20 cm or more, a soil sampling auger can be used.
Garden trowels, narrow-bladed shovels or spades are also useful especially when sampling
cropped areas or areas with rocky soil (Figure 1). Remember to clean all equipment and
footwear of adhering soil before leaving a sampling site to reduce the risk of spreading
nematodes to uninfected areas.
Sample size
The goal of a nematode sampling would be to capture information about the presence of
nematode species occurring in one or more landscapes, ecosystems or geographic regions.
The number of samples to be taken will depend on the breadth and depth of the sampling;
where sampling breadth denotes the relative number of sites from which samples are
collected, and sampling depth denotes the extent of taxonomic characterization per
sampling site. Apportioning resources between sampling breadth and depth entails a choice
between extensive or intensive nematode sampling. An extensive sampling maximizes the
number of sampling sites and characterizes a limited number of taxa per site.
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Figure 1. Examples of tools for soil sampling to diagnose nematode infestations
A B
C D
Figure 2. Recommended soil sampling patterns; A. and B: Patterns for perennial plants
(When sampling tree crops, collect sub-samples from around the drip-line, towards and on
alternate sides of the trunk) C: Pattern for annual crop or fallow field; D: Sampling pattern
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for diagnosing nematode problems (Take soil cores from the margin of affected areas and
only from the roots of affected plants that are still alive).
The size of the sampling unit will vary from crop to crop and is largely determined by the
value of the crop. Thus, sampling can be more intensive on high value horticultural and
ornamental crops than on low value field crops. A sample should consist of 10 or more
sub-samples representative of the area being sampled. Any number of sub-samples can be
combined to form a composite sample. If an entire composite sample can not be processed,
sub-sample by mixing the soil or plant tissue sample very gently but thoroughly by hand,
avoiding excessive handling to prevent mechanical damage to the nematodes. The size of
the sub sample depends on the extraction procedure used but should be at least 100ml of
soil and 50g of plant material.
Sampling pattern
The sampling pattern will depend on the purpose of sampling: sampling to predict a
problem or sampling to diagnose a problem. When sampling for predictive nematode
assays (Figure 2), take samples before the desired crop is in the ground. For annual crops,
this is usually soon after (or just before) harvest of the existing crop and several months
before planting the next crop. Sampling close to harvest ensures that nematode populations
are at their peak and that the assay will be a good indicator of any potential problems
An intensive sampling limits the total number of sampled sites but maximizes the number
of characterizations per site. The rationale for sampling will determine whether an intensive
or extensive approach is more appropriate.
For perennial crops, plan to collect samples well before planting so that there will be time
to treat the fields if necessary. It is very difficult to manage nematodes on an established
crop. In high-value established landscapes like golf courses, however, it can be prudent to
sample for nematodes on a regular basis so that management can be scheduled for off-peak
seasons.
Additional information required for predictive sampling includes:
 Current crop and cultivar (or the crop and cultivar to be planted in the case of
pre-plant samples)
 Details of the site (e.g. area, soil type, variability of soil, previous cropping
history).
 Standard of crop management, particularly with regard to irrigation and
nutrition.
 Likely importance of other pests and pathogens of roots.
 Details of previous nematicide use and the responses that were obtained.
When sampling to diagnose a problem, use a combination of agronomic tests when trying
to diagnose whether an existing plant growth problem might be due to nematodes. Collect a
soil sample with roots for nematode problem diagnosis as well as a comparison sample
from a nearby area where growth is more nearly normal. Because most problems have more
than one cause, submit samples for soil nutrient and plant tissue analyses as well. Package
each different kind of sample separately. You can send matching samples from the same
area for soil testing or plant analysis. You can collect samples for nematode problem
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diagnosis any time plants are actively growing and the soil is in good working condition.
Take soil from the root zone of plants that are affected but still alive. Never sample beneath
dead plants. Diagnoses are improved if adequate background information is provided. The
following details are particularly important:
 Crop and cultivar
 Previous crop
 Area involved
 Description of symptoms and their distribution
 Soil texture, soil depth and variability of soil
 Frequency of irrigation and/or rainfall
 Previous nematode control methods used and details of responses obtained
For larger cropped areas, divide the field into 1 hectare units and make a grid pattern that
covers the entire 1 ha unit. The length of intervals between sampling points on the grid will
depend on the sampling precision that you require. Very close intervals, e.g. 2m x 2m will
reflect the nematode distribution more precisely than, for example, 10m x 10m. Collect a
sub-sample from at least 20 points throughout the fields. Collect separate samples for areas
with different soil types, different cropping histories, or different management objectives.
Sampling depth
The depth of sampling has to consider the depth of rooting of the crop being sampled. For
most crops, a sampling depth of 20 cm is adequate; for deep rooted perennial crops,
different depths can be sampled, for example, 15, 30, 60, 100 cm.
Sampling time
Soil and root samples can be taken and reliably processed as needed, whenever the soil is
not dry. For best results, collect samples before planting a field but fairly close to planting
time, as the nematodes present at this time can generally be related to yield in annual crops.
Samples can also be collected between mid season and harvest as nematode numbers
increase towards harvest. For established perennial crops, collect samples during growth
flushes.
Care of samples
Caution! Collect samples in sturdy plastic bags and close the bags firmly. Place above
ground plant materials and soil samples in separate bags but place the roots together with
soil in the same bag; the roots should be covered with soil. Do not moisten soil samples
that were collected from dry soil. Transport samples in a cool-box or other insulated
containers, preferably with ice packs. Put samples in a cool place or refrigerate until
submission. If you do not want to extract straightaway, most nematodes in the samples will
survive storage at 4oC for several weeks. Do not allow samples to dry as the nematodes will
die before the sample arrives in the laboratory. Temperatures above 35oC will also kill
many nematodes. Submit samples to the nematode laboratory promptly and if possible
process within one week from collection!
Direct examination of plant material

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Nematodes can usually be seen by examining small amounts of gently washed plant tissue
such as roots, leaves, stems or seeds with a stereomicroscope. The plant tissue is examined
in water in an open Petri dish or large watch glass and teased apart with strong mounted
needles. Nematodes released from the tissue will float out and can be collected with a
handling needle or fine pipette.
Since nematodes are translucent and difficult to see in plant tissues, staining helps to make
them more visible. The plant tissue may be cleared in diluted sodium hypochlorite bleach
before examination. The strength and time of bleaching will depend on the plant material
and type of bleach. Rinse out the bleach before transferring the plant material to a glass vial
or beaker with acid fuschsin solution (875ml of lactic acid, 63ml of glycerol, 62 ml of water
and 0.1g of acid fuschsin). Boil the plant material in the solution for about 30 seconds in a
microwave or on a hotplate (should be in a ventilated area to avoid lactic acid fumes).
Several small samples can be stained in one operation by wrapping each in a piece of
muslin cloth.
The plant material is allowed to cool before washing off the excess stain in running tap
water. Thereafter, the plant material is transferred to a solution of equal volumes of glycerol
and water acidified with a few drops of lactic acid. After several hours to a few days,
depending upon the type of material, the differentially stained nematodes can be examined
in the largely unstained plant tissue.
B. Extracting live nematodes

Extracting nematodes from Soil
An endless range of extraction tools and techniques have been developed mostly because
no single technique will efficiently extract all sizes and kinds of nematodes. Some are
rarely used, others have been adapted to suit local conditions and requirements and
availability of equipment.
Modified Baermann Technique - Extraction tray method (Whitehead and Hemming,

1965)
This is the simplest method which was adapted from the Baermann Funnel Technique. This
method is not quantitatively reliable as it selects against slow and non-moving nematodes,
but it is extremely cheap, relatively fast, very good at yielding large numbers of live active
worms and it extracts live, active nematodes from soil and plant material in a clear
suspension. The three components required are: (1) a large plastic tray, (2) a wide mesh and
(3) reasonably strong kitchen paper (Figure 3). The larger the tray, the more material can be
extracted in one run. A handy size and type is e.g. a dissection tray or a photographic
developing tray. The mesh can be a single layer of thick plastic or rubber netting, or
alternatively it can consist of a plastic-coated metal grid covered with thin plastic gauze. It
should NOT contain any bare metal, because this may release toxic ions.
Place the mesh in the tray, cover its bottom and sides with a single sheet of kitchen paper
(one or two layers) and spread a thin, uniform layer of soil out over the paper. Gently pour
clean water (tap water may contain nematodes!) into the tray until the soil is wet but not
submerged. Avoid spilling soil particles along the sides or through the filter paper - they
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will cloud up the extract and make it difficult to see the nematodes. Leave the tray until the
next day. Actively moving nematodes will sooner or later crawl down through the kitchen
paper and sink to the bottom of the tray. A large proportion of the sampled nematodes will
thus collect on the bottom of the tray overnight, and the next morning you should have
plenty of nematodes ready for further treatment.
Concentrate the suspended nematodes by pouring the water out of the tray over a fine sieve
(mesh 25 m or less) and washing the nematodes off into a small beaker with about 20-30
ml water. If you do not have a sieve, pour the extract water into a sufficiently large beaker
and allow the nematodes to settle (takes about 30 minutes). Then pour off most of the top
water and transfer the remaining water to a smaller beaker. Repeat until you have 20-30 ml
suspension left. If you have sampled on a good spot, you should now have hundreds or
even thousands of nematodes ready for further processing.
This method is called Active method and it is not effective for inactive nematodes such as
Trichodorids, Longidorids and Criconematids. Therefore, to be consistent, it is advisable to
supplement it with other techniques – the Passive methods. There are two major methods –
Elutriation and Sieving. Elutriation involves separating nematodes from soil by an up-
current of water, the strength of which is such that the nematodes are held in suspension
whilst the heavier soil particles sink. With these techniques, both active and sluggish
nematodes are extracted from soil, results are quickly available but they involve fairly
complicated apparatus, require considerable expertise, and depend upon a plentiful supply
of clean water at high pressure, thus limiting their usefulness.
Sieving Technique
The sieving technique is also known as the bucket-sieving method. Although crude, it is
widely used as it enables the extraction of large numbers of both active and inactive
nematodes in a relatively short time. Equipment required includes two plastic buckets
(about 5L), sieves of 15 – 20 cm diameter made with wire mesh, preferably stainless steel
of aperture size of 2mm, 720, 250, 125, 90, 63, 45,25 m, respectively and tall 100 ml
beakers for the residue from sieves (Figure 4).
Usually only three or four sets of sieves will be used for a particular sample, with the sieves
selected to match the size of nematode it is hoped to extract, and to suit the type of soil
involved. In general, sieve openings should be no greater than 1/10 of the nematode length.
Most adults of large nematodes (e.g. Anguina, Belonolaimus, Hirshmanniella, Longidorus
and Xiphinema) are caught on a 250m aperture sieve, adults of average sized nematodes
(e.g. Aphelenchoides, Ditylenchus and Hemicycliophora) on a 90m aperture sieve, and
many juveniles and small adults (e.g. Criconemoides, Paratrichodorus, Paratylenchus,
Pratylenchus and Radopgolus) on a 63m aperture. A 45 m or even 25m aperture sieve
is used to recover small juveniles (e.g. Meloidogyne, Heterodera, and most others). Ready
made sieves are expensive. Caution! Use sieves singly, never stack them and never
attempt to work samples through them all simultaneously, as this may reduce the efficiency
of recovery. Fine sieves are easily clogged, but this can partially be avoided by pouring the
suspension on a sieve inclined at an angle of about 30o to the horizontal. However the
number of nematodes recovered on the sieve will be reduced. Gently patting the underside
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of the sieve into the water bucket below and lifting it in and out a few times will help to
clear it.
Procedure
1. Mix the soil sample thoroughly and place a known volume of soil in a bucket
and fill to about ¾ with water. Dry soils should be soaked for a few hours. The
mixture is stirred to free nematodes from the soil and suspend them in the water.
Flocculating agents such as Separan NP10 (12.5g/ml) might be used to help
break up soil aggregates in heavy clay soils.
2. Let the mixture settle for 30 – 60 seconds and decant over a 2mm aperture sieve
into another bucket. Avoid pouring the sediment. Add less water to the
sediment in the first bucket and repeat this step 2 – 3 times to increase nematode
recovery. Any sediment left in the first bucket is discarded and the bucket
washed out. The sieve is rinsed over the second bucket.
3. The contents in the second bucket are stirred, allowed to settle for about 10
seconds and then poured through a 710 m aperture sieve into the clean bucket,
leaving behind heavy soil particles to which more water is added and the
process repeated, if desired. Collect the residue nematodes on the sieve by
directing a gentle stream of water on the upper surface of the sieve so as to wash
out small nematodes and eggs. Transfer the nematodes on the sieve into a
beaker using a gentle stream of water leaving behind any heavy particles.
4. Repeat the process using 250, 125 and 90m aperture sieves and collecting the
residues as described above. The residues of each sieve can be pooled in one
beaker or kept separate in different beakers. If the contents of the beakers
appear cloudy, it is because the residue on the sieve was inadequately rinsed. If
necessary, the contents should be poured back onto the sieve and rinsed again
over the bucket containing the remaining suspension before proceeding to the
next sieve in the series. The contents of the collecting beakers are allowed to
settle for 1-2 hours and the supernatant liquid is carefully decanted or siphoned
off leaving about 20ml in the bottom (some nematodes tend not to settle and
may require refrigerating the supernatant liquid). The material can be
transferred to a viewing dish and examined. If the suspension still contains a
significant amount of debris, further processing by centrifugal floatation or
modified Baermann techniques will result in an almost clean nematode
suspension. However, sluggish and inactive nematodes can be lost.
If you use 3 same sieves in series, you can be sure that you will recover about 100% of your
nematodes. For example if the soil sample had 10 nematodes, 1st Sieve will retain 7, 3 will
escape through. The 2nd sieve will retain 70% of the 3 nematodes ~ 9 nematodes recovered
and 30% will escape through (~ 1 nematode). The 3rd sieve will retain 70% of the 1
nematode ~ 100% recovery.
1. Sieving/Centrifugation method
This is an extension of the sieving method.
1. After decanting some of the supernatant liquid in step 4 above, transfer the
supernatant to four 50ml centrifuge tubes.
2. Centrifuge for 7 minutes at 1750rpm
3. Decant supernatant from tubes and discard
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4. Add sugar solution (450g/l water) to tubes

5. Shake tubes, centrifuge for 3 minutes at 1750 rpm
6. Pour suspension through the 45 m sieve
Rinse the residue from sieve for examination
Extraction of Nematodes from plant materials

Baermann Funnel Technique and its modifications
The Baermann funnel technique uses a funnel of 10 – 15 cm with rubber tubing attached to
the funnel stem and closed with a spring or screw clip. The funnel is placed in a suitable
support and almost filled with tap water. Plant material containing nematodes is chopped
into small pieces (< 1cm length), placed in a muslin cloth or nylon gauze, which is folded
to enclose the material and then gently submerged into the water in the funnel.
Plastic tray
Wide mesh
Soil kitchen paper
Figure 3: - Extraction tray method
Figure 4: The sieving technique (bucket-sieving method)

Nematodes emerge from the tissues and sink to the bottom of the funnel stem. After 24 –
48 hours, fully open the clamp and rapidly withdraw 5 – 10 ml of water containing the
nematodes and transfer it to a shallow viewing dish for examination. When this technique
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is used in its original form, nematode recovery is low (about 20% of other methods)
because of anaerobic conditions that develop due to bacterial decay of submerged organic
matter and lack of oxygen at the base of the funnel stem. This technique is handy and, in
order to improve its efficiency, it has been modified in several ways to become the standard
method for extraction of nematodes from plant tissues and soil. For example,
1. Modified Baermann funnel technique: This technique uses a supporting mesh to

hold the plant material partly submerged in water to avoid anaerobic decomposition.
Supporting gauze-mesh is fixed at the bottom of a plastic ring (cut from Perspex,
polythelene or vinyl tubes). Wet-strength facial tissue or milk filter is placed on the
supporting mesh and materials from which active nematodes can be extracted, such
as chopped plant material, thin layers of soil or residues obtained by sieving or
maceration, are then placed onto the facial tissue. Water is added to the funnel so
that the facial tissue is partly submerged in water. After 24 – 28 hours, collect
nematodes as described above.
2. The other modification is to use a shallow tray, dish or bowl to further improve
aeration and reduce the number of nematodes remaining on the wall of the funnel.
Similar to the above, a facial tissue is placed on a support and the chopped plant
material or soil is placed on it. The support and the material for nematode
extraction is placed in a tray (or dish, bowl, etc) filled with tap water. Small feet
(for example cut pieces of polythelene) can be attached to the support ring to give a
space of about 2mm between the base of the support and the collecting tray. The
material on the sieve should be moist (but not flooded) and it may be kept moist by
placing another facial tissue on top or using a relatively large tissue and folding it
over the material (Figure 5).
Do not pour water over the sample to avoid washing debris through the tissue. After 24
– 48 hours (sometimes less), the support with the sample is gently removed and the
contents on the tray transferred to a beaker. Any nematodes remaining on the tray can
be rinsed into the beaker using a spray bottle. The sample can be re-extracted if
necessary. Oxygenation (and hence nematode recovery) can be improved by adding 1 –
3 % H2O2.
Maceration techniques
Maceration can be used for extracting active nematodes as well as immobile stages of the
sedentary nematodes from plant tissues (e.g. corms, bulbs, cloves, storage roots, crowns,
leaves and small plants). The plant material is chopped into lengths of < 1 cm and then
placed in about 100 ml of water and macerated in an electric blender. The maceration time
required depends on the type of blender and the type of plant material. Maceration needs to
be continued long enough to give nematodes easy way out from the tissues but not to
damage or render them immobile. The resulting suspension can be extracted using the
modified Baermann techniques described above.
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Figure 5: Baermann Funnel Technique (top) and one of its modifications
Bioassays for detecting and quantifying nematodes

Because of the aggregated distribution pattern of nematodes, the chance of detecting a
small population of nematodes (for example 1 juvenile per 2 L soil) is relatively low when
small volumes of soil are processed. Difficulties in detecting low but economically
important populations of some nematodes can be overcome to a certain extent with
bioassay procedures. For example, to detect and quantify root-knot nematodes, samples
are collected as described previously but at least 1L of soil must be retained. Samples of 1-
5 L of soil are added to pots, a susceptible tomato seedling is planted and the plants are
grown in a warm environment (ideally 22-30°C). If information on the presence or absence
of the nematode is all that is required, plants are carefully washed from soil after 4-8 weeks
and roots observed for signs of galling. High gall ratings indicate a high population of root-
knot nematode . Bioassays can also be used to quantify the root-knot nematode population,
provided plants are harvested before a second generation of juveniles invades roots. Since
reproduction can begin in 4-5 weeks under ideal conditions (i.e. temperatures of 25-28°C),
plants are best removed from soil 3-4 weeks after planting. Provided nematode populations
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are low, (i.e. less than about 100 root-knot nematodes/litre), a single gall will generally
represent a single nematode. Thus, counting the number of galls on the root system will
give an indication of the number of root-knot nematodes in the potted soil volume. When
counting galls, roots should be floated in water in a tray with a dark coloured base, as this
enables small galls to be seen more easily with the naked eye.
C. HANDLING NEMATODES
Extracted nematodes can be examined directly under a microscope to the genus level using
viewing dishes or counting slides or can be processed further to slides. All or part of
extracted nematode suspension (depending on nematode density) can be transferred to an
open counting slide1 and examined directly under a microscope to genus level (at times up
to species level) and the nematode populations can be counted using hand held tally
counters or a bank of counters. However, for nematode identification to the species level, it
is advisable that temporary or permanent slides are made. The nematodes must first be
killed, fixed and properly mounted.
Killing and fixing nematodes

A few specimens can be killed by transferring them to a drop of water on a glass slide,
which is then heated over a small flame for a few seconds or by placing the slide on a hot
plate at 65 – 70OC so that the nematodes assume their heat relaxed shape. The specimen
can be examined directly under the microscope or can be transferred to a fixative or fixed
on the slide by adding an equal amount of double strength fixative.
A number of fixatives are commonly used for preserving nematodes. Most of these contain
formalin and should be handled with due care (rumour has it that a substantial percentage
of Nematologists died from cancers induced by a lifetime of inhaling formalin vapours!).
After fixation, nematodes are usually transferred to glycerine via a dehydration procedure,
because this will stop further denaturising and decay, while also resulting in highly
transparent specimen ideal for observation through high-powered light microscopes. Stains
are not commonly used, because these will not only highlight certain features but also mask
other structures.
Below, two easy and quick methods for fixing and processing nematodes are presented.
Prior to this, however, it is important to stress that you must at all costs avoid contraction of
the nematodes prior to death. For example, never fix live nematodes with cold fixative.
They will not be killed instantaneously, but (apart from suffering a gruesome death) will
contract and coil to a degree that often makes them useless for detailed study.
To make sure nematodes are nicely stretched upon fixation, you must kill them
instantaneously, either by using hot fixative or by heat-killing them prior to adding fixative.
1
there are various types of counting slides developed by various nematologists with varying capacities which range from
a petri dish or watch glass with a grid, which can be marked on the inside of its base to guide searching of nematodes, to
multi-chambered counting slides which allow examination of several samples on one slide
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Both procedures result in what is known euphemistically as "heat relaxation", relying on a

knock-out heat shock to instantly relax the muscles of the nematodes. Note also that it may
sometimes be advisable to starve the nematodes for a few days prior to killing and fixing,
because well-fed nematodes can contain so many intestinal granules that other organs
remain obscured even after transfer to glycerine.
The best way to kill nematodes that are collected in a small volume of water (e.g. from an
extraction tray or from cultures) is to transfer them to a glass vial and put this in a 70-90OC
water bath. Stir the vial for 20-30 seconds and check under a stereomicroscope that the
nematodes are all motionless and stretched out. Make sure they are not boiled - this disrupts
cellular structure. After heat-killing, fix the nematodes with hot fixative under a flow hood
(hot fixative will be more chemically active and you should avoid inhaling it at all times).
Fixatives
1. Formalin: 8ml Formalin (40% formaldehyde) topped with distilled water up to
100ml. CaCO4 powder can be added to neutralize the free formic acid that can cause
darkening and granulation of tissue
2. Formal acetic (FA) or formal propionic (FP) 4:1: 10 ml Formalin (40%

formaldehyde), 1ml glacial acetic acid (or propionic acid), 2 ml glycerol, topped
with distilled water up to 100ml. Addition of glycerol means that the nematodes
can be brought directly from fixative to glycerol by slow evaporation. Also it
means that the nematodes stored in vials will eventually end up in glycerol should
the fixative evaporate (see processing and mounting nematodes below).
3. TAF: 7ml Formalin (2 – 4% formaldehyde) + 2% glycerol, 2 ml tri-ethanol-amine

(to neutralize the free formic acid that can cause darkening and granulation of
tissue), 91 ml water.
FA 4:1 and FP 4:1 are probably the most widely used fixatives that also allow long term
preservation. TAF is a commonly used fixative, as nematodes fixed in TAF retain their
lifelike appearance in it for several hours, but it is not good for long term preservation as
some degeneration of the nematode cuticle can occur. Processing nematodes further to
glycerol improves preservation.
Fixation of plant tissue and soil
In some (if not most cases) plant tissues and soil samples will be processed for nematodes
within a few days after sampling. In order to prevent population changes during extended
storage (and also avoid quarantine restrictions applicable to live material), roots, shoots and
soil can be fixed for storage and subsequent examination by adding to them an equal
volume of hot (65-70OC) double-strength fixative, in a sufficiently large plastic bottle with
screw-cap, which is closed and shaken thoroughly. Alternatively, fresh plant material can
be put directly into hot lactoglycerol; this softens tissues and is particularly helpful in the
recovery of root-knot nematode females from roots.
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Note that dead nematodes don't wriggle, so you cannot extract them with an extraction tray.
If you fix a plant tissue/soil sample, you will therefore need to use more complicated
extraction devices such as Cobb sieves, centrifuge, etc.
Processing and Mounting Nematodes

In fixed nematodes, much of the internal body contents, especially the gonad structure, may
be obscured by the granular appearance of the intestines. Specimens can be cleared by
processing with lactophenol (Caution: phenol fumes are dangerous to health!).
Lactoglycerol (equal amounts of lactic acid, glycerol and distilled water) or glycerol, to
which a stain (0.05% acid fuchsin or 0.05% methyl blue) is preferred.
Fixing with formalin-glycerine and transferring to glycerine through ethanol

(Seinhorst, 1962)
Prepare double-strength Formalin – Glycerine fixative containing 8% formalin and 2%
glycerine in distilled water. Transfer live nematodes to a small glass vial and allow them to
settle to the bottom. Draw off surplus water until they are left in about 2 ml water. Kill the
nematodes by stirring the vial 20-30 seconds in a 70 -90OC water bath, check they are all
dead and stretched, and then add an equal volume of 65-70OC fixative. Stir, and then leave
the vial for a day to allow the fixative to penetrate and act on all tissues.
Take the vial with Formalin – Glycerine – fixed nematodes and draw off as much fixative
as possible without losing nematodes (if the vial has a narrow opening transfer the
nematodes to a cavity block). Fill the vial or block with a solution made from 20 ml of 96%
ethanol, 1 ml glycerol and 79 ml distilled water (to the brim if in a cavity block, to about 5
mm high if in a vial). Place the block or vial on a platform inside a closed glass jar
containing an excess of 96% ethanol (1/10 volume of the glass jar). Leave overnight in an
incubator at 35-40 C. This will allow all water in the suspension with the nematodes to be
replaced with ethanol.
The next day, take the vial or block out of the glass vessel and leave it open in the 35 –
40OC incubator for 2 – 3 hours, to evaporate about half of the ethanol (if necessary cover
partly to prevent complete evaporation). Refill with 5% glycerin-95% ethanol solution,
leave for another 2 – 3 hours, and refill one last time before leaving the vial or block
overnight in the incubator at 35 – 40OC. By the next day, the nematodes will be
impregnated in pure glycerine and ready for mounting in slides, or for storing without fear
of desiccation.
Note that the nematodes processed to glycerol are very soft and should be handled
carefully. The entire Formalin – Glycerine – ethanol procedure takes only three days and
usually results in well-fixed nematodes that will not decay for decades. Transferring
through ethanol dissolves cuticular lipids, however, and may result in a finely wrinkled
cuticle that will show up as such under the scanning electron microscope. If you want to
avoid this, try the slightly slower method below.
Fixing with TAF and transferring to glycerine through evaporation
15
Prepare double-strength TAF fixative containing 8% formalin and 2% triethanolamine in

distilled water. Transfer live nematodes to a small glass vial and allow them to settle to the
bottom. Draw off surplus water until they are left in about 2 ml water. Kill the nematodes
by stirring the vial 20-30 seconds in a 70-90OC water bath, check they are all dead and
stretched, and then add an equal volume of 65-70OC fixative. Stir, and then leave the vial
alone for a day to allow the fixative to penetrate and act on all tissues.
Take the vial with TAF – fixed nematodes; transfer the nematodes to a cavity block (this
could be easier for subsequent manipulation under a stereo microscope). Draw off as much
fixative as possible without losing nematodes, and then fill the vial or block with a solution
of 5% glycerine in distilled water (to the brim if in a cavity block, to about 5 mm high if in
a vial). Place the block or vial in an incubator at 35-40OC and cover it nearly completely,
leaving a narrow slit for slow evaporation. Leave until a substantial amount of water has
evaporated. Refill with 5% glycerine, and then leave at least two more days in the incubator
until all water has evaporated. Check the degree of dehydration by transferring two or three
specimens to a drop of pure glycerine, if their cuticle collapses, they are not yet completely
dehydrated - return everything to the incubator for another day or two. The entire
TAF/evaporation procedure takes four to six days and often results in perfectly fixed
nematodes.
In comparison with Seinhorst slow method, this method is recommended for better
instantaneous preservation and for Scanning Electron Microscope (SEM) material, but less
suitable for long-term preservation.
Mounting Nematodes
Temporary Slides
Temporary slides with freshly killed/fixed specimens mounted in TAF can be made to view
some important features of nematodes. Place the specimens and supports (e.g. glass fibre
or beads) in a small drop of fixative and put a coverslip. Blot off excess fixative with tissue
paper, seal the cover slip with Vaseline or nail varnish and observe under a microscope.
Observation of detailed morphology of live nematodes can also be done by making a

temporary slide. Add 1 – 2 drops of hot 4 – 5% agar on a glass slide, and immediately
flatten this agar with another glass slide having spacer strips of thick plastic tape. Carefully
remove the top slide when the agar has set. Add a drop of water on the agar, transfer the
nematode to it and place a coverslip on top. The pressure between the coverslip and the
hard agar will slow down the worm sufficiently to observe it with oil immersion
magnification. If you want to immobilize it almost completely, smear some Vaseline on the
rims of the coverslip, place it on top of the agar and nematode, and very carefully press
down the rims of the coverslip until the nematode is trapped but not squashed. A complete
Vaseline seal will also prevent desiccation. It is usually possible to recover the live
nematode from such a slide after studying it.
Permanent Slides
16
Once nematodes have been fixed and transferred to glycerine, permanent mounts can be
made.
Step by step procedure
1. Preparing a glass slide: Fill a Petri dish with paraffin granules, melt them at about
60OC and allow the paraffin to set into a solid layer. Take a 10 cm long cross-cut
metal tube with smooth, thin rim and slightly smaller diameter than the coverslips
you use (e.g. a 16 mm diameter tube for 18 mm diameter coverslips) and heat one
end in a flame. When the other end of the tube is beginning to get hot in your hand,
push the heated end down vertically in the paraffin so that it gets covered by
melting paraffin, and then press this end down vertically on the middle of a glass
slide. Lift the tube, and you should leave behind a complete 3-4 mm thick ring of
setting paraffin. Transfer a very small drop (can be applied using a syringe fixed
with a small needle) of anhydrous glycerol (heated for 4 hours at 35 – 40OC in an
oven) to the centre of this wax ring on a slide, leaving a spot of 4-5 mm on the slide.
Repeat this for as many slides as you wish to make.
Note: Getting the proportions of wax and glycerine right is important: too little
paraffin and too much glycerine will result in an incomplete seal, too much wax and
too little glycerine will result in nematodes being covered or trapped by paraffin.
2. Transferring nematodes: Pick out the nematode specimens you want with a needle
and transfer them to the glycerine drop in the centre of a wax-ringed glass slide.
You can usually mount up to ten of them per slide; more will too often result in
specimens overlapping or ending up in paraffin. After transferring the required
number to a slide, put it under the stereo microscope and using a handling needle
arrange all nematodes in the centre so that they touch the side of the slide and are
not floating, making sure none overlap with one another. Place three glass supports
around the nematodes.
3. Sealing and shuffling: Drop a coverslip over the wax ring and glycerine drop, and
put the slide on a moderately hot plate (or a mesh or metal plate above a small
flame). Allow the paraffin to melt around the glycerine drop, and allow all air to
escape from under the coverslip. Then put the slide back under the stereo
microscope, and check that no nematodes are overlapping. If nematodes are
overlapping, gently push the coverslip in the required direction to dislodge one of
the overlapping nematodes. If the paraffin has set by now, return the slide to the hot
plate. You can also re-heat and gently push the coverslip sideways to turn
specimens over. Once set, the paraffin will both act as a seal and a separating layer
between the coverslip and the glass slide, and ideally your slide will contain just a
small circular central area with glycerine and nematodes.
17
If some specimens are covered by smudges of paraffin under the coverslip, and/or
the paraffin is too thick to observe specimens with high power objectives, put the
slide back on the hot plate and allow the wax to heat and spread out further so that it
forms a thinner layer. If you want to pick out specimens for transfer to another slide
or for use in SEM or cross-sections, gently open the coverslip with a scalpel or thin
needle while keeping track of your specimen(s) under the stereo microscope. This
may go better if you first heat the slide gently (e.g. leave it on the lamp housing of
the microscope for a few seconds). Make sure you do not allow the glycerine to boil
during any of these operations, and never push down on the coverslip while the wax
is molten.
When satisfied with the arrangement of your nematodes on the slide, you can
permanently seal off the coverslip with colourless nail varnish.
The Noble Art of Picking Worms

While you can cheat and use pipettes or micropipettes for most culture procedures, there
are no easy ways out when you need to make slides: you have to learn to pick nematodes
with an eyelash, a hair, a fine needle or other picking device. Picking worms off fairly dry
agar shouldn't be too difficult (unless you have a very agile species), although you may
need to use an inconveniently thick or flattened pick if you do not want to damage the agar
surface. Picking worms out of a suspension is a different matter, however.
Half the trick is getting a good picking device, e.g. a fine and rigid insect needle tapped
against the table to bend its tip to a minute hook, or a hand-picked hair from the most
recalcitrant and bushy moustache available in your lab. Mount one of these on a handle and
prepare yourself for a few memorable hours at the stereo microscope.
The other half of the trick is getting the hang of suspended-nematode-dynamics while
working at a stereo microscope. Select the specimen of your choice in a Petri dish or cavity
block with nematodes in liquid (water, glycerine or whatever is appropriate for the
procedure at hand). Try to pull this nematode free from the bottom with a short twitch of
the pick, and tease it up to the surface of the liquid until it is more or less horizontal, using
one hand to move the pick and the other to change focus. Then position the tip of the pick
for the final twitch. For curled worms, try to catch them by the crook of the curved part(s)
of their body. Straight worms are more fun (or more trouble), because these will often only
come out if you position the pick just below them and at a slight angle to their body axis.
With luck, the viscosity and surface tension of the liquid will make the worm stick to the
pick instead of pulling it back down. Small straight worms are the best (or worst) because
they only have a small surface for viscosity to act upon, and will obstinately refuse to cling
to your pick. Teasing a straight, 0.3 mm long nematode out of pure glycerine is a
particularly nerdy way of spending long winter evenings, or challenging friends and
enemies!
The easy way (using Pipettes)
18
A generally faster and safer method of picking single specimens from a suspension consists
of using a pipette. This will especially be easier if you are not at all used to nematode-
fishing.
Take a fine Pasteur pipette, heat it near the tip above a flame until the glass softens, and
pull off the tip smoothly. Break off under stereo microscope the drawn-out end of this glass
tip until you have an opening of appropriate size, i.e. about as wide as one to one-half the
body length of the nematodes you are going to handle. Fit the pipette with a mouth tube.
Gently warm the pipette over a slow flame and while the pipette is still warm, suck up
single nematodes. This technique is especially efficient for handling very small nematodes.
Further Reading
Bloemers, G.F. & Hodda, M. (1995) A method for extracting nematodes from tropical
forest soil. Pedobiologia 39: 331-343.
Hooper, D.J., Hallmann J. and Subbotin, S. (2005). Methods of Extraction, processing and
detection of plant and soil nematodes. in: Luc, M.; Sikora, R.A. & Bridge, J. (eds.) Plant
parasitic nematodes in subtropical and tropical agriculture. Second edition. CAB
International, Wallingford: 53-86.
Southey, J.F. (1986 (ed.). Laboratory methods for work with plant and soil nematodes.
MAFF, London.
19
CHAPTER 2
BIOLOGY OF PLANT PARASITIC NEMATODES

Waceke Wanjohi
Kenyatta University,
Department of Plant and Microbial Sciences (PMS)
P. O. Box 43844 – 00100, Nairobi, Kenya.
Objective: Develop an understanding of the general biology of plant parasitic nematodes

and its implication to nematode control and management
1. Introduction
Plant parasitic nematodes (PPN) are essentially aquatic and spend a greater part of their life
cycle in the soil. They infect all plant organs where they feed ectoparasitically or
endoparasitically using their stylet. A few are semi- endoparasitic where only the anterior
part of the nematode penetrates the root and the posterior part remains outside the root.
Endoparasitic nematodes are relatively more damaging than the ectoparasites. Infected
plants in general exhibit stunting, chlorosis, wilting and reduced yield, in addition to several
below-ground symptoms. Such symptoms are often confused with other soil problems,
including compaction and nutrient deficiencies, and nematode damage is often overlooked.
Besides nematodes causing diseases on their own, they interact with other nematodes and
pathogens to cause disease complexes frequently resulting in more severe diseases.
Most PPN exhibit parthenogenetic (males absent, very rare or non – functional) and
amphimictic (males and females are separate) type of reproduction. Eggs are deposited
singly or in masses either in the soil or within plant tissues.
Most PPN have four larval stages between the egg and adult, with intervening moults. A
life cycle from egg to egg can be completed within 3-4 weeks under optimum
environmental conditions; temperature is a key factor in determining the duration of the life
cycle. Eggs hatch under favourable environmental factors or in response to root exudates.
Passive dispersal aided by several agents plays a significant role in the dissemination of
nematodes. In a number of genera, eggs are the survival stages, being protected either in a
gelatinous matrix (root knot nematodes) or within the cyst (cyst nematodes). Some PPN
enter a reversible anhydrobiotic state to survive desiccation.
20
2. Nematode pathology
Plant parasitic nematodes are biotrophic parasites which obtain nutrients from the
cytoplasm of living root, stem and leaf cells for development, growth and survival.
Nematodes have evolved diverse parasitic strategies and feeding relationships with their
host plants. They possess a hollow and a protrusible feeding structure, the stylet and a
pharynx, which has undergone morphological and physiological adaptations to suit the
feeding relationships. Depending on the species, they feed from the cytoplasm of
unmodified living plant cells or have evolved to modify root cells into elaborate feeding
cells as in root knot nematodes (RKN). The nematodes use their stylet to pierce and
penetrate the cell wall of a plant cell, inject gland secretions through the stylet orifice into
the cell and withdraw and ingest nutrients from the cytoplasm. Nematodes that enter root
tissue also use their stylet to cut openings and/or inject secretions to dissolve (intracellular
migration) or weaken (intercellular migration) the cell wall or middle lamella.
In general, all PPN damage plants by direct mechanical injury using the stylet during
penetration and/or by secretion of enzymes into the plant cells while the nematode is
feeding. The physical presence of endoparasitic nematodes inside the host also affects the
functioning of the host. As a result of nematode feeding, the architecture and extent of the
root system is altered, so that it is less efficient at taking up nutrients and water from soil.
The extent of nematode damage depends to a large extent on the inoculum density (level of
infestation). Low or moderate numbers of nematodes may not cause much injury but large
numbers severely damage or kill their hosts.
Feeding relationships
The feeding relationships between nematodes and susceptible plants are diverse and the
amount of tissue destruction and the degree of plant response are often related to the type of
feeding relationship. While some PPN are ectoparasites, others are endoparasites or semi-
endoparasites.
Ectoparasites
Ectoparasites feed from root tissue by inserting their stylet from outside the root (Plate 1a –
b). The group consists of several morphologically divergent families that have evolved
different feeding strategies. As a rule, species that have a short stylet feed on epidermal
cells e.g. Tylenchorhychus dubius and those with long stylets feed on deeper tissues e.g.
Belonolaimus, and Dolichodorus spp. Ectoparasites are either migratory or sedentary.
Migratory ectoparasites
These nematodes have the most primitive mode of parasitism; they remain outside of the
root and use their protrusible stylet to feed either on epidermal cells or cells deeper within
the root. With the exception of species of a few genera that feed on root tips, e.g.
Belonolaimus, nematodes with this type of feeding strategy generally cause little obvious
21
tissue damage. The migratory ectoparasites remove cytoplasm from the parasitized cell,
frequently causing their death and then move to another cell to repeat the feeding process.
Dorylaimid migratory ectoparasites include Trichodorus spp, Xiphinema index and
Longidorus elongatus while Tylenchid migratory ectoparasite includes Tylenchorhychus
dubius. Psilenchus, Tylenchus and Atylenchus feed only on root hairs.
Sedentary ectoparasites
These nematodes feed from a single site or plant cell for a prolonged period of time while
remaining outside the root. Sedentary ectoparasites such as Criconemella xenoplax use a
single feeding cell as a nutrient source for several days before the nematode moves on to
establish another feeding cell. Feeding by C. xenoplax causes little tissue damage compared
to Hemicycliophora arenaria and some Helicotylenchus species (migratory ecto-
endoparasites) which induce terminal galls when feeding on the root tips. Hoplolaimus and
Telotylenchus are semi-endoparasites feeding internally and externally on plant root tissues.
Endoparasites
Endoparasitic nematodes invade the root tissue with part or all of their body. Some feed
soon after entering the root while others feed only after migrating to a preferred feeding site
(e.g. the cortex and the xylem parenchyma cells) (Plate 1c )
Migratory endoparasites
Migratory endoparasites such as Pratylenchus, Hirschmanniella and Radopholus spp,
which have a small but robust stylet enter the root and periodically feed as they migrate
intracellularly through the root tissue. They primarily inhabit the cortical tissue of the root
and retain their mobility and feed on the tissues as they move. This causes extensive
destruction of root tissue along the path of the migrating nematode.
Sedentary endoparasites
Some endoparasitic nematodes become sedentary (Meloidogyne, Globodera, Sphaeronema
and Heterodera) and feed from a single cell or a group of cells for a prolonged period of
time. For this sustained feeding, the sedentary parasites have evolved very specialized and
complex feeding relationships with their host plants. The nematodes invade roots as
vermiform second stage juveniles (J2) and development depends upon the modification of
the phenotype and the function of the specific root cells to form specialised feeding cells
that become permanent source of nutrients for the nematodes (Plate 1d). They modify the
root cells of susceptible hosts into elaborate feeding cells including modulating complex
changes in cell morphology, function and gene expression. When feeding commences, the
juvenile‟s body grows and it becomes saccate and immobile. In this feeding strategy,
destruction of root tissue is usually limited to cells around the feeding site and the
nematode. Root-knot nematodes induce giant cells which are enlarged multinucleate cells
that are full of cytoplasm and metabolically very active, whereas cyst nematodes form
similar cells but are as a result of cell wall breakdown and the formation of a syncytium
(Plate 1d). Both types of feeding cell create a transfer cell that enables the rapid transfer of
nutrients across the cell to support the development of the female nematode and the
production of large numbers of eggs. While the migratory endoparasites primarily inhabit
the cortical tissue of the root, the sedentary endoparasites pass through the cortex and
invade the vascular cylinder where they induce feeding sites and become sedentary. Unlike
22
the sedentary endoparasites which have a fixed feeding site (nurse cells or syncytia) and
lose their mobility and become obese, the migratory endoparasites feed on the tissues as
they move, retain their mobility and their vermiform shape. Other sedentary endoparasites
include Trophotylenchulus obscurus, Tylenchulus semipenetrans, Verutus volvingentis,
Cryphodera utahensis, Rotylenchulus reniformis etc.
While the J2 is the infective stage in root-knot, cyst and potato cyst nematodes, all stages of
ectoparasites and most migratory endoparasites are infective. In Rotylenchulus spp the
immature female is the infective stage.
Adaptations to mode of feeding
Evolutionary adaptations of nematodes for plant parasitism led to the development of the
protrusible stylet as well as marked morphological and physiological modifications of the
oesophagus to form elaborate secretory glands which are the principal source of the
secretions involved in plant parasitism (Plate 2 a-c). These glands enlarged considerably as
nematodes evolved from free-living nematodes to obtain nutrients from plants.
Tylenchids are well adapted for plant parasitism. In addition to the stylet, they have a well
developed oesophagus with a muscular metacarpus containing a triradiate pump chamber
and three large and complex specialised secretory gland cells in the pharynx. During
feeding, rapid maximum dilation of the pump chamber creates the suction necessary for the
nematode to ingest nutrients from the feeding cell through the lumen of the stylet and the
oesophagus and force their passage into the intestines. The one dorsal and two subventral
glands produce secretions involved in plant parasitism. The dorsal gland cell has a long
cytoplasmic extension that extends anteriorly through the metacarpus to terminate into an
ampulla, a collecting reservoir for secretory granules in the oesophagus near the stylet
knobs. In contrast, the subventral gland cells have short cytoplasmic extensions that
terminate in ampullae at the base of the pump chamber in the metacarpus. A sclerotized
duct and an elaborate valve with a membrane-delineated end sac connects the ampulla to
the oesophageal lumen. The valve controls the release of secretions into the lumen of the
oesophagus. The secretions are synthesized in the nuclear region of the glands cells and
transported along the microtubules in the cytoplasmic extension to accumulate near the
valves in the ampulla prior to their contents being secreted. The association of the neural
processes and the neurosecretory cells with the gland cytoplasmic extension and ampulla
indicates that the regulated secretion is controlled by the nervous systems.
During parasitic development of sedentary endoparasitic species (Meloidogyne spp;

Heterdera spp and Globodera spp), the oesophageal gland cells undergo distinct
morphological changes. In Meloidogyne spp. the sub-ventral gland cells are the most active
glands in infective J2s but as they establish a feeding relationship with the host tissue and
increase in body width, the dorsal gland increases and the subventral glands decrease in
size. Therefore in adult females, the dorsal gland cell predominates and becomes the
functional secreting gland while the sub-ventral are greatly reduced indicating a changing
role for the oesophageal gland cells and their secretions at different stages of the nematode
life cycle.
23
The secretions play a key role in nematode penetration, migration through the root,
modification and maintenance of root cells as feeding sites, formation of feeding tubes and
/or digestion of host cell cytoplasm to facilitate nutrient acquisition by the nematode.
During parasitism of the plant cell, the nematode‟s stylet penetrates the cell wall but does
not pierce the plasma membrane, which becomes invaginated around the stylet tip. The
secretions may be deposited outside the plasma membrane or injected directly into the
cytoplasm of the host cell through perforation in the plasma membrane at the stylet orifice.
The secretions by sedentary parasites transform root cells into metabolically active feeding
sites by formation of feeding tubes within the cytoplasm of the parasitized cell. They
modify directly or indirectly gene expression to induce profound morphological,
physiological and molecular changes in the host cell to enable them to function as a
continuous source of nutrients for the nematodes parasitic stages. Removal of nutrients
from feeding cells by sedentary nematodes may involve direct withdrawal of the
cytoplasmic components (Criconemella xenoplax) or withdrawal facilitated by a feeding
tube produced by the parasite (Meloidogyne, Heterodera, Globodera and Rotylenchulus
species).
3. Symptomatology
Symptoms may vary according to nematode parasitic habits and host–parasite relationships,
and other factors such as host age and physiological conditions. The symptoms may be seen
both above and below- ground.
Above ground symptoms

Symptoms associated with root nematodes are a direct result of the impaired ability of root
systems to take up water and nutrients and thus are essentially similar to symptoms of any
root damage that interfers with the physical support and water and nutrient absorption
systems. They are thus often similar to mineral deficiencies, inadequate or excessive water
supply and generally poor soils. Symptoms are more pronounced if the plants are already
affected by other adverse conditions or are attacked by other pathogens. Plants growing
under highly favourable conditions may be heavily attacked by nematodes but show few
above ground symptoms.
The most universal above ground symptoms of a nematode disease are
i. Stunting –the reduction of growth rate, reduction in amount of foliage and

progressive death (die-back) of plants. The stunted and chlorotic plants are
distributed in circular to oval areas of variable size in the field but patches of
damaged plants may be elongated if infested soil is moved in the direction of
cultivation (Plate 3a-b)
ii. Chlorosis (yellowing of leaves), poor yield, early senescence, premature dropping
of fruits and flowers, fruit malformation. Wilting due to the effect on the functions
of roots is also a common symptom of nematode infected plants.
iii. Other above ground symptoms are associated with specific nematode species. For
example, leaves with dark green spots, angular or cuneiform in shape, with
interveinal discoloration and necrosis are associated with Aphelenchoides
ritzemabosi on chrysanthemum leaves while twisting and white tips of leaves of rice
24
are associated with Aphelenchoides besseyi (Plate 3c). Yellowing and collapse of
palm trees followed by a rapid death and a red necrosis in the vascular bundles on
the stem forming a red ring in coconut and oil palm is due to infection by
Bursaphelenchus cocophilus (Plate 3d). Galls in stems, leaves and seeds of cereals
and grasses are caused by Anguina spp. (Plate 3e). Toppling of banana plants
especially during fruit bearing is due to Radopholus similis (Plate 3f) Twisting of
leaves and raised yellow lesions on stems and leaves on onions and narcissi are by
Ditylenchus dipsaci), twisted panicles and empty grains by Ditylenchus angustus on
rice, yellowing and rapid death of pine trees (Bursaphelenchus xylophilus) and
distorted apical growth and crimpling of leaves and inflorescence (Aphelenchoides
besseyi and Aphelenchoides fragariae in strawberry) and so forth.
Below-ground symptoms
i. Reduced absorption of water and mineral nutrients by the secondary roots.
ii. Quantitative and qualitative changes in root exudates such as reduced concentration
of amino acids, increased concentration of sucrose, and a disappearance of glucose,
threonine, serine, histidine and citric acid in plants infected by some root-knot
species.
iii. Water movement is affected thus influencing the water potential of leaves, stomatal
conductivity, transpiration and root conductivity. Root conductivity can be reduced
in beans infected by M. hapla, and in potato plants infected by Pratylenchus
penetrans. Globodera rostochiensis causes mineral deficiency (N, P, K and Mg) by
affecting ion absorption and reduced water absorption due to poor root
development. Calcium concentration may increase due to reduced potassium uptake
and dehydration or through the disruption of the endodermis by the nematodes.
iv. Reduction of root system especially the secondary feeder roots
v. Abnormal development of roots (Plate 4a-h)
 Overall root galling (Meloidogyne spp. and Nacobbus aberrans)
 Ulcerations (ulcers/ lesions). Sharply demarcated necroses in different layers
of the plant tissues. This results from reactions of phenolic substances in the
plants to the secretions discharged by nematodes. Roots with longitudinal
necrotic areas are typical of Pratylenchus spp, Radopholus spp;
Hirschmaniella spp. infection (Plate 4g)
 Dry rots usually develop from infestation of fleshy parts of the plants
(tubers, root vegetables, stolons) eg by Ditylenchus dipsaci on onions (Plate
4h.), D. destructor on potatoes, R. similis on banana rhizomes. The
nematodes cause dry rots in association with secondary invading
microorganisms.
 Excessive branching of secondary roots (M. hapla, Pratylenchus spp. N.
aberrans. Localized proliferation of lateral roots (Some Meloidogyne spp
and Heterodera spp). Parasitism of young roots stimulates formation of
lateral roots. The lateral roots are also infected and the entire root system
becomes dendroid and reticulate in appearance. In Heterodera spp. this
causes the “root-beard” disease
 Swollen, hooked root tip galls (Subanguina spp, Xiphinema spp,
Meloidogyne graminicola). Retardation of growth on the root tip. The root
25
system is dwarfed and thickened in appearance (Trichodorus, Longidorus,

Xiphinema spp,). Roots ending in rounded galls (Longidorus spp. and
Hemicycliophora spp). Stubby roots, suppression of root growth
(Trichodorus spp and Paratrichodorus)
4. Nematode disease complexes
Plant parasitic nematodes can be the sole pathogens or may interact with other plant
pathogens or nematodes to cause a disease complex. Nematodes frequently form disease
complexes with wilt-inducing and root-rot fungi. Infection by Meloidogyne spp.,
Pratylenchus and Rotylenchulus reniformis nematodes, for example, increases the
incidence and severity of Fusarium, Verticillium spp, Pythium spp. wilt in a number of
plants. Meloidogyne spp., Pratylenchus, Anguina and Ditylenchus interact with disease
causing bacteria, Clavibacter Pseudomonas and Agrobacterium to increase the severity of
the disease. In nematode infested soils, tomatoes were attacked by Ralstonia solanacearum
but in nematode free soils they remained healthy. In general, nematodes serve as vectors of
other plant pathogens, and/or wounding agents that increase the susceptibility of the host.
In general plant parasitic nematodes enhance host susceptibility leading to increased rate of
development and severity of wilt and fungal rot diseases. The root exudates from root knot
nematode (RKN) infected plants stimulate the fungal pathogens and suppress
actinomycetes, which are antagonists of the wilt fungus (Fusarium spp.). The physiological
change of RKN- infected plants also enhances penetration by the fungus and wilt
development. Root knot nematode infection increases the pathogenicity of the wilt fungus
and consequently the severity of the disease. The nematodes establish their feeding sites in
the xylem parenchyma cells, bringing about significant changes in the morphology,
anatomy and biochemistry of the plant. Giant cells induced by RKN remain in a state of
high metabolic activity through continuous stimulation by the nematode. The high
concentrations of sugars, hemi - cellulose, organic acids, free amino acids, proteins and
lipids benefits the fungal pathogens. The giant cells remain in a perpetual juvenile state
which delays maturation and suberisation of other vascular tissues and thus fusarium
successfully penetrates and establishes in the xylem elements. Inhibition of phytoalexins by
the nematodes implies loss of resistance to the wilt fungus. Inhibition of tyloses formation
by the RKN on tomato is a possible mechanism for increased wilt. Tyloses formed in the
xylem vessels by the expansion of xylem parenchyma through the pits do not develop from
xylem parenchyma cells which are transformed into giant cells or physiologically altered
adjacent cells.
Nematodes are important vectors of viruses. Xiphinema and Longidorus transmit

nepoviruses. Trichodorus and Paratrichodorus vector the tobraviruses. Xiphinema index
and X. americanum, for example transmits grapevine fan leaf and tomato ring spot viruses,
respectively. Longidorus elongatus transmits the raspberry ringspot virus, Trichodorus
cylindricus, the tobacco rattle virus and Paratrichodorus minor, the pepper ringspot virus.
The nematodes transmit viruses in a non-circulative manner being the ingestion-egestion
type of transmission. The nematode acquires/ingests the virus particles while feeding on the
virus infected plant, retains the particles at specific sites within the nematode and egest
them by dissociation from the sites of retention. The virus retention sites differ among the
26
nematode genera. In Xiphinema, for example, the site is the cuticular lining of the
odontophore, the oesophagus and the oesophageal pump while in Longidorus, the site is the
inner surface of the cuticular ondontostyle and its guiding sheath. In Trichodorus and
Paratrichodorus the site is in the lining of the oesophagus from the most anterior region to
the cardia (oesophageal-intestinal valve) but not associated with the onchiostyle. All these
surfaces are shed during moulting and therefore the virus particles do not pass from one
stage of the nematode to another during development.
Bacteria can be transmitted by nematodes externally on their body surfaces or internally

within their alimentary canal. Anguina tritici is closely associated with Clavibacter tritici
causing the yellow ear rot or “tundu” in India. The bacterium is associated with the body
surface of the juveniles inside the seed galls. Ditylenchus dipsaci transmits Clavibacter
michiganense pv. insidiosum which causes wilt in alfalfa. The bacterium is carried on the
body of the nematodes into the crown buds and is placed in conducive infection courts.
Nematodes have also been reported to transmit fungal pathogens. Anguina tritici is a vector
of spores of Dilophospora alopecuri which attacks aerial parts of cereals. The nematode
while moving between the leaf sheaths to reach the growing point takes the fungal conidia
and deposits them on a growing point. Further, the nematode by feeding on the tender
leaves helps in the penetration and establishment of the fungus.
All PPN wound plants either by a simple micro-puncture or by rupturing or separating

cells. They may thereby either introduce a pathogen on or within their bodies or aid the
entry of a pathogen already present on the plant cell surface. Wounding is particularly
important for bacteria as they enter plants mainly through wounds. Besides creating
wounds the nematodes also modify the host tissue to enrich the substrate nutritionally to the
advantage of the bacteria. For example Meloidogyne modifies tomato tissues and makes
them more susceptible to bacterial canker (Clavibacter michiganense) and bacterial wilt
(Ralstonia solanacearum) especially when the nematodes are inoculated before the
bacterium.
5. Mode of Reproduction
There are mainly three methods of reproduction in nematodes.

i. Amphimixis is also referred to as bisexual reproduction, cross fertilization or
gonochorism. The sexes are separate and the females produce oocytes which are
fertilized by the sperms from the males. Examples; Anguina tritici, some
Meloidogyne species and the mycophagous species, Aphelenchus avenae. In this
type of reproduction, the males and females are present in equal numbers,
copulation is required and females do not produce sperms.
Reproduction using amphimixis is also retained as a facultative ability among some

parthenogenetic species and hermaphrodites.
In amphimictic species of Hoplolaimina, the spermatheca is usually round and axial

and contains elongated sperm, each consisting of a granulated nucleus and a hyaline
27
tail portion; the eggs do not mature if no sperm are present in the spermatheca to
fertilize them. In parthenogenetic species, the spermatheca is a small and empty sac.
ii. Parthenogenesis also referred as autotoky is common where males are very rare,
absent or non-functional and are not involved in reproduction. Copulation is
optional and females reproduce without sperms.
There are two main kinds, meiotic and mitotic parthenogenesis. The 2 types differ
in whether or not a first meiotic division takes place in the oocytes. Meiotic
parthenogenesis is in Rhabditis, Meloidogyne, Heterodera, Pratylechus and some
Longidoridae as well as Aphelenchus avenae. Mitotic parthenogenesis occurs in
Rhabditis, Meloidogyne, Helicotylenchus, Pratylenchus.
iii. Hermaphroditism (automixis or self-fertilization) is as in Caenorhabditis elegans,

in which a single gonad produces both the oocytes and sperms. This together with
amphimixis is common in rhabditids and is reported in other free-living nematodes
and plant parasites, including criconematids and in several predatory species. It is
indicated by the occurrence of hermaphrodite females whose spermathecae contain
sperms in the absence of males in the population. The hermaphrodite is
morphologically female but has a syngonic (an ovotestis) usually acting
protandrically (sperm produced first). After the initial sperm production and their
storage in the spermatheca, the gonad switches to oocyte production. A sperm will
fertilize any oocyte produced subsequently, thereby achieving automixis. This will
continue until the sperm supply is exhausted after which egg -laying may stop in
some species.
6. Life cycle and Hatching
The life histories of most plant parasitic nematodes are in general quite similar in that all
have four larval stages. Eggs may be laid singly or stuck together in masses in a gelatinous
matrix secreted by the females. Some females (Heterodera spp.) die and the cuticle tan to
form cysts. Many Heterodera spp also produce a proportion of their eggs in a gelatinous
matrix (egg mass) attached to the cyst. In RKN all the eggs are laid in an egg sac which
may be buried partially within the host-derived root gall which Meloidogyne spp. induce
during feeding. Egg masses are also produced by the semi-endoparasitic nematodes such as
Rotylenchulus reniformis. Egg sacs and cysts serve to protect the eggs from desiccation and
natural enemies. The eggs are typically oval without spines, plugs or excrescences. The
uterine egg is semi-fluid and passes through the vagina and vulva with ease. Most
Tylenchida are oviparous but a number of entomoparasitic genera have ovoviviparous
species in which the eggs hatch in the uterus. Occasionally, in oviparous females, the eggs
hatch in the uterus and kill the mother a phenomenon called “endotokia matricida”.
In most species, the egg shell has 3 layers, the outer lipoprotein layer derived from the
vitelline layer of the fertilised oocyte and retains the membrane structure. The middle
chitinous layer is usually the thickest and provides the eggshell with its structural strength.
It has a chitin microfibril core (providing tensile strength) with a collagen-like protein coat
(providing the rigidity). The innermost lipid layer represents the main permeability barrier
28
of the eggshell. It consists of two or three lipoprotein membranes although in Heterodera

schachtii the layer is tetralaminate.
The juvenile within the egg develops to adult through four moults, the first moult normally
occurring within the egg. The egg develops into a first stage juvenile (J1). The juvenile
coils several times within the egg shell and lies still. The J1 grows in size and undergoes the
first moult within the egg and then hatches as a J2. The J2 is fully developed except that it
lacks reproductive organs and is small in size (Plate 5a-b).
Summary of life cycle
Egg

J1 – moults in egg; hatches from egg in Anguina tritici

J2 – hatches from egg; infective and dauer stage in A. tritici

J3

J4 – dauer and infective stage in D. dipsaci

Adult
The J2 undergoes a second moult and becomes a J3 and the J3 undergoes a third moult to
become a J4. The J4 undergoes a fourth moult and differentiates into adult females and
males and then matures. A life cycle from egg to egg can be completed within 3-4 weeks
under optimum environmental conditions. In Longidorus spp. the life cycle takes 2 yrs
while in Ditylenchus dipsaci it takes 19-23 days.
Hatching
In general given favourable environmental conditions (suitable temperature, oxygen
availability and soil moisture levels and in the absence of physiological barriers such as
diapause, hatching in most species occurs without requiring specific ques. In a few species,
the juvenile hatches when the conditions of the external medium meet particular
requirements. For example, in R. reniformis some Meloidogyne, Globodera and Heterodera
species, hatching is stimulated by the root secretions from host plants. In certain plant
parasitic nematode species, the parasitic life cycle is synchronized closely with that of the
host with the aid of environmental and host derived stimuli, to maximize the reproductive
success of the nematode.
In PPN, the J1 moults within the egg and the resulting J2 hatches. Each egg contains a
single juvenile, which hatches by cutting the egg-shell with its stylet by striking it with
intermittent rhythmic blows or by rupturing the egg-shell with its tail tip as in Heterodera
iri, or through normal rupture of the egg-shell due to juvenile enzymatic secretions and
movement.
29
The eggs of the cyst nematodes survive in the soil in round (Globodera) or lemon-shaped
(Heterodera) cysts each containing several hundred eggs. There are small openings at the
neck and the vulval ends of the cyst through which the hatched juveniles escape.
Once hatched the J2s of Globodera rostochiensis and G. pallida can survive for <2 weeks
without feeding, Meloidogyne javanica and Tylenchulus semipenetrans can persist in the
field for months.
7. Nematode dispersal
Nematodes can be dispersed actively or passively.

Active dispersal
Nematodes do not move very far or very quickly by their own locomotory power in the
soil. Active nematode migration mostly occurs in the rhizosphere as they are attracted to
root exudates. Meloidogyne spp., for example, are attracted to an area just behind the root
tip while others such as Pratylenchus are attracted to the root-tips and sometimes further
back. The root tip is the region of high metabolic activity from which numerous substances
diffuse, some of which act as attractants (gibberellic acid, glutamic acids, tyrosine, amino
acids and carbon dioxide), some as repellants and some neither.
The best known example of active migration above ground is by Aphelenchoides spp. (e.g.
A. ritzemabosi) which moves up the wet external plant parts and then invades the leaves of
the host plant.
Passive dispersal
Dispersal by water
Water is a frequent means of passive dispersal. This may include surface run-off such as
overland flow, streams, rivers, irrigation canals, percolation and interflow. Infiltration and
percolation of water accounts for some downward nematode dispersal but the distance
varies with soil properties and precipitation. Dispersal of Radopholus similis, for example,
is aided by percolation. Interflow is lateral underground movement of water where
percolation water is forced laterally when it comes in contact with an impervious soil layer.
Above ground nematodes can be splashed to plants by falling rain or overhead irrigation.
Dispersal by wind
Wind blowing on bare soils or on low-growing plants or young plants can disperse
nematodes e.g. Heterodera schachtii, G. rostochiensis, Criconemoides, Helicotylenchus,
Meloidogyne, Pratylenchus, Tylenchorhynchus spp. etc.
Phoretic dispersal
This includes the involvement of another animal to aid dispersal. Insects are important in
dispersing nematodes that attack the aerial parts of plants. Nematodes can pass through the
digestive tract of animals and remain infectious in some instances, being dependent on the
mobility and the speed of the vector and the survival capacity of the nematode. For
30
example, the palm weevil, Rhynchophorus palmarum disperses Bursaphelenchus

cocophilus (red-ring of coconut palm). The J3 is deposited onto the palm tree when the
weevil is ovipositing her eggs. Bursaphelenchus xylophilus (pine wilt) is transmitted by
Monochamus alternatus; a cerambycid beetle. In this instance, the nematode‟s 4th stage
dauer juvenile is carried on the outside of the beetle under the elytra and in the trachea and
deposited into the pine as the beetle feeds.
Planting materials
Nematodes can spread through planting materials such as seeds, vegetative propagating
materials (tubers, corms, bulbs), seedlings and rootstocks. Nematodes spread this way can
lead to serious losses in the mature crop or in subsequent crops if nematode build -up is not
checked.
Dispersal through seeds: Anguina tritici in wheat (Triticum aestivum)
Ditylenchus dipsaci in beans (Vicia faba) and onions (Allium cepa)
Aphelenchoides arachidis and Ditylenchus africanus in ground nuts (Arachis
hypogaea).
Tubers: Scutellonema bradys, Pratylenchus coffeae, Radopholus similis, Meloidogyne
spp in yams (Dioscorea spp).
Bulbs: Ditylenchus dipsaci in onions, garlic and narcissi
Rhizomes: Radopholus similis in ginger (Zingiber officinale) and tumeric (Curcuma
domestica).
Corms: Radopholus similis , Pratylenchus coffeae, P. goodeyi and Helicotylenchus
multicintus
Seedlings/transplants: Nematodes that are associated with seedlings in nurseries are
transferred to the field during transplanting.
Dispersal by other means

Aphelenchoides besseyi can be spread by contacts between aerial parts of adjacent plants.
Leaf drop or wind – blown leaves can disperse foliar inhabiting nematodes such as
Ditylenchus dipsaci and Aphelenchoides spp.
Man‟s activities also aid in the dispersal of nematodes. E.g. transport of infected plant
materials between different countries (international dispersal) or parts of the countries
(local spread) or by farm implements between fields and cultivations within fields.
8. Nematode Survival
Survival strategies allow endurance of harsh environmental extremes and synchrony with
seasonal hosts.
There are a number of ways in which nematodes can increase their chances of survival in
adverse environmental conditions. They include;
31
1. Migrating to escape unfavourable environmental stress. For example, when surface

soils are dry the nematodes may migrate to deeper layers of soil. An exodus of
nematodes from senescing plants to younger plants
2. High reproductive capacity; example Ditylenchus dipsaci reproduces within the host
and builds up high levels of infection. When the host senesces, the nematodes
accumulate as 4th stage juveniles which persist in the soil until they infect a new
host (J4 is the infective stage). The J4 are resistant to desiccation and are capable of
anhydrobiosis (Plate 6a)
3. Producing survival stages within the life cycle of the nematode is also a survival
strategy. Examples: Eggs, infective larvae, cysts, dauer larvae etc as in cyst and
root-knot nematodes, J2 of Anguina spp., J3 of several species of Aphelenchoides
and J4 of D. dipsaci are capable of surviving long drought periods.
4. Life cycle synchronization. The nematodes develop mechanisms for synchronizing
their life cycle with the availability of food or hosts e.g diapause as in Globodera
rostochiensis, arrested development and dauer larval formation.
5. Continue activities but modify physiology and behaviour in order to continue living
under the changed conditions.
Dormancy
The nematodes respond to adverse conditions by entering into dormancy which includes
diapause and quiescence. Dormancy which can occur at most stages of the nematode‟s life
cycle, is the slowing down or stopping of the nematodes‟ metabolic activities and changing
their physiology or structure accordingly while the unfavourable conditions prevail. It
ensures that the consumption of energy reserves is kept to a minimum until such a time as
they are required for the migration to the host and subsequent invasion or penetration.
Diapause is a state of arrested development whereby hatching or activity does not occur
until specific requirements have been satisfied even if suitable environmental cues are
available. Diapause enables the J2s to overcome cyclic long-term conditions which are
unfavourable for hatching and infection e.g winter/summer and/ or the absence of the host
and is common in temperate species.
Diapause unlike quiescence is temporarily irreversible and requires other “triggers” to

break the dormancy even when all the environmental factors are favourable. For example
host-derived stimulus/hatching factor serves as triggers in some nematodes. Thus diapause
is associated with a decrease in metabolism and it is not a direct response to unfavourable
conditions (unlike quiescence or crytobiosis) but rather to stimuli that heralds the seasonal
onset of such conditions. For example, photoperiod- induced diapause in the eggs of
Globodera rostochiensis.
Diapause may also include a dauer = “enduring” juvenile formation, for example in RKN
and cyst nematodes (Heterodera and Globodera spp.). The length of time that a nematode
remains in diapause in nature is for the most part, fixed and is a reflection of the length of
the hostile season to which the nematode is subjected and with which it has developed
synchrony.
32
Diapause allows nematodes to arrest development either before or after exiting the host, the
length of the dormancy being dictated by (and in synchrony with) seasonal rhythms,
enabling eggs or juveniles to withstand harshness of the external environment. Diapause is
therefore not only a means of survival but it is also a means whereby activity of the
nematode is made to coincide with that of the host thus increasing the chance of infection.
Diapause thus enhances fitness of those nematodes which possess it. Its main function is to
act as a timing mechanism to coordinate nematode activity with that of its host in seasonal
but otherwise unpredictable environments.
Diapause in PPN is categorized into 2 distinct types; obligate and facultative diapause.
Obligate diapause- initiated by endogenous factors which are programmed into the life
cycle of the nematode and which may or may not be modulated by the host and is
terminated by predetermined periods of specific environmental conditions. E.g
Meloidogyne naasi and Heterodera avenae
Facultative diapause- initiated by exogenous environmental factors and is terminated by

either endogenous factors whose duration is dependent on prevailing physical
environmental conditions or by the presence of the host. Though more common in
Heterodera spp where it provides long-term hatching delays while root- diffusate gives
more precise short-term hatching signals, as also evidenced in Globodera and Meloidogyne
spp. The ability of Meloidogyne infestations to persist in fallow soil or through periods of
drought resides in the ability of females to produce dormant eggs (embryonic diapause)
where eggs do not hatch immediately after they are laid.
Quiescence= “slowed ageing” is a spontaneous reversible response to unpredictable

unfavourable environmental conditions that results in reduced but measurable metabolism.
It occurs at any time of the year and release from quiescence occurs when favourable
conditions return.
Although quiescence is usually a facultative response occurring only when stress is present,
it can also be an obligatory part of the nematode‟s life cycle and hence referred to as
crytobiosis/ anabiosis. Obligate quiescence occurs when the environmental cue affects a
specific receptive stage of the nematode life cycle, e. g quiescence of unhatched J2s of
several cyst nematodes in the absence of root diffusates while facultative quiescience is
nematode stage non-specific.
Crytobiosis
Crytobiosis = “suspended animation” takes place if the stress persists or increases and
therefore metabolism ceases altogether or it is not measurable. In this situation, all
reversible life processes are suspended for a considerable time due to unfavourable
conditions.
Alterations in metabolism can be triggered by adverse environmental conditions such as

i. dessication/dehydration= anhydrobiosis
ii. Low and high temperature =cryobiosis and thermobiosis
iii. Lack of oxygen = anoxybiosis
33
iv. Osmotic shock (high solute concentration = osmobiosis

and are reversible once the stress is removed.
Anhydrobiosis- adaptation to desiccation

Both quiescence and crytobiosis induced by dehydration stress constitute anhydrobiotic
survival. In anhydrobiosis, the metabolism comes reversibly to a stand still. Water is an
essential component that constitutes 76% of the volume of animals, it‟s the medium for
biochemical reactions and is essential in the structure of biological macromolecules and
membranes. In general, therefore, most animals die when they lose more than 15-20% of
their body water. Dehydration stress is thus a most important challenge to nematodes since
they are basically aquatic in nature. For nematodes parasitising the aerial parts of the plants
anhydrobiotic survival is a routine part of their life cycle, involving for the most part
specific juvenile stages that are renown for their abilities to survive in this state.
The ability of nematodes to survive depends on their inherent ability to withstand

desiccation and the stage of nematode development; some nematodes survive better under
some moisture conditions than others. Tylenchorhynchus martini survives best in soils
between 40-60% field capacity and survival is lowest at 11% and at saturation. The
optimum soil moisture range is between 40 – 80% of field capacity. Dry soils do not allow
movement and saturated soils lack oxygen and also inhibit free movement. Excessive
moisture kills nematodes through starvation, suffocation or toxins produced by some
anaerobes such as Clostridium spp. against Tylenchorhynchus martini.
For PPN to enter successfully into anhydrobiosis, they must experience slow and controlled
rates of evaporative water loss. Few nematodes are capable of withstanding rapid water loss
and prolonged periods of dehydration. Anhydrobiotic PPN can however be divided into 2
broad categories; slow dehydration strategists (SDS) and fast dehydration strategists (FDS).
Slow dehydration strategists need a slow rate of water loss to successfully enter
anhydrobiosis. They include Pratylenchus thornei, Helicotylenchus dihystera, Scutellonema
cavenessi and Aphelenchus avenae.
Fast dehydration strategists can survive immediate exposure to low relative humidity and
include the anguinids and Ditylenchus dipsaci associated with aerial parts of the plants.
They are able to tolerate rapid and repeated cycles of dehydration and rehydration.
Anguinids induce galls in the host inflorescence which may provide a barrier to water loss
so that their first experience of desiccation may involve a slow rate of water loss. They
however will survive fast rates of water loss on subsequent rehydration and desiccation.
The unhatched infective juveniles of many cyst nematodes are FDS since the egg shell and
the cyst wall may ensure the necessary slow rate of water loss. Fast dehydration strategists
can survive rapid loss of water from their surroundings but themselves have mechanisms
for ensuring the slow loss of water from their body through behavioural and physiological
mechanisms.
34
Species Longevity whilst desiccated
Anguina tritici 32 years

Ditylenchus dipsaci 23 years
Globodera rostochiensis 8 years
Scutellonema cavenessi 1 month
Pratylenchus thornei 1 day
Helicotylenchus dihystera 1 day
Mechanisms of anhydrobiotic survival

They include
i. Adaptations to reduce the rate of water loss (slow dehydration)
ii. Behavioural adaptations
iii. Biochemical and physiological adaptations that help maintain structural and
functional integrity at the cellular level
i. Reducing the rate of water loss

Nematodes have structures that aid anhydrobiotic survival by forming a barrier around the
organism. They include the lipid membranes in eggs, the matrix in egg sacs or cuticles of
juveniles and adults. The restricted permeability of the nematode egg shell and cyst ensure
a slow rate of water loss and may ensure enclosed unhatched J2 survive anhydrobiotically.
The permeability of the egg shell of G. rostochiensis decreases as it dries. A lipid layer is
the main permeability barrier of the egg shell.
The presence of a gelatinous matrix (egg sac) besides the egg shell presents a barrier and
allows slow drying of egg masses of the RKN. The gelatinous matrix is reduced to a tough
granular mat during drying hence providing the first line of defence in providing a barrier
to water loss. When the soil pores around the egg sac drain, the moisture content of the
gelatinous matrix around the eggs decreases only slightly thus maintaining a high level of
moisture inside.
Adults of Rotylenchulus reniformis have a sheath which plays a role in controlling water
loss during desiccation.
Many PPN remain within the senescing plant host tissue and this provides a physical barrier
to slow evaporative water loss.
Induction of galls in the inflorescences by anguinids (Anguina tritici, A. agrostis) is a true

crytobiotic anhydrobiosis. Survival in anguinids is necessary because in order to infect new
hosts, these nematodes must spend sometime exposed to the harsh dehydration regimes
associated with soil - air, plant – air interfaces before being protected by the plant host itself
during the endophytic phase of the life cycle. In anguinids, anhydrobiotic survival involves
slow dehydration within the host itself as an initial phase ensuring the development of
resistant J2. Slow drying enables anhydrobiotes to prepare themselves metabolically for the
onset of anhydrobiosis. It also involves a modification of hosts inflorescence induced by the
35
adult nematodes themselves resulting in the formation of galls around the developing mass
of juveniles. Slow drying is assured by virtue of mutual protection and by the physical
barrier presented by the gall itself. The J2‟s of the two nematodes develop tightly packed
aggregates (not preceded by swarming within the gall). Coiling of individual nematodes
outside of the mass also occurs thus enhancing anhydrobiotic survival. Finally an exodus of
nematodes from senescing plants to younger plants has survival importance.
In Ditylenchus dipsaci, the resistant J4 are able to control water loss to a much greater
extent than other stages in the life cycle because of changes in the permeability
characteristics of their cuticle. The nematode is also adept at clumping, coiling and using
plant tissue to slow rates of drying. Both J4 and to a lesser extent J3 stages have an intrinsic
ability to control water loss through a decrease in permeability of the cuticle as it dries. The
nematode also exhibits behavioural adaptations to dehydration.
Behavioural adaptations to dessication

These are mainly exhibited by FDS and include clumping and coiling.
Clumping
Clumping is often preceded by swarming which is a coordinated/synchronized movement
by a mass of nematodes to specific parts of the host or from one location to another. The
swarms therefore consist of nematodes showing synchronized movements and are involved
in migration and dispersal and may be followed by aggregation where nematodes form
dense masses. As dehydration progresses, the nematodes actively coil before becoming
inactive. Coiling may involve the whole of the nematode mass or just those on the
periphery. Swarming occurs in bacteriophagous and mycophagous nematodes and PPN.
The clumps formed may aid desiccation survival but swarming itself is not triggered by
desiccation. Swarming may be triggered by accumulation of toxic waste products, the
exhaustion of food supplies and / or the senescence of a host plant. In Ditylenchus dipsaci
J4 swarm from senescing host tissue and form clumps/aggregates commonly referred to as
“eelworm wool” and are coiled between scales of infected bulbs (tulips & narcissus) or
inside bean pods (Plate 6a). The formation of clumps aids the control of water loss since
nematodes on the outside will dry first and form a layer that slows down the rate of water
loss from nematodes deeper in the aggregation. This is called the egg-shell effect.
Aggregation also occurs in Aphelenchus avenae and anguinids which accumulate in
infected host inflorescences and induce them to form galls.
Coiling is associated with a wide range of nematodes as a desiccation survival mechanism,

which reduces the rate of water loss by reducing the exposed surface area of the nematode.
Rotylenchulus reniformis coiled nematodes survive desiccation at 80% and 40% relative
humidity. Coiling may follow or occur in conjunction with swarming and aggregation or
the nematodes coil individually. The primary stimulus for coiling behaviour appears to be
the restriction of lateral movement that occurs in an evaporating water film. Nematodes will
also coil in response to increases in temperature and osmotic stress.
Biochemical adaptations to dessication
36
Biochemical changes also occur during entry into anhydrobiosis, for example, elevated
levels of the disaccharide trehalose. Trehalose stabilises membranes during desiccation and
rehydration by attaching to the polar head groups of the phospholipids and preventing
phase changes that may cause the membrane to become leaky; (“water replacement
hypothesis”). Trehalose also stabilises tissues via vitrification by acting as a high viscosity,
low molecular mobility medium. Trehalose also prevents protein denaturation oxidative
damage and browning reactions as a free radical scavenging agent and as an inert energy
source as in A. avenae. There are also desiccation induced heat- tolerant proteins as in A.
avenae.
Physiological adaptations include change in cuticular permeability and a decrease in

surface area by the shrinkage of the body of the nematode as in D. dipsaci J4 which
narrows or deepens the grooves of the cuticular annulations.
Cryobiosis and thermobiosis – adaptation to temperature changes

All organisms have an optimum temperature at which their metabolism and hence growth
and activity are greatest. Temperature is a major factor limiting distribution of PPN world
wide and in different soil depths. Meloidogyne incognita and M. javanica are found in
warmer parts of the world unlike M. hapla and can therefore survive better in high and low
temperatures, respectively. Nematodes are poikilotherms; their body temperature follows
that of the environment. The optimum soil temperature range for activity is 16-290C, above
and below this range, nematodes become inactive and temperatures below 40C and above
400C may be lethal. Nematodes exhibit a range of responses to temperature changes. The
metabolic rate decreases as the temperature decreases due to the lower kinetic energy
imparted to reactions but proteins are not denatured by the low temperature and so the
effects are potentially reversible.
Cryobiosis- Nematodes are more likely to survive very low temperature (< 00C) than high
temperature (>500C) since the adverse effects are potentially avoidable or reversible.
Nematodes can extend the survivable limits of both high and low temperature by triggering
protective responses. Some nematodes enter cryobiosis surviving temperatures below that
at which metabolism ceases with some surviving temperatures as low as minus 800C. As
the temperature falls, nematodes display cold stupor, cold coma and freezing. If the
temperature of an animal is below the temperature at which its body fluids freeze (melting
point of their body fluids), it is at risk of freezing. Nematodes survive these conditions by
being either freeze tolerant (surviving ice formation within their bodies) or freeze averse
(preventing ice nucleation and maintaining their body fluids as a liquid at temperatures well
below their melting point, by super cooling). In freeze tolerant nematodes, the temperature
at which it freezes (the super cooling point) is well above the temperature at which the
animal dies (the lower lethal temperature-LLT) and usually close to its melting point, for
example Ditylenchus dipsaci and Aphelenchoides ritzemabosi. In freeze averse nematodes,
the super cooling point is considerably below its melting point and fairly close to the LLT
and it dies once it freezes. In these nematodes such as G. rostochiensis, they avoid freezing
by preventing ice nucleation and allowing super cooling.
Other adaptations involved in freezing tolerance and cold tolerance include; the production
of sugars (trehalose and glycerol) and polyols as cryoprotectants or anti-freezes, avoidance
37
of membrane transitions which may be involved in non-freezing injury and production of

stress proteins.
Thermobiosis-A nematode exposed to increasing temperature is first likely to display heat

stupor, where movement becomes disoriented or normal processes/ activities are disrupted,
then heat coma, where movement and activity ceases, followed by death at the thermal
death point.
Nematodes may tolerate heat through the production of heat-shock proteins (stress proteins
and molecular chaperones) that both stabilize the target proteins (protecting them against
denaturation) and reactivate damaged proteins. For example, embryogenesis and hatching
of the eggs of M. javanica and M. naasi can be slowed or stopped at elevated temperature
and readily resumed once the temperature stress is removed.
Anoxybiosis – survival at low oxygen levels

Conditions associated with low oxygen tensions are generally a feature of the soil
environment caused by an increased level of soil saturation (up to and including flooding)
which restricts the rate at which oxygen can diffuse from air into the interstitial spaces of
the soil and by the microbial activity placing demands on the available oxygen. Poor
aeration induces cryptobiosis in juveniles enhancing anoxybiotic survival. Eggs of PPN and
juveniles of many soil –dwelling nematodes are able to tolerate low oxygen tension by
reducing normal oxidative metabolism levels and show some capacity for facultative
anaerobiosis by switching to fermentative as opposed to oxidative respiratory pathways
during anoxia. Examples include Caenorhabditis spp, eggs of many Meloidogyne spp and a
majority of free-living mycophagus nematodes. This constitutes a physiological adaptation.
Certain stages of PPN e.g. J2 of Anguina sp. and all stages of Anguina amsinckiae must
endure protracted periods of low oxygen tension and eventually anoxia as the plant
senesces and the galls dry. Response to anaerobiotic conditions depends on developmental
stages and species involved. Trichodorus christiei, Xiphinema americanum are more
sensitive to anaerobiosis than Tylenchulus semipenetrans and M. incognita.
Osmobiosis- survival at high salt levels

A majority of PPN and free living nematodes are osmoconformers, showing limited ability
for osmotic or ionic regulation. High osmotic stress may be produced by drying of the soil,
addition of fertilizers etc. Osmobiosis (which protects a nematode from the effects of
fluctuations in the external physical environment) is evidenced in cyst nematodes
(Globodera rostochiensis and Heterodera goettingiana). The high concentration of
trehalose in the perivitelline fluid of the eggs of Globodera rostochiensis imposes an
osmotic stress which maintains the enclosed J2 in a state of quiescence. Hatching stimulus
results in an increase in permeability of the egg shell, which allows the trehalose to diffuse
out of the egg and for water to enter to remove the osmotic stress. Trehalose is a common
component of the perivitelline fluid (in the perivitelline space of the egg) of a variety of
nematode eggs that do not require specific biotic factors (hatching factors) to break
dormancy. Trehalose acts as a cryoprotectant or antifreeze.
38
In hyposmotic conditions water enters the body and will have to be removed while in
hyperosmotic conditions water will be lost and the nematode stressed. The water lost may
induce a state of osmobiosis which enables the nematode to survive the stress period.
References
Gaugler R. and Anwar B. L. (eds.) 2004. Nematode behaviour. CABI Publishing,
Oxfordshire, UK. 419pp
Lee D. L. (ed.) 2002. The biology of nematodes. Taylor and Francis, London UK. 635 pp
Luc, M., Sikora R. A. and Bridge J. 2005. Plant Parasitic Nematodes in Subtropical and
Tropical Agriculture CABI Publishing, Oxfordshire, UK. 871 pp
Perry R. N. and Wright D. J. (eds.) 1998. The Physiology and Biochemistry of Free – living
and Plant parasitic nematodes. CABI Publishing, Oxfordshire, UK. 438pp
Wajid K. M. (ed.). 1993. Nematode Interactions. Chapman and Hall, London UK. 377pp
39
CHAPTER 3
DIAGNOSIS AND MORPHOLOGY OF PLANT

PARASITIC NEMATODES
Zibusiso Sibanda
Formerly Nematology Department Kutsaga Research Station
PO Box 1909 Harare. Zimbabwe
1. INTRODUCTION
This chapter provides a basic introduction to the diagnosis of plant parasitic

nematodes, particular reference being paid to those groups found in eastern and southern
Africa. The information present in this chapter is a summary of a more comprehensive
training manual that was prepared for the First Gatsby Regional Nematology Training
Course. The detailed manual can be made available on request on CD.
Illustrations have been adapted from the published work of other nematologists and
their invaluable contribution is acknowledged.
COMMON MORPHOMETRIC ABBREVIATIONS
There are a number of abbreviations for the various characters measured and ratios
calculated when compiling morphometric profiles of nematodes. Some of these values are
of more use than others and so an understanding of their potential variability and reliability
is of paramount importance.
The most commonly used abbreviations are as follows:
L = Total body length (head to tail tip).

L‟ = Body length from head to anus/cloaca (used when tail is very long
and therefore subject to greater variation and the likelihood of
breakage).
a = Total body length divided by maximum body width.
b = Total body length divided by oesophageal length (the oesophagus is
defined as head end to oesophago-intestinal junction, i.e. not to the
posterior tip of the overlapping gland lobes).
b' = Total body length divided by distance from anterior end of body to
posterior end of oesophageal gland lobes.
c = Total body length divided by tail length.
c' = Tail length divided by anal/cloacal body width.
V = Position of vulva from anterior end expressed as percentage of body
length. Superior figures refer to the extent of anterior and/or
posterior gonad or uterine sac, also as percentage of body length.
40
V' = Position of vulva from anterior end expressed as percentage of

distance from head to anus (i.e., use L‟, not L).
T = Distance between cloaca and anteriormost part of testis expressed as
percentage of body length.
m = Length of conical part of stylet as percentage of whole stylet length.
o = Distance of dorsal oesophageal gland opening behind stylet knobs as
percentage of stylet length
MB = Distance of median bulb from anterior end expressed as a percentage
of total oesophageal length.
Caudal ratio A = Length of hyaline tail divided by its proximal width.
Caudal ratio B = Length of hyaline tail divided by its width at a point 5µm from its
terminus.
µm = One thousandth of a millimetre (micron).
The following additional parameters are routinely used in the taxonomy of criconematids:
R = Total number of body annules.

Rex = Number of annules from anterior end to excretory pore.
RV = Number of annules from posterior end to vulva.
RVan = Number of annules from vulva to anus.
Ran = Number of annules from posterior end to anus.
2. GENERAL MORPHOLOGY
It is not the object of this module to provide any depth of detail concerning
nematode morphology or classification. Nonetheless, it is important to understand some of
the fundamentals of nematode morphology in order to more fully understand the biology of
nematode-plant interactions.
Nematodes are lower invertebrate animals. They are highly diversified and perhaps
the most numerous multicellular animals on the earth. Like insects, they are found in
almost all types of biotypes and occur in unimaginable numbers and in a wide variety of
shapes and sizes. Nematodes are generally free-living in marine, freshwater or soil
environments, but a large number of species are parasitic on different kinds of plants and
animals. The parasitic species are of considerable agricultural, clinical and veterinary
importance as pests of plants and parasites of Man and livestock respectively. Nematodes
are found at the bottom of lakes, rivers and at enormous depths in the oceans. Some
species can withstand temperatures constantly below freezing point while others live in the
waters of hot springs.
By 1930 some 4,500 species of nematode had been described. This rose to 9,000 by
1950, and the present-day number of known species of nematodes is well over 15,000. The
estimated number of existing species ranges from 500,000 to several million, but the truth
is that nobody has much of an idea of their total number. This means that the majority of
41
nematode species are not yet known and the taxonomy and phylogeny of this group are thus
still very much in the early stages.
Nematode morphology is only visible to us after special procedures for extraction
and fixation, and by means of complex instruments of observation. These procedures and
instruments completely determine our knowledge of nematode morphology, because they
impose a strict limit on the level of detail with which we can study nematodes. They may
even lead us to substantial errors through the incorrect interpretation of the artefacts which
they can generate.
All these factors boil down to the following essentials: describing the morphology
of nematodes requires a great degree of jargon, because of their diversity and uniqueness,
as well as a great degree of caution and training in microscopical observation. The
following paragraphs will introduce you to the major descriptive terms in nematode
morphology. Although real experience can only be gained by examining fixed and live
nematodes for yourself, knowledge of the terminology should at least allow you to identify
the main organs and structures in the nematode body, and will thus set you on your way to
chart the endless diversity of this phylum.
Fig. 1. – Anatomy of a plant-parasitic nematode : Rotylenchoides variocaudatus

(Dropkin, 1980).
42
3. PRINCIPLES OF SYSTEMATICS2
Taxonomy (Greek taxis = arrangement; nomos = law) deals with the naming and
recognizing of taxa, and systematics (Greek systema = whole consisting of parts) is a
method of arranging taxa into a hierarchical system of classification. Unfortunately, these
two terms are often confused, even by people who should know better. The fashionable
term 'biosystematics' is usually used to embrace both concepts.
The classification of animals is a basic part of zoology. It serves the purpose of
recognizing, and hence distinguishing, animals with respect to each other.
Classification at present is based on taxonomic characters, mainly from
morphology and comparative anatomy. These taxonomic characters have different values
and significance at different grades and categories. For example, there are characters of
specific, generic and familial values. Determination of their value, or weighting, is done
through using various methodologies as will be discussed here.
The monothetic concept in classification is based on the use of a single character or
feature as against the polythetic concept which involves the use of several characters. The
monothetic concept has often led to ambiguities in classification.
Rules of Zoological Nomenclature
The taxonomy and classification of animals are governed by rules listed in The
International Code of Zoological Nomenclature (ICZN). The Code covers categories from
subspecies to superfamily only. It does not cover ordinal, class, or Phylum ranks, nor does
it cover the infra-subspecies categories such as form, variety, strain, biotype, etc.
Species are based on populations of individuals which can be studied, but genera
and higher categories are more abstract, often with extremely vague boundaries and limits.
The name of a species is binominal, i.e. it is a binomen consisting of two words, generic
and specific. The genus- and species-group names should appear in italics or in a type face
different from that of the text. All the names in suprageneric categories are uninomina, i.e.
made up of one word and not italicized. All the names of taxonomic categories except
species and subspecies names begin with a capital letter.
In Tylenchida, taxa are recognized and classified mainly on morphological
characters. It is only recently that ecological, biochemical, cytological and embryological
characters have been introduced, but still in a very limited manner. The
characterization of taxonomic groups involves morphological, biochemical and cytogenetic
taxonomical methods while determining their rank is done by three broadly categorized
methods:
(1) evolutionary systematics

(2) cladism
(3) phenetics
2
After M R Siddiqi
43
A. Morphological Taxonomy:
Morphological characters provide more than 90% of the data used in taxonomy and
classification of plant and soil nematodes. These characters show considerable variation
and care must be taken in using them. Morphometric and allometric measurements often
vary greatly under the influence of geographical and ecological conditions. Techniques,
types of equipment, methods of observation and personal skills also result in variations.
Some characters used to differentiate species of Pratylenchus, e.g. stylet knob
shape, length of outer margins of cephalic framework, the shape of the spermatheca and tail
tip, tend to vary through host influence.
B. Biochemical Taxonomy:
Gel electrophoresis has been widely used as a technique in the identification of

plant-parasitic species. Evans (1971) was able to distinguish between Ditylenchus
destructor and D. myceliophagus populations through the study of their esterase patterns as
revealed by gel electrophoresis. Different protein band profiles were obtained by using disc
electrophoretic separation of soluble proteins and enzymes of some species of Ditylenchus,
Heterodera and Meloidogyne (Dickson et al., 1971). However, these techniques have been
used only in a limited manner, although the scope is considerable. Immunological
techniques involving DNA hybridization and isoenzyme analysis have been applied to
certain aspects of plant nematode taxonomy, although specialized biochemical knowledge,
skills and instrumentation are required to use such techniques. The current way forward
seems to be via molecular techniques, particularly those employing polymerase chain
reaction (PCR) amplification of DNA.
C. Cytogenetic Taxonomy:
Cytological analysis can be utilised in the differentiation of species and pathotypes.

Chromosome number has been used in the differentiation between some species of
Meloidogyne and between Radopholus similis similis and R. s. citrophilus, although in the
latter case chromosome number has been shown to be more variable than previously
thought and therefore useless as a race marker. Chromosome number also differs between
nematode groups, for example, Criconematoidea and Hoplolaimidae have a basic
chromosome number of n = 5; Dolichodoridae including Belonolaiminae have n = 8.
1. Evolutionary systematics
The traditional evolutionary approach involves the reconstruction of phylogeny by using
past histories of animals as revealed by the study of their fossils and comparative
morphology/anatomy.
2. Cladism
The cladistic analysis is a kind of interpretation of similarities in and between taxa. The
similarities are established by in-group and out-group comparison of two types of
characters or character states.
44
3. Phenetics
This approach tries to classify organisms on over-all similarities in their structure
without bothering to know how they have come about. The classification of Tylenchida
has been based almost entirely on structural similarities which have seldom been
supplemented with ecological, ethological and physiological data. For this order, the
phenetic approach will continue to have a major role to play since identification and
classification will have to be based largely on morphological data.
The use of a large number of characters, as in numerical taxonomy, has yielded
consistent results in developing hierarchical classification. Sokal & Sneath (1963, p. 264)
state "One of the attractive properties of taxonomies based on large numbers of characters
is their robustness under different statistical treatments. They consider all characters of
equal weighting. Key characters, however, are recognisable after a careful over-all
comparison has been made.
REFERENCES AND ADDITIONAL READING
Blackith, R.M. and Blackith, R.E. 1976. A multivariate study of Tylenchus Bastian, 1865
(Nematoda, Tylenchidae) and some related genera. Nematologica 22, 235-259
Cobb, N.A. 1920. One hundred new nemas. Contrib. Sci. Nematol. 9, 217-343
Coomans, A. 1983. General principles for the phylogenetic systematics of nematodes. In:
Concepts in Nematode Systematics. (Ed. Stone, A.R., Platt, H.M. & Khalil, L.F.). London
& New York: Academic Press, pp. 1-10
Dickson, D.W., Huisingh, D. and Sasser, J.N. 1971. Dehydrogenases, acid and alkaline
phosphatases, and esterases for chemotaxonomy of selected Meloidogyne, Ditylenchus,
Heterodera and Aphelenchus spp. Journal of Nematology 3, 1-16
Evans, A.A.F. 1971. Taxonomic value of gel electrophoresis of proteins from

mycetophagous and plant parasitic nematodes. Int. J. Biochem. 2, 72-79
Fortuner, R., Maggenti, A.R. and Ehittaker, L.M. 1984. Morphometrical variability in
Helicotylenchus Steiner, 1945: 4. Study of field populations of H. pseudorobustus and
related species. Rev. Nematol. 7, 121-135
Hennig, W. 1966. Phylogenetic Systematics. Urbana, USA: Univ. Illinois Press, 263pp
Hooper, D.J. 1969. Identification of plant and soil nematodes. In: Nematodes of Tropical
Crops (Ed. Peachey, J.E.). St Albans, UK: Comm. Bur. Helminth., Tech. Commun. No. 40,
pp. 37-66
45
International Code of Zoological Nomenclature, 1985. III Edit. International Trust for
Zoological Nomenclature, c/o Commonwealth Institute of Entomology, 56 Queen's Gate,
London SW7 5JR
Jones, F.G.W. 1965. Parasitism in plant nematodes. In: Plant Nematology (Ed. Southey,
J.F) Tech. Bull. (MAFF) No. 7, London, UK: HMSO, pp.30-34
Mai, W.F. and Lyon, H.H. 1975. Pictorial key to genera of plant-parasitic nematodes.
Fourth edition, revised. Ithaca & London: Comstock Publishing Associates, Cornell
University Press, 219 pp
Mayr, E. 1969. Principles of Systematic Zoology. New York, USA: McGraw-Hill, 428pp
Moss, W.W. and Webster, W.A. 1970. Phenetics and numerical taxonomy applied to
systematic Nematology. Journal of Nematology 2, 16-25
Platnick, N.I. 1977. Cladograms, phylogenetic trees, and hypothesis testing. Syst. Zool. 26,
438-442
Simpson, G.G 1961. Principles of Animal Taxonomy. New York, USA: Columbia
University Press, xii + 247pp
Sokal, R.R and Sneath, P.H.A. 1963. Principles of Numerical Taxonomy. San Francisco,
USA: Freeman & Co., 359pp
Triataphyllou, A.C. 1970. Cytogenetic aspects of evolution of the family Heteroderidae.

Journal of Nematology 2, 26-32
Triantaphyllou, A.C. 1983. Cytogenetic aspects of nematode evolution. In: Concepts in

Nematode Systematics. (Eds Stone, Platt, H.M. & Khalil, L.F.) Academic Press, pp. 55-71
Williams. T.D. 1968. Plant-parasitic nematodes. In: Pathologist's Pocketbook. Kew, UK:
Commonwealth Mycological Institute, pp. 119-136
4. SYSTEMATIC SCHEME
The following simplified scheme will be followed throughout this manual. It

should not be interpreted as the only valid scheme and can be modified according to
personal requirements/preferences/whim. Only a small proportion of the genera listed will
be dealt with in detail and many will not be mentioned further as they are not of importance
in eastern and southern Africa.
The scheme is based on the published works of Siddiqi (1986; 2000) and Hunt
(1993) and updated where necessary. Synonyms and subgenera are not listed in order to
46
enhance clarity. The relevant section of the scheme is repeated for convenience as each
major group is considered in the manual.
Major nematode genera of economic importance, together with confusable genera,
in eastern and southern Africa are printed in bold type.
5. ORDER TYLENCHIDA
This order contains the majority of plant parasitic nematodes and is usually divided into
three suborders. These are the Tylenchina, Criconematina and Hexatylina, the majority of
plant parasites being in the two former groupings. In some schemes there is a fourth
suborder, the Aphelenchina, although in this manual the aphelenchs are regarded as
belonging to a separate order. In practical respects it matters little which scheme is
embraced, the higher classification of nematodes being somewhat arbitrary, particularly at
present with the advent of molecular-based phylogenies.
SUBORDER TYLENCHINA
Diagnosis: Tylenchida. Males with normal oesophagus and stylet (stylet occasionally
reduced in some Hoplolaimoidea). Phasmids present, sometimes enlarged, scutellum like
and migratory (Hoplolaiminae), in Tylenchoidea papilla-like phasmids or phasmid-like
structures present in submedian position on body, in female near vulva, just dorsal to the
lateral fields. Cuticle often distinctly annulated, sometimes marked with longitudinal striae
or grooves, but annules never retrorse or with scales, spines, appendages or a double
cuticle. Cephalic region generally distinctly annulated. Stylet variable in length; conus
normally as long as shaft, if elongate then shaft more than 10µm long; knobs variable in
shape and size but never large and anchor-shaped as in Criconematoidea of Criconematina,
occasionally absent (e.g. Psilenchinae). Precorpus (= procorpus) cylindroid or fusiform;
postcorpus or median bulb muscular with valve plates but not amalgamated with precorpus
into a large muscular cylinder, or a non-muscular fusiform swelling. Isthmus slender.
Oesophageal glands forming a basal bulb or extending over intestine. Oesophago-
intestinal valve (= cardia) 3-celled, reduced in forms with overlapping glands. Nerve ring
circum-oesophageal. Excretory pore generally in oesophageal region.
Female: Slender, or obese (spherical, kidney- or lemon-shaped) and may turn into a cyst in
some sedentary Hoplolaimoidea. Usually didelphic, amphidelphic (becoming didelphic-
prodelphic in Heteroderidae and Meloidogynidae). Posterior reproductive branch
secondarily reduced or represented by a postvulval uterine sac (e.g. Pratylenchinae,
Trophurinae), but in Tylenchoidea as a rule lacking, except for a uterine sac. Vulva a large
transverse slit, rarely oval (e.g. some Merliniinae), equatorial or submedian, shifted to lie
subterminally or terminally in swollen females. Ovaries generally paired, normally
outstretched and with serially arranged oocytes, rachis absent (except in obese females).
Males: Oesophagus and stylet normally developed (except some Hoplolaimoidea).

Testis single (except some Meloidogyne spp. in which testes are paired, probably due to sex
47
reversal; the female primarily being didelphic) outstretched. Spicules paired, rarely
dissimilar, cephalated, arcuate, distally round to pointed, never setaceous or U-shaped.
Gubernaculum simple or modified; fixed or protrusible.
Juveniles: Similar to female in most details occasionally with reduced oesophagus and
stylet (e.g. Rotylenchulus) due to rapid development from egg to adult without feeding.
Bionomics: Algal and moss feeders, and parasites of lower and higher plants; attacking
subterranean parts, not parasitic in above-ground parts of plants or in animals.
Key to superfamilies of Tylenchina
1. Phasmids not detectable on tail; phasmid-like

structures present much anterior to tail
region, dorsal to lateral fields, in female
near vulva; tails generally filiform .............................................Tylenchoidea
Phasmids present in or near tail region (except

for migratory scutella of Hoplolaiminae),
in lateral fields, not near vulva; tails generally
not filiform, if filiform then with distinct
phasmids ..........................................................................................................2
2. Subventral oesophageal glands enlarged, usually

extending past the dorsal gland; sexual
dimorphism in anterior region manifest .................................Hoplolaimoidea
Subventral oesophageal glands not enlarged, not

extending past the dorsal gland; sexual
dimorphism in anterior region not manifest ..........................Dolichodoroidea
5.1. SUPERFAMILY TYLENCHOIDEA
The classification of this superfamily is fairly complex and open to differing points
of view. Although there are many nominal genera, it is not usually necessary to identify the
group much beyond a general catch-all grouping such as Tylenchus, sensu lato (i.e. in the
'broad sense' and taken to include all the related genera).
Most of the genera of this superfamily are associates of algae, moss, lichen and
plant roots. They are not root parasites of any agricultural significance and so will be dealt
with only superficially. They are, however, commonly encountered in large numbers in
washings from soils rich in organic matter and so need to be recognised and separated from
the potentially more damaging plant parasitic nematodes.
48
These nematodes are considered to be fairly primitive in form, being a conservative

group with many ancestral characteristics (e.g. weak feeding apparatus, poorly developed
corpus, filiform tail, etc.).
FAMILY TYLENCHIDAE Örley, 1880
Diagnosis: Tylenchoidea. Small to moderately large (0.3-1.3mm), not abnormally

slender nematodes. Lateral field with 2, 3 or 4 incisures. Labial disc inconspicuous or
absent; a perioral disc present in Discotylenchus. Stylet small (6-21µm), conus usually
less than half total stylet length, rarely with distinct lumen; basal knobs or swellings small,
rounded, or absent (e.g. Neopsilenchus, only subventral knobs absent in Irantylenchus);.
Orifice of dorsal gland close to, or at some distance (Boleodorinae, Thadinae) from, stylet
base. Precorpus cylindrical. Post corpus or median bulb fusiform or oval, not filling body
width, muscular and valvate, or non-muscular, non-valvate; may be absent. Isthmus
slender. Basal bulb with glands abutting intestine, with cardia at base. Vulva a transverse
slit; lateral membranes rarely present (Aglenchus). Postvulval uterine sac one body width
or less offset (except Thadinae), often elongated and directed anteriorly. Ovary single,
anteriorly outstretched, oocytes usually in a row. Testis outstretched, tip rarely reflexed.
Spicules small, slender, arcuate. Gubernaculum linear or trough-shaped, fixed. Tails
generally filiform, similar in both sexes. Bursa adanal, simple, not lobed.
Note: The important characteristics of this group include:

1) weakly sclerotised head skeleton
2) stylet and basal knobs weak
3) basal bulb not overlapping intestine
4) genital tract monodelphic, prodelphic
5) „phasmid‟ present near vulva, not usually within lateral field
6) tail elongate-filiform in both sexes
7) bursa adanal
The body is generally vermiform (worm-like), with the posterior portion slender and
tapering sharply to pointed or rounded terminus. The vulva is positioned in the posterior
section of the body, although due to the attenuated tail of some genera, the vulval ratio can
be greatly affected by variations in development of, or damage to, the filamentous portion.
This is resolved by using V' (dividing the length from head to vulva by the length from
head to anus  100). On death the body can be slightly curved ventrally (Discotylenchus)
to open C-shaped (Tylenchus).
The oesophagus is very variable in shape. The procorpus can be with or without
median bulb. There is usually a long isthmus and basal bulb. The basal bulb can overlap
the intestine, but the gland nuclei remain within the bulb. The head shape is variable and
can be used to distinguish genera and species. The stylet is usually delicate and short
(average for the family is about 13µm).
The genital tract is monodelphic, prodelphic with a post-vulval uterine sac of
variable length usually present. The length of the uterine sac is important in identifying
49
genera and species. The area around the vulva is important (the presence or absence of
cuticular flaps, angle at which the vagina exits via the vulva).
The female tail shape is variable between genera, but is usually attenuated and
tapering to a pointed terminus. The male tail is similar in shape to that of the female,
except for the presence of spicules, gubernaculum and bursa. The spicules are tylenchid in
shape, with a simple gubernaculum and the bursa is adanal. The presence of males in
populations is dependant on species - some are parthenogenetic and others amphimictic.
Bionomics: Associates of algae, mosses, lichens and plant roots, but generally not root
parasites of any significance.
Key to major subfamilies of Tylenchidae
1. Amphidial apertures prominent, posterior to level of cephalic

papillae, partially covered by a cuticular flap ..............................Boleodorinae
Amphidial apertures rarely prominent, anterior to cephalic
papillae, not covered by a cuticular flap ...........................................................2
2. Lateral field with 2 incisures (single ridge) .................................Duosulciinae

Lateral field with 3 or 4 incisures (2 or 3 ridges), rarely obscure ....................3
3. Crustaformeria generally tricolumellate; spermatheca offset; tails

elongate filiform ............................................................................Tylenchinae
Crustaformeria quadricolumellate; spermatheca axial; tails short,
conoid to subcylindrical .....................................................................Thadinae
Subfamily TYLENCHINAE Örley, 1880 (Marcinowsky, 1909)
Diagnosis: Tylenchidae. Body about 0.3-1.3mm long. Cuticle finely striated, or coarsely
annulated (but smooth in Polenchus). Lateral field with 3 or 4 incisures. Cephalic
region continuous or slightly offset. Stylet 6-21µm long, conus shorter than or rarely as
long as shaft, with tip appearing solid; knobs distinct (except Irantylenchus). Orifice of
dorsal oesophageal gland at about a quarter or less of stylet length behind stylet base.
Muscular median oesophageal bulb present or rarely absent (Sakia). Basal bulb offset
from intestine. Vulval lips simple, or modified (Aglenchus). Post vulval uterine sac
present (except Aglenchus). Ovary single, outstretched, occasionally reflexed at the tip.
Testis outstretched; sperm small, rounded. Spicules and gubernaculum typical of the
family. Bursa adanal, simple. Tail generally filiform in shape.
Type genus:
Tylenchus Bastian, 1865
Other genera:
50
Tylenchus Bastian, 1865

Coslenchus Siddiqi, 1978
Filenchus Andrássy, 1954 (Meyl, 1961)
Key to major genera of Tylenchinae
1. Median oesophageal bulb absent or represented by a

non-muscular, non-valvate swelling .........................................................Sakia
Median oesophageal bulb present, muscular, valvate ..................................... 2
2. Subventral knobs of stylet absent ................................... ............Irantylenchus

Subventral knobs of stylet present .. .................................................................3
3. Cephalic region with a perioral disc .........................................Discotylenchus

Cephalic region without a perioral disc . ..........................................................4
4. Longitudinal ridges outside lateral fields present ... ........................Coslenchus

Longitudinal ridges outside lateral fields absent ............................................. 5
5. Cuticle non-annulated .......................................................................Polenchus

Cuticle annulated .. ...........................................................................................6
6. Postvulval uterine sac absent;

male cloacal lips tubular . .................................................................Aglenchus
Postvulval uterine sac present;
male cloacal lips not tubular .. ..........................................................................7
7. Tail ventrally arcuate or hook-like ...................................................Tylenchus

Tail not ventrally arcuate or hook-like .............................................Filenchus
Genus TYLENCHUS Bastian, 1865

syn. Tylelenchus Bastian, 1865 (= officially rejected name)
Aerotylenchus Fotedar & Handoo, 1979
Areotylenchus Fortuner, 1984 (junior objective synonym of Aerotylenchus)
Diagnosis: Tylenchinae. Small to medium sized (0.4-1.3mm), ventrally curved upon

relaxation. Cuticle moderately thick (1-2µm), distinctly annulated. Lateral fields each
with four incisures. Cephalic region continuous, annulated; framework with light or no
sclerotization. Stylet 8-21µm long, with conus comprising between one third and half of
stylet length and round, posteriorly sloping basal knobs. Median oesophageal bulb oval,
muscular, anterior to middle of oesophagus and with refractive valve plates; basal bulb
pyriform. Cardia distinct. Excretory pore usually opposite basal bulb. Vulva a transverse
slit, usually at 60-70% of body length, lips not modified; epiptygma or lateral membranes
absent. Vagina generally at a right angle to body axis. Postvulval uterine sac about a
body width or less long. Spermatheca round to oval, offset. Ovary outstretched. Tail
ventrally arcuate, often hooked, regularly tapering to a pointed or minutely rounded
51
terminus. Bursa adanal, margins crenate. Spicules cephalated, arcuate, 13-25µm long.
Gubernaculum simple, fixed. Cloacal lips slightly raised, anterior one pointed, posterior
usually rounded; not tubular. Hypoptygma absent.
Type species:
Tylenchus davainei Bastian, 1865
Other species:
Many other species have been named, but only a fraction of these are regarded as
valid, the remainder being species inquirendae et incertae sedis.
Andrássy I. (1979). The genera and species of the family Tylenchidae Örley, 1880
(Nematoda). The genus Tylenchus Bastian, 1865. Acta zool. hung. 25: 1-33.
(Nematoda). The genus Aglenchus (Andrássy, 1965) Meyl, 1961, Miculenchus Andrássy,
1959, and Polenchus gen. n. Acta zool. hung., 26: 1-20.
(Nematoda). The genera Malenchus Andrássy, 1968. Acta zool. hung., 17: 1-47.
(Nematoda). The genera Cephalenchus (Goodey, 1962) Golden, 1971 and Allotylenchus
gen. n. Acta zool. hung., 30: 1-28.
Geraert E. and Raski D.J. (1987). A reappraisal of the Tylenchina (Nemata) 3. The family
Tylenchidae Örley, 1880. Revue de Nématologie 19: 143-161.
Golden A.M. (1971). Classification of the genera and higher categories of the order
Tylenchida (Nematoda) In: Zuckerman B.M., Mai W.F. & Rohde R.A. (eds.) Plant
parasitic nematodes Vol. 1, London & New York, Academic Press : 191-232.
Maggenti A.R., Luc M., Raski D.J., Fortuner R. and Geraert E. (1987). A reappraisal of the
Tylenchina (Nemata) 2. The sub-order Tylenchina. Revue de Nématologie 10: 135-142.
Raski D.J., Koshy P.K. and Sosamma U.K. (1982). A revision of the sub-family
Ecphyadophorinae Skarbilovich, 1959 (Tylenchidae: Nematoda). Revue de Nématologie 5:
119-138.
Raski D.J. and Maggenti A.R. (1983). Tylenchidae: Morphological Diversity in a Natural
Evolutionary Group. In: Stone A.R., Platt H.M. & Khalil L.F. (eds) Concepts in Nematode
Systematics, London & New York, Academic Press : 131-142.
52
Siddiqi M.R. (1971). Structure of the oesophagus in the classification of the superfamily
Tylenchoidea (Nematoda). Indian Journal of Nematology 1: 24-43.
Siddiqi M.R (1986). Tylenchida: Parasites of plants and insects. CAB International,
Wallingford, UK, 645pp.
Siddiqi M.R (2000). Tylenchida: Parasites of plants and insects. 2nd edition, CAB
International, Wallingford, UK. (in press).
5.2. SUPERFAMILY DOLICHODOROIDEA
Diagnosis: Tylenchina. No marked sexual dimorphism in anterior region. Cuticle

prominently annulated, not showing distinct outer and inner layers. Lateral fields each with
2-6 incisures reducing towards extremities, except Belonolaimus which has one incisure.
Phasmids small. Cephalic framework mostly with high arches and conspicuous extensions
projecting posteriorly, with light to heavy sclerotization; annules generally distinct, basal
annule not indented; labial disc prominent in some members of Tylenchorhynchinae and
Belonolaiminae which have very long stylets. Stylet long (over 100µm) to short (about 10-
12µm), with distinct basal knobs (knobs absent in Psilenchidae). Orifice of dorsal
oesophageal gland near stylet base. Corpus with cylindrical precorpus (= procorpus) and a
muscular round to oval postcorpus (= metacorpus) having refractive valve plates.
Isthmus slender. Basal (terminal) oesophageal bulb present, usually containing the
oesophageal glands. Occasionally, the dorsal gland enlarges and overlaps anterior
end of intestine, while subventral glands remain small and anterior to the dorsal gland
and may or may not overlap intestine. Oesophago-intestinal valve or cardia 3-celled,
well-developed, but reduced in forms with overlapping glands. Female reproductive
system didelphic, amphidelphic, secondarily becoming pseudo-monoprodelphic by the
reduction of the posterior branch in Trophurinae. Vulva a transverse slit, rarely round or
oval, median or postmedian, with or without epiptygma; lateral vulval membranes absent
(except in subgenus Dolichorhynchus). Vagina at right angle to body axis, cuticularized in
Dolichodorinae. Ovaries outstretched, oocytes in one row except in region of
multiplication. Female tail rarely less than two anal body widths long, variously
modified being conoid, cylindroid, subclavate or elongate-filiform, similar to that of the
male in Psilenchidae. Male monorchic, with outstretched testis; sperm small to moderately
large, rounded, with little cytoplasm. Bursa enveloping entire tail or, in Psilenchidae,
adanal. Spicules symmetrical, cephalated, ventrally arcuate, with or without distal flanges
(vela), tip broadly rounded or pointed, with terminal or subterminal pore. Gubernaculum
simple or modified, fixed or protrusible. Juveniles essentially similar to female. Migratory
ectoparasites of roots.
53
FAMILY DOLICHODORIDAE
The Dolichodoridae comprises a large number of diverse species found throughout

the world although perhaps most common in cooler, more temperate soils. All the species
are ectoparasitic on the roots of plants and some species have been shown to be of
economic importance on certain crops. In this scheme the lateral line character is
maintained as a reliable and easily ascertained means of identifying groups (= genera) of
similar nematodes. Personal preference, however, may dictate an alternative scheme
involving several large genera, each embracing a large number of species, the number of
lateral lines still being utilized, but as a means of division into smaller, more manageable
groups of species.
Characters used in the identification of Dolichodoridae
(i) Body size and death posture: large sized (1.5mm or longer) forms include
Dolichodorus, Belonolaimus, Macrotrophurus while medium sized (0.7 - about 1mm) and
small sized (under 0.7mm) constitute almost all the rest of the dolichodorids. The body
shape is always elongate-cylindrical tapering towards both ends, it is never obese. The
death posture is usually straight to arcuate, but is characteristically spiral in
Neodolichodorus and some Amplimerlinius.
(ii) Cuticle: The cuticle is at least 2 layered, marked by transverse striae making
coarse or fine annules (striae absent in Macrotrophurus). The lateral regions are thickened
and marked by longitudinal incisures except in Belonolaimus which has a single lateral
groove on each side running from head to tail tip. Lateral fields may or may not be
areolated, i.e. traversed by the transverse striae, and the areolation may be complete or
incomplete. Longitudinal striae may be present all along the body (Tessellus and
Scutylenchus) or may be restricted to the anterior end.
The character of the presence of either 6, 5 or 3 incisures in the lateral field has been
used to distinguish Merliniinae, Quinisulcius, Uliginotylenchus and Divittus, respectively.
Triversus and Dolichodorus were differentiated from Tylenchorhynchus and
Neodolichodorus, respectively also by 3 versus 4 incisures.
The presence of longitudinal cuticular ridges outside the lateral fields has been used
to diagnose the genera Trilineellus, Mulkorhynchus, Neodolichorhynchus and
Prodolichorhynchus.
Localized thickening of the cuticle at the tail terminus has been used to characterize
the genera Trophurus, Macrotrophurus, Paratrophurus, Amplimerlinius and Bitylenchus.
(iii) Cephalic region: Shape (low, high, offset, continuous, smooth or striated) and
degree of sclerotization of the cephalic frame-work are important characters. Several
genera do not have a distinct labial disc, but Dolichodorus, Belonolaimus, Sauertylenchus
have a rounded labial disc.
54
(iv) Stylet: A short stylet (under 35µm) with conus about as long as the shaft is a
common feature, although some genera have a very long stylet. Basal knobs are always
present, their shape having diagnostic value. A long stylet with the conus much longer than
the shaft is found in Dolichodorus, Neodolichodorus, Macrotrophurus and Hexadorus. As
a rule, a long stylet has the conus tubular until its tip and a short stylet has a conus that
appears to be solid in its anterior third because the aperture is located a little behind the tip.
(v) Oesophagus: Most dolichodorids have a tylenchoid oesophagus with a muscular

median bulb, a slender isthmus and a glandular, non-overlapping basal bulb separated from
the intestine by a cardia. Belonolaiminae and Telotylenchinae, however, have the glands
lobe-like and extending over the anterior end of the intestine; the dorsal gland forming most
of the lobe.
(vi) Intestine: A simple tube with the lumen indistinct. It extends into the tail
cavity in some groups (e.g. in Bitylenchus, Trophurus, Dolichodorinae), but never does so
in Merliniinae. Serpentine canals or fasciculi may or may not be present.
(vii) Tail: One of the most important characters is the size and shape of the female
tail. The male tail is always conical and enveloped by a bursa. The tail is relatively long
when compared to the anal body width and this is a simple and relatively reliable method
for distinguishing members of this group from the hoplolaimids.
(viii) Female reproductive system: The shape and position of vulva,

presence/absence of epiptygma, sclerotization of vagina and whether the spermatheca is
axial or offset and spherical or lobed when distended with sperm are all important
characters. As a rule, dolichodorids are didelphic with two equally developed reproductive
branches (i.e. amphididelphic), but in Trophurus the posterior branch is rudimentary (i.e.
pseudomonodelphic).
(ix) Male reproductive system: The testis is single and outstretched and produces
small rounded sperm. Shape and size of spicules, gubernaculum and bursa are all important
characters. The spicules have a broadly rounded tip with a large central depression in
Merliniinae, whereas most other dolichodorids have spicules with a narrow tip and large
distal flanges. The gubernaculum is trough-like and fixed in Merliniinae and rod-like and
protrusible in Telotylenchinae and many other groups.
Family DOLICHODORIDAE Chitwood in Chitwood & Chitwood, 1950
Diagnosis: Dolichodoroidea. Small to large sized (generally about 0.5-1.5mm), straight,

usually arcuate, or more curved. Cuticle strongly annulated (except in Macrotrophurus).
Lateral fields each with 1-6 incisures. Phasmids pore-like, caudal. Cephalic region
annulated, round. Stylet well developed, knobbed; conus not much shorter than the shaft.
Oesophageal glands generally either in a basal bulb abutting intestine, or the dorsal
gland is enlarged, forming most of the lobe extending over intestine; subventral glands
not enlarged. Didelphic, amphidelphic, rarely (in Trophurinae) pseudo-monoprodelphic.
55
Vulva median or submedian, transverse slit-like, rarely transversely oval (e.g. Nagelus);
lips simple or modified. Vagina sometimes (Dolichodorinae, Belonolaiminae)
cuticularized. Tails dissimilar between sexes.
Key to subfamilies of Dolichodoridae
1. Female tail elongate-conoid with narrow, mucronate,

spicate or finely drawn out tip except Neodolichodorus
in which tail is obtusely rounded, under 2 anal body
widths long and with terminal cuticle not abnormally
thickened; bursa trilobed .................................................................................2
Female tail not elongate-conoid or with narrow, mucronate,
spicate or finely drawn out tip (except rarely in some Merlinius
and Triversus spp.), if under 2 anal body widths long, then
with terminal cuticle abnormally thickened; bursa not
trilobed (except Dolichorhynchus in which only the terminus
is trilobed) ........................................................................................................3
2. Stylet very long (over 50µm), conus much longer than the
shaft; labial disc conspicuous ..................................................Dolichodorinae
Stylet not very long (under 40µm), conus about as long as
the shaft; labial disc inconspicuous ..............................................Meiodorinae
3. Cephalic region usually 4-lobed; lateral lip areas reduced;

amphidial apertures longitudinal slit-like; oesophageal
gland lobe-like, extending over intestine, oesophago-
intestinal junction less than one body width behind
median bulb .............................................................................Belonolaiminae
Cephalic region not usually 4-lobed; lateral lip areas not reduced,
amphidial apertures pore-like or transversely oval; oesophageal
glands in a bulb abutting intestine, if forming a lobe extending
over intestine (e.g. Telotylenchinae) then oesophago-intestinal
junction more than one body width behind median bulb .................................4
4. Posterior ovary degenerate or absent ...........................................Trophurinae

Posterior ovary normal .....................................................................................5
5. Lateral field with 6 incisures; male with hypoptygma (paired

papillae on posterior anal lip) ........................................................Merliniinae
Lateral field with 3-5 incisures; male without hypoptygma . ...........................6
6. Amphidial apertures conspicuous, postlabial; stylet over
80µm long, conus much longer than the shaft ........ ...........Macrotrophurinae
Amphidial aperture inconspicuous, labial; stylet under
50µm, conus not much longer than the shaft ................................................... 7
56
7. Dorsal oesophageal gland forming a long lobe over intestine,

its nucleus located behind oesophago-intestinal junction ....... Telotylenchinae
Dorsal oesophageal gland not forming a long lobe over intestine,
its nucleus located in front of
oesophago-intestinal junction ......................................... Tylenchorhynchinae
Subfamily DOLICHODORINAE Chitwood in Chitwood & Chitwood, 1950
Diagnosis: Dolichodoridae. Large sized (1.5-3mm). Lateral field with 3 or 4 incisures,

areolated. Cephalic region offset, distinctly annulated, framework strongly cuticularized;
labial disc distinct; stylet very long (over 50µm) with conus markedly longer than
shaft, and prominent knobs. Intestinal fasciculi and postrectal sac present. Vagina vera
cuticularized. Ovaries paired. Female and juvenile tails long, filiform to spicate or, if
obtuse to mammillate then less than 2 anal body widths long and with annulated terminus.
Male tail short, conical, bursa trilobed with 2 large lateral flap-like lobes and a small
terminal lobe; spicules with or without distal flanges; gubernaculum large, protrusible or
fixed.
Type genus:
Dolichodorus Cobb, 1914
Other genus:
Neodolichodorus Andrássy, 1976
Key to genera of Dolichodorinae
1. Lateral field with 3 incisures; female tail spicate or filiform ..... Dolichodorus
Lateral field with 4 incisures; female tail obtuse
or mammillate ........................................................................Neodolichodorus
Genus DOLICHODORUS Cobb, 1914
Diagnosis:. Dolichodorinae. Body straight to arcuate when relaxed. Lateral field with 3
incisures, completely areolated. Phasmids just postanal. Amphidial apertures in the form
of longitudinal slits. Cephalic region prominently 4-lobed with a large offset labial disc.
Cuticularization of basal plate of framework massive. Stylet exceptionally elongate (54-
170µm), stylet guide (stoma) elongate-tubular. Precorpus swollen with convoluted lumen
when stylet is retracted. Median and basal bulbs well developed, joined by a short slender
isthmus. Excretory pore well anterior to hemizonid. Vulval lips not modified. Vaginal
sclerotization in lateral view symmetrical. Female tail convex-conoid anteriorly then
conoid-spicate to filiform. Postrectal intestinal sac only filling convex-conoid region of
tail. Bursa large, trilobed. Spicules robust, with large flanges; gubernaculum also robust,
protrusible.
57
Type species:
Dolichodorus heterocephalus Cobb, 1914
Fortuner, R. and Luc, M. (1987). A reappraisal of Tylenchina (Nemata). 6. The family

Belonolaimidae Whitehead, 1960. Revue de Nématologie, 10, 183-202.
Luc, M. and Fortuner, R. (1987). A reappraisal of Tylenchina (Nemata). 5. The family

Dolichodoridae Chitwood, 1950. Revue de Nématologie, 10, 177-181.
Siddiqi, M. R. (1986). Tylenchida: Parasites of Plants and Insects. CAB International,

5.3. SUPERFAMILY HOPLOLAIMOIDEA
Diagnosis: Tylenchina. Small to large nematodes (about 0.5-2mm). Sexual dimorphism

in cephalic region present, and may also be present in stylet, oesophagus and body shape,
although indistinct in some Pratylenchidae. Cuticle with distinct outer and inner layers,
strongly annulated; longitudinal striae may be present but longitudinal ridges outside lateral
fields absent. Lateral fields, each with 4 incisures reducing towards extremities,
occasionally 3, 2, 1 or none. Cephalic framework well developed, strongly cuticularized
and refractive, generally less developed in males. Labial disc present. Cephalic sensilla
not on surface. Amphidial apertures pore- or slit-like, just below labial disc. Deirids
absent (except Pratylenchoides). Phasmids small, with pore-like apertures, or large,
scutellum-like, always lateral in position, in or near anal region, near tail terminus or much
anterior to anus at different levels; absent in Aphasmatylenchinae. Stylet well developed,
2-5 times maximum width of lip region; protractors tubular around stylet; conus about as
long as shaft, knobs prominent. Orifice of dorsal oesophageal gland close to or at some
distance from stylet base. Oesophageal glands not enclosed in a basal bulb, but lobed,
overlapping intestine (except Pararotylenchus and some Pratylenchoides spp. in which
they form a pseudobulb). Subventral glands enlarged, equal to or usually larger than
dorsal gland; nuclei of one or both subventral glands lying posterior to that of the
dorsal gland. Postcorporal or median bulb always well developed (except in some males
with degenerate oesophagus), muscular, with refractive valve plates. A cellular cardia
absent, but oesophago-intestinal junction provided with a small cuticular valvula.
Intestinal cell walls and lumen usually indistinct; rectum and anus distinct.
Female reproductive system basically didelphic, amphidelphic; posterior branch
may be reduced or represented by a uterine sac only. Vulva a transverse slit, lips usually
not modified, in swollen females may be located subterminally or terminally; epiptygma
present or absent; lateral membranes, if present, not conspicuous. Ovaries outstretched,
reflexed or coiled in obese forms. Tails dissimilar between sexes (except some
Pratylenchidae). Female tail generally short (less than two anal body widths) but may vary
58
to become elongate-conoid, absent in some swollen females. Bursa enveloping tail,

subterminal, or absent; usually without phasmidial pseudoribs. Male tail usually short and
with a distinct hyaline terminal portion. Spicules paired, similar or dissimilar, cephalated,
straight to arcuate, with or without distal flanges, independently protrusible. Gubernaculum
fixed or protrusible, with or without terminal titillae; telamon (= capitulum) present in
several genera.
Type family:
Hoplolaimidae Filipjev, 1934
Other families:
Heteroderidae Filipjev & Schuurmans Stekhoven, 1941
Meloidogynidae Skarbilovich, 1959
Pratylenchidae Thorne, 1949
Key to families of Hoplolaimoidea
1. Mature female round, pear- or lemon-shaped behind neck, with

anus terminal or nearly so; male with stylet larger than that
of female and tail non-bursate, very short or absent, develops by
metamorphosis (except Meloidodera); third- and fourth-stage
juveniles often swollen……...............................................................................2
Mature female not round, pear- or lemon-shaped, with anus not
terminal; male with stylet equal to or smaller than that of female and
tail bursate, not very short (except Verutus), does not develop by
metamorphosis; third-;and fourth-stage juveniles normally not swollen …......3
2. Excretory pore in mature female opposite or anterior to median bulb;

labial sensilla of female and juveniles on surface; male with large lip cap
and large transverse slit-like amphidial apertures;
gall-inciting ..............................................................................Meloidogynidae
Excretory pore in mature female behind median bulb; labial
sensilla of females and juveniles on surface; male with small
lip cap and with small oval to round amphidial apertures;
notgall-inciting ……....................................................................Heteroderidae
3. Juveniles and females with low arched cephalic framework,

ecto-or endoparasites of roots ……............................................Pratylenchidae
Juveniles and females with high arched cephalic framework,
Ecto- or endoparasites of roots…............................................Hoplolaimidae
59
FAMILY HOPLOLAIMIDAE
A family containing a wide range of genera usually distinguished initially by their

short tails, strong stylets, distinct head sclerotization and high, conoid head region. It
includes the group of nematode genera commonly referred to as "spiral nematodes" because
of their coiled postures when relaxed. Some genera in the family are amongst the most
commonly found and widely distributed nematodes in both temperature and tropical
climates. Species of these genera include a number of economically important plant
parasites.
The main diagnostic characters of the family Hoplolaimidae are:
i) female and male tails short, usually less than 2 anal body-widths long
ii) head high, conoid; sclerotization distinct
iii) strong stylet, usually greater than 20µm long, with well developed stylet knobs
iv) oesophageal glands lobed, normally overlapping intestine (exception
Pararotylenchus)
v) male tail usually with enveloping bursa (exception Rotylenchulus)
vi) phasmids usually distinct, pore-like or large (scutella) in differing positions
(exception Aphasmatylenchus)
vii) slight or marked morphological differences between sexes, some genera with
swollen or saccate females
viii) vulva usually median or post-median with paired, opposed reproductive tracts
(exceptions Rotylenchoides, Acontylus)
Family HOPLOLAIMIDAE Filipjev, 1934 (Weiser, 1953)
Diagnosis: Hoplolaimoidea. Small to moderately large (usually 0.6-1.5mm); generally

vermiform. Sexual dimorphism manifest in cephalic region. Lateral fields each with 4
incisures, rarely reduced or absent (e.g. Basirolaimus). Deirids absent. Phasmids either
small, with pore-like apertures near, or a little anterior to, anus or large, scutellum-like
near anus or much anterior to it anywhere on body behind oesophageal region; absent in
Aphasmatylenchus. Cephalic region elevated, high arched, framework strongly
cuticularized. Oesophageal glands overlapping intestine (except Antarctylus). Ovaries
paired or rarely single. Female tail short, rounded (about one body width or less long), or
conical (over 2 anal body widths, e.g. Antarctylus) with protoplasmic portion less than 2
body widths long and a distinct hyaline terminal portion. Male tail short (except
Aphasmatylenchus). Bursa large, enveloping tail; flaps crenate, sometimes indented at
tip, usually lacking phasmidial pseudoribs. Spicules robust or slender, straight to
arcuate, with distal flanges which may be reduced. Gubernaculum large, protrusible or
fixed; telamon (= capitulum) present or absent.
Nominotypical subfamily:
Hoplolaiminae Filipjev, 1934
60
Other subfamilies:
Aphasmatylenchinae Sher, 1965
Rotylenchinae Golilev, 1971
Rotylenchoidinae Whitehead, 1958
Key to subfamilies of Hoplolaimidae
1. Phasmids present …….....................................................................................2

Phasmids absent ……......................................................Aphasmatylenchinae
2. Phasmids scutellum-like ……....................................................Hoplolaiminae

Phasmids pore-like ….......................................................................................3
3. Cephalic region large, generally offset and with indented basal annule;
orifice of dorsal oesophageal gland less than one-fourth stylet length behind stylet
base …...............................Rotylenchinae
Cephalic region small, generally continuous and with smooth
basal annule; orifice of dorsal oesophageal gland more than one fourth stylet length
behind stylet base .............................Rotylenchoidinae
Subfamily HOPLOLAIMINAE Filipjev, 1934
Diagnosis: Hoplolaimidae. Cephalic region usually offset, first and basal annules marked
with longitudinal indentations. Lateral fields sometimes reduced or absent. Phasmids
enlarged, scutellum-like near anus or aberrantly placed on body. Stylet knobs robust,
compact tulip-shaped or offset, round to anteriorly flattened or concave. Dorsal
oesophageal gland orifice about one-fourth or less of stylet length behind stylet base.
Didelphic, amphidelphic; ovaries outstretched. Female tail short, rounded. Spicules robust,
usually flanged distally. Gubernaculum also robust, usually protrusible.
Type genus:
Hoplolaimus Daday, 1905
Other genera:
Aorolaimus Sher, 1963
Peltamigratus Sher, 1964 (= Aorolaimus for some authors)
Scutellonema Andrássy, 1958
Basirolaimus Shamsi, 1979 has been regarded as a separate genus, but Siddiqi
(2000) now regards it as a subgenus of Hoplolaimus. In the key below I key the group out
separately, but it can be regarded as being Hoplolaimus for generic purposes if so desired.
In either event, the crucial character of six oesophageal gland nuclei needs to be observed
to enable species identification..
61
Key to genera of Hoplolaiminae
1. Stylet knobs tulip-shaped, each with 1-3 anteriorly directed tooth like projections
…………………………………………………………..2
Stylet knobs not tulip shaped, without tooth like projections………................3
2. Dorsal oesophageal gland uninucleate; lateral fields well developed, each with 4
incisures on most of body except in H. pararobustus; basal head annule divided
into small squares ..................................Hoplolaimus
Dorsal oesophageal gland quadrinucleate; lateral fields poorly developed or absent,
each with 1-3 or no incisures at places; basal head annule not divided into small
squares .....................Basirolaimus
3. Scutella in or near anal region, close to or opposite each other

.....................................................................Scutellonema
Scutella much anterior to anus, separated from each other….........................4
4. One scutellum prevulval, another postvulvar ............…........Aorolaimus

Both scutella postvulvar ........................................................Peltamigratus
Subfamily HOPLOLAIMINAE Filipjev, 1934
Genus HOPLOLAIMUS Daday, 1905

syn. Basirolaimus Shamsi, 1979
Nemonchus Cobb, 1913
Hoplolaimoides Shakil, 1973
Diagnosis: Hoplolaiminae. Majority of species large, 1-2mm in length, body lying in a

slightly curved position when relaxed, head terminus rounded. Lateral field with 1 to 4
incisures, may be areolated. Head sclerotization very distinct and with a massive
cuticular framework. Lip region offset from body, with marked annulations and
longitudinal striations; basal annule may be divided into small squares. Stylet knobs
massive, tulip-shaped with 1-3 anterior tooth-like projections. Oesophageal glands
overlapping intestine, mainly dorsally, with 3 or 6 nuclei (3 in Hoplolaimus; 6 in
Basirolaimus). Vulva median with paired, opposed reproductive tracts. Epiptygma
indistinct. Female and male tails short, rounded; less than 2 anal body-widths long. Large
phasmids (scutella) present, not opposite one another: one anterior to vulva (between
vulva and oesophagus) and the other posterior to vulva (between vulva and anus).
Spicules massive, may be dimorphic with distal flange of variable size. Gubernaculum
large, protrusible.
Type species:
Hoplolaimus tylenchiformis Daday, 1905
62
Bionomics: Hoplolaimus feed as migratory ectoparasitic, semi-endoparasitic, or

endoparasitic nematodes causing root destruction. Some are known to cause serious plant
damage and yield loss, including H. colombus on soybean and cotton, H. indicus on rice, H.
galeatus on tree seedlings, and H. seinhorsti on cowpea and cotton.
Note: Some authorities accept the genus Basirolaimus which differs from
Hoplolaimus mainly, and most consistently, in the presence of six nuclei in the oesophageal
gland lobe. The six nuclei are not formed by a doubling of the normal three nuclei, but
comprise four nuclei in the dorsal gland and one each in the subventral gland lobes. Siddiqi
(2000) now regards the genus as a subgenus of Hoplolaimus.
Genus SCUTELLONEMA Andrássy, 1958
Diagnosis: Hoplolaiminae. Similar to Aorolaimus and Peltamigratus. Most species less

than 1mm in length, body lying in curved, spiral, or C shape position when relaxed, tail
terminus mainly rounded. Lateral field with 4 incisures. Head sclerotization well
developed; lip region usually offset from body, with annulations and with or without
longitudinal striations. Stylet knobs well developed, rounded or cup-shaped.
Oesophageal glands overlapping intestine mainly dorsally. Vulva median with paired,
opposed reproductive tracts. Female and male tails short, less than 2 anal body-widths long.
Differs from other genera in the group by having the enlarged phasmids or scutella
located opposite or nearly opposite one another on the tail or near anus. Bursa
enveloping tail; usually not indented or lobed.
Type species:
Scutellonema bradys (Steiner & LeHew, 1933) Andrássy, 1958
Other species:
The genus consists of about 50 species, most found in tropical or subtropical
countries.
Bionomics: Scutellonema spp, are migratory ectoparasites or endoparasites, feeding on

roots and tubers. S. bradys is a serious pest of yams (Dioscorea spp.) causing "dry rot
disease" of tubers; S. cavenessi is reported to damage groundnuts, S. clathricaudatum can
reduce yields of vegetables, and S. brachyurus is recorded as a pest of guar bean and is
pathogenic to tobacco.
Subfamily ROTYLENCHOIDINAE Whitehead, 1958
Key to genera of Rotylenchoidinae
1. Oesophageal glands extending over intestine
63
mostly ventrally and ventrolaterally ..................................................................2

Oesophageal glands extending over intestine
mostly dorsally and dorsolaterally ..................................................Rotylenchus
2. Posterior ovary non-functional or absent ...................................Rotylenchoides

Posterior ovary functional .........................................................Helicotylenchus
Genus ROTYLENCHOIDES Whitehead, 1958
Diagnosis: Rotylenchoidinae. Small nematodes around 0.5mm in length, body lying in

open C shape position when relaxed. Lateral field with 4 incisures. Well developed head
sclerotization. Lip region continuous with body, annulated. Stylet strong with well
developed stylet knobs, rounded or cup-shaped. Orifice of dorsal gland less than half
stylet length behind base of stylet. Oesophageal glands overlapping intestine mainly
ventrally. Vulva posterior at 74-92%, with single functional anterior reproductive
tract and short postvulval sac or undifferentiated genital branch. Female and male
tails short, less than 2 anal body-widths long. Small, pore-like phasmids at level of anus.
Tail terminus rounded to sharply pointed.
Type species:
Rotylenchoides brevis Whitehead, 1958
Other species:
The genus consists of 7 species.
Note: According to some authorities this genus should be regarded as a synonym of

Helicotylenchus as there is some variation in the genital tract character.
Genus ROTYLENCHUS Filipjev, 1936

syn. Calvatylus Jairajpuri & Siddiqi, 1977
Diagnosis: Rotylenchinae. Body length of most species between 0.5-1.5mm, body in spiral
or C shape when relaxed. Lateral field with 4 incisures. Well developed head
sclerotization; lip region offset or continuous with body, annulated. Strong stylet and well
developed rounded to cup-shaped stylet knobs. Orifice of dorsal oesophageal gland
opening located less than 25% of the stylet length behind the knobs. Oesophageal
glands overlapping intestine mainly dorsally. Vulva median to post median with paired,
opposed reproductive tracts. Female and male tails short, less than 2 anal body-widths
long. Phasmids small and pore-like, opposite one another, on tail or near anus. Female
tail terminus variable, rounded, curved dorsally or sometimes with small ventral projection.
Spicules robust, distally flanged. Titillae and telamon may be present on gubernaculum.
Bursa not lobed or indented terminally.
Type species:
Rotylenchus robustus (de Man, 1876) Filipjev, 1936
64
Bionomics: Feed mainly as ectoparasites or migratory semi-endoparasites. Relatively little

known of economic importance. R. robustus can cause damage to peas, carrots, lettuce and
spruce seedlings; R. buxophilus can reduce growth of tree seedlings.
Genus HELICOTYLENCHUS Steiner, 1945
Diagnosis: Rotylenchoidinae. Similar in general respects to Rotylenchus, although

usually less robust. Species vary in size from 0.4-1.1mm, the body lying in a loose or tight
spiral when relaxed. Lateral field with 4 incisures. Lip region not offset from body,
annulated; no longitudinal indentations on annules. Stylet strong with well developed
stylet knobs, rounded or cup-shapes. Dorsal oesophageal gland opening 25% or more of
the stylet length behind stylet knobs. Oesophageal gland overlapping intestine mainly
ventrally. Vulva median or postmedian with paired, opposed reproductive tracts. Female
and male tails short, less than 2 anal body-widths long. Small pore-like phasmids on tail or
near anus. Tail terminus variable, normally more curved dorsally, sometimes with ventral
projection. Male tail short, conical with a distinct terminal hyaline portion. Bursa
enveloping tail tip, rarely subterminal. Gubernaculum fixed, trough or rod shaped.
Type species:
Helicotylenchus dihystera (Cobb, 1893) Sher, 1961
Other species:
The genus is very speciose and contains over 180 nominal species. These
nematodes are amongst the most commonly found in both temperate and tropical soils.
Bionomics: Helicotylenchus spp. feed as migratory endoparasites or semi-endoparasites, H.

multicinctus is an economically important root parasite of bananas in many parts of the
world; H. dihystera has a very wide host range and is reported to cause root necrosis of
soybean roots and reduced growth of sugarcane, H. mucronatus has been found causing
extensive root and tuber necrosis of bananas and sweet potato. Different species often
occur in very large populations in the soil, sometimes associated with reduced growth and
yields of crops.
Subfamily ROTYLENCHULINAE Husain & Khan, 1967
Diagnosis: Hoplolaimidae. Small sized (usually 0.5mm or less long). No marked sexual
dimorphism in body shape in young females and males; only mature female saccate or
kidney-shaped. Cephalic region high, continuous, with or without distinct annules.
Cephalic sclerotization, stylet and median oesophageal bulb well developed in juveniles
and females, regressed in males. Male stylet weaker than that of female. Oesophageal
glands in juveniles elongated, overlapping intestine mostly ventrally or laterally.
Didelphic, ovaries reflexed or coiled in a mature female. Young female and male tails
similar in being elongate-conoid and having a long hyaline terminal portion; tail persists
in mature swollen female. Bursa present, low. Juvenile tails tapering to a round tip, 2-3
anal body widths long, hyaline terminal portion smaller than that in young female and in
male.
65
Type genus:
Rotylenchulus Linford & Oliveira, 1940
Other genus:
Senegalonema Germani. Luc & Baldwin, 1984
Key to genera of Rotylenchulinae
1. Dorsal oesophageal gland orifice at less than half stylet length

behind stylet base; young female with outstretched ovaries;
bursa enclosing tail tip ................................................................Senegalonema
Dorsal oesophageal gland orifice at more than half stylet
length behind stylet base; young female with ovaries having
double flexures; bursa not enclosing tail tip ................................Rotylenchulus
Genus ROTYLENCHULUS Linford & Oliveira, 1940
Diagnosis: Rotylenchulinae. Marked sexual dimorphism. Juveniles, males and young

females vermiform, arcuate to spiral upon relaxation. Mature female kidney-shaped,
with an irregular, less swollen neck, a postmedian vulva and a short pointed tail. Cuticle
annulated. Lateral fields each with 4 incisures, non-areolated, obliterated in mature female.
Cephalic region high, continuous. Stylet in juveniles and female 2-3 times cephalic region
width long. Orifice of dorsal oesophageal gland usually about one stylet length behind
stylet base. Subventral glands in the normal position, dorsal gland shifted laterally to
subventrally, former much longer than the latter.
Immature female: Vermiform, migratory, (has been mistaken for adult stage and formed
the basis for the proposal of new genera, Spyrotylenchus, Leiperotylenchus). Ovaries
paired, with double flexures. Tail elongate-conoid, with prominent hyaline terminal
portion.
Male: Stylet and oesophagus regressed. Tail similar to that of young female; bursa
subterminal, low, not quite projecting beyond tail contour in lateral view (hence
mistakenly reported absent in R. stakmani (= R. reniformis)). Spicules slender, lacking
distal flanges. Gubernaculum fixed. Cloacal lips pointed, not forming a tube; hypoptygma
absent. Juveniles tail more rounded terminally and with shorter terminal portion than that
of female.
Type species:
Rotylenchulus reniformis Linford & Oliveira, 1940
66
The genus consists of about 10 spp. with R. reniformis being the most widespread
and of most economic importance. Most species are confined to tropical or subtropical
areas.
Bionomics: The migratory juveniles, males and immature females are found in soil. The
immature female is the parasitic, infective stage invading roots, penetrating the epidermis
and cortex intracellularly and becoming embedded to approximately 1/3 of its length with
its head at the endodermis of the stele. The nematode becomes a sedentary semi-
endoparasite and the body swells, normally to a kidney shape. Feeding on the endodermis
produces cellular changes, but galls are not formed. Eggs are laid in a gelatinous matrix
surrounding female body on surface of root.
Rotylenchulus reniformis has a very wide host range with over 200 plant species
being recorded as hosts. It is an economically important plant parasite causing damage and
reduced yields to many crops including cotton, sweet potato, tomato, pigeon pea, coffee,
mung beans.
Alternative key to genera of whole group
1. Mature female swollen or saccate............................................................................14

Mature female vermiform...........................................................................................2
2. Vulva median or postmedian with two reproductive tracts........................................3

Vulva posterior with single reproductive ract..........................................................13
3. Phasmids enlarged and prominent on tail or along body...........................................4

Phasmids small, on or near tail...................................................................................7
4. Both phasmids posterior to vulva...............................................................................5

One phasmid anterior to vulva, one posterior to vulva...............................................6
5. Phasmids opposite one another,

on or near tail..........................................................................................Scutellonema
Phasmids widely separated between
vulva and anus.......................................................................................Peltamigratus
6. Stylet knobs with prominent anterior projections;

stylet 33-52µm, body normally lying in slight
curve when relaxed..................................................................................Hoplolaimus
Stylet knobs normally rounded without prominent
projections; stylet 23-36µm, body normally in
loose spiral or C shape when relaxed........................................................Aorolaimus
7. Oesophageal glands overlapping intestine.................................................................8

Oesophageal glands not overlapping intestine..................................Pararotylenchus
8. Oesophageal glands overlapping intestine

dorsally or laterally.....................................................................................................9
67
Oesophageal glands overlapping intestine

mainly on ventral side........................................................................Helicotylenchus
9. Opening of dorsal oesophageal gland

(d.o.g.) less than ½ stylet length
behind stylet knobs....................................................................................................10
Opening of d.o.g. ½ or more stylet length
behind stylet knobs...................................................................................................12
10. Tail tapering to almost pointed terminus, greater

than 2 anal body widths in length..............................................................Antarctylus
Tail less than 2 anal body widths in length...............................................................11
11. Oesophageal glands extending over intestine as a

short overlap on dorsal side......................................................................Rotylenchus
Oesophageal glands in long lateral overlap of intestine
filling body cavity (immature female)..................................................Senegalonema
12. Tail tapering to rounded terminus, normally greater

than 2 anal body widths in length; slender stylet
usually less than 20µm (immature female)..........................................Rotylenchulus
Tail not tapering, less than 2 anal body widths in
length; stylet greater than 20µm.................................................................Orientylus
13. Short and variable oesophageal overlap of intestine

usually ventral, mature female vermiform..........................................Rotylenchoides
Long dorsal oesophageal overlap of intestine
(immature female)........................................................................................Acontylus
14. Posterior vulva, single reproductive tract, female

swollen but not saccate.................................................................................Acontylus
Postmedian vulva, two opposed reproductive tracts,
Females saccate........................................................................................................15
15. Dorsal oesophageal gland opening normally greater

than one stylet length behind stylet knobs (vermiform immature
vulvate females present).......................................................Rotylenchulus
Dorsal oesophageal gland opening less
than one stylet length behind stylet knobs............................................Senegalonema
Fortuner, R. (1987). A reappraisal of Tylenchina (Nemata). 8. The family Hoplolaimidae

Filip‟ev, 1934. Revue de Nématologie, 10, 219-232.
68

Siddiqi, M. R. (2000). Tylenchida: Parasites of Plants and Insects. 2nd edition, CAB
International, Wallingford, UK.
FAMILY PRATYLENCHIDAE
The Pratylenchidae includes some of the most important and damaging nematodes
of crops in the tropical and subtropical regions. The family contains a number of genera,
some of which are morphologically very similar, thereby making the taxonomy of the
group rather complex. Most genera are migratory endoparasites of roots and tubers and
thus remain vermiform. However, there are two genera, Achlysiella and Nacobbus, where
the female becomes obese and, as a consequence, is restricted to one site within the root
tissue. These specialized genera are dealt with separately in the schedule on obese
nematodes.
The Pratylenchidae is a family under the superfamily Hoplolaimoidea and is
characterized by:
i) the low, flattened or rounded head

ii) the strong sclerotization of the head skeleton
iii) the relatively short, stout stylet which is almost usually about 2.5 head
widths long
iv) the subventral oesophageal glands usually extending in tandem beyond
the dorsal gland
v) the relatively long, conoid or subconoid tail (c' >·2).
Of these genera, three will be considered in more detail, viz. Pratylenchus,

Hirschmanniella, and Radopholus. Nacobbus, having swollen females, is, for pragmatic
purposes, treated separately in the section 'Obese Nematodes'.
Family PRATYLENCHIDAE Thorne, 1949 (Siddiqi, 1963)
Diagnosis: Hoplolaimoidea. Vermiform nematodes. Cuticle prominently annulated.

Lateral fields each with 4-6 incisures, very rarely areolated behind oesophagus. Deirids
absent (except Pratylenchoides). Phasmids pore-like, on tail well behind anus, extending
into bursa as pseudoribs except in Hirschmanniella. Female: Amphids pore-like,
indistinct, near oral opening which is surrounded by 6 labial pits. Cephalic region low,
anteriorly flattened to broadly rounded, annulated; framework strongly cuticularized; labial
disc inconspicuous, dumb-bell shaped (with SEM) in type-genus. Stylet strong, length
69
not exceeding 3 cephalic region widths (except rarely in Hirschmanniella); conus about
as long as posterior part; knobs large, rounded. Orifice of dorsal gland close (usually 2-
3µm) to stylet base. Precorpus slender. Postcorpus strongly muscular, with prominent
valve plates. Isthmus short. Oesophago-intestinal junction indistinct, with refringent
valvula. Oesophageal glands extending over intestine; subventral glands asymmetrical,
extending past the dorsal gland, three gland nuclei usually lying in tandem. Vulva a
transverse slit, submedian to posterior but not subterminal; lips not modified; lateral
membranes absent. Vagina directed inward. Didelphic, amphidelphic or pseudo-
monoprodelphic. Ovaries outstretched; posterior ovary degenerate in pseudo-monodelphic
forms. Spermatheca large, usually rounded, with small round sperm when impregnated.
Tail conoid, subcylindrical to elongate-conoid, about twice or more anal body width
long, with round to pointed tip which may bear a mucro (Hirschmanniella), generally with
inconspicuous hyaline terminal portion. Male: Stylet and oesophagus similar to those in
female or reduced as in Radopholinae. Tail elongate-conoid, bursa terminal or subterminal.
Testis single, outstretched, spermatocytes in one or two rows. Spicules similar, cephalated,
arcuate, pointed with subterminal opening on dorsal or ventral side. Gubernaculum simple,
fixed, or complex with telamon or titillae; protrusible. Hypoptygma present or absent.
Juveniles resemble females in having similar anterior region and tail. Obligate migratory
endoparasites of roots.
Pratylenchinae Thorne, 1949
Other subfamilies:
Hirschmanniellinae Fotedar & Handoo, 1978
Radopholinae Allen & Sher, 1967
Key to subfamilies of Pratylenchidae
1. Oesophageal glands overlapping intestine

mostly ventrally; without sexual dimorphism
in the anterior region .........................................................................................2
Oesophageal glands overlapping intestine mostly
dorsally; with sexual dimorphism in the
anterior region ....................................................................Radopholinae
2. Tails similar between sexes; phasmids

near terminus .....................................................................Hirschmanniellinae
Tails dissimilar between sexes; phasmids
not near terminus ..................................................................Pratylenchinae
70
Simplified key to major genera of Pratylenchidae
1. Oesophageal glands overlapping the intestine dorsally; male with knob-like

head and degenerate oesophagus ................................................Radopholus
Oesophageal glands overlapping the intestine ventrally, or mostly so; male
with low rounded or flattened head and normal oesophagus ........................2.
2. Female with median vulva and two genital tracts
(amphididelphic) ...............................................................Hirschmanniella
Female with posterior vulva (V = 70%) and a single genital tract
(monoprodelphic) ......................................................................Pratylenchus
Subfamily PRATYLENCHINAE Thorne, 1949
Diagnosis: Pratylenchidae. No marked sexual dimorphism in anterior region. Small

sized (under 1mm long). Lateral field with 4-6 incisures. Cephalic region low, usually
flattened, with 2-3 annules. Stylet generally less than 20µm long. Oesophageal glands
lobe-like, about 3 body widths or less long, mostly on ventral side of intestine. Ovaries
paired or single. Female tail 3 anal body widths or less long, subcylindrical to conoid,
lacking a mucro. Male tail conoid, arcuate, completely enveloped by a bursa. Spicules
with subterminal pore on dorsal or ventral side. Gubernaculum fixed. Hypoptygma
generally present. Endoparasites of roots causing typical lesions, rarely attacking aquatic or
marsh plants.
Type-genus:
Pratylenchus Filipjev, 1936
Other genus:
Zygotylenchus Siddiqi, 1963
Genus PRATYLENCHUS Filipjev, 1936
Diagnosis: Pratylenchidae. Small nematodes (less than 1mm long) dying slightly curved
ventrally on application of gentle heat. No marked sexual dimorphism in form of
anterior region. Head region low, flattened, usually appearing as a flat, black cap under
the stereomicroscope. Lip region divided into 2, 3 or 4 annules and continuous with the
body contour; strongly cuticularized. Stylet 20µm or less in length (i.e. about twice, or
slightly more, of the head width), moderately cuticularized and with rounded or anteriorly
concave knobs. Oesophagus equally developed in both sexes, median bulb well developed;
oesophageal gland lobes overlapping the intestine ventrally. Female: vulva well
posterior, usually at 70-80% of body length; genital system with a single anteriorly
directed tract and a variable post-vulval section which may show some differentiation, but
is never functional (mono-prodelphic); spermatheca oval or round and usually filled with
sperm in bisexual species; tail sub-cylindrical or more or less conoid with a broad to
71
narrowly rounded or truncate terminus which may be smooth or annulated. Male: tail
short, dorsally convex-conoid; bursa extending to tail tip; spicules slender, arcuate.
Type species:
Pratylenchus pratensis (de Man, 1880) Filipjev, 1936
Other species:
Almost 100 species have been described. They are often distinguished by rather
subtle characters, thus making this a difficult genus to approach for the non-specialist.
Bionomics: Migratory endoparasites with all stages found in the root cortex. Low soil
populations can be associated with high root populations. The nematodes feed mainly on
cortex cells and form cavities containing 'nests' or colonies of nematodes of all stages.
Discolouration of affected tissues is usually pronounced. Above ground symptoms of
attack include chlorosis and stunting. Some species reproduce sexually while others are
parthenogenetic. The life-cycle can be completed in three to four weeks and the nematodes
can survive in the absence of host plants for several months. Most of the important species
are polyphagous, although P. goodeyi may be restricted to banana. The major species are
P. brachyurus, P. coffeae, P. goodeyi, P. penetrans and P. zeae
Distribution: P. brachyurus, P. coffeae and P. zeae are widely distributed in tropical

areas; P. penetrans mainly in cooler regions of the tropics; P. goodeyi on banana in Crete
and the Canary Islands and in the cooler, montane areas of Cameroon, Ethiopia, Kenya,
Tanzania, Uganda and Burundi.
Subfamily RADOPHOLINAE Allen & Sher, 1967
Key to genera
1. Posterior ovary normal .............................................................................2

Posterior ovary degenerate or absent .................................................................3
2. Deirids absent; all gland nuclei behind

oesophago-intestinal junction ................................................Radopholus
Deirids present; at least one gland nucleus
opposite or anterior oesophago-
intestinal junction .................................................................Pratylenchoides
3. Female cephalic region conoid-rounded; stylet robust,
20µm or longer, generally with tulip-shaped knobs;
male with asymmetrical cephalic region and a
subterminal bursa .........................................................................Hoplotylus
Female cephalic region broadly rounded;
stylet not robust, less than 20µm long with
knobs not tulip-shaped; male with symmetrical
72
cephalic region and a terminal bursa ................................................................4
4. Subventral oesophageal glands partially extending on

ventral side of intestine; male
oesophagus not degenerate ...................................................Apratylenchoides
Subventral oesophageal glands not extending on
ventral side of intestine; male
oesophagus degenerate .......................................................Radopholoides
Genus RADOPHOLUS Thorne, 1949
Diagnosis: Pratylenchidae. Small nematodes (less than 1mm long) dying more or less
straight or slightly curved ventrally on application of gentle heat. Marked sexual
dimorphism in form of anterior region: female head region low, rounded, continuous or
slightly offset. Male cephalic sclerotization, stylet and oesophagus reduced; female
cephalic sclerotization strong, stylet and oesophagus well developed. Median bulb in
female oesophagus well developed and oesophageal glands overlapping the intestine
mostly dorsally.
Female: vulva median, with two functional and equally developed genital tracts
(amphididelphic); spermathecae rounded and with sperm in bisexual species; tail elongate,
conoid (about 60µm long in R. similis).
Male: tail elongate, conoid, ventrally arcuate; bursa not reaching to tail tip in R. similis
and most other species; spicules slender, arcuate.
Type species:
Radopholus similis (Cobb, 1893) Thorne, 1949
Other species:
Over 20 species have been placed in this genus, but some belong to other genera
such as Achlysiella or Zygradus. Achlysiella is discussed in more detail in the section
Obese Nematodes.
Bionomics: Migratory endoparasites of root and corm/tuber tissues. In roots, the feeding
activities are restricted to the cortex causing cavitation, discolouration and severe damage,
thus allowing secondary invasion by other micro-organisms. The adult male is non-
feeding. The major species is R. similis which currently has two recognised host races or
biotypes. R. similis attacks banana and many other plants. R. similis citrophilus (recognised
as a separate species by some authorities on differing chromosome count and minor
morphological details) was proposed for a citrus race, but is no longer recognised at
specific or subspecific level. The importance of another citrus attacking species, R. citri,
has yet to be demonstrated under field conditions.
73
Distribution: The majority of species have been described from the Australasian region.
However, R. similis is now found world-wide in tropical regions and occurs virtually
everywhere that banana is grown. The citrus race, formerly known as R. similis citrophilus,
is recorded from Florida. Radopholus citri was described attacking citrus in Java and may
represent a threat to other citrus growing areas if introduced.
REFERENCES
Café Filho, A. C. and Huang, C. S. (1989). Description of Pratylenchus pseudofallax n. sp.

with a key to species of the genus Pratylenchus Filipjev, 1936 (Nematoda: Pratylenchidae).
Revue de Nématologie, 12, 7-15.
Luc, M. (1987). A reappraisal of Tylenchina (Nemata). 7. The family Pratylenchidae

Thorne, 1949. Revue de Nématologie, 10, 203-218.

FAMILY MELOIDOGYNIDAE
The Meloidogynidae (colloquially known as 'root knot' nematodes) comprises a

diverse and widespread group of nematodes amongst which are a number of economic
importance. Typically they are sedentary, semi-endoparasites, the females swelling
enormously and laying large numbers of eggs into a protective gelatinous matrix, The J2 is
the infective stage and after invading a root becomes sessile and initiates the development
of specialized trophic cells to nurture the subsequent stages. The nematode also releases
chemicals which initiate gall formation. Vermiform males are produced in many species,
often when food supply is a restrictive factor. Many species are extremely polyphagous
whereas others are more host specific. The greatest number of species occur in the tropics
and sub-tropics, although a few species have a temperate distribution and a single
generation a year.
Diagnosis: Meloidogynidae. Sexually dimorphic. Mature female sedentary, swollen,

globular or pear-shaped, cuticle thin, white, annulated. Non cyst forming. Tail absent,
anus and vulva terminal surrounded by characteristic pattern of striae on the cuticle
(perineal pattern). Excretory pore anterior to median bulb and near to stylet knobs,
head skeleton hexaradiate. Stylet <25m long, with well developed basal knobs; procorpus
cylindrical followed by spherical metacorpus with well developed musculature and
74
cuticular valve plates; procorpus and metacorpus not amalgamated. Oesophageal glands
overlapping intestine ventrally. Genital tracts paired, elongated, anteriorly directed and
much convoluted. Eggs laid in an external gelatinous matrix. The eggs are usually
unembryonated and not retained in the female body. The first moult occurs within the egg,
the second-stage juveniles hatching and being the infective stage. Female: Sedentary,
swollen, globular or pear-shaped, cuticle thin, white, annulated, tail absent, anus and vulva
terminal surrounded by characteristic pattern of striae on the cuticle (perineal pattern),
excretory pore anterior to median bulb near to stylet knobs, head skeleton hexaradiate.
Stylet <25µm long, well developed basal knobs: procorpus cylindrical followed by
spherical metacorpus with well developed musculature and cuticular valve plates,
procorpus and metacorpus not amalgamated. Oesophageal glands overlap intestine
ventrally. Genital tracts paired, elongated and convoluted. Eggs laid in gelatinous matrix,
usually unembryonated and not retained in the female body. First moult within the egg,
second-stage juveniles hatch and are infective. Male vermiform, migratory, stylet well
developed, <33m; cephalic framework well developed, hexaradiate; short bluntly rounded
tail; no bursa, usually one, but sometimes two testes; paired slender spicules, simple
gubernaculum. Intersexes or sex reversal may occur, particularly in response to nutrient
stress. Second-stage juveniles vermiform, infective, migratory; head skeleton hexaradiate,
stylet slender, knobbed, stylet <23m long. Hyaline region of tail less than half the tail
length. Heat relaxed form straight to arcuate. The 3rd and 4th stage juveniles occur within
the root and are swollen, sedentary, with a blunt terminus and no stylet. They develop
within the cuticle of 2nd stage juveniles, the tail spike of which is retained.
Type species:
Meloidogyne exigua Goeldi, 1892
syn. Heterodera exigua (Goeldi, 1892) Marcinowski, 1909
Other species:
The genus contains a large and ever increasing number (>70) of species with several
new species being proposed each year. Many of the species are becoming difficult to
differentiate using classical taxonomic techniques and biochemical or molecular techniques
are being increasingly employed to try to circumvent the problem. Many of the taxonomic
problems are undoubtedly due to the fact that most species reproduce by mitotic
parthenogenesis.
Note: The genus Hypsoperine is sometimes recognised as being distinct from Meloidogyne
(e.g. Siddiqi, 1986). It differs by having the anus and vulva located on a cone-like
elevation or protuberance on the mature female and in having the excretory pore of the J2
anterior to the hemizonid as opposed to posterior. In addition, the females often lie parallel
to the root axis in contrast to Meloidogyne.
Bionomics: Obligatory, sedentary endoparasites of plant roots inciting gall formation and
a complex trophic system of giant cells in the root cortex. Many species exhibit extreme
polyphagy, thus making control by crop rotation difficult.
75
MORPHOLOGICAL CHARACTERS RECOMMENDED FOR USE IN SPECIES

IDENTIFICATION
Most important characters
J2: tail length

hyaline tail length
tail shape
Mature female: stylet length

stylet shape
excretory pore position as ratio of stylet length
perineal pattern
Male: stylet length

stylet cone length
stylet shape
head shape (light microscope)
distance of dorsal oesophageal gland orifice (DGO)
from stylet base
Supplementary characters
J2: head shape

stylet shape
presence of inflated rectum or not
Female: body shape
Male: head shape

tail shape
spicule shape
CIH Descriptions of Plant Parasitic Nematodes Sets 1-8. CAB International, Wallingford,
UK.
Jepson, S. B. (1987). Identification of root-knot nematodes (Meloidogyne species). CABI

Wallingford, UK. 265 pp.
76
Luc M., Maggenti A.R. and Fortuner R. (1988). A reappraisal of the Tylenchina (Nemata) :
9. The family Heteroderidae Filipjev & Schuurmans Stekhoven, 1941 Revue de
Nématologie 11, 159 - 179.
Sasser, J. N. and Carter, C. C. (1986). An advanced treatise on Meloidogyne. Vol. I,

422pp.; Vol. II, 223pp.

Taylor, A. L. (1987). Identification and estimation of root-knot nematode species in mixed

populations. Florida Department of Agriculture and Consumer Services.
5.4. SUPERFAMILY CRICONEMATOIDEA
Many of the criconematids are commonly referred to as „ring nematodes‟ because of

the pronounced annulation that is so typical of many species. They form one of the most
distinct groups of plant parasitic nematodes, but are not always well represented in
extractions because of their sluggish nature - sieving techniques usually recover far more
than tray methods. Most species are ectoparasitic on plant roots, those with longer stylets
being able to feed deep within the root cortex. In some genera the mature female has a
tendency to become obese. Those nematodes with truly obese females (e.g. Tylenchulus)
are dealt with under the section „Obese nematodes‟ to facilitate comparison with other
nematodes sharing a similar trophic habit. Although not systematically accurate, this
approach does have the advantage of being pragmatic.
The following systematic scheme contains the major generic names commonly
found in the literature. However, there is a long history of flux in the systematics of the
Criconematinae and Macroposthoniinae, a flux that continues in the present day, some
workers splitting the subfamilies into numerous small genera, others preferring a broader
approach and retaining only a few, somewhat loosely defined genera. In this introductory
account the latter course is adopted for simplicity, although some of the other generic
names are mentioned in passing.
The superfamily may be conveniently divided into three families that are easily
differentiated from on another on morphology: the Criconematidae, or ring nematodes; the
Hemicycliophoridae, or sheath nematodes and the Paratylenchidae, or pin nematodes. The
taxonomy of this group may involve features of the juvenile stages (e.g. whether the cuticle
is spined or not), and this can cause problems when material is limited or where there is a
mixed population. Males are degenerate and usually short-lived. They are usually rather
uncommon in extractions, although they can be abundant under certain circumstances. As
a consequence, it is the female morphology that is most utilized for diagnostic purposes, the
presence of males usually being indicated by the presence of sperm in the female
spermatheca.
77
Key to subfamilies of Criconematidae and Hemicycliophoridae
1. Stylet knobs anchor-shaped (anteriorly concave) ………....Criconematidae (2)

Stylet knobs sloping backwards ....................................Hemicycliophoridae (4)
2. Double cuticle present, annules normal ..................…...Hemicriconemoidinae
Double cuticle absent, very heavily annulated
nematodes, annules retrorse ..............................................................................3
3. Juveniles with cuticular scales or spines, adults
usually with scales or spines ..................................…..............Criconematinae
Adults lacking cuticular scales or spines and
juveniles usually lacking such structures ..........….............Macroposthoniinae
4. Females with a double cuticle, the outer being
well-developed and not membranous. Head annules
not separated ........................….........................................Hemicycliophorinae
Females without a double cuticle or, if present,
the outer sheath membranous, much thinner than the
body cuticle and closely adpressed. Head annules
well separated …………………………………………………Caloosiinae
FAMILY CRICONEMATIDAE
Characters of Criconematinae and Macroposthoniinae
The nematodes belonging to these subfamilies are characterized by the following:
i) Small stout bodies with cuticle marked by deep striae forming conspicuous
annules which are either plain or bear scales or spine-like retrorse projections which may or
may not be arranged in longitudinal rows.
ii) The spear or stylet is usually greatly elongated, 45-142µm long, with very long
conus and large, anteriorly directed basal knobs.
iii) The precorpus or procorpus and the median bulb of the oesophagus are not
demarcated but amalgamated.
iv) The isthmus is very short and the basal bulb is small and spathulate.
v) The female gonad is monoprodelphic without a posterior uterine sac.
vi) Males degenerate, stylet absent, lateral fields present, bursa greatly reduced or
absent.
vii) Phasmids absent.
78
Some characters of taxonomic importance
The morphology of this group is markedly at variance with other plant parasites and
this in turn means that the characters used in the delineation and identification of this group
differ somewhat. The major characters are listed below:
1. The length and width of the body.

2. The number of body annules (R).
3. The number of annules between the labial disc and base of stylet knobs (Rst).
4. The number of annules between the labial disc and the excretory pore (Rex).
5. The number of annules from tail terminus to vulva (RV).
6. The number of annules from tail terminus to anus (Ran).
7. The number of annules between vulva and anus (Rvan).
8. The shape, size, number and arrangement of cuticular outgrowths (scales, spines,
etc.) on the annules.
9. Anastomoses of annules may or may not be present.
10. Submedian lobes/lips. Submedian lobes when present, are visible in lateral as well
as dorso-ventral view, but can be observed best in an en face view. There are 4
submedian lobes, their size being variable in different species. These lobes may be
connected dorsally and ventrally. In some species 6 lips (4 submedian lobes and 2
lateral) are present instead of submedian lobes. Often the lateral lips are reduced or
absent.
11. Labial disc. Submedian lobes and labial plates surround a labial disc with an I-
shaped oral opening. This disc may be elevated.
12. Amphids. Amphidial openings are oval or slit-like, observed better in an en face
view.
13. Spear or stylet consists of a very long conical part (conus), a short cylindrical part
and the basal knobs. The basal knobs are always projecting forward (anchor-
shaped).
14. Genital tract. The female genital tract is mono-prodelphic and lacks a posterior
uterine sac. It consists of vulva, vagina, uterus, oviduct and the ovary.
The single ovary consists of the usual germinal and growth zones. Its length
depends on the age of the female and in older females it may extend into the
oesophageal region. The oviduct is a short tubular region that enters the uterus
between spermatheca (if present) and quadricolumella. The vagina is oblique to the
body axis and directed anteriorly.
15. Vulva when viewed ventrally may be open, closed or with an overlapping anterior
lip.
16. Tail is greatly variable, may be conical pointed or conical rounded or short rounded.
If annules are ornamented their size usually increases towards the tail end.
17. Males are rare. The following features are of importance: shape of body upon
killing, body length and width, shape of head, tail, position of excretory pore, length
and shape of spicules, gubernaculum, number of lines in the lateral fields and the
bursa.
79
18. Juveniles have smooth, crenate or spined posterior edges to the annules. Sometimes
early stages show crenate or spined annules but the older stages do not. The number
of annules on the juveniles may be more than those of the adults. Male juveniles do
not possess a stylet.
Subfamily CRICONEMATINAE
Diagnosis: Criconematidae. Cuticle ornamented in juveniles, and usually the adults,

with scales or spine-like outgrowths. These are usually arranged in longitudinal rows.
Many genera have been described in this group, but the major ones are:
Criconema Hofmänner & Menzel, 1914
Bakernema Wu, 1964
Crossonema Mehta & Raski, 1971
Ogma Southern, 1914
Key to genera - adult females
1. Spines or scales arranged in longitudinal rows .........................................Ogma

Spines or scales absent or not arranged in longitudinal rows.............................2
2. Spines or scales absent ......................................................................Criconema
Spines or scales present .....................................................................................3
3. Cuticle outgrowth membranous, hardly discernible
Head not separate and bearing similar appendages
to other annules ................................................................................Bakernema
Cuticle outgrowths stronger, well-defined
Head separate, smooth or fringed ...................................................Crossonema
Subfamily MACROPOSTHONIINAE
Diagnosis: Criconematidae. Cuticle smooth in juveniles and females, lacking spines or

scale-like outgrowths, but may occasionally be finely crenate. The most commonly cited
genera are:
Criconemoides Taylor, 1936

Discocriconemella De Grisse & Loof, 1965
There has been considerable doubt about the type species of Macroposthonia and
Criconemoides and both genera have been synonymized under Criconemella by some
authors whilst others disagreed. It should be noted that there was also an attempt to replace
the use of both Macroposthonia and Criconemella by the name Mesocriconema. A recent
ruling from the International Commission on Zoological Nomenclature re-established
Criconemoides as a valid genus and this name should now be used instead of
Macroposthonia, Criconemella or Mesocriconema.
80
Key to genera - adult females
1. Head annule disc or saucer-shaped .......................................Discocriconemella

Head annule normal, not disc or saucer-shaped .........................Criconemoides
Genus CRICONEMOIDES Taylor, 1936

(after Ebsary, 1982)
Diagnosis: Macroposthoniinae. Female body 0.20-1.0mm. Annules 42-210, smooth to

crenate, anastomoses present or absent. Head annules normal, not discoid. Submedian
lobes present or absent, if present may be fused to varying degrees or separate. Labial
plates present or absent. Vulva open or closed, anterior vulval lip ornamented or not.
Vagina straight or sigmoid. Stylet well developed, 25-129µm. Oesophagus about 30% of
body length. Tail shape variable. Juveniles: annules smooth to crenate. Males: head end
rounded to conoid, lateral incisures 2 to 4, usually 4, bursa distinct.
Type species:
Criconemoides morgensis (Hofmänner in Hofmänner & Menzel, 1914) Taylor,
1936.
Other species:
Many described and mostly difficult to identify. Most species have been placed in
Macroposthonia, Criconemella or Mesocriconema at one time or another although a recent
ruling by the International Commission on Zoological Nomenclature re-instated
Criconemoides as a valid genus, thereby relegating the other generic names to junior
synonymy.
Subfamily HEMICRICONEMOIDINAE
Diagnosis: Criconematidae. Female and juveniles sausage-shaped with rounded, coarse

annules. Cuticle double, but outer sheath usually fairly closely adpressed. Stylet
strong with anchor-shaped basal knobs.
There is only one genus.
Genus HEMICRICONEMOIDES Chitwood & Birchfield, 1957
Diagnosis: With the characters of the subfamily.
Type species:
Hemicriconemoides wessoni Chitwood & Birchfield, 1957
81
FAMILY HEMICYCLIOPHORIDAE
Diagnosis: Criconematoidea. Females and juveniles with a double cuticle, annules not
retrorse and lacking scales or spine-like structures. Stylet strong with rounded basal
knobs. Males lacking a functional stylet and oesophagus. Bursa present.
There are four nominal genera:
Hemicycliophora de Man, 1921

Aulosphora Siddiqi, 1980
Colbranium Andrássy, 1979
Loofia Siddiqi, 1980
Some authors do not accept Aulosphora or Loofia, both being regarded as junior
synonyms of Hemicycliophora.
Key to genera - adults
1. Vulva lips round, low, not modified. Body not recessed behind vulva; spicules
arcuate ..........................................................................................2
Vulva lips modified, pointed, elongate. Body recessed behind vulva; spicules semi-
circular or hooked-shaped .......................................................................3
2. Female head offset by deep groove. Vulva and anus sub-terminal; bursa almost
reaching tail tip ………………………………………….Colbranium
Female head not offset. Vulva and anus not subterminal; bursa short, not almost
reaching tail tip .......... ...............................................................................Loofia
3. Vulval lips less than 3 body annules long, often divergent. Spicules semi-circular in
shape; pre- and post-cloacal regions of bursa in ratio
1:1…........................................................................................Hemicycliophora
Vulval lips longer than 3 body annules, almost parallel. Spicules U- or hook-
shaped; pre- and post-cloacal regions of bursa in ratio 3-
4:1.....................................................................................................Aulosphora
Subfamily HEMICYCLIOPHORINAE
Genus HEMICYCLIOPHORA de Man, 1921

(after Siddiqi, 1980)
Diagnosis: Hemicycliophoridae. Body just behind vulva deeply recessed. Vulva lips
modified, elongate but less than 3 annules long, usually divergent. Female tail elongate,
tapering, filiform, cylindrical or, rarely, hemispherical. Spicules semi-circular. Penial
tube well developed but less than a body width long, directed outward and forward. Body
82
just in front of penial tube recessed. Pre- and post-cloacal regions of bursa almost
equal.
Type species:
Hemicycliophora typica de Man, 1921
FAMILY PARATYLENCHIDAE
The Paratylenchidae or pin nematodes comprises a small group of criconematid

nematodes characterized by:
i) a criconematid-type oesophagus and monoprodelphic genital system

ii) a well developed stylet, which may be very long and slender
iii) fine annulation
iv) male stylet and oesophagus non-functional and degenerate
Identification to species level is a difficult process as the nematodes are so small

and many of the characters are subjective.
Note Gracilacus is sometimes regarded either as a junior synonym or subgenus of

Paratylenchus, the only consistent differences between the two genera being stylet length
(perhaps an arbitrary character) and a tendency in Gracilacus females to become obese
(although this state is not yet confirmed for all species).
Key to subfamilies
1. Isthmus short, bursa absent or very weakly developed ...........Paratylenchinae

Isthmus elongate, bursa present, well developed ......Tylenchocriconematinae
Subfamily PARATYLENCHINAE
Diagnosis: Paratylenchidae. Female body slender or saccate. Stylet well developed and
may be extremely long and attenuate. Female genital tract monoprodelphic, outstretched.
Male bursa absent or very weakly developed.
Key to genera
1. Mature female obese with cuticular ornamentation in the form of minute

refractive tubercles. Tail very short and blunt ................................Cacopaurus
Mature female usually slender, but if obese then without cuticular
ornamentation. Tail not very short and blunt ..................................................2.
2. Female stylet shorter than 38µm .................................................Paratylenchus
83
Female stylet 40µm long or longer ...................................................Gracilacus
Genus PARATYLENCHUS Micoletzky, 1922
Diagnosis: Paratylenchinae. Small nematodes, less than 0.5mm long and usually dying in a
fairly closed 'C' shape. Stylet well developed, but shorter than 38µm long. Vulva well
posterior, genital tract monoprodelphic, outstretched. Tail long, conoid, with a pointed or
finely rounded tip. Male oesophagus and stylet degenerate, bursa absent.
Type species:
Paratylenchus bukowinensis Micoletzky, 1922
Other species:
Many species have been described, but they are very difficult to identify because of
their minute size and the difficulty of assessing some of the character states – even the
number of lateral lines can be difficult to ascertain with certainty.
Bionomics: Often occur in very large numbers and seem to be particularly associated with
woody plants.
Andrássy, I. (1979). Revision of the subfamily Criconematinae Taylor, 1936 (Nematoda).

Opusc. zool. Budapest., 16, 11-57.
Ebsary, B. A. (1982). Bakernema yukonense n. sp. (Nematoda: Criconematidae) with keys

to the species of Criconemella and Discocriconemella. Canadian Journal of Zoology, 60,
3033-3047.
Raski, D. J. and Luc, M. (1987). A reappraisal of Tylenchina (Nemata). 10. The

superfamily Criconematoidea Taylor, 1936. Revue de Nématologie, 10, 409-444.

5.5. SUPERFAMILY ANGUINOIDEA
Diagnosis: Hexatylina. Small to large sized (0.4-3.5mm), in some genera adults may be
obese and sedentary. No marked sexual dimorphism in anterior region. Cuticle with
fine transverse striations, often appearing smooth. Lateral field plain, or with 4, 6 or
more incisures. Deirids usually present. Phasmids absent. Cephalic region low, cap-
84
like, smooth with indistinct or no annulations, generally continuous with body contour;
framework hexaradiate, faintly cuticularized. Oral opening a small round pore
surrounded by 6 labial papillae; amphids indistinct, oval, pore-like, slightly dorso-
sublateral, labial but at some distance from oral opening. Stylet small (under 15µm), with
small rounded knobs. Orifice of dorsal gland close to stylet base. Median oesophageal
bulb present or absent, with or without refractive valve plates. Oesophageal glands tend to
be enlarged, forming a basal bulb, or rarely, the dorsal gland extends over intestine dorsally
and laterally; no stem-like extension at base of basal bulb (except Cynipanguina which
has a different type of extension). A tricellular cardia absent; two anteriormost
intestinal cells often acting as a valve. Gonads single, anteriorly outstretched, may be
reflexed or coiled in swollen adults; germ cells may be arranged about a rachis. Vulva a
large transverse slit, posteriorly located; lateral vulval membranes rarely distinct
(Pterotylenchus). Spermatheca axial, elongated, sac-like (except Pseudhalenchus).
Sphincter valve between oviduct and uterus may be present. A postvulval sac often as long
as or longer than body width present (absent in Diptenchus), may serve as storage for
sperm, often with a small rudiment of posterior reproductive branch. Sperm round, large,
with a prominent translucent cytoplasmic vesicle around the nucleus. Spicules robust,
anteriorly expanded, separate or fused medially, tip often truncate or broadly rounded.
Gubernaculum simple, trough-like, not protrusible, rarely absent (Nothanguina). Bursa
moderately large, usually subterminal, but may extend to terminus (Sychnotylenchidae) or
be adanal. Tails similar between sexes (except when bursa is terminal), usually elongate-
conoid, may be cylindroid or filiform; juvenile tails often elongate-conoid to filiform.
Fungal feeders; parasites of lower (mosses, seaweeds) and higher plants, attacking
stems, leaves, floral parts and seeds, almost always inciting galls; root parasitism known
only for Subanguina radicicola which incites and inhabits galls; also associates of insects
(Sychnotylenchidae) but not parasitic in insects or other animals).
Nominotypical family:
Anguinidae Nicoll, 1935 (1926)
Relationship: Members of this superfamily are closely similar to those of the

Paurodontidae and free-living forms of the Sphaerularioidea (suborder Hexatylina), but
differ in lacking an entomoparasitic phase in their life-history and a stem-like extension
from the oesophageal base (except in Cynipanguina). Traditionally Anguina and
Ditylenchus have been classified in the Tylenchidae, but they differ from the members of
this family in their different origin and evolution from fungal feeding forms and in having
the two anteriormost cells of the intestine modified to act as a valve (a tricellular cardia is
absent in this group), a prominent crustaformeria, large sized sperm with a prominent
cytoplasmic vesicle and an absence of phasmids.
Bionomics: The genera and species of the family Anguinidae are mainly fungal feeders or
parasites of stems, leaves, flower parts and seeds where they usually incite galls. The only
known root parasitic species is Subanguina radicicola, which incites and inhabits root galls
on various grasses.
85
Important characteristics of the family Anguinidae:
1) the cephalic region is low and smooth; framework hexaradiate, sectors almost
equal.
2) the basal oesophageal bulb is either small or large, offset from intestine; the
dorsal gland may become enlarged and extend over intestine as a lobe.
3) Vulva is generally less than 85% of body length; post vulval uterine sac
present or rarely (Diptenchus) absent.
4) Ovary outstretched or reflexed; oocytes may be arranged around a rachis in
obese females.
5) The tail is similar between the sexes; female tail rarely subcylindrical, never
cylindrical or hooked.
6) The bursa varies from adanal to subterminal, never encircling tail tip.
Key to families of Anguinoidea
1. Terminal excretory duct abnormally widened and cuticularized:

parasites of marine plants (algae)....................................................Halenchidae
Terminal excretory duct not abnormally widened and
cuticularized; not parasites of marine plants......................................................2
2. Female tail cylindroid or subcylindroid, dissimilar to that of male;

bursa enveloping tail terminus; associates of insects...........Sychnotylenchidae
Female tail conoid to filiform, rarely subcylindroid,
similar to that of male; bursa not enveloping tail terminus;
very rarely associates of insects........................................................Anguinidae
Family ANGUINIDAE Nicoll, 1935 (1926)
Diagnosis: Anguinoidea. Adults from about 0.4 to 3mm long, slender or obese. Cephalic
region low, smooth; framework hexaradiate, sectors almost equal. Muscular valvate
median bulb present or absent. Basal oesophageal bulb small or large, offset from intestine,
or the dorsal gland may become enlarged and extend over intestine as a lobe. Excretory
duct not abnormally widened or cuticularized. Vulva generally at less than 85% of body
length. Postvulval uterine sac present, or rarely (e.g. Diptenchus) absent. Ovary
outstretched or reflexed; oocytes may be arranged about a rachis in obese females. Tails
similar between sexes, female tail conoid to filiform, rarely subcylindrical, but never
cylindrical or hooked. Bursa variable from adanal to subterminal, never enclosing tail
tip. Fungus feeders or parasites of stem, leaves, flower parts and seeds where they usually
incite galls, not root parasites (except Subanguina radicicola which incites and inhabits
roots galls).
Anguininae Nicoll, 1935 (1926)
86
Key to subfamilies of Anguinidae
1. Postcorpus bulboid, muscular, valvate.............................................Anguininae

Postcorpus not bulboid, muscular, or alvate.....................................................2
2. Female spiral, obese; gubernaculum absent..............................Nothanguininae

Female not spiral, obese; gubernaculum absent.....................Nothotylenchinae
Subfamily ANGUININAE Nicoll, 1935 (1926)
Diagnosis: Anguinidae. Adults slender or obese, straight, arcuate or strongly curved when
relaxed. Precorpus cylindroid. Postcorpus muscular, with distinct valve plates. Basal
bulb enclosing oesophageal glands present, dorsal gland may form a lobe extending over
intestine dorsally or laterally (Pseudhalenchus, Safianema, some Anguina). Vulva lacking
lateral membranes. Postvulval uterine sac well developed (absent in Diptenchus).
Crustaformeria with 4 or more rows of cells. Ovary outstretched or with tip reflexed once
or twice. Gubernaculum present.
Type genus:
Anguina Scopoli, 1777
Key to genera of Anguininae
1. Crustaformeria with more than 20 cells; female generally

obese (partially obese in several Subanguina spp.)............................................2
Crustaformeria with less than 20 cells (generally 16 cells);
female slender, occasionally partially obese......................................................4
2. Oesophageal base with a long digit-like extension.......................Cynipanguina
Oesophageal base without a digit-like extension...............................................3
3. Gametocytes usually arranged about a rachis; female

strongly obese, spirally urved...............................................................Anguina
Gametocytes not arranged about a rachis; female
generally not strongly obese or spirally curved...............................Subanguina
4. Dorsal oesophageal gland forming a long lobe extending over

intestine, with nucleus lying posterior to oesophago-intestinal junction...........5
Dorsal oesophageal gland not forming a long lobe extending over
intestine, with nucleus anterior to oesophago-intestinal junction......................6
5. Lateral field with 4 incisures; spermatheca rounded.................Pseudhalenchus

Lateral field with 6 incisures; spermatheca elongate-saccate.............Safianema
6. Postvulval uterine sac absent.............................................................Diptenchus

Postvulval uterine sac present..........................................................Ditylenchus
87
Genus ANGUINA Scopoli, 1777
Diagnosis: Anguininae. Medium to large sized (1-2.7mm), obese; mature female curved
generally in one to one-and-a-half spirals. Median oesophageal bulb muscular. Basal
bulb in adults enlarged, continuous or offset from isthmus by a constriction, base usually
extending over anterior end of intestine. Ovary with one or two flexures anteriorly due to
excessive growth; oocytes in multiple rows, arranged about a rachis. Crustaformeria a
long tube formed by a large number of cells in multiple irregular rows. Spermatogonia
in multiple rows. Bursa subterminal. Second-stage juvenile generally resistant and is the
infective stage. Obligate plant parasites inciting galls in seeds of cereals and grasses,
stems, leaves and inflorescence of various monocotyledonous plants; type species causes
wheat seed galls (ear cockles); only A. amsinkiae and A. balsamophila parasitize
dicotyledonous plants.
Type species:
Anguina tritici (Steinbuch, 1799) Filipjev, 1936
Bionomics: Within the genus Anguina, there are several species of economic importance.
The second stage juvenile (J2) is the resistant and infective stage and can survive
desiccation for thirty years or more. The life-cycle of an Anguina species can be
exemplified by Anguina tritici. A. tritici (the type species) infects grasses, wheat and
barley. The nematodes initially feed ectoparasitically on the growing points and leaf bases
until they are able to enter the inflorescences as the embryo seeds develop. The nematodes
then develop to adults, which feed and mate inside the grain. The female lays a large
number of eggs (between a few hundred to 32,000). The adults die in the galled grain, the
entire cavity of which is occupied by the J2 stage which gradually desiccates. As a result of
infection by the nematode, the grain is destroyed. The life cycle resumes the following
season when rain softens the gall, allowing the J2 nematodes to revive by absorbing water.
They then migrate in search of germinating host seedlings which they invade. A. tritici can
easily be controlled by efficient seed cleaning techniques which remove the galls from the
grain and thus avoids the possibility of sowing them with seed for the next crop.
Genus DITYLENCHUS Filipjev, 1936

syn. Anguillulina (Ditylenchus) Filipjev (Schneider, 1939)
Diagnosis: Anguinidae. Body usually under 1.5mm long, not curving strongly when
relaxed; mature adults slender. Lateral field with 4 or 6 (occasionally more) incisures
which may be indistinct. Median bulb muscular, with valve plates. Isthmus not marked off
from basal bulb. Basal bulb a thin elastic sac containing oesophageal glands; base of
bulb may extend over intestine but the nuclei of glands remain within the bulb anterior to
the oesophago-intestinal junction. Intestine with 2 normal sized but often hyaline cells at
its anterior end, possibly acting as a valve; lumen of intestine not considerably narrowed in
anterior region. Ovary outstretched, with 1 or 2 rows of oocytes. Vagina at right angle to
body axis or nearly so, not directed forward. Postvulval uterine sac present. Testis
outstretched. Bursa adanal to subterminal. Tails elongate-conoid to subcylindrical or
filiform. Fungal feeders and parasites of higher plants, several species including the type
species are capable of attacking aerial parts.
88
Type species:
Ditylenchus dipsaci (Kühn, 1857) Filipjev, 1936
Bionomics: Many species in this large genus are fungal feeders and are not known to attack
higher plants. There are, however, several important plant parasitic species which are
capable of causing extensive loss to agricultural crops. The most famous species is D.
dipsaci, known as the stem nematode or stem and bulb nematode. This species attacks over
450 plant species and occurs as biological races or biotypes. The fourth stage juvenile (J4)
is the survival stage and in drying plant tissues, the nematodes tend to aggregate, forming
clumps of "eelworm wool". The nematode is a migratory endoparasite of bulbs and the
green parts of plants. It can be devastating on onion and broad bean crops. Another
species, D. angustus, the Ufra nematode, attacks rice and can be particularly damaging on
deep water rice in Bangladesh and Vietnam. The nematodes invade and feed on the
developing rice seedlings and on mature plants are found in the tender parts of the stem,
leaf sheath and flower spike. Both these species have four lateral lines, but D. destructor, a
member of the D. triformis group, has 6 incisures in the lateral field. Members of this
group are mainly fungal feeders, although D. destructor attacks potato tubers and bulbous
iris. D. myceliophagus attacks and destroys the mycelia of cultivated mushrooms.
Brzeski, M. W. (1981). The genera of the Anguinidae (Nematoda: Tylenchida). Revue de

Nématologie, 4: 23-34.
Fortuner, R. and Maggenti, A. R. (1987). A reappraisal of the Tylenchina (Nemata). 4. The

family Anguinidae Nicoll, 1935 (1926). Revue de Nématologie, 10: 163-176
Fortuner, R. and Raski, D. J. (1987). A review of Neotylenchoidea Thorne, 1941 (Nemata :

Tylenchida). Revue de Nématologie, 10: 257-267
Siddiqi, M. R. (1986). Tylenchida: Parasites of plants and insects. CABI International,

5.6. OBESE NEMATODES
Plant parasitic nematodes with obese or swollen females occur in a number of

unrelated families within the Tylenchida, although the phenomenon is best represented, and
reaches its highest evolutionary development, in the Heteroderidae and Meloidogynidae.
Nematodes with obese females usually protect their eggs, either by forming a cyst or cyst-
like structure in which the eggs are retained, or by secreting a gelatinous matrix in which
the eggs are embedded. Where a gelatinous matrix is produced, the number of eggs laid is
often fairly small, the eggs themselves being relatively large and the ensuing juveniles
rapidly moulting to the adult stages without feeding. However, in other nematodes, such as
89
Meloidogyne, large numbers of eggs, perhaps in excess of 500, are laid into the matrix and
the juvenile stages are also parasitic.
Nematodes belonging to the Heteroderidae and Meloidogynidae (ie. the cyst and
root knot nematodes) are dealt with separately in this course, this section concentrating on
some of the other genera commonly found parasitizing the roots of crops. For the sake of
completeness, a list of the families and the obese genera they contain is appended.
Family TYLENCHULIDAE Skarbilovich, 1947
This is a small group of interesting nematodes, the most important genus being
Tylenchulus, although both the distribution and significance of Trophotylenchulus in
tropical soils are certainly underestimated. All genera are sedentary, semi-endoparasitic
root parasites and are often associated with tree crops. Tylenchulus semipenetrans is a
serious pest of citrus plantations and causes the disease known as 'slow decline'. This
nematode has a world-wide distribution and is virtually ubiquitous in citrus growing areas.
It has been disseminated on the roots of infested planting material and is often a serious
pest of established orchards.
Key to genera
1. Adult female spirally coiled; excretory duct directed posteriorly in relation to

the pore...............................................................................................................2
Adult female not coiled, excretory duct directed anteriorly from the pore........3
2. Excretory pore well behind oesophageal region...................Trophotylenchulus
Excretory pore in oesophageal region.............................................Trophonema
3. Adult female with circumoral disc, excretory pore at
less than 65% of body length.......................................................Ivotylenchulus
Adult female without oral disc, excretory pore at
more than 65% of body length.........................................................Tylenchulus
Genus TYLENCHULUS Cobb, 1913
Diagnosis: Criconematoidea. Mature female swelling posteriorly with the bulk of the
body protruding from the root surface and the attenuated anterior section embedded in the
cortex. They are small and range in length from 0.35-0.50mm. Female excretory pore far
posterior at about 68-85% of the body length and located slightly anterior to the vulva, the
duct being directed anteriorly. Female head skeleton weak; stylet slender. Oesophagus of
modified criconematid form, the median bulb being well developed and the non-
overlapping basal bulb distinct. Vulva posterior; genital tract mono-prodelphic and coiled.
Male vermiform with delicate stylet and degenerate oesophagus; bursa absent. Juveniles
vermiform with a posteriorly located excretory pore and a long, pointed tail (they die
straight and strongly resemble Tylenchus under the stereomicroscope).
Type species:
Tylenchulus semipenetrans Cobb, 1913
90
Other species:
T. furcus Van den Berg & Spaull, 1982
Bionomics: Sedentary semi-endoparasites, mainly attacking tree crops. The second stage
female juveniles are the infective stage and penetrate the root cortex. Male juveniles moult
to the adult without feeding. Feeding by the females induces the development of 'nurse'
cells in the cortex, a trophic specialization. These cells eventually disintegrate into a mass
of necrotic tissue and the epidermal layer is sloughed. The eggs are deposited in a
gelatinous matrix which is secreted by the excretory system (hence the posterior excretory
pore). Heavily infested roots will be coated with such egg masses and, as soil particles
adhere to the gelatinous material, the roots have a dirty appearance after gentle washing
when compared to healthy roots. Very large populations of T. semipenetrans can be found
on the roots of infected citrus and immense numbers of vermiform stages in the
rhizosphere. The gradual decline in vigour and cropping ability caused by the citrus
nematode causes the disease known colloquially as 'slow decline'. T. furcus was recorded
on sugar cane and grass roots in South Africa.
Family HOPLOLAIMIDAE Filipjev, 1934
Genus ROTYLENCHULUS Linford & Oliveira, 1940
Diagnosis: Hoplolaimoidea. Sexually dimorphic. Immature female found free in the soil,
vermiform and dying C-shaped when heat-relaxed. Head region rounded to conoid and
continuous with body contour. Head skeleton of medium development, stylet moderately
strong with rounded basal knobs. Dorsal oesophageal gland orifice well posterior to stylet
knobs (0.6-1.9  the stylet length). Glands well developed with a long, more or less lateral
overlap. Vulva usually posteriorly located (58-72%), genital tracts didelphic, each with a
double flexure. Tail conoid with a rounded terminus. Mature female found on roots;
swollen to kidney shaped body protruding from root, anterior part irregular. Vulval lips
protuberant, genital tracts convoluted. Male vermiform, free living in soil. Head skeleton,
stylet and oesophagus reduced, but still conspicuous. Tail pointed, spicules curved, bursa
not reaching tail tip. Juveniles free living in soil and resembling immature female in
general respects.
Type species:
Rotylenchulus reniformis Linford & Oliveira, 1940
Bionomics: The eggs are laid in a gelatinous matrix. On hatching, the juveniles moult to
the immature female or male without feeding. The immature female is the invasive stage,
but only the anterior part penetrates the root tissue, the posterior section remaining in the
soil and swelling ie semi-endoparasitism. About 50 eggs are deposited in the gelatinous
matrix which is produced by specialized vaginal cells. R. reniformis has an extremely wide
91
host range, is almost ubiquitous in tropical and subtropical soils and is reported to cause
damage to a number of crops.
Siddiqi M.R. (1986). Tylenchida: Parasites of plants and insects. CAB International,
6. ORDER APHELENCHIDA
The order Aphelenchida is for nematodes with a tylenchid type oesophagus

comprised of a mouth stylet, cylindrical procorpus, swollen muscular metacorpus (median
bulb) with distinct refractive crescentic thickenings and a terminal glandular region which
usually overlaps the intestine. Some controversy exists over whether the aphelenchs
deserve ordinal status or are merely a suborder of the Tylenchida. The two groups show
substantial differences in fundamental morphological characters, but the weighting of such
differences is open to subjective interpretation. Members differ from Tylenchida in the fact
that the dorsal oesophageal gland duct opens into the anterior part of the median
oesophageal bulb in aphelenchs, not into the lumen of the procorpus as in tylenchs.
Females, except of some insect parasitic forms, remain vermiform; the vulva is in the
posterior third of the body, there is a single prodelphic (anterior) ovary and there is often a
post vulval sac. The mouth stylet is often weak, usually under 20µm long, rarely with well
developed basal knobs, but there are some striking exceptions with stylets greatly in excess
of 100m.
The Aphelenchida is divided into two superfamilies Aphelenchoidea and
Aphelenchoidoidea. The Aphelenchoidea has the two families Aphelenchidae and
Paraphelenchidae, each of which is based on a single genus. Species within this group,
although often associated with diseased plant tissues, seem to be fungivorous and, like
some Aphelenchoides species, occasionally cause damage in mushroom culture.
The Aphelenchoidoidea has more families, the exact number depending on the
author involved. The Entaphelenchidae are obligate insect parasites and the Seinuridae are
mainly predatory. Many of the others, especially Parasitaphelenchidae, Ektaphelenchinae,
Cryptaphelenchinae and Schistonchinae are insect parasites or associates in some cases
particularly of bark boring beetles. However, within Parasitaphelenchidae,
Rhadinaphelenchus cocophilus and Bursaphelenchus xylophilus (Bursaphelenchinae) are
involved in serious disease of palm and pine trees respectively. Acugutturus parasiticus
Hunt, 1980 (the only species in Acugutturinae), an ectoparasite of the American cockroach,
and other ectoparasites of noctuid moths are unique in the Aphelenchida because of the 50-
160µm long stylet. The rarely occurring Anomyctus xenurus (the only species in
Anomyctinae) also has a long stylet (30-35µm) and is possibly a predatory nematode. The
plant parasitic "bud and leaf" nematodes are in the genus Aphelenchoides
(Aphelenchoidinae), but only three species (A. besseyi, A, fragariae and A. ritzemabosi) are
of general importance and a few more are occasionally associated with plant damage.
92
Order APHELENCHIDA Siddiqi, 1980
Diagnosis (modified after Siddiqi, 1980): Soil dwelling or insect associates; trophic habit
mycetophagous, phytoparasitic, predaceous or entomoparasitic. Body small to long
(0.2mm - 2.5mm), vermiform, rarely obese except in some insect parasites. Cuticle thin,
usually finely annulated and with lateral fields marked by 0 to more than 12 incisures.
Cephalic region low, rounded, continuous or offset and with weak or moderate
sclerotization. Stylet always present, length usually 10 - 20µm, but exceptionally up to
185µm; conus usually shorter than the shaft, but much longer in certain insect ectoparasitic
forms. Basal swellings or knobs usually weakly developed or entirely absent.
Oesophagus comprising a narrow, cylindrical procorpus, a strongly-developed, offset,
ovoid to rounded rectangular median bulb with crescentic valve plates and well
developed oesophageal glands forming a dorsally overlapping lobe in all genera apart
from Paraphelenchus where the glands are small and enclosed in a non-overlapping basal
bulb. All three gland orifi (ie. including the dorsal gland orifice) located within the
median bulb. Isthmus usually short or absent. Nerve ring circum-oesophageal or circum-
intestinal. Intestine cellular with distinct lumen. Rectum usually distinct except in some
insect associates. Anus a broad transverse slit with an overhanging anterior lip, but
absent or degenerate in some insect parasites or associates. Vulva posterior at 60 - 98%,
usually in the form of a transverse slit or, exceptionally, an oval pore (Aphelenchus).
Genital tract monoprodelphic, usually outstretched, but occasionally with a double
flexure. Spermatheca axial, if present. Post-uterine sac usually present and may act as a
spermatheca. Male genital system monorchic, outstretched. Spicules typically rosethorn-
shaped with prominent apex and rostrum, or derived therefrom, but elongate and
cephalated in Aphelenchus and Paraphelenchus. Gubernaculum usually absent, but well
developed and elongate in Aphelenchus and Paraphelenchus. Bursa usually absent, but a
true peloderan bursa with supporting ribs is present in Aphelenchus only, although some
genera may have a terminal flap of cuticle (= bursa).
Type genus:
Aphelenchus Bastian, 1865
Key to superfamilies
1. Spicules slender, cephalated. Gubernaculum well developed, elongate; V- shaped in

cross-section. Lateral fields with six or more incisures. Oesophagus with distinct isthmus
and nerve ring circum-oesophageal........Aphelenchoidea
Spicules robust, rosethorn-shaped or derived therefrom; typically with a dorsal and

ventral limb joined by a transverse bar. Gubernaculum absent or, if present, consisting of a
small structure near the distal tip of the dorsal limb of the spicule; never elongate or V-
shaped in cross-section. Lateral fields usually with four or fewer incisures, exceptionally
six. Oesophagus lacking a distinct isthmus which, if visible at all, is a short stump less than
the distance from the valve plates to the base of the bulb in length; nerve ring circum-
intestinal...............................................................................Aphelenchoidoidea
93
Superfamily APHELENCHOIDEA
Fuchs, 1937 (Thorne, 1949)
Diagnosis: Aphelenchina. Cephalic region low, flattened, continuous with body contour.
Lateral fields with six or more incisures. Oesophagus with a distinct isthmus; glands
either in a dorsally overlapping lobe (Aphelenchidae) or restrained within a non-
overlapping basal bulb (Paraphelenchidae). Nerve ring circum-oesophageal. Vulva in
the form of either a transverse oval pore (Aphelenchidae) or a transverse slit
(Paraphelenchidae). Female tail short, sub-cylindroid to conoid and with a broadly rounded
terminus which may be mucronate. Spicules slender, ventrally arcuate; cephalated.
Gubernaculum well developed, elongate; V-shaped in cross-section. Bursa present or
absent; if present then well developed, peloderan and supported by four pairs of ribs
(only three pairs reported by Thorne (1961) in A. eremitus).
Type genus:
Aphelenchus Bastian, 1865
Type family:
Aphelenchidae Fuchs, 1937 (Steiner, 1949)
Other family:
Paraphelenchidae T. Goodey, 1951 (J. B. Goodey, 1960)
Key to families
1. Male with prominent peloderan bursa supported by four pairs of ribs. Female
vulval aperture in the form of an oval pore. Oesophageal glands forming a
long dorsally overlapping lobe....................................................Aphelenchidae
Male lacking a bursa. Vulva in the form of a transverse slit. Oesophageal

glands small, retained within a non-overlapping basal bulb..Paraphelenchidae
Family APHELENCHIDAE Fuchs, 1937 (Steiner, 1949)
Diagnosis: Aphelenchoidea. Soil dwelling or found in decaying plant material. More than
six lateral lines (usually 10 to 12). Vulval aperture in the form of a transverse oval pore.
Oesophageal glands free, forming a dorsally overlapping lobe. Female tail short, cylindroid
with a rounded tip. Male bursa well developed, peloderan in form and with four pairs of
supporting ribs, one pair adanal and the other three postanal.
Genus APHELENCHUS Bastian, 1865
94
Diagnosis: Aphelenchidae. Medium sized to fairly long nematodes (0.5-1.2mm). Heat

relaxed form more or less straight to ventrally arcuate. Cephalic region low, rounded and
slightly offset. Cuticle finely annulated. Lateral fields broad with numerous incisures,
usually in excess of ten, exceptionally fewer (A. sparsus only has four to six, for example).
Stylet slender with slight basal swellings. Procorpus cylindrical, narrowing just before
joining the large, ovoid, median bulb with centrally placed, crescentic valve plates.
Oesophageal glands in a dorsal lobe. Nerve ring just behind the median bulb and more or
less opposite the excretory pore. Vulva posterior at about 70-80% of the body length.
Vulval lips slightly protuberant and body often narrowing sharply just behind the vulva.
Vagina sloping anteriorly. Genital tract monoprodelphic, outstretched. Developing
oocytes mostly in a single row. Post-uterine sac extending for up to half the vulva/anus
distance. Tail short, cylindroid, sometimes slightly ventrally concave, ending in a
broadly rounded terminus. Male, when present, with paired, slender spicules which are
ventrally arcuate and slightly cephalated proximally. Gubernaculum linear. Bursa well
developed, extending from the proximal region of the spicules to the tail tip and
supported by four (exceptionally three) pairs of ribs; one pair preanal and the other three
in a subterminal group. Tail short, conoid, tapering to a narrowly rounded point.
Type species:
Aphelenchus avenae Bastian, 1865
Bionomics: A. avenae is common throughout the world in many soils and rotting plant
tissues, it readily reproduces on a wide range of fungi including cultivated mushroom, but
does not seem to cause primary damage to higher plants.
Family PARAPHELENCHIDAE T. Goodey, 1951 (J. B. Goodey, 1960)
Diagnosis: Aphelenchoidea. Soil dwelling. Usually six to eight lateral lines. Oesophageal
glands small, enclosed in an abutting basal bulb which is amalgamated with the isthmus.
Vulva in the form of a transverse slit. Female tail short, sub-cylindroid and usually with a
mucronate terminus. Male bursa absent, four to five pairs of caudal papillae
Type genus:
Paraphelenchus Micoletzky, 1922 (Micoletzky, 1925)
Superfamily APHELENCHOIDOIDEA
Skarbilovich, 1947 (Siddiqi, 1980)
Diagnosis: Aphelenchina. Cephalic region usually high and offset from body contour.
Lateral fields with four or fewer incisures (very exceptionally six). Stylet often with
basal knobs or swellings. Oesophagus with isthmus rudimentary or absent. Nerve ring
95
circumoesophageal. Oesophageal glands in a dorsally overlapping lobe. Vulva in the

form of a transverse slit. Female tail conoid to a pointed or narrowly rounded terminus
which may be mucronate or otherwise adorned. Spicules robust, rosethorn-shaped or
derived therefrom; usually with a prominent apex and rostrum. Gubernaculum
absent or indistinct; if present it is a small structure located at the distal tip of the dorsal
limb of the spicules and is never elongate, linear or V-shaped in cross-section. Caudal
papillae usually numbering two or three pairs, occasionally as many as five pairs. Bursa
absent around the cloaca, but a small terminal flap of cuticle is present in the
Parasitaphelenchidae.
Type genus:
Aphelenchoides Fischer, 1894.
Type family:
Aphelenchoididae Skarbilovich, 1947 (Paramonov, 1953)
Key to families
1. Stylet of both sexes very long (50 - 180µm), attenuated; the conus constituting
the majority of the stylet length. Ectoparasites of insects..........Acugutturidae
Stylet usually about 10 - 20µm long, never over 35µm long and never attenuate
with the conus constituting the majority of the stylet length.............2.
2. Mature females with swollen body, endoparasitic in the haemocoel of beetles.

Three adult forms: male; immature female; mature parasitic female.
............................................................................Entaphelenchidae
Mature females not swollen or endoparasitic. Only two adult forms in the life
cycle..................................................................................................................3.
3. Females with functional anus and elongate tails more than four anal body widths
long, often becoming attenuate or filiform, but may be more cylindroid with a rounded
or spathulate terminus. Male tail elongate conoid with a spicate
terminus................................................................................Seinuridae
Females with short or medium conoid tails usually less than four anal body
widths long, but if longer and with a filiform or spicate terminus then the female
anus is non-functional............................................................................4.
4. Males with small bursa-like flap of cuticle at tail tip......Parasitaphelenchidae

Males lacking such a bursa................................................................................5.
5. Females lacking a functional anus and rectum, the intestine ending as a blind
diverticulum in the tail region. Stylet typically with a wide
lumen.....................................................................................Ektaphelenchidae
Females with a functional anus and rectum, intestine not ending as a blind
diverticulum. Stylet usually robust, with a narrow lumen and basal knobs or
swellings................................................................................Aphelenchoididae
96
Family APHELENCHOIDIDAE Skarbilovich, 1947 (Paramonov,1953)
Diagnosis: Aphelenchoidoidea. Stylet slender, with narrow lumen and usually with small
basal knobs or swellings. Post-uterine sac usually present. Female tail of medium length,
conoid, with pointed or rounded, often mucronate, terminus. Spicules paired, separate,
rosethorn-shaped or derived therefrom. Gubernaculum absent. Bursa absent.
Key to subfamilies
1. Cephalic region low, rounded, lacking an

obvious oral disc....................................................................Aphelenchoidinae
Cephalic region high, almost spherical, and with
a prominent cuticularized oral disc................................................Anomyctinae
Subfamily APHELENCHOIDINAE Skarbilovich, 1947
Type genus:
Aphelenchoides Fischer, 1894
Key to genera
1. Tail tip bearing four pedunculate tubercles with fringed margins. A vulval flap,
formed by the posterior extension of the anterior lip,
may be present.................................................................................Laimaphelenchus
Tail tip not as above, but may be variously mucronate. Vulval flap never
present........................................................................................................................2.
.
2. Vulva well posterior, at about 78-93% with the mean value in excess of 80%.
Male tail initially short conoid and then narrowing rapidly to a digitiform or
short filiform extension..............................................................................................3.
Vulva more anterior at about 60-75% with the mean value less than 80% and
usually between 65-70%. Male tail conoid, evenly tapering....................................4.
3. Post-uterine sac absent, female tail subconoid, tapering evenly to a pointed

terminus. Apex of spicules greatly elongated and continuing the line of the
dorsal limb. Associates of nitidulid beetles...................................Sheraphelenchus
Post-uterine sac present, well developed. Female tail dome-shaped with a
terminal spike. Apex of spicule small, knob-like. Associates of scolytid
beetles..........................................................................................Ruehmaphelenchus
4. Stylet very robust, about 17µm long and with massive, rounded basal knobs.
Cuticular annules coarse (1.7µm)..............................................................Megadorus
97
Stylet not with the above combination of characters, although it may be robust
and as long as 21-24µm. Cuticular annules fine.......................................................5.
5. Stylet robust, 21-24µm long with well developed knobs. Found associated with
fig wasps or inside figs...........................................................................Schistonchus
Stylet and other characters not as above....................................................................6.
6. Lips higher than wide, stylet short, robust and with rounded knobs. Conus
distinctly shorter than the shaft. Spicules strongly curved and with the apex
and rostrum well developed................................................................Tylaphelenchus
Not with the above combination of characters, spicules rosethorn-shaped, apex
and rostrum low and rounded or absent............................................Aphelenchoides
Genus APHELENCHOIDES Fischer, 1894
Diagnosis: Aphelenchoidinae. Small to long nematodes, usually between 0.4 to 1.2mm in

length. Heat relaxed females die straight to ventrally arcuate whereas the males assume a
'walking-stick' shape with the tail region sharply curled ventrally. Cuticle finely
annulated. Lateral fields often with four incisures but may be two or three. Cephalic
region usually rounded in form and slightly offset. There are six equally sized lips and the
cephalic skeleton is weak. Stylet slender, usually with basal knobs or swellings, often
about 10 - 12µm long and usually less than 20µm long. Procorpus cylindrical, leading to a
well developed ovoid or spherical median bulb with central valve plates. Oesophageal
gland lobe well developed and lying dorsal to the intestine. Nerve ring and excretory pore
posterior to the median bulb, although the excretory pore may be anterior or posterior to the
nerve ring. Vulva postmedian, usually at between 60-75% of the body length, only very
exceptionally more posterior. Genital tract monoprodelphic, typically outstretched, but
may reflex. Post-uterine sac usually present and often containing spermatozoa, but may
be absent. Tail conoid with a variable terminus which may be bluntly or finely
rounded, digitate or bifurcate or with a ventral projection. One or more mucrons of
various shapes may be present. Male tail strongly hooked ventrally to form the
characteristic 'walking-stick' form, conoid in shape and tapering to a variable terminus.
Spicules thorn-shaped, paired and separate. The rostrum and apex are usually well
developed, but may be almost absent. Typically there are three pairs of caudal papillae, one
pair adanal, one pair subterminal and the other in between. Bursa absent.
Type species:
Aphelenchoides kuehnii Fischer, 1894.
syn. A. (Aphelenchoides) kuehnii Fischer, 1894 (Filipjev, 1934)
Bionomics: Although over 100 species have been described in this genus, few have been
implicated as important plant parasites. The most important are A. besseyi on rice and
strawberry, A. fragariae on strawberry, ferns and other ornamentals and A. ritzemabosi on
chrysanthemum, strawberry and many other crops and weeds especially composites. A.
arachidis infests the testa of groundnut seed, A. blastophthorus is occasionally reported on
98
scabious and A. subtenuis from bulbs. All of these, exceptionally A. ritzemabosi, can also
be cultured on fungi as can A. composticola which can cause serious damage to cultivated
mushroom. Some other species often found associated with diseased plants are A.
bicaudatus, A. saprophilus and A. hamatus and these also feed on fungi.
Allen, M. W. (1952). Taxonomic status of the bud and leaf nematodes related to
Aphelenchoides fragariae (Ritzema Bos, 1891). Proceedings of the Helminthological
Society of Washington 19, 108-120.
Anderson, R. V. and Hooper, D. J. (1980). Diagnostic value of vagina structure in the

taxonomy of Aphelenchus Bastian, 1865 (Nematoda: Aphelenchidae) with a description of
A. (Anaphelenchus) isomerus n. subgen. n. sp. Canadian Journal of Zoology 58, 924-928.
Baranovskaya, I. A. (1981). [Plant and soil nematodes (Aphelenchoididae and Seinuridae)]

In Russian. Moscow, Isdatelstvo Nauka 233pp. [See Helm. Abs. B, 51 No. 1208]
Dean, C.G. (1979). Red ring disease of coconut. Tech. Comm. No. 47. Commonwealth
Agricultural Bureau, UK, 70pp.
Franklin, M.T. (1957). Aphelenchoides composticola n. sp. and A. saprophilus n. sp. from
mushroom compost and rotting plant tissues. Nematologica 2, 306-313.
Franklin, M.T. (1978). Aphelenchoides and related genera. In: Southey, J. F. Edit. Plant
nematology. MAFF, RB407; London HMSO pp. 172-187.
Goodey, Y. (1961). Soil and freshwater nematodes. Rewritten by J.B. Goodey. London,
Methuen 544pp.
Hooper, D.J. and Clark, S.A. (1980). Scanning electron micrographs of the head region of
some species of Aphelenchoidea (Aphelenchina: Nematoda). Nematologica 26, 47-56.
Hunt, D.J. (1993). Aphelenchida, Longidoridae and Trichodoridae: Their systematics and
bionomics. CAB International, Wallingford, UK., 372pp.
Husain, S.I. and Khan, A.M. (1967). On the status of the genera of the superfamily
Aphelenchoidea (Fuchs, 1937) Thorne, 1949 with descriptions of six new species of
nematodes from India. Proceedings of the Helminthological Society of Washington 34,
167-174.
Massey, C.L. (1974). Biology and taxonomy of Nematode parasites and associates of bark
beetles in the United States. Agriculture Handbook No. 446 Washington, D.C., United
States Department of Agriculture. 233pp.
99
Nickle, W.R. (1970). A taxonomic review of the Aphelenchoidea (Fuchs, 1937) Thorne,
1949 (Nematoda, Tylenchida). Journal of Nematology 2, 375-392.
Nickle, W.R. and Hooper, D.J. (1991). The Aphelenchinae bud, leaf and insect nematodes.
In: W.R. Nickle Ed. Manual of Agricultural Nematology New York: Marcel Dekker. pp
465-507.
Paramonov, A.A. (1964). [Principles of Phytonematology. Vol. 2. Taxonomy of

phytonematodes] In Russian. Moscow, Izdatelstvo Akad. Nauk SSR 446pp. (See also
printed and published by Indian National Science Documentation Centre for U.S. Dept.
Agric. & Nat'nl Sci. Foundation Washington, 1976. 570pp.
Siddiqi, M.R. (1980). The origin and phylogeny of the nematode orders Tylenchida
Thorne, 1949 and Aphelenchida n. ord. Helminthological Abstracts Series B, Plant
Nematology 49 , 143-170.
Thorne, G. (1961). Principles of nematology. New York, McGraw-Hill 553pp.
Yin, K., Fang, Y. and Tarjan, A.C. (1988). A key to the species in the genus
Bursaphelenchus with a description of Bursaphelenchus hunanensis sp. n. (Nematoda:
Aphelenchoididae) found in pine wood in Hunan province, China. Proc. Helminthol. Soc.
Wash. 55, 1-11.
7. ORDER DORYLAIMIDA
SUPERFAMILY DORYLAIMOIDEA
FAMILY LONGIDORIDAE
Although Longidorus elongatus (de Man) was first described in 1876, it was not
until the last 25 years or so that longidorids received much attention from nematologists.
Since about 1960 many new species have been described and extensive studies have been
carried out on the biology of selected species, usually those implicated in nepovirus
transmission.
Despite the fact that longidorids are large nematodes they tend to be under-recorded
because of the use of unsuitable extraction techniques, the nematodes remaining in the soil
and not actively participating in the extraction process. The best method is probably the
immersion-sieving technique and it is always a good idea to supplement the standard tray
extractions with this method.
Longidorids are root ectoparasites often congregating in large numbers at, or just
behind, root tips where their feeding gives rise to a characteristic galling. The
Longidoridae is the only family in the Dorylaimida known to contain numerous important
plant-parasitic nematodes.
100
Morphology
Longidorids conform to the basic dorylaimoid type, the most obvious difference being the
greatly elongate odontostyle and odontophore and to some extent their long, relatively
slender bodies. The oesophagus is in two parts - a narrow anterior section and an
expanded, muscular, basal portion containing the glands and gland ducts. The female
genital system varies from didelphic to monodelphic or mono-opisthodelphic, with most
variations in between. The vulva is usually median or anterior to median. The male tail
region often curls sharply ventrad on death due to the contraction of the oblique copulatory
muscles. The body cuticle is usually thin but tends to be thicker at the neck region and on
the tail where radial striations may be prominent. Body pores consist of a lateral series and,
especially in the oesophageal region, a ventral and dorsal series may also be present. The
amphidial apertures vary from broad slits to obscure pores but are always lateral and just
behind the lip region.
Family LONGIDORIDAE (Thorne, 1935) Meyl, 1961
Diagnosis: Dorylaimoidea. Long to very long, slender nematodes ranging from 1.5 to
12mm in length. Cuticle smooth. Cephalic region rounded, continuous with body contour
or offset. Lips amalgamated with the usual 6 + 10 circlets of papillae. Amphidial
apertures ranging from small pores to broad transverse slits. Amphids large, pouch-like or
stirrup-shaped. Lateral chords broad with one to three rows of lateral body pores. Dorsal
and ventral series of body pores usually present. Odontostyle greatly elongate, attenuate,
50 - 220µm long. Odontophore elongate, sometimes with three strong, basal flanges.
Junction of cheilostome and stomodaeum marked by a strongly cuticularized guide
ring varying in position from near the lip region to near the odontostyle base. Oesophagus
in two parts: an anterior, narrow, tubular section and posterior, short cylindrical, bulb
containing longitudinal valve plates. Three oesophageal glands - one dorsal and two
ventrosublateral. Female vulva located anteriorly to post-median in position, usually
median. Genital tracts typically amphididelphic reflexed, but may be pseudomonodelphic
or mono-opisthodelphic. Uterus of some species of Xiphinema and Xiphidorus with
various cuticularized structures in the lumen and/or attached to the walls. Spicules large,
dorylaimoid with lateral accessory guiding pieces. Tail shape variable, but generally similar
in each sex.
Type genus:
Longidorus Micoletzky, 1922 (Filipjev, 1934)
Type subfamily:
Longidorinae Thorne, 1935
Other subfamilies:
Xiphidorinae Khan, Chawla & Saha, 1978 (Jairajpuri & Ahmad, 1992)
Xiphinematinae Dalmasso, 1969
Bionomics of Longidoridae: Ectoparasites found in the rhizosphere of a wide variety of

plants. Some species of Longidorus and Xiphinema have been implicated in vectoring
101
certain plant viruses. The direct feeding damage includes root tip galling and stunting of
the root system.
Distribution of Longidoridae: Longidorus is widespread in Europe, Asia, Africa and

North America, but is probably introduced to South America. Paralongidorus is
widespread in Africa and Asia and Xiphinema is found world-wide, but with particular
diversity in southern Africa. Xiphidorus and Paraxiphidorus are restricted to South
America and Longidoroides is found in southern and eastern Africa and India.
Key to subfamilies
1 Dorsal gland nucleus elongate, smaller than those of the ventrosublateral glands
and located at some distance posterior to its orifice.................................................2.
Dorsal gland nucleus round, larger than those of the ventrosublateral glands and
located adjacent to its orifice.............................................................Xiphinematinae
2. Amphidial aperture a minute slit (pore-like), odontostyle with furcate base, guide
ring located near to odontostyle/odontophore junction, male copulatory
supplements few in number (less than 8) and with an hiatus between the adanal
pair and the ventromedian series............................................................Xiphidorinae
Not with the above combination of characters.......................................Longidorinae
The family can be divided into two major subfamilies, the Longidorinae and
Xiphinematinae, using a suite of characters. The major differential characters are:
Character Longidorinae Xiphinematinae
guide ring usually around anterior usually around posterior

part of odontostyle part of odontostyle
amphidial aperture pore-like or slit-like slit-like
odontostyle base simple forked
odontophore base simple heavily flanged
dorsal oesophageal well posterior to its just posterior to its

gland nucleus orifice, small orifice, large
102
The following genera in each subfamily are discussed in more detail:
Longidorinae Longidorus Micoletzky, 1922 (Filipjev, 1934)

Paralongidorus Siddiqi, Hooper & Khan, 1963
(included under Paralongidorus are Siddiqia Khan, Chawla & Saha, 1976 and Inagreius
Khan, 1981. Both these genera fit well under Paralongidorus and differ only in having an
offset head. Inagreius differs from Siddiqia only in having a cup-like bilobed amphidial
pouch.
Xiphinematinae Xiphinema Cobb, 1913.
Key to genera
1. Odontostyle base forked, odontophore base strongly

flanged................................................................................................Xiphinema
Odontostyle base simple, odontophore base may be thickened but not
strongly flanged..................................................................................................2
2. Amphidial aperture pore-like............................................................Longidorus
Amphidial aperture slit-like........................................................Paralongidorus
MAJOR DIFFERENTIAL CHARACTERS
Longidorus and Paralongidorus
1) body length or 'L' can be useful to divide species into broad categories
2) the lip region can be expanded from the body contour; low and rounded and
continuous with the body contour or continuous and more or less truncate
3) tail shape can be almost hemispheroid and bluntly rounded; short conical with a
broadly rounded terminus or short conical with a more sharply tapering terminus
4) odontostyle length
5) position of guide ring in relation to anterior end. Can be expressed as number of
labial widths posterior to head end
6) tail length, c and c' are of importance
7) amphidial pouch shape can vary from saccate to bilobed.
The ratio 'a' can be of use but 'b' is of little importance and 'V' tends to be about the
same for all species.
Xiphinema
1) as for Longidorus, 'L' can be useful to divide species into broad categories, e.g. all
the X. americanum group have short bodies, but can be rather variable
103
2) 'V' is useful as the vulva can be rather anterior in the monodelphic species. 'V' is
fairly constant although geographical variation can be considerable
3) odontostyle length - usually fairly constant within a population, but can vary
considerably between populations. The modern trend is to combine the odontostyle
and odontophore lengths and use the 'total stylet length' as the morphometric
character
4) female genital tract - very important as variations in structure are used to sub-divide
species into a number of groups:
(a) mono-opisthodelphic - a single posteriorly directed tract with no trace of any

anterior branch
(b) pseudo-mono-opisthodelphic - as above but some tissue of the anterior

branch remains. This tissue can be:
(i) a short undifferentiated uterine sac
(ii) a longer organ showing some differentiation but lacking an
ovary
(c) didelphic -
(i) one branch anteriad and one posteriad, both functional
(ii) as above but with a small, non-functional anterior ovary
(d) monoprodelphic - posterior branch absent leaving an anterior genital tract.

Only one species recorded in this group. This is almost certainly an
erroneous observation.
In addition there can be uterine differentiation of a cuticular nature. The true 'Z-
organ' is a spherical, muscular structure separated from the uterine tissues by sphincters and
containing a number of cuticularized pieces. Pseudo-Z organs are cuticular spines or
globular structures, often scattered along the uterine wall and are possibly more widespread
than presently recorded. Presence or absence of these structures can be important but
careful observation is essential for correct determination.
5) tail shape and length are very important. The shape can vary from short and bluntly
rounded right through to elongate and filiform. Ratio c' can vary from 0.5 to 20 and
is a very useful parameter. Tail shape can be divided into the following broad
groups:
(i) tail smoothly rounded, hemispherical

(ii) tail rounded but spatulate or clavate
(iii) tail hemispheroid with a terminal bulge, peg or mucron
(iv) tail short, conical, digitate (c' <2.5)
(v) tail regularly short conical (c' <2.5)
104
(vi) tail long conical (c' 2.5-7.5)

(vii) tail very long, attenuated (c' >7.5)
Other features of the tail, such as the presence or absence of a blind terminal canal
are also used.
6) heat-relaxed form (habitus) - fairly constant, species varying from dying in a fairly
tight spiral through various degrees of curvature to almost straight
7) lip region - whether offset, continuous, expanded etc. Can be useful but also tends
to be rather subjective at times
8) presence or absence of males
9) shape of juvenile tails - may be more useful when full data on juvenile tail shapes
are available.
Subfamily LONGIDORINAE Thorne, 1935
Genus LONGIDORUS Micoletzky, 1922
Diagnosis: Longidorinae. Body long to very long (3 to >10mm) and slender. Heat relaxed
form varying from more or less straight to C-shaped. Lateral chords broad and with one or
two rows of lateral body pores. Cephalic region rounded; continuous or offset. Lips fused
and with the usual 6 + 10 arrangement of papillae. Amphidial apertures in the form of
small, inconspicuous pores which lead back to well developed pouch-like amphid
fovea. Odontostyle elongate, needle-like; not heavily cuticularized. Dilatores buccae
absent. Guiding apparatus with a simple ring usually situated within a couple of
head-widths of the anterior end, but exceptionally further posterior, perhaps at up to 40%
of the odontostyle length. Junction of odontostyle and odontophore simple.
Odontophore about two thirds of the odontostyle in length, moderately cuticularized,
thickening slightly in the posterior region, but lacking basal flanges. Odontostylet
protractor muscles attached to base of odontophore and running parallel to the cephalic
region. Oesophagus comprising a narrow, cylindrical anterior section, which is looped
back on itself when the odontostylet is in the retracted position. There are three glands:
dorsal and two ventrosublateral. The nucleus of the dorsal gland is situated some
distance posteriorly to the orifice and is smaller than the ventrosublateral nuclei. Nerve
ring located around the narrow anterior section of the oesophagus; a second nerve ring,
located more posteriorly, occurs in some species. Hemizonid prominent. Intestine simple,
prerectum well developed and several anal body widths long. Anus in the form of a
transverse slit. Vulva a transverse slit, median in position. Vagina well developed,
muscular, at right angles to the body axis and leading to a substantial ovejector. Genital
tract amphididelphic, reflexed. Tail short, dorsally convex-conoid to a finely rounded
terminus, or broadly rounded. Several pairs of caudal pores present. Tail similar in shape
to that of the female.
105
Type species:
Longidorus elongatus (de Man, 1876) Micoletzky, 1922
Bionomics:
Ecto-parasites of plant roots, particularly soft rooted forms.
Genus PARALONGIDORUS Siddiqi, Hooper & Khan, 1963
Diagnosis: Longidorinae. Long, slender nematodes up to 12mm in length. Heat relaxed

form more or less straight to C-shaped. Cephalic region continuous with body contour
or expanded and offset by a distinct groove. Amphidial apertures in the form of
transverse slits. Amphidial fovea elongate, funnel-shaped. Odontostyle long, attenuate
and may be strongly cuticularized. Junction with odontophore ranges from plain to
markedly furcate. Odontophore usually lacking well developed basal flanges, but showing
distinct development in exceptional species. Guiding ring markedly posterior to the lip
region, but not usually at more than a third of the odontostyle length. Guiding sheath not
extending anterior to the guide ring. Oesophagus comprising a narrow tubular section
anteriorly which expands abruptly into a posterior bulboid section. Nucleus of dorsal
oesophageal gland small, rounded; situated some distance behind its orifice. Nuclei of
anterior ventrosublateral glands more developed than that of the dorsal gland. Female
vulva median to post-median in position. Genital tracts amphididelphic, reflexed. Uterine
spines or other cuticularizations not reported. Tail short, rounded; may be conoid or
hemispheroid. Spicules paired, massive; dorylaimoid in form with accessory guiding
pieces. Tail similar to that of the female.
Type species:
Paralongidorus sali Siddiqi, Hooper and Khan, 1963
Other species:
About 50 other species have been proposed.
Subfamily XIPHINEMATINAE Dalmasso, 1969
Genus XIPHINEMA Cobb, 1913
Diagnosis: Xiphinematinae. Body long to very long, 1.5 to 6mm, and fairly stout. Heat
relaxed form straight, ventrally arcuate, C-shaped or an open spiral. Cuticle smooth.
Lateral chords broad with one or two rows of lateral body pores. Dorsal and ventral series
of body pores may be present, particularly in the oesophageal region. Cephalic region
rounded, continuous or offset. Lips fused with the usual 6 + 10 circlets of papillae.
Amphidial apertures broad slits extending for almost the entire lip width. Amphid
fovea stirrup- or funnel- shaped. Odontostyle elongate, needle-like; heavily
cuticularized. Dilatores buccae present. Guiding apparatus tubular with a strongly
cuticularized posterior ring and, apparently, a lightly cuticularized anterior ring
(really just a fold in the guiding sheath). The guide ring proper is posteriorly located
106
near the odontostyle/odontophore junction. Proximal end of odontostyle appearing

forked at its junction with the odontophore which is strongly developed with three
massive posterior flanges to which the protractor muscles attach. Oesophagus comprising
a narrow, cylindrical anterior section, which is normally looped back on itself, leading to an
expanded cylindroid expansion containing the glands. Dorsal gland nucleus located at
the same level as the orifice, more developed than ventrosublateral nuclei. Nerve ring
around the anterior section of the oesophagus. Hemizonid prominent. Intestine simple,
pre-rectum well developed and several anal body widths long. Anus in the form of a
transverse slit. Vulva located anteriorly to post-median, in the form of a transverse slit.
Vagina well developed, muscular; at right angles to the body axis or posteriorly directed in
some forms with an anterior vulva. Ovejector prominent. Genital tract variable; often
amphididelphic reflexed, but as the vulva migrates anteriorly the anterior branch
progressively regresses, first becoming non-functional, then a remnant and finally
completely absent (= mono-opisthodelphic). Some species display cuticularized
structures in the uterus. Rarely these cuticularizations are found in a Z organ, a
specialized structure with thick walls and circular muscles which is constricted at both ends
by a sphincter. More commonly the cuticularizations take the form of spines or variously
shaped structures in the uterus. Tail form very variable e.g. short hemispheroid, with or
without a digitate process, medium to long conoid, initially conoid and then attenuating to a
filiform terminal section. Male genital tract diorchic, opposed. Spicules paired, massive,
dorylaimoid in form with distal accessory guiding pieces. Tail of similar form to that of the
female.
Type species:
Xiphinema americanum Cobb, 1913
Other species:
Well over 240 species have been described and are currently regarded as valid. The
taxonomy of the genus is increasingly complex, the best approach being the polytomous or
multiple entry key as used by Loof and Luc (1990, 1993). The X. americanum-group is
possibly only accessible via molecular studies.
Bionomics:
Ecto-parasites of plant roots and capable of attacking woody plants. Some species
are virus vectors.
Coomans, A. (1975) Morphology of Longidoridae. In: Nematode Vectors of Plant Viruses.

Edited by Lamberti, F., Taylor, C.E. & Seinhorst, J.W. Plenum Press, pp.15-37.
Hunt, D.J. (1993) Aphelenchida, Longidoridae and Trichodoridae: Their Systematics and
Bionomics. CAB International, 372pp.
107
Lamberti, F. (1975) Taxonomy of Longidorus and Paralongidorus. In: Nematode Vectors

of Plant Viruses. Edited by Lamberti, F., Taylor, C.E. & Seinhorst, J.W. Plenum Press,
pp.71-90.
Loof, P.A.A. and Luc, M. (1990) A revised polytomous key for the identification of
species of the genus Xiphinema Cobb, 1913 (Nematoda: Longidoridae) with exclusion of
the X. americanum-group. Systematic Parasitology, 16: 35-66.
Loof, P.A.A. and Luc, M. (1993) A revised polytomous key for the identification of
species of the genus Xiphinema Cobb, 1913 (Nematoda: Longidoridae) with exclusion of
the X. americanum-group: Supplement 1. Systematic Parasitology, 24: 185-189.
Luc, M. (1975) Taxonomy of Xiphinema. In: Nematode Vectors of Plant Viruses. Edited
by Lamberti, F., Taylor, C.E. & Seinhorst, J.W. Plenum Press, pp.39-52.
Luc, M. & Dalmasso, A. (1975) A 'lattice' for the identification of species of Xiphinema.
In: Nematode Vectors of Plant Viruses. Edited by Lamberti, F., Taylor, C.E. & Seinhorst,
J.W. Plenum Press, pp.53-70.
8. ORDER TRIPLONCHIDA
FAMILY TRICHODORIDAE
The first trichodorid was described by de Man in 1880, when he named Dorylaimus
primitivus, a species which subsequently became a member of Trichodorus, a new genus
erected by Cobb in 1913.
Trichodorids are relatively distinctive under the stereomicroscope because of their
plump, cigar-shaped bodies which are rounded at both ends, but the lack of an easily
recognizable stylet often leads the inexperienced to mistake them for non-parasitic
nematodes. Live trichodorids are very sluggish in their movements and often appear to be
moribund. When heat - relaxed they either die straight or slightly ventrally arcuate with
varying degrees of curvature in the male tail region. The body length varies from 0.5 -
1.5mm, but is usually less than 1mm. The tail is extremely short and rounded in both
sexes, particularly so in the female where the anus is subterminal.
The cuticle is thick and smooth and is lacking in transverse striae when viewed
under the light microscope. It is loose and wrinkly and may swell abnormally in some
species after fixation. The cephalic region is rounded and slightly offset, the outer ring of
papillae sometimes being particularly prominent. The amphidial apertures are located just
posterior to the lip region and take the form of short, gaping, ellipsoid structures about one
third to half the head width in size with the sensilla sac just behind the amphid pouch or
fovea. The excretory pore is small and fairly inconspicuous and is in the oesophageal
region or, rarely, a few body widths posterior. In the female, lateral body pores may be
present or absent and their presence or absence within a body width of the vulva ('advulval'
pores) is regarded as a reliable generic character.
The stoma is cuticularized and forms a simple guiding tube around the anterior part
of the onchiostyle, the proximal portion appearing as a ring or collar. The onchiostyle
108
differs in origin from both the odontostyle of the dorylaims and the stomatostylet of the
tylenchs and aphelenchs. The anterior section consists of a dorsal tooth which is
predominantly solid, but proximally may appear to be hollow. This is formed within the
region of the stoma and is attached posteriorly to an extension which is fused to the dorsal
wall of the pharynx and serves for attachment of the protractor muscles. The onchiostyle is
used as a pick to penetrate cell walls and no food passes through its structure in contrast to
the hollow odontostyle and stomatostylet of the dorylaims and tylenchs/aphelenchs
respectively. Onchiostyle length varies from about 25µm to 150µm.
The oesophagus comprises two parts: a narrow, tubular anterior section and a
posterior, poorly muscular bulboid expansion which accommodates the oesophageal
glands; bulb lumen and orifi of glands obscure. The shape of the oesophagus is noticeably
different to that of dorylaims, the posterior section gradually expanding to form a spathulate
bulb as opposed to the cylindrical form in the latter group
The vulva is median to postmedian (V = 50-60%) in Trichodorus and
Paratrichodorus, but far posterior in Monotrichodorus (V = 75-86%) and Allotrichodorus
(V = 80-90%). The lips are non-protuberant and the vulval aperture takes the form of a
small pore or a slit, either transverse or longitudinal, and often located in a shallow
depression. The vagina may be relatively short and with weakly developed, inconspicuous
musculature, as in Paratrichodorus or extending further into the body with thicker walls
and strong musculature as, for example, in Trichodorus. Distally, at the junction of the
vulva and vagina, there is a cuticularized ring which may be weak or strong in development
and which, in lateral view, appears as two, variously shaped, cuticularized bodies, one
anterior and one posterior to the vaginal lumen.
The male genital tract is monorchic and outstretched. The spicules are paired,
separate and either ventrally arcuate, or modified therefrom, as in Trichodorus, or more or
less straight with the distal tips curved ventrad.
In Trichodorus, where a bursa is lacking, the copulatory muscles extend anteriorly
for several spicule lengths and, on contraction, produce the typical J-shaped heat-relaxed
form. Conversely, in Paratrichodorus the bursa is well developed, the muscles extend for
less than a spicule length and, as a consequence, the heat-relaxed male dies more or less
straight.
A bursa may be absent, as in Trichodorus, well developed as in most
Paratrichodorus or merely represented by a flattening and thickening of the
ventrosublateral cuticle of the cloacal region.
Bionomics: Trichodorids are polyphagous migratory ectoparasites of the roots of perennial

and woody plants. Interest in the biology of trichodorids increased markedly in the sixties
and seventies when they were shown to be capable of transmitting plant viruses.
Root pathology includes browning of epidermal tissues and cessation of root
elongation („stubby root‟ disease). Cortical cells are only fed upon if made accessible by
root cracking or collapse of epidermal tissues as a result of a mass attack. Cytoplasmic
streaming to the feeding site occurs even after onchiostyle withdrawal. Cells can recover if
the nematode abandons a site soon after penetration, thus allowing the possibility of virus
transmission from infected vectors), their major pest status stems from their known ability
to transmit three tobraviruses: tobacco rattle virus (TRV); pea early browning virus (PEBV)
and pepper ring spot virus (PRV). TRV is currently known from Europe, Japan and USA,
109
PEBV only from Europe and PRV only from South America. Virus particles are
selectively adsorbed on the lining of the oesophagus and dissociation occurs as the
nematode saliva is injected into the host , although the root cell must not be killed or
damaged if transmission is to be successful. All juvenile stages can vector the viruses, but
need to be reinfected after each moult as the virus retaining cuticle of the oesophagus is
sloughed. Adult vectors can remain infective for long periods.
Trichodorids appear to be most widespread and abundant in light sandy soils
although this does not preclude their occurrence in heavier soils. They appear to be highly
susceptible to mechanical damage and as a result are usually found below the depth of
cultivation.
SUPERFAMILY TRICHODOROIDEA THORNE, 1935 (SIDDIQI, 1974)
Diagnosis: Body 0.5 - 1.5mm long, plump, females cigar-shaped, males straight or J-
shaped on heat relaxation. Cuticle thick, smooth; may swell abnormally on death.
Amphidial apertures wide, gaping ellipses; sensilla sac separated from the fovea only
by a constriction. Onchiostyle distally solid, dorsally convex, attached to the dorsal
wall of the pharynx. Simple anterior guiding ring present. Oesophagus consisting of a
narrow anterior section expanding into a posterior bulboid section of spathulate or
pyriform shape containing five glands - one dorsal, two anterior ventrosublateral and two
posterior ventrosublateral. Distinct excretory pore present, located either within the
oesophageal region or slightly posterior. Prerectum absent. Vulva small, median to
slightly postmedian or more posterior in position depending on whether the genital system
is amphididelphic, reflexed, or monoprodelphic, reflexed. Uterus a simple tube; oviduct
consisting of two cells. Spermatheca present or absent. Female anus almost terminal.
Male genital tract monorchic, outstretched. Spicules straight or curved with the spicule
protractor muscles forming a capsule around the proximal half of the retracted
spicules. Bursa present or absent. Tail very short and rounded.
Type genus:
Trichodorus Cobb, 1913
Type family:
Trichodoridae Thorne, 1935 (Siddiqi, 1961)
Other families:
Monotypic.
Family TRICHODORIDAE Thorne, 1935 (Siddiqi, 1961)
Diagnosis: Trichodoroidea. With the characters of the superfamily.
Type genus:
Trichodorus Cobb, 1913
110
Keys to genera
Female:
1. Female genital tract amphididelphic.................................................................2.

Female genital tract monoprodelphic................................................................3.
2. Vagina extending halfway into body, musculature well developed; lateral body
pores within one body width of vulva; cuticle not abnormally swollen on
fixation.............................................................................................Trichodorus
Vagina extending for up to a third of the corresponding body width; no lateral
body pores within one body width of vulva; cuticle abnormally swollen on
fixation......................................................................................Paratrichodorus
3. Lateral body pores present near vulva.....................................Monotrichodorus

Lateral body pores absent...........................................................Allotrichodorus
Male:
1. Diagonal copulatory muscles extending far anterior to head of

retracted spicules. Tail region sharply curved
(habitus J-shaped) on heat relaxation...............................................Trichodorus
Diagonal copulatory muscles either within the range of the
retracted spicules or only extending about one body width
anterior to spicule head. Tail region almost straight and not
sharply recurved on heat relaxation...................................................................2.
2. Three ventromedian copulatory supplements more or less within the range of

the retracted spicules, bursa present...........................................Allotrichodorus
Only one or two ventromedian copulatory supplements within the range of the
retracted spicules, bursa present or absent........................................................3.
.
3. Spicule protractor capsule weakly developed, bursa usually prominent, diagonal
copulatory muscles within range of the retracted
spicules......................................................................................Paratrichodorus
Spicule protractor capsule prominent, bursa absent, diagonal copulatory muscles
extending just anterior to retracted spicules..............Monotrichodorus
Genus TRICHODORUS Cobb, 1913
Diagnosis: Trichodoridae. Body plump, cylindrical with rounded ends. Heat relaxed
females die ventrally arcuate, the males J-shaped with the tail region more sharply
curved ventrad. Cuticle not swelling strongly on fixation. Oesophagus consisting of a
narrow anterior section which expands posteriorly to form a spathulate bulb. Bulb usually
non-overlapping, but in some species a ventral overlap develops whereas in others the
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intestine extends dorsally along the bulb to form an overlap. Posterior ventrosublateral
nuclei located anterior to the oesophago-intestinal junction and with the dorsal nucleus
usually at the same level. Vulva a median pore or a transverse or longitudinal slit. Vagina
extending into the body for about half the corresponding diameter. Vaginal
musculature well developed and prominent and sclerotization usually strong. Genital
tract amphididelphic reflexed; spermatheca present, although weakly developed in a few
species. Anus subterminal; tail rounded. Spicules more or less ventrally arcuate, never
straight; either smooth or with various ornamentations, bristles, etc. Gubernaculum
present. Bursa absent (but the lateral cuticle may appear as a bursa T. cylindricus). Tail
short, rounded, with one pair of ventrosublateral papillae and a pair of caudal pores.
Type species:
T. obtusus Cobb, 1913 (species inquirenda)
Other species:
About 50 species have been described and are currently regarded as valid.
Bionomics: Migratory polyphagous ectoparasites of plant roots found mainly in sandy or

sandy-loam soils in temperate regions. In addition to causing stubby root symptoms from
their direct feeding activities, some species are known to be virus vectors and are of
considerable economic importance in Europe and the USA.
Distribution: Almost world-wide (Africa, Asia, Australia, Europe, North America, Central
America), but predominantly in the more temperate regions of Europe and North America.
Some of the present distribution is probably due to nematodes being introduced with non-
indigenous plants.
Genus PARATRICHODORUS Siddiqi, 1974
Diagnosis: Trichodoridae. Body plump, cigar-shaped, heat relaxed form more or less
straight in both sexes. Cuticle swelling strongly after heat relaxation and/or acid-fixation.
Lateral body pores present in about half the species, but typically not present within a
body-width of the vulva (reportedly present in four or five species). Onchiostyle dorsally
convex. Oesophagus consisting of a narrow anterior section expanding posteriorly to form
a bulb which usually overlaps the intestine ventrally. Posterior ventrosublateral nuclei
located near to the oesophago-intestinal junction, the dorsal nucleus usually being near the
oesophageal enlargement. An extension of the intestine along the dorsal side of the bulb
may be present or absent. Vulva a minute median pore or small transverse or longitudinal
slit. Vagina extending into the body for up to one third the corresponding diameter,
musculature weakly developed and inconspicuous; sclerotization poorly developed.
Genital tract amphididelphic, reflexed. Spermatheca present or absent. Anus subterminal,
tail rounded. Males lacking or rare in about 40% of the nominal species. Spicules
normally straight; transversely striated, except at the extremities; gubernaculum present.
Bursa present, but may be inconspicuous. Tail short, rounded.
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Type species:
Paratrichodorus tunisiensis (Siddiqi, 1963) Siddiqi, 1974
Bionomics: Migratory polyphagous ectoparasites of plant roots. In addition to causing

stubby root symptoms from their direct feeding activities, some species are known to be
virus vectors, but information on the vast majority of species is lacking.
Distribution: World-wide, but mainly in tropical and subtropical regions. The genus may
well have been introduced relatively recently to Central and South America with non-
indigenous plants.
Decraemer, W. (1991). Stubby root and virus vector nematodes: Trichodorus,

Paratrichodorus, Allotrichodorus and Monotrichodorus. In: Nickle, W.R. (ed.), Manual of
agricultural helminthology, Marcel Dekker, New York, pp. 587-625.
Decraemer, W. (1995). The family Trichodoridae: Stubby root and virus vector nematodes.
Kluwer Academic Publishers, 360pp.
Hunt, D.J. (1993). Aphelenchida, Longidoridae and Trichodoridae: Their systematics and
bionomics. CAB International, Wallingford, UK, 372pp.
Siddiqi, M.R. (1974). Systematics of the genus Trichodorus Cobb, 1913 (Nematoda:
Dorylaimida), with descriptions of three new species. Nematologica, 19: 259-278.
Siddiqi, M.R. (1980). On the generic status of Atlantadorus Siddiqi, 1974 and Nanidorus
Siddiqi, 1974 (Nematoda: Trichodoridae). Systematic Parasitology, 1: 151-152.
Siddiqi, M.R. (1983). Phylogenetic relationships of the soil orders Dorylaimida,

Mononchida, Triplonchida and Alaimida, with a revised classification of the subclass
Enoplia. Pakistan Journal of Nematology, 1: 79-110.
Siddiqi, M.R. (1992). Studies on plant-parasitic nematode genus Monotrichodorus, with

descriptions of three new species from South America and Paratrichodorus paramirzai sp.
n. from India. Afro-Asian Journal of Nematology, 1: 180-191.
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CHAPTER 4
MANAGEMENT OF PLANT PARASITIC

NEMATODES
J.W. Kimenju, Dept. of Plant Science and Crop Protection,
University of Nairobi, P.O.Box 30197,
Nairobi, Kenya
INTRODUCTION
It is estimated that plant parasitic nematodes account for about 12% yield losses on a
worldwide scale. The losses can be both quantitative and qualitative in nature. In response
to the losses, nematologists and growers have developed several strategies for their control
and management.
The terms “pest control” and “pest management” are often used interchangeably but they
have different meanings. Control can be defined as an effort to kill unwanted organisms at
a particular time or place. It implies a specific act or a few acts within a limited time frame
leading to a marked reduction in either the pest population or the damage caused by the pest
Management, on the other hand can be defined as the act of managing, including the whole
system of prevention and treatment of a pest or disease. Management implies several pest
control tactics in concert over an extended period of time. Control can refer to those
specific tactics which are applied to reduce or eliminate nematode populations while
nematode management can be reserved for those efforts employed to reduce the numbers of
nematodes to non-injurious levels characterized by the use of multiple control procedures.
The main principles guiding nematode management programs are:
(i) Most plant parasitic nematodes have a wide host range.

(ii) Dispersal of plant-parasitic nematodes is usually passive but may be active or
aided by vectors.
(iii) The principal dispersal agents of plant parasitic nematodes are water, man, wind
and arthropods.
(iv) Soil, water and plant residues are the main reservoirs of plant parasitic
nematodes
(v) Knowledge of the factors that affect nematodes is invaluable in their
management.
(vi) Control of nematodes induced diseases is usually directed at inhibition of the
nematodes themselves.
(vii) Control strategies should be more preventive rather than curative and aimed at
preventing build-up of high population densities.
(viii) Sustainable management of plant parasitic nematodes requires that all viable
strategies be combined into integrated pest management packages.
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The knowledge that the initial population density will determine the expected yield is, in
many cases, the reason to reduce the initial population level. Some plant parasitic
nematodes, however, have such a high multiplication rate that even low densities can
cause remarkable damage. For such nematodes one has to try to reduce the number of
infected plants by eliminating the infection sources and by therapy of infected plants. The
reduction of nematode population levels can be obtained by the following methods:
(i) Killing the nematodes by starvation.
(ii) Directly killing the nematodes by a chemical or any other technique applied before
the crop is sown or planted.
(iii) Using chemicals that prevent the nematodes from feeding.
Sustainable management of plant parasitic nematodes can be achieved through an

integration of different tactics that fall into five broad strategies: -
(i) Preventing introduction and spread of nematodes

(ii) Cultural practices, particularly cropping systems, fallowing, resistant cultivars
and organic amendments.
(iii) Physical agents especially heat
(iv) Chemicals (nematicides)
(v) Biological control
PREVENTING INTRODUCTION AND SPREAD OF

NEMATODES
Exclusion is the first control to consider in nematode management. It is easier to deal with
nematodes before they become established in agricultural fields than it is to subsequently
eradicate or manage them. Many nematode species are carried in seed or propagating stock
(endoparasites); others can be present in soil adhering to tools or farm machinery. The
speed at which nematodes can establish themselves in an agricultural field depends on their
initial population and the frequency at which suitable host plants are grown in the same
field. Polyphagous (wide host range) nematodes are usually favoured in most situations
where different hosts are alternated.
Prevention of nematode spread can be considered at different levels; the farm, the nation
and the world. At farm level, a judicious choice of the propagation material must be the
basis for each crop. For this reason the multiplication sites (nurseries) should be
established on unsuspected or disinfested land. If the production of nematode free plants
appears to be impossible, one must apply methods that disinfest the soil such as
thermotherapy and chemotherapy. One of the ways nematodes are disseminated from
one field to another is the soil adhering to farm machinery. In many instances, new
infection sites are found at the points in the field where the farmer starts ploughing.
Cleaning the farm machinery would prevent the fields from infection. Irrigation is
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another way of nematode dissemination. Nematodes are spread by water in both the field
and in greenhouses. Sedimentation of the nematodes in a basin or a reservoir can
reduce their presence in the water and limit their spread. Filtration of irrigation water is
also gaining popularity as a nematode management strategy especially in capital-
intensive production systems.
To protect the individual farmer and to limit the nematode presence to a restricted area,
different countries restrict circulation of contaminated plant material. Legal measures
aiming at the reduction of the infections of dangerous plant parasitic nematodes are
frequent.
At the international level, important phytosanitary problems are governed by quarantine

regulations. Quarantine refers to regulatory actions aimed at preventing or retarding the
introduction, establishment and spread of dangerous pests. Quarantine pests are those of
potential national economic importance that are not yet present in the country or
endangered region or that are present but not widely distributed. Consignments must be
free of quarantine pests (Zero tolerance).
The 10 most frequently cited nematodes in the quarantine list of 125 countries are
(frequencies between brackets): Globodera rostochiensis (51), Ditylenchus dipsaci (23),
Heterodera schachtii (16), Ditylenchus angustus (13), Aphelenchoides fragariae (13),
Ditylenchus destructor (12), Radopholus similes (11), Meloidogyne javanica (11),
Aphelenchoides ritzemabosi (11), and Aphelenchoides besseyi (9).
Exclusion of plant material is confined to plants known to be hosts of pests or of biological

races or strains of pests with extremely high risk for the importing country, and originating
from countries where the pest is known to occur. Soil accompanying plant material is
undoubtedly a serious quarantine risk. It is, therefore, prohibited by most quarantine
regulations.
Spot Treatment
Spot treatment entails marking out spots of high nematode density and treatment of the
same to reduce spread of the nematodes. The strategy is based on the fact that nematodes
are hardly evenly distributed in a field. Treatment of the spots may be achieved through
application of high amounts of organic substrates or chemical nematicides.
CULTURAL PRACTICES
Crop rotation
Crop rotation is a simple nematode management method which also forms part of good
agricultural practices. Plant parasitic nematodes are obligate parasites: they need a host
plant for both their development and multiplication. Each species of phytonematodes has a
range of hosts, which may be wide, but does not include all crop plants. Nematodes
numbers increase on favorable and decline on unfavorable hosts.
In crop rotation for management of a nematode species, susceptible crops are rotated with
immune or resistant crops. The susceptible crop is usually the most profitable and the
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rotation crops less profitable. A rotation should be planned so that the nematode population
is at its lowest level when the principal or the most profitable and most susceptible crop is
planted. For example, tomato is a profitable crop but susceptible to all the common species
of Meloidogyne. After a tomato crop is harvested, the root-knot nematode population in the
soil is usually high. A second tomato crop would be severely damaged. If the nematode
species present is not M. hapla or race 1 of M. arenaria, peanuts can follow a tomato crop
without the risk of damage. While the peanuts are growing, the nematodes cannot
reproduce. Instead, many of the juveniles in the soil die or become non-infective because
of starvation and the attacks of predators, fungi and other natural enemies. Examples of
poor host crops that are commonly used in rotations to suppress root-knot nematodes
include maize, onions, wheat, rhodes grass, and asparagus.
Crop rotation is associated with major limitations. Non-host plants are not always
interesting from the financial point of view and they may be unknown to the farmer.
Sometimes rotation crops require special capital investments, which may not fit in the farm
budget. For all these reasons, rotations must be well tested before their practice can be
extended. Moreover, there is always a risk that rotations can enhance multiplication of
nematodes that were not hazards to the main crop.
Early planting: Plant parasitic nematodes are mainly harmful in the early stages of the
crop, when the root system is not well developed. In the temperate region growers can sow
or plant in cool periods, so the crop gets started before the nematodes are active. By the
time the soil warms and juveniles of cyst nematodes (G. rostochiensis, G. pallida and H.
schachtii) invade growing roots, plants have already accumulated reserves and can
withstand attack. Continued early plantings, however, may select strains of plant parasitic
nematodes adapted to the system. Although tropical conditions are generally unfavourable
for the cyst nematodes, high altitude zones experience temperate-like climate and are
therefore prone to cyst nematode colonization.
Desiccation: Nematodes in an active state are very sensitive to desiccation. When

transferred directly from water to an environment with low relative humidity or high
osmotic pressure, they are killed in a few minutes. On the other hand slow drying of
nematodes usually induces anhydrobiosis, increasing their resistance to adverse
environmental conditions including dryness and toxic substances.
In arid and semiarid areas, 80% mortality can be achieved by rapid desiccation of the soil
over a short period of time. In such areas, ploughing at intervals of 2 – 4 weeks during the
dry season can reduce populations of root-knot nematodes substantially. This exposes eggs
and juveniles in roots and deeper layers of the soil to rapid desiccation and may be
sufficient to significantly increase the yield of a subsequent crop. Intermittent irrigation
during the period of desiccation improves nematode control, owing to the increased
susceptibility of reactivated nematodes to desiccation.
Flooding: Flooding of the soil cuts off its oxygen concentration to practically zero within
one or two days. Carbon dioxide begins to increase, following flooding, due to reduction
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by anaerobic bacteria. Other chemical changes in flooded soil are: denitrification,

accumulation of ammonia, reduction of iron, manganese and sulphates and production of
organic acids, methane and hydrogen sulphide.
Flooding has been practiced as an economical method of nematode control in bananas

grown on peaty clay soil in Surinam. In that country, banana fields become heavily
infested by Radopholus similis after 4-5 years. The banana plants are then destroyed and
the fields are flooded for 4 – 5 months and then replanted with hot-water-treated rhizomes.
Soil disinfestation by flooding is mainly caused by an indirect effect due to an increase of
toxic substances produced by anaerobic microbiological activity in the soil, rather than by
the direct effect of lack of oxygen. Flooding is restricted in adoption because it is only
viable in the valley bottoms and in regions that are endowed with enormous water
resources. It may also be used to explain the general decline in nematode numbers after
excessively wet seasons.
Soil amendments: Many substances can be added to soil to increase organic matter:
manure from domestic animals, sewage sludge from municipal waste disposal facilities,
crop residues after harvest, and cover crops. Products obtained after the processing of
agricultural products are also suitable for incorporation into the soil: cottonseed hulls, and
oil cakes.
Organic soil amendments are known to improve the structure and water holding capacity
of the soil. Plants developing in a substrate rich in organic matter usually grow more
vigorously and thus tolerate damage by harmful organisms including nematodes.
Breakdown of organic matter releases compounds that may be toxic to nematodes. In
particular, decomposing residues of plant tissues release simple organic acids such as
acetic, propionic and butyric acids. These may remain for several weeks in concentrations
sufficient to kill some plant parasitic nematodes with little or no effect on free-living
species. In addition, organic substrates stimulate build-up of indigenous microorganisms
some of which are antagonistic to phytonematodes. For instance, addition of chitin to the
soil, derived from crustaceans, stimulates the growth of actinomycetes, some of which are
antagonistic to plant parasitic nematodes. It also stimulates increase of fungi with enzymes
capable of digesting chitin. Many of these fungi penetrate cyst nematodes to attack
chitinous walls of eggs. During degradation of organic matter by microorganisms, toxic
metabolites that kill nematodes are released.
The lower efficacy of amendments as compared to nematicides is outweighed by their

cheapness and relative availability. However, amendments are usually bulky and need to
be applied in large quantities. That makes them more appealing to small-scale farmers who
have limited financial resources but with access to locally produced materials. The most
widely studied materials in East and southern Africa include crop residues, animal manures
(chicken, cow, goat, and pig manures), compost, green manures and industrial wastes such
as sawdust, sugarcane bagasse , and molasses.
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Antagonistic Plants
Several plant species have been characterized as being antagonistic to plant parasitic
nematodes. The most intensively studied include marigolds, asparagus, sunnhemp, mustard
and neem. The antagonism results mainly from release of root exudates that have
nematicidal properties. Nematode-antagonistic plants have shown high potential in
nematode management when used in sequential or multiple cropping systems. Available
information shows that marigolds and sunnhemp are more effective than fallow in root-knot
nematode suppression. Over 50% decline in juvenile numbers has been reported in fields
left under marigolds for three months. Wide-scale usage of the plants is, however, restricted
because most of the plants have little or no market value. When used as companion crops,
most of them exhibit a strong weed effect resulting in reduced yields of the principal crop.
PHYSICAL METHODS
Like all organisms, nematodes have limited resistance to physical stresses exerted upon
them. Physical control methods exploit this quality. However, nematodes are usually well
protected in host tissues or soil. Therefore physical methods of control require high
energies. Moreover, there is only a small difference in heat tolerance between plants and
nematodes.
Control by heating
Nematodes are very susceptible to heat. Few nematodes resist temperatures higher than
600C for a duration of 30 minutes. Soil disinfestation by means of vapour has been
practiced in intensive crop production systems for almost a century. It is still the most
appropriate way to transfer heat to the soil, because the intensity is relatively low (1000C)
and quantity is high. Heat distribution is relatively good. The steam moves as water vapour
to the point where it is needed and then condenses on soil particles cooler than itself.
Sheet steaming is widely used because of its low capital and labour cost. In this system,
steam is blown under a plastic sheet 0.25 mm thick, anchored at the edges by sand bags or
chains and left for at least eight hours to penetrate into the soil. The drier the soil is,
initially, the more condensed water it can absorb and the deeper the heat can penetrate.
Steam penetration is slower in loam than in sandy soils, therefore one is advised to cultivate
the soil before steaming. To improve heat penetration to the deeper soil layers, a permanent
system can be developed in which the steam is blown in through pipes buried at a depth of
60 cm, and left to move to the soil surface.
Comparing different steaming techniques, Dutch researchers obtained the best results with
„negative pressure steaming‟. With this method, steam is blown under a plastic sheet and
pulled into the soil by negative pressure, created in the soil by an exhaust fan, which sucks
the air out of the soil through buried perforated drainpipes at a depth of 60 cm.
Most plant parasitic organisms are destroyed at a temperature of 600C for 30 minutes.
Higher temperatures induce chemical changes in the soil, which may have adverse effects
on subsequent growth. The release of an excessively large amount of manganese may
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cause toxicity in crops, especially in soils with low pH. Plant growth may also be affected
by an increase in levels of nitrite and ammonium nitrogen after steam sterilization.
Moreover, at high temperatures most of the soil microflora is destroyed, creating a
biological vacuum that may support the rapid development of residue and re-contaminating
organisms.
Soil Solarization
The technique of soil solarization was developed in Israel. The field is tilled to fineness
and irrigated in mid-summer and while soil moisture is still at or just below field capacity, a
clear thin polyethylene sheet is then spread over the soil and its edges buried. The sheet is
left undisturbed for periods ranging from two to nine weeks depending upon several factors
including the level of solar radiation. The polyethylene sheet is removed at the end of the
solarization period and the field is available for normal use. Irrigation can be done simply
by flooding or by sprinklers or underground drip irrigation systems.
The choice of the mulching material is important. The material should have a high
transparency to permit short-wave solar radiation to reach the soil, but it should be
impermeable to long-wave radiation (greenhouse effect) and to water vapour and gases
leaving the ground. Transparent mulching has been found to be more effective than black,
since it transmits most of the incident radiation to the soil, whereas the black polyethylene
tends to heat up.
The heating of soil under polyethylene is generally less near the edges of the plot. Thus it
is preferable to treat larger plots with continuous sealed mulch. The optimum period of
solarization depends on factors such as the quantity of solar radiation available locally, the
soil characteristics, the type of mulching materials, the thermal sensitivity of the target
organism and the available time in the cropping system for solarization.
The principles of solarization include:-

(i) Accumulation of heat in polyethylene mulched soil by transmission of short wave
solar radiation and prevention of loss of long wave radiation from the soil.
(ii) High soil moisture improves the thermal conductivity of the soil.
(iii) Greater thermal sensitivity of hydrated than of desiccated organisms.
(iv) Shift of the biological equilibrium in favour of the natural enemies of plant
pathogens.
(v) Production of toxic gases and release of toxic mineral ions.
(vi) Prolonged exposure to high temperature kills or weakens the pathogen, rendering
them more vulnerable to their natural enemies.
(vii) Water vapour condensing under the mulch reduces heat loss and may concentrate
solar radiation.
(viii) Control of weeds that serve as alternative hosts of nematodes and other pathogens.
Soil solarization can be combined with other methods of control. It has been observed
that control of nematodes with combinations of solarization and either 1, 3-
dichloropropene, ethylene dibromide, metham sodium, ethoprophos or formaldehyde is
better than with any of these treatments alone. Covering of soil with clear polythene is a
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mandatory step after application of metham sodium to reduce escape of gases from the soil.
It should be noted that the polythene is too expensive for use by most small-scale farmers in
East and southern Africa. The method has high potential in treatment of small areas such as
nurseries where disease free planting materials are produced.
Disinfestation by heating with electricity

Under intensive horticultural production in greenhouses, soil may be heated using buried
electrical resistances. Observations have demonstrated that when the soil temperature is
kept at 500C for one hour, root-knot nematodes are effectively controlled. The costs are
reduced when the heating takes place in the warm season. Although the method is restricted
in adoption due to the cost of energy, it has practical value in nurseries producing high
value planting materials especially for export. Such high value produce include
chrysanthemum flowers, carnations and roses.
Field burning
The most primitive way of heating soil is by burning stubble over it. However,
experiments have showed that burning of dry leaf litter, 10 cm thick, killed root-knot
nematodes to a depth of only 9 cm. In some cases, burning of straw and stubble after
harvest may be effective in protecting the next crop from severe attack especially by foliar
nematodes. Burning of crop resides is, however, disastrous because it denies the soil much-
needed organic matter. It also poses environmental hazards, coupled with the risk of fire
spreading to non-target areas. As earlier stated addition of organic substrates into the soil
helps to establish biological control against nematodes and also improves plant growth as a
result of enhanced nutrients and water holding capacity.
Disinfestation of planting material using heat

Hot water treatment has been successfully used in the control plant parasitic nematodes.
The treatment is useful as a preventive control measure in propagating material (seeds,
rootstocks, corms, tubers, rhizomes) that might harbour endoparasitic nematodes. The
chances of success in hot water treatment increases as the difference between thermal
sensitivity of the host and nematodes increases, with the latter being more sensitive. The
smaller the difference is, the more accurately the temperature and time of the treatment
must be controlled. It is good to note that both plants and nematodes are less susceptible to
high temperatures when they are at dormancy or dehydrated. To improve the efficiency of
hot water treatments it is necessary to displace trapped air by presoaking the propagules in
water, since the air acts as an insulator. Presoaking for about two hours before hot water
treatment is a common practice to control Aphelenchoides besseyi in rice seed. Hot water
treatment of rhizomes of bananas to control Radopholus similis is improved by peeling all
necrotic tissue from the corms before treatment.
Control by irradiation
The negative consequences of UV, gamma and X-rays on nematode reproduction, motility
and morphology are well documented. Gamma-rays have multiple effects which may
include sterilization, delayed gonadal growth, delayed egg-hatching and morphological
121
abnormalities. The movements of the nematodes are also known to become sluggish. The
irradiation effects depend on the developmental stage of the nematode and also on both the
irradiation doses and the nematodes species. Eggs may develop normally after a UV-
treatment but the juveniles hatching from irradiated eggs exhibit reduced growth and
reproduction is usually inhibited. Increasing doses of UV cause an increase in mortality of
the second stage juveniles
The use of UV-, X- and gamma rays for soil and substrate disinfestations appears to be
impractical because the soil and substrate acts as a buffer and absorbs most of the liberated
energy. The irradiation of nematode infected plants is also not feasible because plants are
more susceptible than nematodes. Ionizing rays can, however, be applied in disinfestation
of nutrient solutions used in hydroponic-type systems. A small UV-unit can treat about
2500-liter water per hour. The technology is no doubt beyond the reach of small-scale
growers especially the subsistence farmers. It can be recommended for adoption in high-
technology flower production systems.
Resistance
Growing resistant cultivars provides an ideal method for maintaining nematode population
densities below damaging levels. Resistant cultivars have several advantages over other
methods applied nematode management:
(i) It can completely prevent nematode reproduction, unlike some of the alternative
methods of control such as crop rotation.
(ii) Their use requires little or no additional technology and is cost effective.
(iii) It allows rotations to be shortened.
(iv) Adoption of resistant cultivars is not associated with any toxic residues.
For some low-value crops, breeding resistant cultivars is probably the only practical long-
term approach to nematode control. This is also true for high-value crops, which are very
expensive to establish and / or maintain with frequent nematicide treatments.
Besides their resistance to plant parasitic nematodes, resistant cultivars need to be tolerant;
those that are intolerant suffer extreme damage if grown in heavily infested soil. Tolerant
cultivars that are not resistant tend to increase the nematode population densities to
damaging levels.
Resistance
Resistance describes the effects of host genes that restrict or prevent nematode
multiplication in a host species. Tolerance of damage is independent of resistance and
relates to the ability of a host genotype to withstand or recover from the damaging effects
of nematode attack and yield well.
Resistance induced by the nematode depends on the nematode biology. Ectoparasites

generally have a necrotrophic relationship with the plant. The injected saliva liquefies the
cytoplasm that accumulates around the stylet tip and is rapidly ingested, usually killing the
host cell. The nematode then moves to a new cell and repeats the process. Such behaviour
limits the possibilities for effective induced resistance. Migratory endoparasites may have
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a necrotrophic type of feeding; in some species, however, the feeding is partly biotrophic as
it involves the induction of favourable changes in cells adjacent to the feeding site.
Resistant cultivars are available to species within this group, especially the stem and leaf
parasites. Sedentary endoparasites are obligate biotrophs. Nematodes within this group are
the most damaging to plants and resistance to them is widespread.
Tolerance
Cultivars of a crop are regarded as differing in their tolerance if the decrease in their growth
and / or yield due to damage by nematodes differs significantly when they are grown in
uniformly infested soil. Where cultivars of different yield potentials are being compared,
the proportional effect on yield must be compared in adjacent, uniformly uninfested and
infested conditions. Resistance and tolerance are independent attributes of plants but
resistance may confer tolerance especially if it decreases the incidence of nematode attack
or parasitism. Equally, mechanisms of tolerance exist that are independent of resistance.
Tolerance is probably widespread and important in wild plants but is likely to be lost during
crop breeding.
Various mechanisms of tolerance have been suggested. They include: differences in

numbers of roots, compensatory root growth, delayed senescence and enhanced water
uptake. Tolerance is independent of resistance and it appears to be largely nonspecific.
The use of tolerance is a valuable strategy for many crops or situations where alternative
control measures are not available. However, tolerance of cyst and root-knot nematodes
generally needs to be combined with a degree of resistance, otherwise populations will be
increased to densities where even tolerant cultivars are damaged. Susceptible and tolerant
genotypes may, however, be useful in rotations following resistant genotypes as a means of
reducing the rate of selection of virulence.
BIOLOGICAL CONTROL
Many natural enemies attack plant parasitic nematodes in the soil and reduce their
populations. They include bacteria, rickettsia, fungi, protozoa, tardigrades, tubellarians,
nematodes, enchytraeids, mites, and insects. It is important to determine the nature and
extent of such attacks on nematode multiplication in order to establish whether these
enemies can be exploited to reduce damage and increase crop yield.
There are two types of biological control: induced, where the biological control agent has
been applied by man, and natural, where the agents have increased to suppress nematode
multiplication without being specifically introduced. The term suppression is used to
indicate a reduction in the numbers of nematodes not simply a reduction in disease
symptoms; it may be general or specific if only one or two organisms are involved.
Biological control should not be considered as a replacement for the chemical control.
Growers that require rapid kills when their fields are found to be heavily infested have few
123
alternatives to chemical treatment. Biological control is slow acting and cannot fulfill this
requirement. It, therefore, fits very well in an integrated nematode management system.
Introduced organisms that must become established in soil and spread are likely to be
affected greatly by environmental conditions. Control is also density-dependent, with a
greater proportion of nematodes escaping attack when they are at low densities. Hence,
rarely do natural enemies eradicate pests. Equilibrium populations are established that
should be below the economic threshold for damage to the crop.
Natural enemies as biological control agents
Pasteuria penetrans
Pasteuria penetrans is an obligate parasite of some plant parasitic nematodes. Nematodes
become infected in soils when they get in contact with the endospores, which adhere to
their cuticle. For instance, infected second -stage Meloidogyne juveniles enter roots and
begin feeding before the spores germinate. A germ tube penetrates the cuticle and gives
rise to a vegetative microcolony that fragments and proliferates throughout the nematode
body cavity. Eventually females become filled with spores and egg production is
prevented. Pasteuria penetrans is very virulent and has reduced Meloidogyne populations,
in pots, by 99% in only three weeks.
Pasteuria penetrans can survive several years in air-dried

soil apparently without loss of viability and is little
affected by soil conditions or a range of nematicides.
Spores adhere to many plant parasitic nematodes, but
only 20 – 30% germinate, and attachment may not
necessarily lead to infection.
The concentration of spores and the period of nematode
activity in soil, therefore, determines the number of
nematodes killed. P. penetrans spores are non-motile and
are spread in soil chiefly by water movement and by tillage
practices. Although P. penetrans appears to have
considerable potential as a biological control agent, its
commercial exploitation is prevented by lack of methods
for large-scale production.
Figure 1. Root knot nematode juvenile encumbered with spores of P.penetrans
Nematode trapping fungi

Fungi that produce adhesive network traps are good saprophytic competitors and grow
rapidly in vitro but are less efficient at trapping nematodes than some that are slower
growing and capture their prey on adhesive knobs and branches or in constricting rings.
124
Most trapping fungi do not seem capable of rapid colonization and are considered poor
competitive saprophytes that do not readily establish when added to soil. A carbohydrate
source other than nematodes is necessary to support mycelial growth.
Different species of trapping fungi vary in their ability to capture nematodes, but there is
little evidence of specificity for particular prey species, and once traps are produced, most
types of nematodes are caught (Fig. 2). The limited trapping activity and its non-specific
nature have meant that these fungi are difficult to exploit as control agents, and control has
been inconsistent. A commercial strain of Arthrobotrys irregularis has been produced on
rye grain and marketed as Royal 350. It is recommended that the fungus be applied at 1.4
t/ha at least 1 month before planting and incorporated into the soil. Reduced root galling
caused by Meloidogyne spp. and increased tomato yield has been reported.
Arthrobotrys robusta, sold under the trademark Royal 300, is used for the control of
nematodes damaging mushrooms (Ditylenchus myceliophagus). It is introduced into the
compost on rye grain at a rate of 1% with the mushroom spawn. In one trial, Royal 300
increased yield by 20% and reduced the final populations by 40%.
A B
A B
C D
Figure 2. Mycelial network of a nematode trapping fungus, Arthrobotyrs oligospora (A),
nematodes trapped by the fungus (B & C) and a nematode trapped in adhesive mycelial
columnar of Monacrosporium sp (D)
125
Endoparasitic fungi of vermiform nematodes

The endoparasitic fungi produce small spores that contain too little energy to initiate
colonization in soil. Hence spores remain dormant until they adhere to a passing nematode,
after which they germinate, penetrate the cuticle and colonize the host. Nematoctonus
concurrens and N. haptocladus belong to this group and are quite effective against
nematodes.
Parasites of nematode eggs
Eggs are more susceptible to infection before second stage juveniles have developed, and if
young females are parasitized their fecundity is decreased. Therefore these fungi are more
effective if they are able to parasitize females and egg masses soon after emergence on
roots. Fungal parasites of eggs appear to be facultative parasites that can be grown readily
in vitro, and their survival in soil is probably not dependent on the presence of nematodes.
Most infection occurs when the females and cysts are on the roots and not when they are
dispersed in soil. Assuming that the most convenient time for applying these fungi is
before planting, they must survive several weeks in soil in sufficient numbers to control the
new generation of nematodes. Application of an energy source with the inoculum‟ is
therefore essential.
Egg parasites do not kill active juveniles. Hence, in heavily infested field, crop damage
may not be decreased after treatment, particularly when the nematode has only one
generation per season. Such treatments may prove difficult for growers to accept and
necessitate the combined use of a nematicide to reduce initial damage.
A considerable range of fungi have been reported to colonize eggs of cyst- and root- knot
nematodes. The best documented are: Paecilomyces lilacinus, Dactylella oviparasitica and
Pochonia chlamydosporia.
Applications of Paecilomyces lilacinus on cereal grains readily colonize soil, and a single
treatment may be sufficient to establish the fungus. Rates of 0.4 t/ha seem to reduce
damage caused by Meloidogyne.
Dactylella oviparasitica and Pochonia chlamydosporia parasitise eggs of root knot and
cyst nematodes and have been associated with the natural control of M. incognita on a wide
range of crops. It is easy to grow these fungi in vitro. They also colonize the rhizosphere
of plants without causing damage.
Factors affecting biological control in soil

Compared to other methods of control, the efficacy of biological control agents is more
likely to be influenced by the environment .Each agent seems to have its own optimum
conditions of particularly pH, temperature and moisture. The biological control is
influenced by the multiplication rate of the nematode and by the time the nematode is
exposed to the antagonist. Endoparasitic nematodes are relatively harder to control by
biological means as they spend a great part of their lives inside the roots.
126
The major restriction on the development of effective biological control of nematodes is the
large bulk of soil that must be treated to ensure contact between host and agent. In
addition, mycostatic factors generally produced by the microbial population in the soil may
prevent the germination or growth of fungal propagules introduced into the soil.
CHEMICAL CONTROL
Chemicals have been used to control nematodes for a long time. In the period up to the
First World War, carbon disulphide, formaldehyde, cyanides and other chemicals were
investigated and proved to be too expensive for general nematode control. Chloropicrin, a
war gas, became available as a surplus after the war. This compound improved crop
production where poor yields resulted from repeated monoculture. Its success stimulated
search for other compounds and the discovery of the nematicidal effect of D –D, a by
product of petroleum refining, and of EDB followed. The use of nematicides is
recommended when:
(i) The nematode population requires an immediate intervention
(ii) Other methods fail to reduce nematode populations to acceptable levels.
(iii) The crop protection should be at maximum eg on export materials.
(iv) Besides nematodes, other crop parasites are to be controlled.
Types of nematicides
Nematicides fall into two groups:
1. Fumigants are volatile liquids that vaporize and dissolve in the soil solution. Their high
vapour pressure distributes the gas in all directions through soil pores. Most of the
fumigants are phytotoxic and directly kill nematodes and their eggs. Two types can be
distinguished:
(i) Halagenated aliphatic hydrocarbons: methyl bromide, ethylene dibromide (EDB)
1,3-dichloropropene mixtures and
chloropicrine.
(i.i) Methyl isothiocyanate precursor compounds: Metham sodium, dazomet, and
methyl isothiocyanate mixtures: solutions in xylol or mixed with, 1,3-
dichloropropane mixtures (Vorlex).
2. Nonfumigants nematicides are often known as nematostats, as they do not kill
nematodes directly but do affect their behaviour. Most of these nematicides have a
systemic action. Different formulations exist and these nematicides can be applied at
sowing or planting and also later. Two chemical groups can be considered:
(i.) Organophosphates: fenamiphos, ethoprophos, thionazin, fensulphotion.
(ii) Carbamates: aldicarb, oxamyl, carbofuran, methoxyl.
It should be emphasized that chemicals are potentially harzardous to the environment, man
and other non-target organisms. Judicious application is strongly recommended with the
aim of reducing the risks associated with them and also the pesticide residues in agricultural
produce. Chemical application can only be economical in production of high value crops
because they are expensive.
127
A number of chemicals have been banned for use, particularly in crops destined for the
export market. Growers are advised to get information about permitted products before
using nematicides on any crops. Details about methods and rates of application, modes of
action can be obtained from various literature.
CONCLUSION
Many strategies have been developed for use in nematode control. Wisdom is, however,
required in selection of the most appropriate strategies for use in different situations.
Selection of nematode control methods should be guided by the economics of crop
production, target nematode species and potential nematode threats, environmental and
food safety regulations. Emphasis should be placed on the fact that no single strategy can
be relied upon to sustainably contain nematode populations. Integrated pest and crop
management approaches encompassing as many feasible strategies as possible should be
adopted.
REFERENCES
Brown R.H. & Kerry B.R. (Eds.). 1987.Principles and practice of nematode control in
crops. Academic Press New York.447 pp.
Decker H. 1989. Plant nematodes and their control (Phytonematology). E.J. Brill, Leiden
540 pp.
Dropkin V. H.1989. Introduction to plant nematology. John Wiley & Sons New York 304
pp.
Luc M., Sikora R.A. & Bridge J. (Eds.). 1990. Plant parasitic nematodes in subtropical and
tropical agriculture. CAB International, Wallingford. 629 pp.
Nickle W. R. 1991. Manual of Agricultural Nematology. Marcel Dekker, Inc. New York
1035 pp.
Stirling G.R. 1991. Biological control of plant parasitic nematodes: progress, problems and
prospects. CAB International, Wallingford.282 pp.
Wajid Khan M. (Ed.). 1993.Nematode interactions. Chapman & Hall, London . 337 pp.
Zuckerman B. M. & Rhode R.A. (Eds.). 1981. Plant parasitic nematodes Volume III.
Academic Press New York. 508 pp.
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NIESA Training Manual

FUNDED BY NIESA and UNIVERSITY OF NAIROBI, CROP PROTECTION

CONTRIBUTORS: J. Kimenju, Z. Sibanda, H. Talwana and W. Wanjohi

This uneven distribution makes collection of truly representative samples difficult as it

Figure 1. Examples of tools for soil sampling to diagnose nematode infestations

Direct examination of plant material

B. Extracting live nematodes

Modified Baermann Technique - Extraction tray method (Whitehead and Hemming,

4. Add sugar solution (450g/l water) to tubes

Extraction of Nematodes from plant materials

Figure 3: - Extraction tray method

Figure 4: The sieving technique (bucket-sieving method)

1. Modified Baermann funnel technique: This technique uses a supporting mesh to

Figure 5: Baermann Funnel Technique (top) and one of its modifications

Bioassays for detecting and quantifying nematodes

Killing and fixing nematodes

Both procedures result in what is known euphemistically as "heat relaxation", relying on a

2. Formal acetic (FA) or formal propionic (FP) 4:1: 10 ml Formalin (40%

3. TAF: 7ml Formalin (2 – 4% formaldehyde) + 2% glycerol, 2 ml tri-ethanol-amine

Processing and Mounting Nematodes

Fixing with formalin-glycerine and transferring to glycerine through ethanol

Fixing with TAF and transferring to glycerine through evaporation

Prepare double-strength TAF fixative containing 8% formalin and 2% triethanolamine in

Observation of detailed morphology of live nematodes can also be done by making a

Step by step procedure

The Noble Art of Picking Worms

The easy way (using Pipettes)

BIOLOGY OF PLANT PARASITIC NEMATODES

Objective: Develop an understanding of the general biology of plant parasitic nematodes

Adaptations to mode of feeding

During parasitic development of sedentary endoparasitic species (Meloidogyne spp;

Above ground symptoms

i. Stunting –the reduction of growth rate, reduction in amount of foliage and

system is dwarfed and thickened in appearance (Trichodorus, Longidorus,

4. Nematode disease complexes

Nematodes are important vectors of viruses. Xiphinema and Longidorus transmit

Bacteria can be transmitted by nematodes externally on their body surfaces or internally

All PPN wound plants either by a simple micro-puncture or by rupturing or separating

There are mainly three methods of reproduction in nematodes.

Reproduction using amphimixis is also retained as a facultative ability among some

In amphimictic species of Hoplolaimina, the spermatheca is usually round and axial

iii. Hermaphroditism (automixis or self-fertilization) is as in Caenorhabditis elegans,

6. Life cycle and Hatching

of the eggshell. It consists of two or three lipoprotein membranes although in Heterodera

Summary of life cycle

Nematodes can be dispersed actively or passively.

example, the palm weevil, Rhynchophorus palmarum disperses Bursaphelenchus

Dispersal by other means

1. Migrating to escape unfavourable environmental stress. For example, when surface

Diapause unlike quiescence is temporarily irreversible and requires other “triggers” to

Facultative diapause- initiated by exogenous environmental factors and is terminated by

Quiescence= “slowed ageing” is a spontaneous reversible response to unpredictable

Alterations in metabolism can be triggered by adverse environmental conditions such as

iv. Osmotic shock (high solute concentration = osmobiosis

Anhydrobiosis- adaptation to desiccation

The ability of nematodes to survive depends on their inherent ability to withstand

Species Longevity whilst desiccated

Anguina tritici 32 years

Mechanisms of anhydrobiotic survival

i. Reducing the rate of water loss

Induction of galls in the inflorescences by anguinids (Anguina tritici, A. agrostis) is a true

Behavioural adaptations to dessication

Coiling is associated with a wide range of nematodes as a desiccation survival mechanism,