TERM PAPER

“MICROBIAL PHYSIOLOGY AND

METABOLISM”
Topic: - PAECILOMYCES LILACINUS

D.O.S:-19-11-10

SUBMITTED BY:
Mr:- SHASHI SHARMA
Roll no :- B-15
Reg. No:-11006142
Section:-P8003
 TABLE OF CONTENT

1. ABSTRACT

2. INTRODUCTION
3. DIVERSITY
4. IMAGES
5. TAXONOMY
6. DESCRIPTION
7. LIFE CYCLE
8. PRODUCT DETAILS
9. SOIL APPLICATION
10. PLANT PARASITIC NEMATODES
11. METHODS FOR NEMATODE CONTROL
12. SOLID STATE FERMENTATION
13. WEST INDIAN JOURNAL
14. BIOCONTROL AGENT
15. METHOD OF APPLICATION
16. FREQUENCY OF APPLICATION
17. DOSAGE
18 .REFERENCES
 ABSTRACT

GA Paecilomyces lilacinus is an agent for the potential biological control of soil

nematodes. Arbitrarily primed PCR was used t o fingerprint the genomes of 28 isolates
of this fungus. Most (72%) of the isolates originated from soil of different regions of
Brazil. Fourteen 10-mer oligonucleotide primers of arbitrary sequence revealed 293
scorable binary characters. Distinct genotypes were obtained for each isolate. Cluster
analysis showed a high level of variability among these genotypes. The similarity
among pairwise
comparisons of the isolates varied from 84.3% to F6%, with a mean of 6305%. No
clearly defined phenetic groups were identified by cluster or multivariate analyses. No
correlation with geographical origin or host was detected. In addition, PCR with four
pairs of consensus tRNA gene primers was performed on a subsample of 12 P. lilacinus
isolates, three P. farinosus isolates, two P. fumosomseus isolates, and one isolate of P.
amoenomseus. An inferred phylogeny based on 112 binary characters obtained by
tRNA-PCR showed a monophyletic group which contained most of the P. lilacinus
isolates. In contrast, three isolates of P. farinosus were not in a monophyletic group
under the inferred phylogeny. These results suggest that tRNA fingerprinting could
provide a valuable tool which could be used to develop the molecular taxonomy of
Paecilomyces, as morphological characteristics of asexual structures cannot entirely
resolve species.
Avail from us superior quality Paecilomyces lilacinus which are basically filamentous
fungus, used to kill harmful root-knot nematodes. These are formulated by our experts
as per the varied requirements of the client. These belong to the family of
Trichocomacea.
 INTRODUCTION

Paecilomyces lilacinus is effective in plant parasitic nematodes. At GreenMax AgroTech,

Pacilomyces lilacinus is available as talc based formulation with high density spore population. This
product is also available as combined formulation with Beauveria bassiana. Paecilomyes lilacinns is a
typically soil-borne hyphomycete that has been isolated in different parts of the world, especially in
warmer regions (Domsch & Gams, 1980). P. lilacims has remarkable versatility and can survive on
diverse substrates and infect different hosts (Samson, 1974). It can live as a mycoparasite that
colonizes sclerotia of several species of fungi (Gupta e t al., 1993). Also, it has been reported to infect
eggs and cysts of nematodes (Carneiro, 1992). Although the activity of P. lilacintls on insects is
infrequently reported, there is one report of the use of this fungus to control populations of
Nilaparvata ltrgens in rice fields (Rombach e t al., 1986). Recently, an isolate of P. lilacinns has been
studied as a potential biological agent to control eggs of both the nematode Meloidogyne javanica and
the corn root worm, Diabrotica speciosa, in Brazil (Tigano-Milani e t al. , unpublished data). P.
lilacintrs was the most frequently isolated species, when a survey of selective isolation of
entomopathogenic fungi was conducted on soil samples from different regions of Brazil (Chase e t al.,
1986 ; Tigano-Milani e t al., 1993). To study the efficacy of this potential biopesticide, it will be
necessary to characterize the apparently large endemic, soil-borne population. Environmental release
will also require means for positive identification of strains. Genetic markers can be used to study
populations of P. lilacinas. Markers will be required to evaluate the efficacy of treatments, to study
population dynamics and to allow characterization of genetic variation in P. lilacinns. Until now, P.
lilacintrs has been characterized mainly by virulence tests or biochemical characters (Samson, 1974;
Carneiro, 1992). However, it is difficult to associate variability of these phenotypic characteristics
with overallgenetic and phylogenetic relationships. Environmentally sound and economically feasible
alternatives for pest control are nowa subject of numerous studies due to the development of
resistance to pesticides intargeted pathogens, as well as the withdrawal of commercial pesticides from
the marketdue to environmental and public health concern. Plant-parasitic nematodes, especially root-
knot nematodes (RKN), are major pests of several economically important crop plants, causing severe
yieldloss. Methyl bromide, the most widely usedsoil fumigant against nematodes, has beenbanned in
most developed and developing countries, because of the serious threats it poses to the environment.
So there is an increasing demand all over the world forecofriendly nematicides. Biological control
agents (BCAs), such as fungi, offer great scope for field application, but the development of a viable
bioprocess for its commercial production is not an easy task. Paecilomyces lilacinus (Thom) Samson
is a known soil hyphomycete, and it parasitizesRKN eggs and females showing great nematicidal
activity. Growth physiology of filamentous fungi is an important factor considering their production
as BCAs. The German manufacturer Prophyta produces a commercially patented strain of P. lilacinus,
which is under continuousresearch worldwide. Field applications of BCAs are mainly accomplished
by means of fungal conidiospores, which must be virulent and viable for long periods of storage. Solid
state fermentation (SSF) offers many advantages for large scale and cost effective production of
conidiospores. The present review deals with the relevance of controlling RKN in agriculture, and the
application of the BCA P. lilacinus produced under SSF. Existing methods for controlling nematodes
are discussed with emphasis on biological control research and practices using the spores of P.
lilacinus.

DIVERSITY

Paecilomyces lilacinus is a common saprobic, filamentous fungus.It has been isolated from a wide
range of habitats including cultivated and uncultivated soils,forests,grassland,deserts, estuarine
sediments and sewage sludge. It has also been found in nematode eggs, and occasionally from females
of root-knot and cyst nematodes. In addition, it has frequently been detected in the rhizosphere of
many crops. The species can grow at a wide range of temperatures – from 8°C to 38°C for a few
isolates, with optimal growth in the range 26°C to 30°C. It also has a wide pH tolerance and can grow
on a variety of substrates. P. lilacinus has shown promising results for use as a biocontrol agent to
control the growth of destructive root-knot nematodes.
 Divergent phialides and long, tangled chains of elliptical conidia borne from more
complex fruiting structures characteristic of Paecilomyces lilacinus; magnification
460X.

Scientific
Classification

Kingdom: Fungi

Phylum: Ascomycota

Class: Ascomycetes

Order: Eurotiales

Family: Trichocomaceae

Genus: Paecilomyces

Species: P. lilacinus
IMAGE OF PAECILOMYCES
LILACINUS :-
Paecilomyces lilacinus

Taxonomy

P. lilacinus was classified with the Fungi Imperfecti or Deuteromycetes, fungi for which perfect (i.e.,
sexually reproducing) states have rarely been found. Paecilomyces lilacinus is classified in the section
Isarioidea, for which perfect states have not been found. Many isolates of P. lilacinus have been identified
from around the world and it is accepted that variation exists within the species

Description

P. lilacinus forms a dense mycelium which gives rise to conidiophores .These bear phialides from the ends
of which spores are formed in long chains. Spores germinate when suitable moisture and nutrients are
available. Colonies on malt agar grow rather fast, attaining a diameter of 5–7 cm within 14 days at 25°C,
consisting of a basal felt with a floccose overgrowth of aerial mycelium; at first white, but when
sporulating changing to various shades of vinaceous. The reverse side is sometimes uncolored but usually
in vinaceous shades. The vegetative hyphae are smooth-walled,hyaline, and 2.5–4.0 µm wide.
Conidiophores arising from submerged hyphae, 400–600 µm in length, or arising from aerial hyphae and
half as long. Phialides consisting of a swollen basal part, tapering into a thin distinct neck. conidia are in
divergent chains, ellipsoid to fusiformin shape, and smooth walled to slightly roughened. Chamydospores
are absent.

Life cycles
P. lilacinus is highly adaptable in its life strategy: depending on the availability of nutrients in the
surrounding microenvironments it may be entomopathogenic, mycoparasitic, saprophytic, as well as
nematophagous.

SCOPE
Brandname:Gmaxbioguard:- :-

ProductDetails:
Technical:
Mother culture of Pacilomyces lilacinus was sourced from Project Directorate of Biological Control (PDBC),
Bangalore. Pacilomyces lilacinus, mass multiplied from virulent and pure mother culture is supplied as talc based
formulation. The product will have a minimum population of 1x 107 (CFUs)/g.

Formulation:
Talc carrier based product. The product has minimum shelf life of one year from the date of manufacture.

Composition:

• Pacilomyces lilacinus 1% (w/w)

• Sticking agent – CMC – 1%.
• Inactive Ingredients 98.0% (w/w)
o (Moisture 35%, talc 63%)

Packing:-
The product is available in attractive one kg laminated poly pouches. The packing is moisture and proof and
well tolerates transportation and handling. The product is also supplied in bulk packing of 50kg/25 kg sizes in
HDPEbags.

DESCRIPTAION ABOUT THE PRODUCT:-

Paecilomyces lilacinus is a common saprophytic, entomopathogenic, mycoparasitic, saprophytic, as well as
nematophagous, filamentous fungus. It is parasitic on nematodes infecting eggs, juveniles, and adult females of
root-knot and cyst nematodes. The species can grow at a wide range of temperatures – from 8°C to 38°C for a few
isolates, with optimal growth in the range 26°C to 30°C. It also has a wide pH tolerance and can grow on a variety
of substrates. P. lilacinus has shown promising results for use as a biocontrol agent to control the growth of
destructive root-knot nematodes. Paecilomyces protects the roots against plant parasitic nematodes, specifically
root-knot nematodes (Meloidogyne spp.), Banana nematodes (Radopholus similis) reniform nematode
(Rotylenchulus reniformis), and citrus nematodes (Tylenchulus semipenetrans). These nematodes infect
horticultural crops of economic importance.
 PLANT-PARASITIC NEMATODES

Nematodes are roundworms that belong to the phylum Nematoda. They are the
most abundant creatures on earth, occupying different ecological niches and
living as parasites of humans, animals and plants. Parasitic nematodes can cause
a large-scale multiplication and invasion of their host. Phytoparasitic nematodes
can devastate several economically important crops, causing significant losses in
yield. These nematodes are obligate parasites, and they have developed
different parasitic strategies and relationships with their hosts to attain enough
nutrients for development and reproduction. The products of nematode parasitic
genes can be expressed as morphological structures (e.g., stylet), which allow
researchers to assess the level of parasitism in a particular host plant, where
nematodes can develop critical physiologicalfunctions
in the interaction with their host.

METHODS FOR NEMATODE

CONTROL

Since 1950, the control of phytoparasitic nematodes has been based on chemical
pesticides, although several of them are being withdrawn from the market due to
issues related to the environment lic health. Methyl bromide was widely used
against nematodes, but now it has been withdrawn from the market because of
its adverse effects on the ozone layer. Nematodes also developed resistance
against most of the known pesticides, and this triggered worldwide research for
new alternative agents and methods for nematode control. Possible control
measures change with climate conditions, socio-economical situation of the
country, crop economy, availability of chemical pesticides, resistant cultivars,
and the suitability of agricultural practices.

Resistant plants:-

Plants are resistant to nematodes when they have a reduced level of

reproduction. Nematode resistance genes are present in several crops, and are
an important component of various multiplication programs in tomatoes,
potatoes, cotton, soybean, and cereals. Resistance to nematodes can be either
broad with action against several species of nematodes or narrow against only
selected specific biotypes113. Several resistance genes, dominant or
semidominant, were identified, cloned, and subjected to various studius.

Crop rotation:-

Important method for maintenance and improvement of soil fertility, and for
enhancing yield. In crop rotation, various crops are followed in a certain order in
the same soil. With the same succession of crops reproducing in a regular time
cycle, rotations can be biennial, triennial, and so on. Crop rotation is a very good
strategy that can always be adopted against nematode species with narrow
ranges of plant-host, which is not the case of Meloidogyne sp. However, the order
of plants and the time intervals between susceptible crops depend on the
nematode species.
Chemical control:-

Plant-parasitic nematodes are more vulnerableas juveniles (J2) in soil, when

searching for the roots of host plants. Once an endoparasitic nematode species
penetrates a root, chemical control is more difficult as compounds have to be
non-phytotoxic. There are several nematicides that can be used effectively
against nematode pests of many annual crops, but there appears to be little
progress for management of nematodes in many susceptible perennial crops
without repeated application of nematicides36. There are two kinds of chemical
products that can be utilized against plant parasitic nematodes: soil fumigants
and nematicides. Their application to soil depends on the form of the
formulation, it can be by injection, spraying, mechanical means, or through
irrigation pipes ucts are usually applied before planting and, in the case of
pesticides, they are applied at the time of planting. Fumigants are highly
effective against nematodes, their efficacy is related to their high volatility at
ambient temperatures. All fumigants have low molecular weights, and are
available as gases or liquids. As they volatilize, the gas diffuses through the
spaces between soil particles where the nematodes are killed. The most widely
used fumigant is methyl bromide, which is mainly applied for high valued crops,
such as strawberries and tomatoes, and in lesser amount to grains and
commodities. However, methyl bromide has been banned in developed countries
since 2005. In developing countries, substances with methyl bromide will be
withdrawn from field application by the end of 2015.Other fumigants, such as
chloropicrin, dazomet and meta sodium showed good activity against nematodes
when applied.
.

Biological control :-

An eco-friendly pest management strategy that utilizes deliberate introduction of

living natural enemies to lower the population level of a target pest. These
enemies are commonly referred to as BCAs, which must demonstrate some
characteristics for success in the field, including ability
for rapid colonization of the soil, persistence, virulence, predictable control below
economic threshold, easy production and application, good viability under
storage, low cost of production, compatibility with agrochemicals, and safety. In
nature, it is observed that many natural enemies, such as viruses, bacteria,
rickettsias, fungi, and others, can attack plant parasitic nematodes, but in the
search for suitable BCAs more attention has been given to fungi and bacteria.
Biological control can be either natural (i.e., when a natural population of a
particular organism inhibits the growth and development of nematodes), or
induced (i.e., when BCAs have been introduced artificially). There are two
approaches for introduction: microbial pesticide application for rapid control of a
pest, and the introduction or mass release of a biocontrol agent to provide long
lasting control. The suppression can be specific or non specific, when only one or
two organisms are involved. Researchers have made several attempts to utilize
bacteria for nematode control.

GROWTH PHYSIOLOGY OF
FILAMENTOUS FUNGI

Spore production of filamentous fungi is an important stage in its reproduction.

Spore production consists of the formation and liberation of conidiospores. Life-
cycle of imperfect fungi comprises five steps, which are dormancy of the spore,
germination, development of apical mycelium, and conidiogenesis. Normal
development of the mycelium and suitable conidiogenesis are the main
conditions required for a successful sporogenesis. The conidiospore production is
directly related to the quantity and nature of carbon and nitrogen sources
available in a culture media, and it depends on several other factors including
method of inoculation, media salinity, carbon/nitrogen ratio, aeration, water
content, among others85. Conidiospores are characterized by a low water activity,
absence of cytoplasmic movements, and reduced metabolic activity. Under
favourable conditions, spore germination takes place through the formation of a
vegetative tube, which will be the base of a future mycelium. A spore is
considered as germinated when the length of the longest germ tube is greater
than the dimension of the swollen spore. Different techniques, other than
microscopic examinations can be used to assess spore germination. Gompertz
equation and logistic function can be used for analyzing germination data.
Determination of optimal culture conditions for the large-scale production of
conidiospores of filamentous fungi, which are used as BCAs, is highly significant
for commercial applications. There are several studies carried out to enhance
conidiospore production for BCAs.

SOLID STATE FERMENTATION

SSF can be defined as the growth of microorganisms in a moist solid substrate in

the absence of liquid water. The water content in the moist solid substrate must
be adequate to support growth and metabolism of microorganism. SSF can be
carried out in two types of matrices, either in a natural substrate acting as solid
substrate and a source of nutrients or a nutritionally inert support which must be
impregnated with a liquid nutritive media. The most widely used substrates are
of amilaceous or lignocellulosic origin. Several materials are utilized as inert
supports for SSF, such as sugar cane bagasse, amberlite, vermiculite,
polyurethane foam, and polystyrene beads. SSF has several advantages over
SmF, but the choice of the method should depend on the physiology of the
microorganism and the end product. Comparative evaluations of SSF and SmF
indicated several advantages of SSF processes: simplicity of culture media;
absence of liquid residues;
reduction of contamination due to low water content; culture conditions mimic
the natural environment; ease of aeration (humid or dry) because of porosity of
the material; direct utilization of the fermented material; easy downstream
processing because of high yields; and easy to dehydrate and dry the fermented
product in situ. There are certain disadvantages associ ated with SSF processes
which include: excess of heat generation and subsequent difficulties in heat and
mass transfer, problems with the control of fermentation parameters (e.g., pH,
water content), difficulty in biomass estimation and pre-treatment of the
substrate, among others. SSF processes simulate the living conditions of many
higher filamentous fungi. Hence SSF is the cultivation method of choice for
biotechnological processes, where it is required to consider morphological and
metabolic differences in substrate- penetrating and aerial hyphae (e.g.,
production of conidiospores). There is a lack of information about the influence of
physico-chemical and nutritional parameters on the physiology and kinetics of
growth and sporulation of P. lilacinus under SSF, and the methods for estimation
of biomass. As filamentous fungi grow, hyphae penetrate into the solid matrix
becoming impossible to separate substrate from the mycelium, and thus making
difficult a direct measurement of biomass. Indirect methods for estimation of
biomass are available through the analysis of biomass components, such as
glucosamine,nucleic acids, and proteins. Several aspects should be considered
when
selecting biomass components for assay:
1) They should be major components in the microorganism;
2) They should have little or no influence from the substrate; and
3)They must be consistently present throughout development.
The most important method so far to assess biomass consists of measuring the
production of CO2 and the consumption of O2 by the microorganism during
fermentation. This permits the estimation of biomass and specific growth of the
microorganism inside the reactor through correlations between biomass
synthesis and oxygen consumption. .Important factors in SSF Factors affecting
SSF are purely based on the type of microorganism that is employed in the
process. The microorganism in a SSF process can be either natural microbiota of
the substrate or a pure culture. Ensiling and composting are two methods that
utilize the natural microbiota. Pure cultures are mainly used for the production of
fungal conidiospores, secondary metabolites, antibiotics, and other high value
product. There are several groups of microorganisms which can profusely grow in
solid substrates; however, filamentous fungi are well known for their capacity to
grow in substrates of relatively low moisture content due to their physiological,
enzymological, and biochemical properties. The growth of filamentous fungi
takes place combining the apical extension of hyphae and the generation of new
hyphae by mycelial ramification. This allows fungal growth within the solid matrix
to form a solid structure.
The penetration of hyphae into the substrate enhances the access to available
nutrients,
promoting suitable metabolic activity83.In SSF, the quantity of water present in
the media is a function of the substrate water retention capacity. This quantity
should be sufficient for the growth of microorganisms, without destroying the
solid structure or reducing the porosity of substrate or support. The water
content in the substrate influences the morphology of the microorganisms, and
serves as a carrier for enzymes, nutrients and metabolites, as well as in the
solubilization of oxygen. High moisture content of the substrate can lead to
reduced porosity of the solid matrix, weak oxygen diffusion and a high risk for
bacterial contamination. Low moisture levels result in limited growth of the
microorganism, as the distribution of available nutrients in the substrate is not
uniform. The control and monitoring of the gaseous environment in aerobic SSF
is a critical factor for the growth of microorganisms, which depends on the air
flow rate through the substrate and the rate of O2 consumptionAeration provides
O2 for aerobic growth and fungal metabolism, and it also helps to control
moisture and temperature, and to eliminate CO2 and some other volatile
metabolites. Any adverse change in the gaseous environment may significantly
affect the levels of production of biomass and enzymes. The initial pH of the
substrate in SSF is usually adjusted to support optimal growth of the
microorganism. The pH value varies according to metabolic activity of the
microorganisms. Acid production during fermentation or the formation of urea
tend to decrease the pH. Sudden and drastic change of the substrate pH can be
avoided using a solution of mineral salts with bufferingcapacity, as suggested by
Raimbault and Alazard.

SSF and the production of Biological Control

Agents (BCAs) :-
The most important aspect to be considered when selecting a BCA for
commercial application is the availability of a cost-effective production and
stabilization technology for manufacturing an effective formulation. BCAs are
mainly applied as spores and SSF offers several advantages: 1) The production of
aerial fungal spores are more tolerant to UV radiation; 2) Higher spore stability;
3) Spore resistance to drying; and 4) Higher spore germination rates for longer
storage periods. These better characteristics can be attributed to the presence of
a hydrophobic rodlet layer formed during the production process1. Another
advantage of SSF for production of BCAs is the utilization of agricultural by-
products as substrate for fermentation. The generation of high
amounts of agricultural residues causes serious environmental problems
worldwide, so SSF allows efficient utilization and value addition. The use of
agricultural byproducts for SSF leads to a less expensive process for the
production of BCAs on a large scale. SSF processes are usually cost effective, and
they require reduced labour. In many cases, the fermented substrate can be
used for field application, and thus most technical difficulties associated with
downstream
processing and product formulation aren ruled out. The products of SSF are
usually
air dried or rotavapor dried to be used directly. Roussos et al. studied different
methods for the conservation of fungal spores produced under SSF, and found
that temperature has a significant effect on spore viability in long term storage.
The mass production of fungal spores must be achieved for any commercial
application of BCAs. Therefore, further indepth studies for scaling up SSF
processes are needed, such as the design and development of automated SSF
bioreactors.
Bioreactors employed in SSF processes should provide adequate environmental
conditions for the maximal growth and activity of microorganisms. SSF
bioreactors have been studied for the commercial production of biopesticides,
metabolites, fermented foods, and other product. Several problems in scaling up
SSF processes have been found, such as variations in biomass production, high
inoculum level, substrate sterilization, heat generation due to microbial
metabolic activity, on-line monitoring of aeration, or pH. Bioreactor design for a
particular SSF process should consider: the type of substrate or support to be
used, particle size and mechanical resistance of the substrate, oxygen transfer,
nature of the gaseous phase between the particles of substrate or support,
morphology of microorganisms, and a suitable sterilization process. SSF
bioreactors can be classified in accordance with the quantity of substrate utilized
in the process. They are divided into two categories: laboratory scale (gkg
capacity) and pilot or industrial scale (kg-tons). Another type of classification is
based on the design of fermenters, which may provide agitation or aeration
devices. Laboratory scale bioreactor comprises simple devices, such as petri
dishes, Erlenmeyer flasks, jars and Roux bottles, which are mainly utilized for
screening of microorganisms and substrates. These small bioreactors cannot
provide aeration and agitation controls. Autoclavable plastic bags are also useful
and commonly used for the production of fungal inoculum. The utilization of
plastic bags for the production of fungal spores from Pochonia chlamydosporia
has been reported (substrate: rice and corn grains) for application as a BCA
against nematodes111. Column type bioreactors are well studied as they provide
on-line information of the microorganism’s respiration. These reactors monitor
respiration and other gaseous exchanges, and are mainly used for process
optimization studies. Column bioreactors can also monitor and control aeration,
so they are used as a model for designing and manufacturing several other types
of reactors. The substrate can be cooled by evaporation, and heat generation can
be minimized through convection and heat exchange by glass walls with the help
of a water bath. Reactors designed on continuous agitation are called rotating
drum reactors. These are perforated drums with a horizontal paddle mixer.
Rotating drum reactors were designed to increase contact between the reactor
wall and solid media, as well as to facilitate oxygen transfer. However, they have
several disadvantages, such as agglomeration of substrate, difficulties in
temperature regulation, low oxygen transfer, and alterations of substrate
structure due to intense agitation. Using a similar device from the Zymotis
bioreactor for reducing heat and providing high air flow, a novel bioreactor was
designed and patented by the German company Prophyta. This new bioreactor is
exclusively used for the production of the BCA P. lilacinus, strain PL-251. It has
perforated plates where heat exchangers are located at the bottom, and the
substrate on the top. The flow of sterile air is facilitated by perforated plates.
Another reactor patented by Durand et al was used for the production of fungal
conidiospores in biological control. This reactor has a capacity of 50 L, and it is
fitted with a planetary agitation device and controls for temperature, relative
humidity, and sterilization. Scaling-up is still a bottleneck for the widespread
commercial application of SSF. However, the development of rational and
computer-controlled processes during last decades brought about advances in
SSF, namely: the modelling of microbial growth on solid substrates, and energy
and mass transfer in different types of bioreactors. New methods are now
available for measuring SSF parameters, such as water activity and biomass
production, as well as statistical tools for process optimization. These
breakthroughs in SSF will certainly promote the commercial production and
application of BCA.

WEST INDIAN JOURNAL

Paecilomyces lilacinus fungemia in a Jamaican

neonate:=

Invasive opportunistic fungal infections have increased in frequency over the last two
decades, while Candida sp remains the most common cause of fungemia (1, 2). This
report describes the fourth documented case of Paecilomyces lilacinus fungemia, the
first such case to be reported at the University Hospital of the West Indies (UHWI).

A 35-week gestation female infant, weighing 3.5 kg was born to a 31-year old mother
with a normal antenatal history at a hospital in rural Jamaica. At birth, the infant was
jaundiced and was diagnosed as having Downs Syndrome. The infant subsequently
developed a low grade pyrexia and was transferred to the UHWI on day thirteen.
Significant findings included jaundice, pyrexia (100oF), abdominal distention and
cardiac murmurs consistent with a ventricular septal defect and patent ductus
arteriosus. A diagnosis of sepsis with necrotizing enterocolitis was made. Results of a
full septic screen were negative. A nasogastric tube was inserted for oral feeds and
intravenous ampicillin, cloxacillin, gentamicin and metronidazole were administered
through a peripheral intravenous catheter. These antibiotics were administered for
seven days with resolution of the fever and abdominal distention, and incomplete
resolution of jaundice. Oral feeds were subsequently commenced. The infant remained
mildly icteric and again developed abdominal distention and fever on day 14, when a
second septic screen was done. An ultrasound investigating the infants abdominal
distention revealed multiple abscesses, aspirates from which were culture positive for
Citrobacter diversus and Escherichia coli. A new antibiotic regimen was commenced
with intravenous rocephin, metronidazole and ampicillin. The infant however remained
febrile.

Results of the two sets of blood cultures taken from the peripheral vein on day 14 at
the UHWI were positive for a fungus initially identified as Penicillium spp. This isolate
was subsequently referred to Mayo Clinic, Minnesota, USA, for further identification.
Antimicrobial treatment was changed to include intravenous amphotericin B, amikacin
and ceftazidime. Amphotericin B was given for 18 days. The isolate was later confirmed
by Mayo Clinic to be Paecilomyces lilacinus The patient responded favourably to
treatment with amphotericin B and was discharged on maintenance fluconazole.

Paecilomyces lilacinus is a ubiquitous fungal saprophyte and an uncommon human

pathogen which is known to infect a variety of organs with varying degrees of morbidity
and mortality (1, 3, 4). Identification of the fungus is difficult as it is closely related to
the genus Penicillium (1). Prolonged use of antibiotics and invasive procedures,
including peripheral intravenous catheters predispose to acquisition of opportunistic
pathogens, such as Paecilomyces. Although Paecilomyces lilacinus has been reported to
be resistant to amphotericin B and fluconazole, the infant herein presented responded
favourably to this management (5).

Human pathogenicity:-

P. lilacinus is an infrequent cause of human disease. Most reported cases involve patients with
compromised immune systems, indwelling foreign devices, or intraocular lens implants. Research of
the last decade suggests it may be an emerging pathogen of both immunocompromised as well as
immunocompetent adults.

Biocontrol agent:-

Plant-parasitic nematodes cause significant economic losses to a wide variety of crops. Chemical
control is a widely used option for plant-parasitic nematode management. However, chemical
nematicides are now being reappraised in respect of environmental hazard, high costs, limited
availability in many developing countries or their diminished effectiveness following repeated
applications.

Control of plant parasitic nematodes :-

P. lilacinus was first observed in association with nematode eggs in 1966 and the fungus was
subsequently found parasitising the eggs of Meloidogyne incognita in Peru. It has now been isolated
from many cyst and root-knot nematodes and from soil in many locations. Several successful field
trials using P. lilacinus against pest nematodes were conducted in Peru. The Peruvian isolate was then
sent to nematologists in 46 countries for testing, as part of the International Meloidogyne project,
resulting in many more field trials on a range of crops in many soil types and climates Field trials,
glasshouse trials and in vitro testing of P. lilacinus continues and more isolates have been collected
from soil, nematodes and occasionally from insects. Isolates vary in their pathogenicity to plant-
parasitic nematodes. Some isolates are aggressive parasites while other, though morphologically
indistinguishable, are less or non-pathogenic. Sometimes isolates which looked promising in vitro or
in glasshouse trials have failed to provide control in the field.

Enzymes :-
Many enzymes produced by P. lilacinus have been studied. A basic serine protease with biological
activity against Meloidogyne hapla eggs has been identified. One strain of P. lilacinus has been shown
to produce proteases and a chitinase, enzymes that could weaken a nematode egg shell so as to enable
a narrow infection peg to push through.
Egg Infection :-

Before infecting a nematode egg, P. lilacinus flattens against the egg surface and becomes closely
appressed to it. P. lilacinus produces simple appressoria anywhere on the nematode egg shell either
after a few hyphae grow along the egg surface, or after a network of hyphae form on the egg. The
presence of appressoria appears to indicate that the egg is, or is about to be, infected. In either case,
the appressorium appears the same, as a simple swelling at the end of a hypha, closely appressed to
the eggshell. Adhesion between the appressorium and nematode egg surface must be strong enough to
withstand the opposing force produced by the extending tip of a penetration hypha When the hypha
has penetrated the egg, it rapidly destroys the juvenile within, before growing out of the now empty
egg shell to produce conidiophores and to grow towards adjacent eggs.

Crops :-
Eggplant,Potato,Chilli,Tomatoes,Cucumbers,
flowers, Orchards, Vineyards Ornamentals in
greenhouses, lawns, nurseries and landscape.

Target Pests :-
Plant parasitic nematodes in soil, Examples include Meloidogyne spp.(Root knot nematodes);
Radopholus similis (Burrowing nematode); Heterodera spp. and Globodera spp. (Cyst nematodes);
Pratylenchus spp. (Root lesion nematodes); Rotylenchulus reniformis (Reniform Nematode);
Nacobbus spp.(False Root knot Nematodes).

Method of application :-

Paecilomyces lilacinus is a naturally occurring fungus found in many kinds of soils

throughout the world. As a pesticide active ingredient, Paecilomyces lilacinus is applied to
soil to control nematodes that attack plant roots. In laboratory studies, it grows optimally at
21-32 degrees C,and does not grow or survive above 36 degrees C. It acts against plant root
nematodes by infecting eggs, juveniles, and adult females.
Suspend Paecilomyces lilacinus in sufficient water (500g/100L) to achieve uniform
application. Apply at the rate of 100-200 g per cubic metre (loose) of greenhouse potting mix,
soil or planting beds.

Paecilomyces lilacinus can be applied through low pressure watering nozzles such as fan
nozzles or other watering systems (drip system) after filtering with filters. Agitate to maintain
suspension. For best effect, treat potting mix several days before use for seeding or
transplants.

Soil application:
For one acre mix about 3 kgs of Gmax Bioguard with 100 kgs of compost, keep the mixture
under shade with sufficient moisture content (30%) for one week time and broadcast in the
field.

Pit application:
For plantation crops like Banana, sprinkle 25 grams of Gmax Bioguard in the pit before
planting. After planting, about 25 grams of Gmax Bioguard can be mixed with compost and
sprinkled around the tree trunk in the soil.

Frequency of application :-
Two to three applications in vegetables ornamentals and 4-5 applications in lawns and
landscape crops are recommended. In the case of high infestation multiple applications are
 recommended. Applications during early stages of plant growth protect the plant during
critical stages of development.

Dosage:-
Soil application: 5 kg /ha along with any organic fertilizer (without pathogenic contaminants).
Seed treatment: @ 4-5 gm per kg of seeds as per standard wet treatment.
Seedling treatment: @ 100 g/l prior to planting.

(Sohwing normal xylem (NX) strands with hyphae(H) of Paecilomyces lilacinus).

References:-
1. Samson, RA (1974). "Paecilomyces and some allied hyphomycetes". Studies in Mycology
(Baarn: Centralbureau voor Schimmelcultures) 6: 119.
2. Anderson, Traute-Heidi; Domsch, K. H.; Gams, W. (1995). Compendium of Soil Fungi. Lubrecht
& Cramer Ltd.
3. Marti GA, Lastra CC, Pelizza SA, García JJ (November 2006). "Isolation of Paecilomyces
lilacinus (Thom) Samson (Ascomycota: Hypocreales) from the Chagas disease vector, Triatoma
infestans Klug (Hemiptera: Reduviidae) in an endemic area in Argentina". Mycopathologia 162
(5): 369–72.
4. Fiedler, Z. and Sosnowska, D. (2007) Nematophagous fungus Paecilomyces lilacinus (Thom)
Samson is also a biological agent for control of greenhouse insects and mite pests. BioControl
52, 547–558
5. Gupta, S.C., Leathers, T.D. and Wicklow, D.T. (1993) Hydrolytic enzymes secreted by
Paecilomyces lilacinuscultured on sclerotia of Aspergillus flavus. Appl Microbiol Biotechnol 39,
99–103.
6. Saberhagen C, Klotz SA, Bartholomew W, Drews D, Dixon A (December 1997). "Infection due
to Paecilomyces lilacinus: a challenging clinical identification". Clin. Infect. Dis. 25 (6): 1411–3.
7. Westenfeld F, Alston WK, Winn WC (June 1996). J. Clin. Microbiol. 34 (6): 1559–62.
8. O'Day DM (January 1977). "Fungal endophthalmitis caused by Paecilomyces lilacinus after
intraocular lens implantation".
9. Jatala, P; Kaltenbach R and Bocangel M. "Biological control of Meloidogyne incognita acrita
and Globodera pallida on potatoes". Journal of Nematology 11: 303.
10. Stirling, GR (1991). Biological Control of Plant Parasitic Nematodes. UK: CABI Publishing.
pp. 282.
11. Stirling, GR; West LM. "Fungal parasites of root-knot nematode eggs from tropical and sub-
tropical regions of Australia". Australasian Plant Pathology 20: 149–154.