Veterinary Parasitology 149 (2007) 111116

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A comparison of modifications of the McMaster method for the

enumeration of Ascaris suum eggs in pig faecal samples
A. Pereckiene a,*, V. Kaziunaite a, A. Vysniauskas a, S. Petkevicius a,b,
A. Malakauskas b, M. Sarkunas b, M.A. Taylor c
a
Laboratory of Parasitology, Veterinary Institute of Lithuanian Veterinary Academy, Instituto 2, LT-08662 Kaisiadorys, Lithuania
b
Department of Infectious Diseases, Lithuanian Veterinary Academy, Tilzes str. 18, LT-47181 Kaunas, Lithuania
c
Veterinary Surveillance, Central Science Laboratory, Sand Hutton, York YO41 1LZ, UK
Received 14 March 2006; received in revised form 13 April 2007; accepted 23 April 2007

Abstract
The comparative efficacies of seven published McMaster method modifications for faecal egg counting were evaluated on pig
faecal samples containing Ascaris suum eggs. Comparisons were made as to the number of samples found to be positive by each of
the methods, the total egg counts per gram (EPG) of faeces, the variations in EPG obtained in the samples examined, and the ease of
use of each of the methods. Each method was evaluated after the examination of 30 samples of faeces. The positive samples were
identified by counting A. suum eggs in one, two and three sections of newly designed McMaster chamber. In the present study
compared methods were reported by: IHenriksen and Aagaard [Henriksen, S.A., Aagaard, K.A., 1976. A simple flotation and
McMaster method. Nord. Vet. Med. 28, 392397]; IIKassai [Kassai, T., 1999. Veterinary Helminthology. Butterworth
Heinemann, Oxford, 260 pp.]; III and IVUrquhart et al. [Urquhart, G.M., Armour, J., Duncan, J.L., Dunn, A.M., Jennings,
F.W., 1996. Veterinary Parasitology, 2nd ed. Blackwell Science Ltd., Oxford, UK, 307 pp.] (centrifugation and non-centrifugation
methods); Vand VIGrnvold [Grnvold, J., 1991. Laboratory diagnoses of helminths common routine methods used in Denmark.
In: Nansen, P., Grnvold, J., Bjrn, H. (Eds.), Seminars on Parasitic Problems in Farm Animals Related to Fodder Production and
Management. The Estonian Academy of Sciences, Tartu, Estonia, pp. 4748] (salt solution, and salt and glucose solution); VII
Thienpont et al. [Thienpont, D., Rochette, F., Vanparijs, O.F.J., 1986. Diagnosing Helminthiasis by Coprological Examination.
Coprological Examination, 2nd ed. Janssen Research Foundation, Beerse, Belgium, 205 pp.]. The number of positive samples by
examining single section ranged from 98.9% (method I), to 51.1% (method VII). Only with methods I and II, there was a 100%
positivity in two out of three of the chambers examined, and FEC obtained using these methods were significantly ( p < 0.01) higher
comparing to remaining methods. Mean FEC varied between 243 EPG (method I) and 82 EPG (method IV). Examination of all
three chambers resulted in four methods (I, II, V and VI) having 100% sensitivity, while method VII had the lowest 83.3%
sensitivity. Mean FEC in this case varied between 239 EPG (method I) and 81 EPG (method IV). Based on the mean FEC for two
chambers, an efficiency coefficient (EF) was calculated and equated to 1 for the highest egg count (method I) and 0.87, 0.57, 0.34,
0.53, 0.49 and 0.50 for remaining methods (IIVII), respectively. Efficiency coefficients make it possible not only to recalculate and
unify results of faeces examination obtained by any method but also to interpret coproscopical examinations by other authors.
Method VII was the easiest and quickest but least sensitive, and method I the most complex but most sensitive. Examining two or
three sections of the McMaster chamber resulted in increased sensitivity for all methods.
# 2007 Elsevier B.V. All rights reserved.

Keywords: Pigs; Ascaris suum; Faecal egg counts; McMaster; Comparison

* Corresponding author. Tel.: +370 37363559; fax: +370 37363559.

E-mail address: petkevicius@lva.lt (A. Pereckiene).

0304-4017/$ see front matter # 2007 Elsevier B.V. All rights reserved.
doi:10.1016/j.vetpar.2007.04.014
112 A. Pereckiene et al. / Veterinary Parasitology 149 (2007) 111116

1. Introduction strate the presence of nematode eggs and permit a

Faecal examination is an important tool for
monitoring worm infections in pig units and for
maintaining effective worm control programmes.
Described faecal examination methods are either
qualitative or quantitative. Qualitative methods provide
information on the species present, whereas quantitative
methods provide an indication of the levels of
infections. Both can be an important tool in determining
the health status of a herd and determining appropriate
treatments and control measures. Examination of faeces
for parasitic eggs may vary from a simple direct smear
to more complex methods involving centrifugation and
use of flotation fluids (MAFF, 1986). Flotation methods
involve separating eggs from faecal debris using a
variety of flotation solutions with specific gravities
which float worm eggs to the surface of the suspension.
The most widely used and standard quantitative
technique is the McMaster method (Gordon and
Whitlock, 1939; Whitlock, 1948). However, there are
many modifications reported in the literature. Reported
methods differ in the weight of faeces examined; the
flotation solution used (chemical salt, level of saturation
and volume); the flotation time; the presence or absence
of a centrifugation step; the design and number of
McMaster counting chambers; the counting method and
multiplication factors employed; and whether any
correction factors are used to allow for faecal
consistency (MAFF, 1986; Dunn and Keymer, 1986;
Cringoli et al., 2004).
Given wide range of modifications of McMaster
method, parasitological laboratories have to decide
which method to choose as the most appropriate method
for the purpose intended. Some modifications of
McMaster method are designed primarily to demon-
quantitative determination of their concentration in
faeces. The use of a centrifugation step facilitates
examination by removal of fine particles and colouring,
and increases the ease of egg identification (MAFF,
1986). For low faecal egg counts particularly, it may be
necessary to choose the most efficient, stable, precise,
and reliable method. Problems can however, occur
when analyzing and comparing results of different
laboratories on the same samples. There is a need to
provide some degree of standardisation of the numerous
modifications of the McMaster method (Cringoli et al.,
2004; Coles et al., 2006), particularly where these are
used in the determination of the presence of anthel-
mintic resistance by faecal egg reduction test (Taylor
et al., 2002; Coles et al., 2006).
The aim of this study was therefore to evaluate the
efficiency, sensitivity and complexity of various
published modifications to the McMaster method and
seek to provide a system of standardisation allowing the
comparison of egg count results between laboratories.
2. Materials and methods
Seven published modifications of the McMaster
method (Table 1), as described in the literature, were
compared. The modifications include different weight
of faeces and volume of water, presence or absence of a
centrifugation step, centrifugation time and speed,
different flotation solution, flotation time, and number
of sections counted.
For the study 3.5 kg of fresh faeces were taken from
pigs naturally infected with ascarids (commercial pig
breeding company). Faeces were thoroughly mixed for
even distribution of helminth eggs and 30 samples were
prepared for each of the McMaster modification. Each

Table 1
Summary of published methods for the McMaster faecal egg counting technique
McMastermethod Author Amount of Volume of Centrifugation Flotation solution Flotation time Sensitivity
faeces (g) water (ml) (specific gravity) in chambers (EPG)
I Henriksen and 4 56 7 min at 1200 rpm NaCl + sugar (1.27) 23 min 20
Aagaard (1976)
II Kassai (1999) 3 42 3 min at 1500 rpm NaCl (1.2) 3 min 50
III Urquhart et al. 3 42 2 min at 2000 rpm NaCl (1.2) 23 min 50
(1996)
IV Urquhart et al. 3 42 None NaCl (1.2) 23 min 50
(1996)
V Grnvold (1991) 4 56 None NaCl (1.2) 23 min 50
VI Grnvold (1991) 4 56 None NaCl + glucose (1.27) 23 min 50
VII Thienpont et al. 2 60 None NaCl (1.2) 23 min 100
(1986)
EPG: eggs per gram of faeces; NaCl: sodium chloride; rpm: revolutions per minute.
 A. Pereckiene et al. / Veterinary Parasitology 149 (2007) 111116 113

were then obtained by counting the ascarid eggs in the

first, second and third chambers separately or in
combination, to give mean faecal egg count (FEC)
values expressed as eggs per gram of faeces (EPG).
The investigation was carried out pursuant of the
Law of the republic of Lithuania regulating animal care
and use for scientific experiments.

3. Statistical analysis

Fig. 1. McMaster chamber design: (A) ruled lines of the chamber; (B)
supporting strips; (C) platforms; (D) three sections.
sample was weighed and 2, 3 and 4 g alloquots
according to the requirements of each method
modification were prepared. All samples were placed
into polythene bags and kept at +4 8C until examination.
Flotation solutions were made up fresh as required and
kept at a room temperature until use. Faecal samples
were stored only for 5 days to prevent influence of egg
hatching on EPG counts.
For microscopic examination, a modified McMaster
chamber with three sections (manufactured in Lithua-
niaCertificate No. 21-37-483) was used (Fig. 1). The
chamber includes the inclusion of beads to protect the
optics of microscope during the microscopic examina-
tion, and to prevent the flow of flotation solution on the
microscope stand. Before examination all samples were
thoroughly mixed to allow for even distribution of eggs
and filling the compartments of McMaster chamber
avoiding formation of bubbles, which may distort the
feacal egg count (FEC) results (Kassai, 1999). Each
filled McMaster chamber was left to stand for 35 min
before microscopic examination, as recommended by
Roepstorff and Nansen (1998). For positive samples, the
number of helminth eggs was counted in one, two and
three sections of the McMaster slide. Feacal egg counts
The difference between the modifications of
McMaster methods was evaluated using one-way
analysis of variance test (ANOVA) and Tukeys
multiple comparison posttest. The SAS programme
was used for analysis (Version 8.2, SAS@ Institute Inc.).
As FECs based on the McMaster method, are usually
performed on two chambers, for the purposes of this
study, an EF was derived from the EPG in 2 chambers
only. The highest EF was equalled to unity (1). This
value was given to the method which yielded the highest
EPG. The egg counts obtained by other methods were
then divided by the highest EPG. In this way the EF of
each method was derived.
4. Results
Sensitivity of the seven methods counting one, two
or three sections is shown in Table 2. The sensitivity by
examining single section ranged from 98.9% (method
I), to 51.1% (method VII). Examining two sections,
methods I and II were 100% sensitive while the method
VII had lowest sensitivity (74.4%). Examination of all
three sections resulted in four methods (I, II, V and VI)
having 100% sensitivity, while method VII had the
lowest (83.3%) sensitivity.
Mean EPG counts for all the seven methods are
shown in Table 3. The first modification of the

Table 2
Efficiency of McMaster method modifications based on number of positive samples in one, two or three chambers
McMaster Number of samples positive for Ascaris suum eggs (n = 30)
method
In one section In two sections In three sections
First Second Third Mean % First + First + Second + Mean % First + second + Mean %
positive second third third positive third positive
I 30 29 30 98.9 30 30 30 100 30 100
II 30 28 30 97.8 30 30 30 100 30 100
III 25 25 26 84.4 29 29 28 95.6 29 96.7
IV 19 20 18 63.3 24 26 25 83.3 28 93.3
V 23 20 22 72.2 28 28 28 93.3 30 100
VI 20 25 22 74.4 29 28 27 93.3 30 100
VII 18 14 14 51.1 22 23 22 74.4 25 83.3
114 A. Pereckiene et al. / Veterinary Parasitology 149 (2007) 111116

Table 3
Mean egg counts and efficiency coefficients for McMaster methods based on counts from one, two or three chambers
McMaster Mean number of A. suum eggs per 1 g of faeces (standard error)
method
In one chamber In two chambers In three chambers Efficiency
* (first + second + third)* coefficients
First Second Third Mean First + First + Second + Mean
second third third
I 264  22 221  25 244  19 243 a  12 243  19 255  16 233  16 244 a  6 239  14 a 1
II 240  20 187  28 210  25 212 a  15 213  19 225  17 197  21 212 a  8 212  17 a 0.87
III 130  18 130  18 150  23 137 b  7 130  13 143  15 140  17 138 b  4 137  13 b 0.57
IV 83  14 93  17 70  12 82 c  7 88  12 77  10 82  10 82 c  3 81  9 c 0.34
V 133  22 123  25 127  23 128 b  3 130  18 130  19 125  18 128 b  2 128  16 b 0.53
VI 90  17 140  19 133  22 121 b  16 113  15 112  14 137  16 121 b  8 121  13 b 0.49
VII 133  22 113  27 120  28 122 b  6 123  18 127  18 117  18 122 b  3 122  14 b 0.50
*
Indices indicate significant differences ( p < 0.05).

McMaster method gave the highest EPG in all cases and
the IVth modification the lowest EPG. Efficiency
coefficients based on mean EPG counts are shown in
Table 3.
Comparable EPG were obtained with the methods I
and II, and there was no significant difference between
them. The remaining methods gave considerably lower
EPG counts, which were significantly ( p < 0.01) lower
compared to I and II.
5. Discussion
The use of nematode egg counting techniques that
provide a quantitative determination of the number of
eggs in faeces is required to determine levels of worm
infections. However, it is important to have in mind that
a number of factors can influence faecal egg count
results. The factors to consider are: the biology of the
worms (species, prepatent period, fecundity, worm
numbers); the influence of the host physiological state
on worm numbers and populations (nutrition, immu-
nity); host management factors (stocking densities;
treatments); the time of year and the laboratory method
used to enumerate faecal egg counts (Lyndal-Murphy,
1993).
Faecal egg counting gives an estimation of worm
numbers and levels of worm infestation, and as such it
should be used as an adjunct to an overall approach to
determine the health status of a herd. Veterinarians or
herd health advisors should therefore consider the
results of FEC along with these other factors when
advising on appropriate control procedures. An obvious
consideration is that eggs are produced only by female
worms and FECs, therefore, do not reflect the total
worm burden present in an animal, which may also
include both males and immature stages. The fecundity
of nematode species varies considerably, some species
producing only small numbers of eggs, while ascarid
worms, such as Ascaris suum, are among the most
productive (Roepstorff and Nansen, 1998). It has also
been reported that different nematode species are more
productive at different times of the day, and at differing
times of the year (Thienpont et al., 1986). Hyostrongy-
lus rubidus and Oesophagostomum spp. in pigs, for
example, have lower fecundity in winter than in summer
(Fossing et al., 1995). Immunity of older animals may
disturb the seasonality (Roepstorff, 1991; Roepstorff
et al., 1996) and it has been determined that the most
widespread intestinal nematode species in pigs are
unevenly distributed in different age groups (Pattison
et al., 1980). Despite this, older resistant animals may
carry the same worm burdens as younger less resistant
individuals, but of lower fecundity (Thienpont et al.,
1986).
FECs can also be influenced by physiological
changes in the animal, for example, the effect of both
pregnancy and lactation and the observed phenomenon
of periparturient rises in egg counts (Lyndal-Murphy,
1993). It has been observed that the incubation period of
A. suum in fattening pigs can be prolonged and the
majority of pigs become infected at the beginning of
fatting period or during the growth period, but only a
small number of animals are identified as infected by
coproscopic examination (Connan, 1985). An addi-
tional consideration is that it has been suggested that
only FECs of 200 EPG for A. suum should be regarded
as positive (Roepstorff, 1997) because smaller FECs
may represent the passage of unembryonated eggs
through the intestine as a consequence of coprophagia.
However, such eggs fail to develop within the pigs
intestines (Boes et al., 1997, 1998; Roepstorff et al.,
1998). Such erroneous results may be a factor of age
 A. Pereckiene et al. / Veterinary Parasitology 149 (2007) 111116
(Roepstorff and Nansen, 1994), or the conditions and
husbandry systems under which the pigs are kept (Boes
et al., 1997).
It has been already indicated that McMaster methods
differ in their efficiencies, sensitivities and complexities
when determining FECs. Methods I, II and III, which
each requires centrifugation step, can be considered
more complex and labour intensive. The remaining
methods (IVVII), without centrifugation step, are
considered as more simple and require less time to
perform. However, the results obtained with the later
methods are less reliable because of higher amount of
debris within the McMaster chamber that hinders
microscopic examination and increases probability of
error. Methods IIII are therefore more appropriate for
quantitative determination of FECs, comparing with
methods IVVII. This can be seen in the method
sensitivities in detecting positive samples, the quanti-
tative FECs, and the EF values. The use of EF values not
only makes it possible to recalculate and unify results of
faeces examination obtained by any method, but also to
potentially compare coproscopical examinations by
other authors. The present results indicate that methods
I and II are the most efficient in identifying the number
of positive samples, and in resultant higher sensitivities
based on FECs, which are significantly higher com-
paring to the remaining methods. Method III is
essentially similar to method II differing only in the
speed and time of centrifugation and when used in the
coproscopical examinations of horse faeces demon-
strated an equally high performance (Vysniauskas et al.,
2005). These methods also differ in the amount of
faeces and the flotation solutions used. In method I, the
faeces were also soaked in water for 30 min, which
appears may increase dispersion of faeces in solution.
Of the remaining non-centrifugation methods, method
VII was the least effective method in detecting positive
samples, whilst method IV was the least sensitive based
on FECs.
For all of the methods, the sample dilution ratio is
1:15 except method VII where the dilution ratio is 1:31.
Though this modification does not need centrifugation,
is simple and time-sparing its reported sensitivity (100
EPG) is much less than other methods (Table 1).
Another factor to consider, is that flotation solutions
differ in their specific gravities (Table 1) and their
ability to float helminth eggs of differing sizes and
weights. Ascarid eggs, as with trematode eggs, are
generally much larger and heavier compared with
trichostrongylid eggs and generally require solutions
with a higher specific gravity (MAFF, 1986). Solutions
containing sugar (sucrose) or glucose have a higher
115
specific gravity (1.27) than salt solutions (1.2) and are
therefore more generally effective for Ascaris eggs.
They are more viscous and easier to control while
filling, and disperse more evenly in the chamber
forming no air bubbles, which can distort the FEC
results. Saturated salt solutions also have the disadvan-
tage of crystallizing more readily.
6. Conclusions
A number of modifications to the McMaster method
used for determining helminth egg counts in faeces
have been reported in the literature. Those methods
that incorporate a centrifugation step offer greater
sensitivity for the quantitative determination of FECs
when examining pig faeces for ascarid eggs. Sensi-
tivity is further improved through the use of flotations
solutions with higher specific gravities. Examining
two or three sections of a McMaster chamber increases
sensitivity for all methods. Consideration should also
be given to the calculation of efficiency coefficients
which offer the potential, not only to recalculate and
unify results of FECs obtained by any method, but also
to interpret coproscopical examinations by other
authors.
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