Veterinary Parasitology 148 (2007) 236–246

www.elsevier.com/locate/vetpar

Gastrointestinal protozoa in non-human primates of

four zoological gardens in Belgium
Bruno Levecke a,*, Pierre Dorny a,b, Thomas Geurden a,
Francis Vercammen c, Jozef Vercruysse a
a
Department of Virology, Parasitology & Immunology, Faculty of Veterinary Medicine, Ghent University,
Salisburylaan 133, 9820 Merelbeke, Belgium
b
Department of Animal Health, Institute of Tropical Medicine, Nationalestraat 155, 2000 Antwerp, Belgium
c
Veterinary Department, Royal Zoological Society of Antwerp, Koningin Astridplein 26, 2018 Antwerp, Belgium
Received 10 May 2007; received in revised form 10 June 2007; accepted 14 June 2007

Abstract
Gastrointestinal parasites are important infectious causes of diarrhoea in captive non-human primates (NHP). However,
prevalence data of gastrointestinal parasites in zoological gardens are scarce. Therefore, a cross-sectional survey was conducted to
estimate the occurrence of gastrointestinal parasites in NHP of four zoological gardens in Belgium. Between August 2004 and April
2006, 910 faecal samples were collected from 222 animals housed in 39 groups. The 31 species involved were representatives of
prosimians, New World (NW) monkeys, Old World (OW) monkeys and apes. Because individual sampling was impossible, a
statistical simulation was performed to estimate a sufficient sample size. All samples were microscopically examined after an acetic
acid–ether concentration. Differences in host species susceptibility were examined by non-parametric tests. Entamoeba spp. (44%)
and Giardia spp. (41%) were the most prevalent species. Other parasites detected were Endolimax nana (36%), Chilomastix mesnili
(21%), Balantidium coli (13%), Trichuris spp. (10%), Iodamoeba bütschlii (5%) and Strongyloides spp. (5%). Parasites for which a
significant difference in susceptibility at the level of host taxonomy was noted were Entamoeba spp. ( p < 0.001) and C. mesnili
( p < 0.05). Samples containing Entamoeba spp. were the most prevalent in OW monkeys ( p < 0.0083). Samples collected from
OW monkeys contained the highest number of parasite species ( p < 0.0083).
# 2007 Elsevier B.V. All rights reserved.

Keywords: Gastrointestinal protozoa; Non-human primates; Zoological gardens; Prevalence; Host species susceptibility

1. Introduction et al., 1993; Verweij et al., 2003a), Giardia spp. (Peisert

et al., 1983; Hamlen and Lawrence, 1994; Kalishman
Infections with gastrointestinal parasites are wide- et al., 1996), Cryptosporidium spp. (Gómez et al., 1992;
spread among non-human primates (NHP). However, as Kalishman et al., 1996; da Silva et al., 2003) and
a consequence of regular deworming and hygienic Balantidium coli (Nakauchi, 1999) are frequently
measures helminth infections are uncommon in captive reported in captive NHP, and are considered as
NHP (Gómez et al., 1996; Verweij et al., 2003a). In important causes of gastro-enteritis in NHP. Infection
contrast, protozoa such as Entamoeba histolytica (Pang by these gastrointestinal parasites may cause watery
diarrhoea, hemorrhagic dysentery, extra-intestinal
pathologies, such as liver abscesses, and even death.
* Corresponding author. Tel.: +32 9 264 7404; fax: +32 9 264 7496. E. histolytica, the causative organism of invasive
E-mail address: [email protected] (B. Levecke). intestinal and extra-intestinal amebiasis, is of major

0304-4017/$ – see front matter # 2007 Elsevier B.V. All rights reserved.
doi:10.1016/j.vetpar.2007.06.020
 B. Levecke et al. / Veterinary Parasitology 148 (2007) 236–246 237

concern. Clinical outbreaks and deaths caused by this was founded in 1843 and is a typical urban zoo of about
parasite are frequently reported (Loomis et al., 1983; 10 ha where most of the NHP are accommodated in
Beaver et al., 1988; Marquez-Monter et al., 1991; indoor enclosures. At study site B (12 ha), study site C
Verweij et al., 2003a). Giardiasis and cryptosporidiosis (55 ha) and study site D (40 ha) the animals are kept on
are of less clinical importance, but are considered as a a large verdant outdoor enclosure. At all sites animals
cause of diarrhoea or failure to thrive in young animals are kept indoor when the temperature drops below
(Miller et al., 1990; Kalishman et al., 1996). B. coli is 10 8C.
probably harmless in most of the NHP species.
However, according to Lee et al. (1990) and Hänichen 2.2. Animals and husbandry
et al. (1995) care should be taken when great apes are
involved. From August 2004 to April 2006, 222 animals
Because of the importance of E. histolytica (Stauffer belonging to 31 NHP species and seven families were
and Ravdin, 2003), Giardia spp. (Thompson, 2000) and studied (Table 1). The families involved were two of
Cryptosporidium spp. (O’Donoghue, 1995) in humans, prosimians (Lemuridae, Galagonidae), two of New
the role of NHP as potential reservoirs for zoonotic World (NW) monkeys (Cebidae and Atelidae), one of
transmission should not be underestimated. Transmis- the Old World (OW) monkeys (Cercopithecidae) and
sion between animals and humans in association with both families of the apes (Hylobatidae and Hominidae)
clinical outbreaks in animal caretakers has been (Groves, 2001). The animals are housed in 39 groups of
reported in various studies (Miller et al., 1990; Hamlen 2–20 individuals (median of four animals). Apart from
and Lawrence, 1994). six groups where two NHP species are mixed, all groups
Although the above-mentioned studies have shown consisted of one species. Of all sites, study site A lodges
the clinical importance of protozoa for both NHP and the largest and most diverse population of NHP. This
animal caretakers, studies to quantify the prevalence site includes half of the animals and 75% of the species
and importance of these parasites in zoological gardens included in this study representing all seven families. A
are scarce. Most studies describe clinical outbreaks and total of 37 animals (11 species) were examined at study
were based on a limited number of animal and/or site B, 38 animals (5 species) at study site C and 17
parasite species (Teare and Loomis, 1982; Loomis et al., animals (3 species) at study site D.
1983; Lee et al., 1990). Moreover, the strategies used to Overall, the median (25th quantile (25Q); 75th
collect faecal samples may thwart the prevalence quantile (75Q)) of the stocking density was 13.5 m2 per
results. animal (7.5; 51.1). The lowest values were found at
In this study, the prevalence of gastrointestinal study site A (median of 8.3 m2 per animal).
protozoa in NHP of four Belgian zoological gardens
was estimated based on a new sampling strategy. In 2.3. Sampling strategy
order to examine all animals within a group a statistical
simulation was performed to estimate a sufficient Due to the group housing of semi-wild NHP
sample size. The 31 species involved were representa- individual sampling was impossible. However, a
tives of NHP of four Belgian zoological gardens and statistical simulation in R (version 2.4.0, The R
included members of Lemuridae, Galagonidae, Cebi- Foundation for Statistical Computing) was used to
dae, Atelidae, Cercopithecidae, Hylobatidae and estimate the sample size needed to examine all animals
Hominidae. A second objective was to examine within a group with a probability of at least 95%
differences in host species susceptibility to gastro- (Appendix A). Because little is known about the
intestinal protozoan infections. defecation behaviour of the different NHP species, an
equal probability of sampling was assumed in this
2. Materials and methods simulation (p1 = p2 =    = pj = 1/n_NHP). To correct
for this unknown variability, a sampling method with
2.1. Study sites replacement was performed (group  Mult(n_sim, p)).
The estimated sample sizes in function of the group
This study was conducted at four Belgian zoological size obtained by this simulation are presented in
gardens. These included the Antwerp Zoo, the Animal Table 2.
Park Planckendael, the Olmen Zoo and the Park For each particular group all samples were picked up
Paradisio. Because of confidentiality, a letter was from the ground during consecutive days until the
randomly assigned to the four study sites. Study site A estimated sample size was approximated.
238 B. Levecke et al. / Veterinary Parasitology 148 (2007) 236–246

Table 1
Animal species and husbandry conditions of NHP kept at four Belgian zoological gardens
Groups Number of Stocking density
animals (animal/m2)
Common name Scientific name
Study site A
Ring-tailed lemur Lemur catta 9 12.0
Red ruffed lemur Varecia rubra 2 7.5
Red ruffed lemur Varecia rubra 4 3.5
Brown greater galago Otolemur crassicaudatus 2 60.0
Goeldi’s marmoset Callimico goeldii 2 3.7 a
Golden-headed lion tamarin Leontopithecus chrysomelas 2
Common marmoset Callithrix jacchus 1 3.3 a
Golden-headed lion tamarin Leontopithecus chrysomelas 2
Common marmoset Callithrix jacchus 2 4.0 a
Golden-headed lion tamarin Leontopithecus chrysomelas 2
Pygmy marmoset Callithrix pygmae 10 1.2
Emperor tamarin Saguinus imperator 2 7.5
Black-headed spider monkey Ateles fusciceps 5 13
Celebes crested macaque Macaca nigra 2 19.9
Mandrill Mandrillus sphinx 14 17.9
Hamadryas baboon Papio hamadryas 20 8.1
Mantled guereza Colobus guereza 7 4.3
Hamlyn’s monkey Cercopithecus hamlyni 8 8.5
Northern plains gray langur Semnopithecus entellus 5 7.9
Javan lutung Trachypithecus auratus 4 3.7
Javan lutung Trachypithecus auratus 6 11.4
Siamang Hylobates syndactylus 4 12.0
Western lowland gorilla Gorilla gorilla 1 78.7a
Mountain gorilla Gorilla beringei 2
Common chimpanzee Pan troglodytes 10 15.7
Bornean Orangutan Pongo pygmaeus 2 83.0
Study site B
Black-and-white ruffed lemur Varecia variegata 6 31.0a
Ring-tailed lemur Lemur catta 3 19.5a
Brown lemur Eulemur fulvus 1
Tufted capuchin Cebus apella 5 44.4
Common squirrel monkey Saimiri sciureus 6 2.5
Common marmoset Callithrix jacchus 2 1.0
Black crested mangabey Lophocebus aterrimus 5 9.3
Black crested gibbon Hylobates concolor 1 14.0a
Red-cheeked gibbon Hylobates gabriellae 1
White-handed gibbon Hylobates lar 4 44.5
Common chimpanzee Pan troglodytes 3 161.7
Study site C
Ring-tailed lemur Lemur catta 8 253.0
Red ruffed lemur Varecia rubra 4 257.4
Black-and-white ruffed lemur Varecia variegata 4 127.2
Common squirrel monkey Saimiri sciureus 20 51.1
Siamang Hylobates syndactylus 2 NA
Study site D
White-handed gibbon Hylobates lar 4 132.2
Northern white-cheeked gibbon Hylobates leucogenys 4 32.3
Bonobo Pan paniscus 9 347.9
NA: not available.
a
Both NHP species are housed in the same enclosures.
 B. Levecke et al. / Veterinary Parasitology 148 (2007) 236–246 239

Table 2 thoroughly mixed after which it was transferred onto a

Estimated sample sizes and probabilities in relation to different group glass microscope slide and covered with a cover glass.
sizes
Each sample was examined at a 400 magnification for
Group size Samples size Probability (%) the presence of eggs, larvae, trophozoites and/or cysts.
(95% confidence interval)
2 6 96.8 (96.5; 97.2) 2.5. Statistical analysis
3 11 96.6 (96.2; 96.9)
4 16 96.4 (96.0; 96.8)
5 21 95.6 (95.1; 96.0)
‘Prevalence’ is defined as the percentage of groups of
6 27 95.6 (95.2; 96.0) NHP infected with a particular parasite species and
7 33 95.8 (95.4; 96.2) ‘diversity’ as the number of parasite species within the
8 38 95.3 (95.0; 95.8) same group. ‘Proportion’ describes the proportion of
9 45 95.4 (95.0; 95.8) samples containing a particular parasite species within
10 50 95.2 (95.0; 95.6)
11 58 95.5 (95.1; 95.9)
the same group and ‘multiple infections’ the median
14 76 95.4 (95.0; 95.8) number of parasite species found in these samples.
20 116 95.4 (95.0; 95.8) Entamoeba spp. was treated as one species. The
Kruskal–Wallis test was performed to test for differ-
ences in proportion and mixed infections between host
2.4. Coprological examination species (SAS 9.1.3, SAS Institute Inc., Cary, NC, USA).
The host species’ classes involved were prosimians,
An acetic acid–ether concentration method on faeces NW monkeys, OW monkeys and apes. The level of
was used to demonstrate the presence of gastrointestinal significance was set at P < 0.05. In addition, a
parasites in NHP (Thienpont et al., 1986). Half a gram Bonferroni pair wise comparison procedure was
of faeces was suspended in 5 ml acetic acid (5%) and performed. For this, the Wilcoxon test was employed
strained through a brush wire sieve to remove debris. and the level of significance was set at 0.0083 (SAS
The fat in the resulting filtrate was removed by 9.1.3, SAS Institute Inc., Cary, NC, USA).
emulsifying the sample with 5 ml of ether followed
by centrifugation at 1500 rpm for 2 min. The resulting 3. Results
supernatant (ether, debris and acetic acid) was discarded
and two drops of diluted iodine (1:100) were added to In total, 910 individual samples were collected. The
the remaining sediment. The stained sediment was number of times a group was sampled varied between 1

Table 3
Prevalence and diversity of parasites in NHP at four Belgian zoological gardens
Number of Entamoeba E. nana I. bütschlii Giardia C.mesnili B. coli Trichuris Strongyloides Diversity
groups spp. (%) (%) (%) spp. (%) (%) (%) spp. (%) spp. (%) (25Q–75Q)
Prosimians 9 33 11 0 44 11 0 0 0 0–2
Study site A 4 50 0 0 25 25 0 0 0
Study site B 2 50 50 0 100 0 0 0 0
Study site C 3 0 0 0 33 0 0 0 0
NW monkeys 10 40 40 0 40 0 0 0 0 0–2
Study site A 6 17 17 0 33 0 0 0 0
Study site B 3 67 67 0 33 0 0 0 0
Study site C 1 100 100 0 100 0 0 0 0
OW monkeys 9 100 44 22 22 55 33 33 0 1–5
Study site A 8 100 50 12 25 50 37 37 0
Study site B 1 100 0 100 0 100 0 0 0
Apes 11 64 45 0 54 18 18 1 18 1–3
Study site A 4 100 25 0 75 25 25 0 25
Study site B 3 67 0 0 33 33 0 33 0
Study site C 1 100 100 0 100 0 0 0 0
Study site D 3 0 100 0 33 0 33 0 33
39 59 36 5 41 21 13 10 5 1–3
240 B. Levecke et al. / Veterinary Parasitology 148 (2007) 236–246

infections
and 11 occasions and the median number of faecal

Mixed
samples produced by the groups measured 1 per animal,

1
0
0
0

0
0

0
0
0
ranging from 0.15 to 8. The median (25Q; 75Q)
probability of sampling all animals within a group was

Strongyloides
96% (93–97).

spp. (%)
3.1. Prevalence and diversity of parasites

0
0
0
0

0
0

0
0
0
A total of eight parasite species were identified, two

Trichuris
protozoan and two nematode species. Prevalence and

spp.
diversity of gastrointestinal parasites are reported in

0
0
0
0

0
0

0
0
0
Table 3. Infections with trematodes and cestodes were not

B. coli
detected. In 33 out of the 39 groups, infection with at least

(%)
one parasite species was detected. The most prevalent

0
0
0
0

0
0

0
0
0
gastrointestinal parasites were: Entamoeba spp. (59%),

I. bütschlii
and Giardia spp. (41%). Other parasites detected were
Endolimax nana (36%), Chilomastix mesnili (21%), B.

(%)
coli (13%), Trichuris spp. (10%), Iodamoeba bütschlii

0
0
0
0

0
0

0
0
0
(5%) and Strongyloides spp. (5%). All parasite species

Giardia spp.
were detected in no less than two study sites. Giardia spp.
and E. nana were the only parasite species observed at all
the four study sites. Groups of OW monkeys harboured
(%)

94

16
27

15
0
0
0

0
0
the most parasite species. In these groups the median
number of parasite species detected was three. The
C. mesnili

smallest diversity was observed in NW monkeys and

prosimians (median of one parasite species).
(%)

25
0

0
0

0
0

0
0
0
3.2. Proportion and multiple infections
E. nana
(%)

Tables 4–7 provide an overview of the results.

0
0
0
0

0
7

0
0
0
Overall, there was a large variation in proportion of
Entamoeba

parasite infections. For the most prevalent parasite

spp. (%)

species, the proportion ranged from 0 to 100% for

Entamoeba spp., and from 0 to 94% for Giardia spp. For
29

24
0
1
0

0
0
Entamoeba spp., the highest proportion was found in 0
Probability

OW monkeys. The median (25Q; 75Q) of proportion in

these host species measured 87.5% (80.0; 96.0). In Both NHP species are housed in the same enclosure.
(%)

Apes, Entamoeba spp. occurred in 12.5% (0; 62.5) of

83
87
96
93

93
94

96
96
96

the samples. Prosimians and NW monkeys were the

Proportion and multiple infections in prosimians
Sample

least infected with this parasite species. Entamoeba spp.

size

was not detected in 75% of these host species. Samples

15 b
34

16

25

40
16
16
4

containing cysts of Giardia spp. were mainly found in

apes. The median proportion in these NHP species was
Group

Black and white ruffed lemur.

size

4% (0; 38.8). The least proportion of this infection was

9
2
4
2

6
3
1

8
4
4

observed in OW monkeys, in which no cysts were found

in 75% of the groups. The less common parasites were
B & W ruffed lemura

B & W ruffed lemura

Brown greater galago

mainly found in OW monkeys. Overall, the median of

Ring-tailed lemur

Ring-tailed lemur

Ring-tailed lemur
Red ruffed lemur
Red ruffed lemur

Red ruffed lemur

the multiple infections measured 1, ranging from 0 to 3.

Brown lemur

OW monkeys harboured the most species of endopar-

Study site A

Study site B

Study site C

asites. In these groups the median (25Q; 75Q) number

Table 4

of infections detected was 1 (1; 2), 1 in apes (0; 1) and 0

b
a

in both NW monkeys (0; 1) and prosimians (0; 0).

Table 5
Proportion and multiple infections in NW monkeys
Group Sample Probability Entamoeba E. nana C. mesnili Giardia I. bütschlii B. coli Trichuris Strongyloides Multiple

B. Levecke et al. / Veterinary Parasitology 148 (2007) 236–246

size size (%) spp. (%) (%) (%) spp. (%) (%) (%) spp. (%) spp. (%) infections
Study site A
Goeldi’s monkey 2 16 a 96 0 0 0 0 0 0 0 0 0
Golden-headed lion tamarin 2
Common marmoset 1 4a 44 0 0 0 0 0 0 0 0 0
Golden-headed lion tamarin 2
Common marmoset 2 12 a 86 8 0 0 0 0 0 0 0 0
Golden-headed lion tamarin 2
Pygmy marmoset 10 5 0 0 80 0 20 0 0 0 0 1
Emperor tamarin 2 4 87 0 0 0 0 0 0 0 0 0
Black-headed spider monkey 5 20 94 0 0 0 45 0 0 0 0 0
Study site B
Tufted capuchin 5 21 96 5 0 0 5 0 0 0 0 0
Common squirrel monkey 6 14 69 7 7 0 0 0 0 0 0 0
Common marmoset 2 6 97 0 67 0 0 0 0 0 0 1
Study site C
Common squirrel monkey 20 127 97 1 56 0 9 0 0 0 0 1
a
Both NHP species are housed in the same enclosure.

241
242 B. Levecke et al. / Veterinary Parasitology 148 (2007) 236–246

infections
Multiple
No significant effects were found of study sites on
proportion of infections (x2(3) = 0.80–6.0, p = 0.84–
0.11) and multiple infections (x2(3) = 4.2, p > 0.2),

2
3
2
1
2
1
1
1

1
neither for stocking density (R = 0.17 to 0.001,
Strongyloides

p = 0.32–0.99; x2(3) = 0.77, p > 0.8).

spp. (%)

3.3. Differences in host species susceptibility

0
0
0
0
0
0
0
0

0
Both proportion and multiple infections (x2(3) = 19.9,
Trichuris
spp. (%)

p = 0.0002) differed between host species. Parasites for

0

0
3
0
0
0

0
20

16

which a significant difference in susceptibility at the

level of taxonomy was noted were Entamoeba spp.
B. coli

(x2(3) = 21.4, p < 0.0001) and C. mesnili (x2(3) = 9.3,

(%)

2
0
0
0
0
0

0
70
52

p < 0.05). However, a marginal p-value was also found

for I. bütschlii (x2(3) = 6.8, p = 0.08), Trichuris spp.
I. bütschlii

(x2(3) = 6.8, p = 0.08) and B. coli (x2(3) = 6.4, p = 0.09).

Samples containing cysts of Entamoeba spp. were more
(%)

37

30
0
0
0

0
0
0

prevalent in OW monkeys than in prosimians (jZj = 3.6,

p < 0.0005), in NW monkeys (jZj = 3.7, p < 0.0005)
spp. (%)
Giardia

and in apes (jZj = 2.7, p < 0.0083). OW monkeys had

a marginal significantly higher number of infected
0
7
2
0
0
0
0
0

samples with C. mesnili than NW monkeys (jZj = 2.6,

p = 0.0099). Samples collected from OW monkeys
C. mesnili

contained the highest number of parasite species

(%)

(prosimians: jZj = 3.5, p < 0.0005; NW monkeys:

0
49
23

11

13
0

0
0
8

jZj = 3.2, p < 0.005; apes: jZj = 2.8, p < 0.0083). No

E. nana

significant differences were observed between the other

host species.
(%)

60
56
42

54
0

0
0
0

4. Discussion
Entamoeba
spp. (%)

In the present study, the majority of the examined

30

100
99
96
80
83
94
64
88

groups were infected with at least one parasite species.

All the parasite species identified in our study have
Probability

previously been described in both captive and free

ranging NHP (Gómez et al., 1996; Ashford and
(%)

99
93
89
93
92
86
94
93

97

Wrangham, 2000; Legesse and Erko, 2004). Protozoa

were most prevalent, which confirms the findings of
Sample
Proportion and multiple infections in OW monkeys

other studies performed in captive NHP (Gómez et al.,

size

10

102
75

30
35
16
14
24

23

1996; Verweij et al., 2003a). The occurrence of these

parasites can be explained by the simplicity of their
Group

lifecycle, because they need no intermediate hosts and

size

2
14
20
7
8
5
4
6

are immediately infective when excreted. Moreover, the

low infective dose and the short prepatent period,
Northern plains gray langur

obviously ease transmission (Tanyuksel and Petri, 2003;

Celebes crested macaque

Black crested mangabey

Thompson and Monis, 2004). The most prevalent

Hamadryas baboon

Hamlyn’s monkey

parasite species in this study were Entamoeba spp. and

Mantled guereza

Giardia spp. More than 40% of the examined groups

Javan lutung
Javan lutung

were infected with one of these endoparasites. However,

Study site A

Study site B
Mandrill

a large variation in proportion and multiple infections

Table 6

was observed between the examined groups. This

variation may be caused by differences in host species
Table 7
Proportion and multiple infections in apes
Group Sample Probability Entamoeba E. nana C. Mesnili Giardia I. bütschlii B. coli Trichuris Strongyloides Multiple
size size (%) spp. (%) (%) (%) spp. (%) (%) (%) spp. (%) spp. (%) infections

B. Levecke et al. / Veterinary Parasitology 148 (2007) 236–246

Study site A
Siamang 4 15b 94 27 0 0 47 0 0 0 7 1
Western lowland gorilla a 1 3b 100 67 0 0 67 0 0 0 0 1
Mountain gorilla a 2
Common chimpanzee 10 49 94 8 8 2 39 0 29 0 0 1
Bornean orang-utan a 2 8 100 62 0 0 0 0 0 0 0 1
Study site B
White-handed gibbon 4 16 96 12 0 0 12 0 0 0 0 0
Black crested gibbona 1 3b 100 0 0 0 0 0 0 33 0 0
Red-cheeked gibbona
Common chimpanzee 3 11 97 100 0 45 0 0 0 0 0 1
Study site C
Siamang 2 10 99 60 10 0 20 0 0 0 0 1
Study site D
White-handed gibbon 4 18 97 0 6 0 0 0 0 0 0 0
Northern w-c gibbonc 4 26 99 0 12 0 4 0 11 0 42 1
Bonoboa 9 22 100 0 41 0 0 0 0 0 0 0
a
Individual samples.
b
Both species are housed in the same enclosure.
c
Northern white-cheeked gibbon.

243
244 B. Levecke et al. / Veterinary Parasitology 148 (2007) 236–246

susceptibility, since OW monkeys were at higher risk of are more appropriate (Verweij et al., 2003b). Among
infections, including those with Entamoeba spp. and other pathogenic parasites that cannot be ruled out in the
harboured the highest number of parasite species. present study was Cryprosporidium spp. (Miller et al.,
Differences in host species susceptibility have been 1990; Muriuki et al., 1997; Gómez et al., 2000), because
hypothesized and might be explained by differences in no appropriate detection technique was used.
behaviour, because the majority of the OW monkeys are Although all parasites found are recognised zoonotic
ground dwellers (Beaver et al., 1988; Munene et al., pathogens, the high prevalence of Giardia warrants
1998). special attention. Zoonotic assemblages (G. duodenalis
Using the sample strategy described in this study, assemblage A and B) have been described in NHP
more insights were gained in the epidemiology of (Thompson et al., 2000; Graczyk et al., 2002; Nizeyi
gastrointestinal parasites in NHP of zoological gardens. et al., 2002, Vitazkova and Wade, 2006). However,
First of all, it is likely to result in more accurate differentiation of the assemblages of G. duodenalis
estimates of the group’s prevalence, because all animals based morphological features is impossible, and there-
were examined with a high probability and none of the fore additional prevalence data based on molecular
samples were pooled. Moreover, it allowed a more techniques are needed to confirm this reservoir function
powerful risk factor analysis, since proportion within of NHP involved in this study. In conclusion, the results
groups could be estimated. of this study demonstrated that gastrointestinal protozoa
Of all parasites identified in this study, Giardia spp., B. and multiple infections are highly prevalent in NHP of
coli, Trichuris spp. and Strongyloides spp. might cause Belgian zoos. There was a large variation in proportion
gastrointestinal enteritis in NHP (Lee et al., 1990; and multiple infections between groups of NHP, which
Hamlen and Lawrence, 1994; Hänichen et al., 1995; might be explained by differences in host species
Kalishman et al., 1996). However, due to the study design susceptibility. The used sample strategy is likely to
the clinical importance of these infections could not be result in more accurate epidemiological data. This study
studied more in depth, in particular for E. histolytica. E. also emphasizes the need for molecular diagnostic tools
histolytica has been previously described in various in NHP to evaluate the clinical importance and zoonotic
species involved in this study (Loomis et al., 1983; risk of these infections.
Beaver et al., 1988; Verweij et al., 2003a; Mätz-Rensing
et al., 2004), but differentiation between other Enta- Acknowledgements
moeba spp. (E. coli, E. hartmanii, E. polecki-like) is
difficult (Verweij et al., 2001, Kebede et al., 2003) and This work received financial support from the Centre
even impossible when E. dispar or E. moshkovskii are of Research and Conservation of The Royal Zoological
involved (Diamond and Clark, 1993, Tanyuksel and Petri, Society of Antwerp, which is supported by the Flemish
2003). For this purpose molecular diagnostic techniques Government (Dehousse grant).
 B. Levecke et al. / Veterinary Parasitology 148 (2007) 236–246 245

Appendix A
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