Veterinary Parasitology 163 (2009) 115–122

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Veterinary Parasitology
journal homepage: www.elsevier.com/locate/vetpar

Zoonotic parasites in fecal samples and fur from dogs and cats
in The Netherlands
Paul A.M. Overgaauw a,*, Linda van Zutphen b, Denise Hoek c, Felix O. Yaya d,
Jeroen Roelfsema c, Elena Pinelli c, Frans van Knapen e, Laetitia M. Kortbeek c
a
RnA BV, Yalelaan 2, 3584 CM, Utrecht, The Netherlands
b
Quantitative Veterinary Epidemiology Group, Wageningen Institute of Animal Sciences, Wageningen University, The Netherlands
c
National Institute for Public Health and the Environment, Department of Parasitology, Laboratory for
Diagnosis and Screening of Infectious Diseases, Bilthoven, The Netherlands
d
Free University of Amsterdam, Department of Biomedical Sciences, The Netherlands
e
Institute for Risk Assessment Sciences, Division of Veterinary Public Health, Utrecht University, The Netherlands

A R T I C L E I N F O A B S T R A C T

Article history: Pets may carry zoonotic pathogens for which owners are at risk. The aim of the study is to
Received 18 September 2008 investigate whether healthy pets harbour zoonotic parasitic infections and to make an
Received in revised form 22 March 2009 inventory of the interactions between pet-owners and their companion animals in the
Accepted 26 March 2009 Netherlands. Fecal and hair samples were collected from healthy household dogs and cats
in Dutch veterinary practices. Owners were interviewed about interaction with their pets.
Keywords: The samples were investigated by microscopy, ELISA, and PCR.
Zoonosis
From 159 households, 152 dogs (D) and 60 cats (C), information and samples were
Toxocara
collected and examination for several zoonotic parasites was performed. Toxocara eggs
Giardia
Cryptosporidium were found in 4.4% (D) and 4.6% (C) of the fecal samples and in 12.2% (D) and 3.4% (C) of the
Hygiene fur samples. The median epg in the fur was 17 (D) and 28 (C) and none of these eggs were
viable. From 15.2% of the dog and 13.6% of the cat feces Giardia was isolated. One canine
and one feline Giardia isolate was a zoonotic assemblage A (12%). Cryptosporidium sp. were
present in 8.7% (D) and 4.6% (C) of the feces.
Fifty percent of the owners allow the pet to lick their faces. Sixty percent of the pets visit
the bedroom; 45–60% (D–C) are allowed on the bed, and 18–30% (D–C) sleep with the
owner in bed. Six percent of the pets always sleep in the bedroom. Of the cats, 45% are
allowed to jump onto the kitchen sink. Nearly 39% of the dog owners never clean up the
feces of their dog. Fifteen percent of the dog owners and 8% of the cat owners always wash
their hands after contact with the animals. Close physical contact between owners and
their pets is common and poses an increased risk of transmission of zoonotic pathogens.
Education of owners by the vet, specifically about hygiene and potential risks, is required.
ß 2009 Elsevier B.V. All rights reserved.

1. Introduction
In The Netherlands, the number of pets has increased
during the last decade, of which 1.8 million (M) dogs, 3.3 M
cats, and 0.9 M rabbits are mostly responsible. Stray cats
* Corresponding author. Tel.: +31 342 419798; fax: +31 342 419794.
E-mail address: p.overgaauw@planet.nl (Paul A.M. Overgaauw).
are common; however, stray dogs are hardly present in the
Netherlands. The number of households with pets
increased from 50% in 1999 to 55% in 2005 (Dutch Council
for Animal Affairs, 2006).
As pets are increasingly considered a member of the
family, physical contact is very common. Companion
animals enhance the psychological and physiological
well-being of the human. Dogs play an important role in
the development and in the treatment of behavioral

0304-4017/$ – see front matter ß 2009 Elsevier B.V. All rights reserved.
doi:10.1016/j.vetpar.2009.03.044
116 P.A.M. Overgaauw et al. / Veterinary Parasitology 163 (2009) 115–122
problems of children, the well-being of the elderly, and
decrease work leave through illness and visits to the doctor
(Beck and Meyers, 1996). Next to the benefits of pets for the
human population, there are also potential health hazards
associated with the ownership of a pet. Besides the risk of
bites, scratches and allergies, several infections can be
transmitted to the human as zoonosis. Significant parasitic
zoonoses of dogs and cats prevalent in The Netherlands are
Toxocara, Giardia, Cryptosporidium and Toxoplasma (Buijs,
1993; Overgaauw, 1997a; Valkenburgh et al., 2007). These
parasites have an oro-fecal transmission cycle and humans
can be infected either by fecal contamination of food, water
or the environment (gardens, sandpits and playgrounds) or
by direct contact (Good et al., 2004; Hill et al., 2000;
Robertson and Thomson, 2002; Smith et al., 2007). Direct
contact with dogs and cats that harbour a patent Toxocara
infection is usually not considered a risk, since the eggs need
to mature several weeks before they are infective (Over-
gaauw, 1997b; Overgaauw and Van Knapen, 2000, 2004).
Recent studies, however, indicate the fur of these pets as
another important source of embryonated Toxocara eggs
(Wolfe and Wright, 2003; Roddie et al., 2008; Aydenizöz-
Özkayhan et al., 2008). G. duodenalis is known to exhibit
species complexity as it has been isolated from humans and
animals, including pets (Eligio-Garcı́a et al., 2005). Compa-
nion animals are most often infected with host-specific
Cryptosporidium sp. from which C. felis and C. canis
infrequently are reported to infect humans (Pedraza-Dı́az
et al., 2001; Fayer, 2008; Wielinga et al., 2008; Xiao and Feng,
2008), despite close and widespread contact. Therefore,
their role in the transmission of human cryptosporidiosis
appears quite limited (Xiao and Fayer, 2008).
In the present study, the results are presented from the
examination of several zoonotic parasites in fecal and fur
samples from healthy household dogs and cats. In addition,
an inventory of the interactions between pet-owners and
their companion animals in the Netherlands is reported.
2. Materials and methods
2.1. Questionnaires
Ten veterinary clinics in urban and rural areas in different
provinces in The Netherlands were visited weekly during
the period of March–May 2007. Owners of clinically healthy
dogs and cats were asked to participate in the study and then
to fill in a questionnaire. In this way information was
collected of the owners, the animals, the physical relation-
ship between pet-owners and their pets, and management
factors such as nutrition (raw vs. commercial food),
defecating location cat (outdoor, cat litter box, or both),
cleaning frequency of the litter box, walking environment
dog (urban, rural and dog exercising area), and deworming
frequency. Finally information regarding fur length, body
weight, living environment, location, breed, and cat outdoor
or indoor access was questioned.
2.2. Egg recovery technique from hair
With the pet-owner consent, fur samples were
collected by combing the dorsum (lumbal and sacral
area) and flanks, representing the areas where owners
normally pet their animals. Combs were cleaned and
every time disinfected with a halogenated organic
compound (Neosabenyl1) after use. At the moment of
analysis, there was no standardized and validated method
published for the isolation of Toxocara eggs from dog and
cat hair. Therefore the following method was developed
and validated using T. canis eggs available at the
Department of Parasitology. Different numbers (8, 39,
and 100) of Toxocara eggs were added separately to hair
samples from a Toxocara-negative dog. To recover back
the Toxocara eggs, the spiked hairs were washed and
shaked vigorously as described below. The overall
recovery rate for this method was 95%.
The hair sample was weighted and samples with a
weight less than 0.0025 g were not examined, as they were
considered too small. A soap solution of three drops
(approximately 0.2 ml) of the detergent Tween 20 (Merck,
Germany) in 40 ml sterile distilled water was prepared to
wash the hair by shaking vigorously and allowing to stand
for a minimum of 10 min. The floating hair was transferred
with a pair of tweezers to another tube and washed again
with 40 ml phosphate buffer solution (PBS). The suspen-
sion was allowed to stand for a minimum of 10 min and the
hairs were discarded. The suspensions from the two tubes
were centrifuged at 800  g for 10 min and the super-
natant decanted until approximately 1 ml. The sediments
from the tubes were re-suspended and transferred to
combine the pellet suspension in a tube using a sterile
pipette. The tubes were flushed once with drops of PBS and
combined. This tube was centrifuged again at 800  g for
10 min and the supernatant again decanted until approxi-
mately 1 ml. The sediment was re-suspended and trans-
ferred to a 1.5 ml Eppendorf tube. Finally, this tube was
centrifuged at 800  g for 10 min and supernatant
discarded till approximately 100 ml. The sediment was
re-suspended and the whole volume examined using a
light microscope under 400 magnifications. Positive
samples for Toxocara eggs were preserved in 0.05 M
sulphuric acid solution to allow embryonation, as
described by De Savigny (1975), for a viability test at a
later point in time.
2.3. Confirmation of isolated Toxocara eggs from the fur
The Toxocara eggs found in the fur were identified using
a PCR with primers NC2 and Tcan1 or NC2 and Tcat1
(Jacobs et al., 1997).
2.4. Determination of the viability of isolated Toxocara eggs
The hair solution samples with Toxocara eggs stored in
0.05 M sulphuric acid were retrieved from storage at room
temperature after 6 weeks. The suspension of Toxocara
eggs was centrifuged at 800  g for 10 min and the
supernatant removed. The suspension was examined by
light microscopy at 100–400 magnification. The eggs
were classified as non-viable (empty, not intact egg wall),
unembryonated or fertilized (intact egg wall, with con-
tents), embryonated (with cell divisions), or infective (with
larva).
 P.A.M. Overgaauw et al. / Veterinary Parasitology 163 (2009) 115–122
2.5. Fecal examination
Fecal samples were provided in a feces container by the
owner. The samples were sent to the laboratory within
24 h of collection and stored at 4 8C until analysis.
An ELISA was used (ProSpecT, Alexon-Trend, USA),
according to the manufacturer’s instruction, to determine
the presence of Giardia specific antigen (GSA 65 protein) in
fecal samples, because with microscopy alone it is possible
to miss Giardia cysts from a single fecal sample (Mank
et al., 1997).
An ether-sedimentation flotation-centrifugation (Rid-
ley and Hawgood, 1956) method was used to identify
nematode parasite eggs and cysts in the fecal samples. The
samples were examined by light microscopy with 100–
400 magnifications.
A modified (cold method) Ziehl–Neelsen staining
(Henriksen and Pohlenz, 1981) was performed on a light
smear from the Ridley’s concentration, after air drying
overnight, to identify oocysts of Cryptosporidium. The slide
was examined by light microscopy with oil immersion lens
at 1000 magnification.
2.6. Genotyping of Giardia and Cryptosporidium
Immediately after a positive ELISA for Giardia in fecal
samples, the cysts were isolated from feces using
Dynabeads that are covered with antibodies directed
against Giardia cysts. DNA was isolated from the cysts after
freeze/thawing using the Puregene Kit (BIOZYM). Isolation
of Giardia cysts and subsequent DNA isolation was
performed to identify the genotype by PCR and direct
sequence (Van der Giessen et al., 2006).
Genotyping was performed with 18S small subunit
gene (Van der Giessen et al., 2006). The 18S gene
amplification was performed using the primers 18S-1
and 18S-A in a 50 ml reaction mixture containing 10 mM
dNTP, 10 buffer II, 25 m/M Mgcl2, 5 U/ml Ampli Taq
(Applied Biosystem, The Netherlands). The PCR was carried
out with the following condition: one cycle of 94 8C for
5 min, followed by 40 cycles of 94 8C for 1 min, 55 8C for
1 min and 72 8C for 3 min.
Purified (Qiagen/Westburg, Leusden, The Netherlands)
PCR products were sequenced in both directions using the
BigDyeTerminator kit (Applied Biosystems, Nieuwekerk a/
d IJsel, The Netherlands) and an ABI automated sequencer.
DNA sequences from this study were compared with DNA
sequences from the reference strain from Zoopnet (RIVM,
The Netherlands). Multiple alignments were made using
ClustalW. PCR on Cryptosporidium positive fecal samples
was performed on the 18S small subunit gene according to
Wielinga et al., 2008. DNA was isolated on three separate
occasions: once with the NucliSens kit (BioMerieux
Benelux B.V. Boxtel), and two times with the Hi Pure Kit
(Roche, Almere, The Netherlands).
2.7. Statistical analysis
For the statistical analysis, three steps in SAS (SAS 9.1.3,
ß 2002–2004, SAS Institute Inc., Cary, NC, USA) were
performed. First, a descriptive analysis was performed to
117
determine the prevalence of the studied pathogens. The
distribution of the animals over the classes of all manage-
ment factors was determined, as well as the prevalence of
the pathogens per class of each management factor.
Secondly, a univariable logistic regression analysis was
done with binomially distributed outcome data to
determine effects of the management factors on infection
with the studied parasites. In the model, the dependent
variable was the presence or absence of infection, and the
independent variable was the management factor. Odds
ratios and their 95% confidence intervals have been
calculated using exact logistic regression.
3. Results
3.1. Sampling and questionnaires
With the pet-owner consent, from 159 households in
total 148 dog and 59 cat fur samples were collected, and
from 92 dogs and 22 cats fecal samples were provided by
the owner.
Questionnaire results show that half of the owners
allow the pet to lick their faces. Sixty percent of the pets
visit the bedroom; 45% of the dogs and 62% of the cats are
allowed on the bed, and 18% and 30% of the dogs and cats,
respectively, are allowed to sleep with the owner in bed. Of
the cats, 45% are allowed to jump onto the kitchen sink.
Most cats are allowed to go outside, but 32% stay indoors
and accordingly use the litter box for defecation. From the
outdoor cats, 76% (32/41) also use the litter box. Fifty-five
percent of owners clean the litter box more often than
twice a week, while 39% of the dog owners never clean up
their dogs’ feces. Forty-five percent of the dogs and 40% of
the cats are dewormed more than twice a year. Fifteen
percent of the dog owners and 8% of the cat owners always
wash their hands after contact with the animals.
3.2. Hair analysis
The results of the analysis of all fecal and fur samples
from in total 224 investigated dogs and cats, originating
from 159 pet-owners, are presented in Table 1. The average
weight of the fur samples, collected after thorough
combing, was 0.129 g for the dog and 0.037 g for the cat
(range: 0.0026–0.4725). In the fur of 18 dogs (12.2%) and
two cats (3.4%) Toxocara eggs were found. The average
number of eggs per sample was 3.5 (1–31) and recalculat-
ing to eggs per gram fur (epg) resulted for the dogs in a
mean epg of 94 (3.8–1065) and a median epg of 17. For the
cat a mean as well as median epg of 28 (22.9–32.5) was
found. The eggs in five examined samples were fertilized
(25%), except one egg (from the sample of 31 eggs) and
measured from 65–80 mm. Out of the 20 samples there
was sufficient remaining material of only five samples to
perform PCR and these eggs were confirmed as Toxocara.
From the remaining samples not enough material could be
extracted to perform PCR for the confirmation of these eggs
as Toxocara. The viability of the eggs was examined after 6
weeks culturing in sulphur acid solution. None of these
eggs were viable. Positive canine hair samples for Toxocara
eggs originated from different breeds and fur length: seven
118 P.A.M. Overgaauw et al. / Veterinary Parasitology 163 (2009) 115–122

Table 1
Percentage of hair and fecal samples positive for zoonotic parasites in dogs and cats.

Dogs % 95% CI Cats % 95% CI Total

Submitted animals 161 63 224

Hair samples 148 59 207

pos. Toxocara 18 12.2 7.4–18.5 2 3.4 0.4–11.7 20

Fecal samples 92 22 114

pos. Toxocara 4 4.4 1.2–10.8 1 4.6 0.1–22.8 5
pos. Giardia 14 15.2 8.6–24.2 3 13.6 2.9–34.9 17
pos. Cryptosporidium 8 8.7 3.8–16.4 1 4.6 0.1–22.8 9

short, nine medium and two long hair breed. The average
age of the dogs was 6.5 years (0.5–13). From nine dogs with
egg-positive hair samples, the feces were also investigated.
These all tested negative for intestinal parasites. From the
two cats, one had no outdoor access.
3.3. Fecal analysis
Based on the fecal analysis with the Ridley concentra-
tion method, four dogs and one cat were positive for T.
canis (Table 1). Fourteen dogs and three cats were positive
for Giardia and eight dogs and one cat for Cryptosporidium.
One dog showed a double infection of Giardia with T. canis
and two other dogs Giardia with Cryptosporidium. All three
dogs with multiple infections originated from the same
household. Next to these animals there was one other dog
double infected with Cryptosporidium and Giardia. There
were no Toxoplasma cysts found in the cat feces.
3.4. Genotyping of Giardia and Cryptosporidium
The Giardia fecal examination with Ridley’s concentra-
tion method resulted in 17 positive samples. With the
enzyme linked immunoabsorbant assay (ELISA) 19 posi-
tive (16 dogs and 3 cats) were found. DNA isolates of these
19 samples were amplified by PCR for 18S rDNA. The 18S
PCR was positive for 16 samples. From 15 positive samples
(13 dogs and 2 cats) sequence analysis was performed.
The genotype was determined from the 18S DNA
sequences alignment on ClustalW and the analysis of five
single nucleotide polymorphisms (SNPs). With the 18S
Table 2
Giardia genotype (assemblage) and 18S sequence polymorphisms of 15
dog and cat samples.
sequences the isolates were grouped into assemblage A
(two samples), assemblage C (seven samples), assemblage
D (three samples), assemblage C/D (one sample), assem-
blage F (one sample), and one sample with an unknown
assemblage. The unknown assemblage had two ambiguous
nucleotide bases/codes (Y and M) (Table 2).
Genotyping the Cryptosporidium positive samples,
determined after the Ziehl–Neelsen staining, proved to
be impossible. PCR yielded no results.
3.5. Risk factors
With logistic regression no significant difference was
found for Giardia, Cryptosporidium and Toxocara (in feces)
infection between dogs and cats.
Only for Toxocara eggs in the fur, an almost significant
higher prevalence in dogs compared to cats was found
(p = 0.08). Toxocara eggs were four times more present in
dog than in cat fur.
Some management factors concern only dogs or cats. A
subsequent logistic regression analysis was performed to
determine effects of management factors on Toxocara in
fur, Toxocara in feces, Toxascaris, Cryptosporidium, and
Giardia infection. Unfortunately the number of animals
with an infection was too low to perform valid statistical
analyses except for Toxocara in fur. In a univariate analysis
it was found that dogs off the leash or both on and off the
leash when outdoors have significantly more often (OR 8.7)
Toxocara eggs in their fur than dogs that are always on the
leash (p = 0.03). For the variables fur length and body
weight of the dogs there was no significant effect found
(p > 0.25).
4. Discussion

No. Species Assemblage 18S (SNPs) sequence Patent Toxocara infections in dogs and cats are
1. Dog A CCTGAT considered as a public health risk because of their zoonotic
2. Dog D TATTTT potential. In the literature the presence of stray dogs, the
3. Cat F CCTGAG
growing popularity of pet dogs and cats, the limited open
4. Dog C CATCTT
5. Dog C CATATT
green areas, pica and geophagia by children, and the close
6. Dog D TATTTT physical contact between owner and pet are given as a risk
7. Dog C CATCTT of transmission (Overgaauw, 1997a; Smith et al., 2006).
8. Dog C CATATT The prevalence of Toxocara infections in household dogs
9. Dog C CATCTT
and cats, based on fecal egg output, in our study is in
10. Cat A CCTGAT
11. Dog C CATCTT agreement with the earlier reported 3–5% in The Nether-
12. Dog C/D YATYTT lands (Overgaauw, 1997c) and Belgium (Claerebout et al.,
13. Dog C CATCTT 2005). In stray animals or animals in animal shelters,
14. Dog unknown YMTTAT
prevalences may be higher, up to 9% in dogs (Le Nobel et al.,
15. Dog D TATTTT
2004) and 28% in cats (Robben et al., 2004).
 P.A.M. Overgaauw et al. / Veterinary Parasitology 163 (2009) 115–122
Table 3
Comparable studies of Toxocara fur infection in dogs and cats.
References Species
Hasslinger et al. (1973) Private/stray cats
Wolfe and Wright (2003) Private/farm/shelter dogs
Regosz (2007) Private dogs
Private cats
Stray cats
Roddie et al. (2008) Adult stray dogs
Puppy stray dogs
Aydenizöz-Özkayhan et al. (2008) Private dogs
This study Private dogs
Private cats
a
Not determined.
Regarding Toxocara eggs in hair, a few other studies in
dogs were carried out in the UK and Ireland (Wolfe and
Wright, 2003; Roddie et al., 2008), France (Regosz, 2007),
and Turkey (Aydenizöz-Özkayhan et al., 2008). Cat fur was
investigated in Germany (Hasslinger et al., 1973) and
France (Regosz, 2007). An overview of these studies is
presented in Table 3. Because the mean epg is easily
influenced by outliers, the median epg (if determined) is
also presented.
We developed a practical and validated egg recovery
technique from hair, without the use of time-consuming
sieves. The overall recovery rate, however, is high. With
this technique Toxocara eggs were found in the hair of 12%
of the investigated dogs and 3% of the cats. An explanation
of the significantly higher number of Toxocara eggs in the
dog fur could be explained by the fact that dog (playing)
behavior results in more soil contact. Cats, moreover, lick
their fur more intensively and thus may find it easier to
remove any present eggs. In dogs the eggs were found in all
fur types and nearly all dogs were adults. There was no
simultaneous Toxocara infection in the feces. Therefore, the
presence of eggs in the fur does not necessarily mean that
this is always a result of self-contamination. This is also the
conclusion for the one cat with contamination of its hair,
but without outdoor access. A possible explanation may be
that the eggs were acquired through transmission for
instance via the shoes of the owner. The presence of eggs in
house dust was established in a survey among dog
breeders (Overgaauw and Boersema, 1998).
The presence of Toxocara eggs in the fur, especially in
puppies and stray dogs, is an indication that direct contact
with these animals may put humans at risk for developing
an infection of clinical toxocarosis. There are two condi-
tions that have to be fulfilled before an infection can
develop.
First, the eggs need to be embryonated to be infective.
None of the eggs found in our study were viable, probably a
result of UV-light influence and/or lack of humidity. Also,
the fact that the eggs were probably obtained from the
environment may be an explanation. Development to an
infective stage will last 3–6 weeks to several months,
depending on soil type and climatic conditions (time of the
year), such as temperature and humidity. In the other
studies a low percentage of the eggs were embryonated.
119
Nr. tested Prevalence Median epg %Embryonated
a
17 1 (6%) ND ND
60 15 (25%) 1 4%
29 0 – –
9 0 – –
7 1 (14%) 10 ND
75 42 (56%) 7 0.1%
25 25 (100%) 793 0.3%
51 11 (22%) 10 8%
148 18 (12%) 17 0%
59 2 (3%) 28 0%
Secondly, ingestion of a sufficient number of eggs is
required to stimulate an immune response and to cause
clinical human toxocarosis. Antibody levels in dogs and
mice were found to be strictly dose-related (Glickman and
Schantz, 1981; Kayes et al., 1985; Pinelli et al., 2007) and
this may also be the case in man. In most of the mentioned
studies, hair samples were collected from the perianal area
after thorough combing, and even then not more than one
egg was found per sample. This, together with the fact that
Toxocara eggs are very sticky, and therefore difficult to
remove from the coat of a dog or cat, makes ingestion of
sufficient numbers of eggs unlikely. It is, however,
remarkable in this context to notice that in the study of
Roddie et al. (2008) some cases were presented with very
high number of eggs in the hair of stray dogs, and mainly
their puppies, without significant difference between the
sampled perianal area and the dorsal region.
In some reports it has been suggested that the epg of
Toxocara in fur of infected animals is much higher than in
soil (Wolfe and Wright, 2003, 2004; Roddie et al., 2008).
This is undoubtedly the case, but does this suggest that
dogs and cats infected with Toxocara may infect people by
direct contact and provide a better explanation of the
epidemiology of the disease? Even in the worst case
scenario of highly contaminated fur, e.g. with the highest
Toxocara epg of 300 and an embryonated rate of 4% from
the study of Wolfe and Wright (2003), it is necessary to
ingest more than 4 g of hair, with 12 embryonated eggs per
gram, to ingest 50 infective eggs. As example Fig. 1 shows
the largest hair sample from our study with a weight 0.47 g
to indicate roughly the size of a few grams.
The hair contamination with Toxocara eggs, found in the
studies of Wolfe and Wright (2003) and Roddie et al.
(2008), is used to suggest that this is probably the main
route of transmission to the human and that there lacks
sufficient evidence of a direct link between seroprevalence
in people and soil contamination (sapro-zoonosis). There
are two arguments to disprove this suggestion. Many
studies reported that children are more frequently infected
than adults and that VLM with more severe clinical
symptoms is mainly found in children of 1–3 years of age
(Glickman and Schantz, 1981; Van Knapen et al., 1983,
1992; Schantz, 1989). This can be explained because young
children often play in and have closer contact with
120 P.A.M. Overgaauw et al. / Veterinary Parasitology 163 (2009) 115–122
Fig. 1. The size of the largest sample of hair, 0.4725 g. Ruler in centimeters
(photo: Denise Hoek).
potentially contaminated soil in yards and sand-pits. In
addition, they put their fingers into their mouth, some-
times eat dirt (pica), and are not very hygienic (Holland
et al., 1995). Around 40% of patients with Toxocara ocular
larva migrans showed a history of pica (Schantz et al.,
1979). Such a high percentage of children of this age are
not likely to have intense direct contact with (stray) dogs
and cats to retrieve enough embryonated eggs from the fur
to cause clinical disease.
Another argument is the fact that some surveys did not
support a relationship of human toxocarosis and exposure
to dogs or cats in the household (Buijs, 1993; Woodruff
et al., 1978). The relationship of the presence of (young)
dogs in households and toxocarosis (Schantz et al., 1979,
1980; Worley et al., 1984) or the presence of Toxocara
antibodies (Glickman et al., 1981; Holland et al., 1995) has
been described, but did not provide a reliable indication
that these dogs or cats were the sources of the infections.
Although in the present study anti-Toxocara antibodies
were not measured in dog owners, we do not consider
stroking a dog or cat a risk for acquiring a Toxocara
infection. However, the results from different studies
reveal the need for sufficient deworming schedules,
especially in puppies.
Giardia has been isolated from humans and animals.
This has raised the issue as to whether G. duodenalis is a
zoonosis. In a study in the Netherlands among gastro-
enteritis patients from general practitioners, Giardia was
found in 5.4% of cases and in 3.3% of controls. There was no
correlation between protozoal diarrhea and the ownership
of pets (De Wit et al., 2001). Assemblage A has been found
in humans, livestock, cats and dogs and assemblage B in
humans, dogs, and cats. Both assemblages seem to be
related with clinical symptoms in humans (Homan and
Mank, 2001). Other genetic assemblages within the G.
duodenalis group appeared to be confined to specific
animal hosts, like cats (assemblage F) and dogs (assem-
blages C and D). In a study in the Netherlands, human and
animal assemblages A and B Giardia-isolates could be
identified. However, phylogenetic analysis revealed dif-
ferent sub-clustering for human and animal isolates,
where host-species-specific assemblages (C–G) could be
identified (Van der Giessen et al., 2006). In our study 14% of
the cat fecal samples and 15% of the dog samples tested
positive for G. duodenalis. Most isolates were of the
assemblages C and D showing species specificity. The
human isolate assemblage A detected in one dog and one
cat sample is zoonotic and the first Dutch detection; until
now, genotyping of Giardia isolates from fecal samples
specific for household pets (dogs and cats) was not
previously reported in The Netherlands. Studies done in
the past on the genotype of Giardia in Dutch human
patients and some animal isolates suggested that the
genotype of G. duodenalis is species-specific, except for the
human genotypes which have been isolated from other
animals (Ten Hove et al., 2007; Van der Giessen et al.,
2006). These findings were in agreement with earlier
findings by Monis and Thompson (2003). In Belgium, 6.4%
of 273 privately owned healthy dogs tested positive for
Giardia infection; the infection was significantly more
prevalent in dogs younger than 1 year. Genotyping
resulted in assemblage A in 19/23 dogs (83%) and
assemblage D in 17% of the dogs (Claerebout et al.,
2005). Further research is needed to establish that pets can
be the source of human infection.
The knowledge of the clinical presentation caused by
Cryptosporidium sp. is changing because it is possible to
type different strains: C. parvum and C. hominis. The latter
spread only between humans and the major reservoir of C.
Parvum is domestic livestock (Hunter and Thomson, 2005).
However, human cryptosporidiosis in relation with dogs or
cats (C. canis and C. felis), is very rare (Xiao and Feng, 2008)
and not found to be a risk factor in England (Goh et al.,
2004). In the Netherlands, of human isolates 1% C. felis was
found (Wielinga et al., 2008).
From the results of the parasitic pathogens, this study
should be considered as an initial one and a successive
study can be performed where infections of human
patients with pets are involved and can be used for further
risk analysis.
Toxoplasma cysts were not found in the cat feces in this
study. Cats only shed Toxoplasma oocysts once during their
lifetime, and only during a limited period of time (2–3
weeks) when they have lost their passive immunity and
have started to hunt or to eat raw meat (ingestion of tissue
cysts). Most of the spontaneous shedding cats found in
surveys are younger than 6 months of age. During the
excretion period they build up enough immunity to resist a
new infection (Dubey, 1995). In the Netherlands 60% of all
1-year-old cats have passed this period, thus they do not
longer play an important role in the epidemiology of
toxoplasmosis (Van Knapen, 1993, unpublished data) the
chance to find a cat that is shedding Toxoplasma cysts is
therefore very limited. A validated PCR for Toxoplasma in
cat feces was not available and is therefore not used.
Regarding the hygiene of owners in their physical
relationship with their pets, Westgarth et al. (2008)
investigated the nature and frequency of contacts between
dogs and owners in a UK area. Similar results were found in
this survey regarding licking the face of the household
members (60%), sleeping in the bedroom (33%) and on the
human’s bed (14%). The close contacts and sharing of beds
were considered to allow transmission of zoonotic diseases
or parasites. The percentages of UK dog owners who said
 P.A.M. Overgaauw et al. / Veterinary Parasitology 163 (2009) 115–122
they pick up their dog’s feces during walks in various
environments are impressive compared with the results
that we found in The Netherlands. Eighty percent or more
of the UK-owners said they do this usually or always in the
street, park area or on public pathways.
5. Conclusions
Nowadays, pets enjoy great freedom when inside,
which enables transmission of zoonotic pathogens to
surfaces and rooms accessible to humans and animals.
Several clinically healthy dogs and cats in this study were
infected with potential zoonotic parasites. The prevalences
may even be underestimated because several parasites
intermittently shed eggs or oocysts. Since no data was
available on the infection status of the owners, no
conclusions can be drawn about the risk for transmission
of these pathogens. Transmission of parasites or protozoa
to the human is only possible if the parasites have been
developed into the infective stage, are pathogenic for
humans (e.g. assemblage A for G. duodenalis), and ingestion
of sufficient numbers of infective agents has taken place.
Acknowledgements
We thank the students Danielle Resink, Janne Teeninga,
Renée van de Ven and Kirsten Peeters for their help in
collecting the materials in the veterinary practices. The
study was funded in part by a research grant from the
Foundation for the Dutch Cat Fancy and Foundation
Kattenzorg, The Hague.
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