Molecular evidence for zoonotic transmission of Giardia duodenalis among dairy farm workers in West Bengal, India

No study in the past has examined the genetic diversity and zoonotic potential of Giardia duodenalis in dairy cattle in India. To assess the importance of these animals as a source of human G. duodenalis infections and determine the epidemiology of bovine giardiasis in India, fecal samples from 180 calves, heifers and adults and 51 dairy farm workers on two dairy farms in West Bengal, India were genotyped by PCR-RFLP analysis of the β-giardin gene of G. duodenalis followed by DNA sequencing of the nested PCR products. The overall prevalence of G. duodenalis in cattle was 12.2% (22/180), the infection being more prevalent in younger calves than in adult cattle. Zoonotic G. duodenalis Assemblage A1 was identified in both calves and workers although the most prevalent genotype detected in cattle was a novel Assemblage E subgenotype. These findings clearly suggest that there is a potential risk of zoonotic transmission of G. duodenalis infections between cattle and humans on dairy farms in India.     

Genotype characterisation of Giardia duodenalis isolates from domestic and farm animals by SSU-rRNA gene sequencing

In order to investigate the genotypes of Giardia duodenalis from domestic and farm animals in Italy, 21 Giardia isolates, 17 from dogs, 1 from cat and 3 from dairy calves, were genetically characterised by SSU-rRNA gene sequencing. Among dogs, 76.5% of isolates showed the dog-specific genotypes (Assemblages C, D and C/D mixed Assemblage) and 23.5% exhibit potential zoonotic genotypes (Assemblage A and A/C mixed Assemblages). The cat isolate belonged to assemblage A, whereas the sequences among the isolates from calves were found to correspond to hoofed-livestock genotype, namely Assemblage E. These findings suggest that infection of humans by zoonotic genotypes from domestic animals could be of low epidemiological significance, although possible. The present study represents the first contribute to the knowledge of G. duodenalis genotypes in domestic and farm animals from Italy.     

Prevalence of Giardia duodenalis genotypes in adult dairy cows

The prevalence of Giardia duodenalis genotypes was determined in adult dairy cows. Fecal specimens were collected from two farms each in Vermont, New York, Pennsylvania, Maryland, Virginia, North Carolina, and Florida. Specimens, cleaned of fecal debris and concentrated using CsCl density gradient centrifugation, were subjected to PCR and DNA sequence analysis. The prevalence of G. duodenalis infection, ranged from 3% to 64%, with an average prevalence of 27% (144 positive cows out of 541 examined). DNA sequence analysis of the 16S rRNA gene revealed the presence of both Assemblage A and Assemblage E, G. duodenalis. Overall, Assemblage E was present in 25% of all animals tested and Assemblage A was present in 2% of the animals. As a percentage of G. duodenalis isolates, Assemblage E represented 94% and Assemblage A represented 6%. Although, most of the cows were infected with a genotype that is not known to be infectious for humans, adult cows on five farms did harbor varying levels of zoonotic Assemblage A, G. duodenalis. Therefore, although adult cows do not appear to be a significant source of human infectious cysts in the environment, the risk from this age group should not entirely be discounted.     

Giardia duodenalis and Cryptosporidium spp. in a veterinary college bovine teaching herd

In a preliminary study, we commonly identified Giardia duodenalis in adult dairy cattle from a veterinary college teaching herd. Therefore, the present study was carried out in order to better understand the potential of adult cattle to act as a source for G. duodenalis infections for students and staff at the veterinary college. Fecal samples were collected bi-weekly from this herd of adult cattle (n=30) over an 8-month period to determine the prevalence of G. duodenalis and Cryptosporidium spp. within the herd. Nested PCR followed by DNA sequencing was then performed on a subset of positive samples in order to better understand the zoonotic potential of these infections. Every cow was sampled between 11 and 18 times, depending on the date the animal joined the teaching herd. In total, 507 fecal samples were collected from 30 different cows and examined for cysts and oocysts using epifluorescence microscopy. G. duodenalis prevalence during the course of the study ranged from 37% (11/30) to 64% (18/28), with a mean of 49%. Cumulative G. duodenalis prevalence was 73% (22/30). Zoonotic G. duodenalis assemblage A genotype was identified in 43% (6/14) of the G. duodenalis-positive samples on which PCR and genetic sequencing were successfully performed. G. duodenalis assemblage E was identified in 57% (8/14) of these samples. Cryptosporidium spp. oocysts were not detected in the feces of any cows during the study period. The presence of the zoonotic G. duodenalis assemblage A in 43% of the sequenced samples indicates that there is a potential risk of infection for students and staff at this research and teaching facility, although the roles of cows as sources of giardiasis in humans remain uncertain. Furthermore, due to the large amount of feces they produce, adult cattle may serve as important sources for G. duodenalis infections in young cattle, or other animals in the facility, despite relatively low numbers of cysts excreted per gram of feces. In contrast, the results of this study indicate that this herd posed a negligible risk of transmitting Cryptosporidium parvum infections to humans.     

The potential for zoonotic transmission of Giardia duodenalis and Cryptosporidium spp. from beef and dairy cattle in Ontario, Canada

The objective of this study was to compare the occurrence and the genotypes and species of Giardia duodenalis and Cryptosporidium spp. in beef and dairy cattle from farms in the Regional Municipality of Waterloo, Ontario, in an effort to determine the potential for zoonotic transmission from these animals. Pooled manure samples were collected from 45 dairy cattle farms and 30 beef cattle farms. The presence of Giardia cysts and Cryptosporidium oocysts was determined by immunofluorescence microscopy, while nested-PCR and DNA sequencing were used to determine genotypes and species. The overall farm prevalence was very high for both Giardia and Cryptosporidium, and was similar for dairy cattle farms (96 and 64%, respectively) and beef cattle farms (97 and 63%, respectively). However, on dairy cattle farms, G. duodenalis and Cryptosporidium spp. were detected in 44% and 6% of total pooled pen manure samples, respectively, with the occurrence of both parasites being generally higher in calves than in older animals. Most Giardia isolates were identified as either the host-adapted genotype G. duodenalis Assemblage E or the zoonotic Assemblage B. Cryptosporidium parvum and Cryptosporidium andersoni were the most frequently identified species in dairy cattle, while the non-zoonotic species Cryptosporidium ryanae and Cryptosporidium bovis were also found. On beef cattle farms, 72% and 27% of the total pooled pen manure samples were positive for Giardia and Cryptosporidium, respectively, with no obvious correlation with age. All Giardia isolates in beef cattle were identified as G. duodenalis Assemblage E, while all Cryptosporidium isolates were identified by sequence analysis as C. andersoni, although microscopic analyses, and subsequent restriction fragment length polymorphism analyses, indicated that other Cryptosporidium species were also present. The results of this study indicate that although Giardia and Cryptosporidium were identified in a higher overall percentage of the pooled beef cattle manure samples than in dairy cattle, firmly established zoonotic genotypes and species were much more common in dairy cattle than in beef cattle in this region. Dairy cattle, and especially dairy calves, may, therefore, pose a greater risk of infection to humans than beef cattle. However, these results may also provide evidence of potential zooanthroponotic transmission (human to animal).     

Genotyping of potentially zoonotic Giardia duodenalis from exotic and wild animals kept in captivity in Brazil

We have studied the variability of glutamate dehydrogenase (gdh) and small subunit ribosomal (SSU) rRNA coding genes of Giardia species in fecal samples isolated from wild and exotic animals in Brazil, and compared with homologous sequences of isolates from human and domestic animals characterized in previous studies. Cysts of Giardia duodenalis were obtained from feces of naturally infected monkeys (Alouatta fusca) (n=20), chinchillas (Chinchilla lanigera) (n=3), ostriches (Struthio camelus) (n=2) and jaguar (Panthera onca) (n=1). Assemblage AI was assigned to the unique isolate of jaguar. All the samples from monkeys, chinchillas, and ostriches were assigned to Assemblage B. There was little evolutionary divergence between the referred isolates and isolates described elsewhere. The Assemblage B isolates identified in this study were closely related to Assemblage BIV isolated from humans. The molecular identification of Assemblages A and B of G. duodenalis isolates from exotic and wild animals demonstrates that such hosts may be a potential reservoir for zoonotic transmission of G. duodenalis.     

Evaluation of the zoonotic potential of Giardia duodenalis in fecal samples from dogs and cats in Ontario

This study determined the distribution and zoonotic potential of Giardia duodenalis assemblage types among canine and feline fecal samples from Ontario. The effectiveness of Giardia assemblage typing methods by sequencing the genes of small subunit ribosomal RNA (ssu-rRNA), β-giardin (bg), glutamate dehydrogenase (gdh), and triose phosphate isomerase (tpi) was evaluated simultaneously. From 2008 to 2010, 118 canine and 15 feline Giardia positive fecal samples were tested. The ssu-rRNA sequencing method typed 64% (75/118) and 87% (13/15) of the Giardia-positive canine and feline samples, respectively. Among the typeable samples, 68% (51/75) of canine samples contained G. duodenalis assemblage D and 31% (23/75) contained G. duodenalis assemblage C (both non-zoonotic assemblage types). Only 1% (1/75) of the typeable canine samples contained a potentially zoonotic assemblage B. In contrast, 100% (13/13) of the typeable feline samples contained potentially zoonotic assemblages A (n = 12) or B (n = 1).     

Genotypic analysis of Giardia duodenalis in domestic cats

BACKGROUND: Giardia duodenalis is an intestinal flagellated protozoan that affects many mammalian species often causing severe diarrheal disease. Several different genotypes have been identified (Assemblages A-G). Most isolates recovered from domestic cats have been assigned to either Assemblage A, the zoonotic form of the parasite, or Assemblage F, identified thus far only in cats. Genotypic variation within G. duodenalis may influence clinical presentation and course of disease. Therefore, host-adapted genotypes may not be responsible for diarrheal disease (eg, Assemblage F in cats). HYPOTHESIS: Multiple Giardia genotypes will be present in domestic cats, including Assemblage F, which will not be correlated with clinical signs. ANIMALS: 250 domestic cats from eastern Mississippi and northwestern Alabama. METHODS: Prevalence survey. Fecal samples evaluated for cysts using a centrifugation concentration technique and a commercially available direct immunoflourescent antibody kit. Giardia isolates were characterized by PCR amplification and sequencing of the glutamate dehydrogenase gene. RESULTS: Both Assemblage A-I (6/17) and Assemblage F (11/17) were identified. Although Assemblage was significantly associated with age and housing, no association was detected between Assemblage and a variety of other factors including the presence of gastrointestinal signs (acute vomiting, diarrhea, and constipation). CONCLUSIONS AND CLINICAL IMPORTANCE: The presence of diarrhea in domestic cats with Giardia cannot be used as a predictor of the presence of zoonotic genotypes in animals within the study area. Although Assemblage A was associated with age and housing, veterinarians should consider any isolation of Giardia from domestic cats as potentially zoonotic.     

Evaluation of immunofluorescence microscopy and enzyme-linked immunosorbent assay in detection of Cryptosporidium and Giardia infections in asymptomatic dogs

The performance of immunofluorescence microscopy (IF) and enzyme-linked immunosorbent assay (ELISA) in canine feces was evaluated. IF and Cryptosporidium ELISA detected 10(5)oocysts/g, while the detection limit for Giardia ELISA was 10(4)cysts/g. The Cryptosporidium ELISA showed 94% specificity but only 71% sensitivity. The Giardia ELISA correlated well with IF (sensitivity 100%, specificity 96%) and was capable of detecting animal specific Giardia duodenalis genotypes. Visual interpretation appeared appropriate for assessment of ELISA results. The proportion of positive samples and possible zoonotic character of Cryptosporidium and Giardia infections in 150 asymptomatic Finnish dogs from the Helsinki area were studied. The overall proportion of dogs positive for Cryptosporidium was 5% (7/150) and that for Giardia 5% (8/150). In dogs < or =12 months old, the corresponding proportions were 17% and 19% (n=36). Sequence analyses of the 18S rDNA gene identified the isolates as Cryptosporidium canis and animal specific genotypes of G. duodenalis (assemblages C-E), indicating restricted risk of zoonotic transmission.     

Giardia duodenalis in small animals and their owners in Germany: A pilot study

Giardia duodenalis is a relevant gastrointestinal protozoan pathogen of humans and animals. This species complex consists of eight genetically different assemblages. Assemblages A and B are pathogenic to humans and pets, thus confer zoonotic potential. The risk of zoonotic transmission has been controversially discussed. The aim of this monocentric cross‐sectional pilot study was to investigate G. duodenalis assemblages in humans and pets living in common households in Berlin/Brandenburg (Germany). Samples from dogs, cats and humans sharing the same households were screened for Giardia infection by antigen‐detecting assays. All human samples were additionally analysed by a Giardia‐specific qPCR. Cyst quantification and sequences of different gene loci (triosephosphate isomerase (tpi), glutamate dehydrogenase (gdh), β‐giardin (bg) and for dogs SSUrDNA) were analysed. A total of 38 households (31 households with dogs and seven with cats) with 69 human individuals participated in the study. Initial antigen‐detecting assays revealed Giardia‐positive results for 13 (39%) canine, one (14%) feline and one human sample. Reanalysis of the human samples by qPCR revealed two more positive specimens (4%). Two of these three samples were identified as assemblage B at all tested loci. Success rate of assemblage typing for pet samples was generally low and comprised mainly the SSUrDNA locus only. Overall, six of 13 Giardia‐positive canine samples were typable (2× A, 1× co‐infection: A and B, 1× C; 2× D). One pair of samples (dog and human) from the same household had a similar but not identical assemblage B sequence at tpi locus. Assemblage A was also detected in the dog specimen, which hampered sequence analysis. In conclusion, although exhibiting limitations due to the sample size, our study highlights the need for better and standardized typing tools to distinguish G. duodenalis strains with higher resolution in order to perform proper case–control studies for a realistic estimation of zoonotic risk.     

Detection of Assemblage A, Giardia duodenalis and Eimeria spp. in alpacas on two Maryland farms

Sixty-one fecal samples were collected from adult alpacas and crias (ages 10 weeks to 10 years) on two farms in central Maryland. The farms raised both suri (silky-haired) and huacaya (crimpy-haired) breeds. Females and crias were housed together on pasture, whereas older/breeding males were maintained on separate pastures. Samples were subjected to a density gradient centrifugation protocol to concentrate parasites and remove fecal debris and were examined by immuno-fluorescent and differential interference contrast microscopy. Oocysts of Eimeria spp. were noted in 14 fecal samples, 6 on MD-1 and 8 on MD-2. Based on oocyst morphometrics two species of Eimeria were present: E. punoensis (19.2 microm x 16.5 microm) and E. alpacae (23.7 microm x 19.5 microm). Five animals shed exclusively E. punoensis, seven shed exclusively E. alpacae, and two had mixed infections. The Eimeria infections were not associated with obvious clinical signs. To determine the presence of Cryptosporidium and Giardia species and genotypes, DNA was extracted from feces and subjected to PCR utilizing specific primers for the ssu-rRNA gene for both parasites. All PCR positive samples were further analyzed by DNA sequencing to identify the species or genotypes that were present. Assemblage A, G. duodenalis was detected in fecal samples from two alpacas on MD-1 and in one alpaca on MD-2. Assemblage E, G. duodenalis and Cryptosporidium spp. were not detected on either farm. Although the prevalence on these two farms was low, alpacas can harbor zoonotic G. duodenalis, and this should be borne in mind by persons interacting with the animals.     

Survey of giardiosis in household and shelter dogs from metropolitan areas of Curitiba, Paraná state, Southern Brazil

Giardia duodenalis is a protozoan parasite that causes a broad range of clinical symptoms varying from none--in asymptomatic carriers--to mild recurring diarrhea consisting of soft, light-colored stools to acute severe diarrhea. In different parts of the world this parasite has raised increased interest due to its possible zoonotic transmission. Among domestic animals, dogs can play an important role in environmental contamination. As there is little information on the frequency of giardiosis in dogs from the Metropolitan Area of Curitiba-State of Paraná, Southern Brazil, the aim of the present work was to evaluate the prevalence of G. duodenalis in two dog populations (household and shelter). To attain the proposed aim, we collected fecal samples from 200 dogs and utilized three diagnostic techniques: Faust's technique (Faust et al. 1939), Benbrook's technique (1963) and polymerase chain reaction (PCR). Faust's technique presented the best results, as it was able to detect a larger number of Giardia cases. Taking Faust's technique as the standard, Benbrook's technique presented 66% sensitivity and PCR demonstrated 69% sensitivity. The shelter dog population showed a 24% occurrence of G. duodenalis while the household population showed a 9% occurrence. Other epidemiological aspects like age, sex, environmental conditions and methodological aspects are discussed in the present article.     

Gastrointestinal Parasites of Urban Dogs in Perth, Western Australia

A study was conducted to determine the prevalence of gastrointestinal parasites in a sample of urban dogs in Perth and the knowledge of their owners about the control and zoonotic transmission of these parasites. Faecal samples (421), collected from dogs originating from five sources, were examined by microscopy and questionnaires administered to dog owners and managers/owners of pet shops. The prevalence of gastrointestinal parasitism was higher in pet shop puppies (51%), than in dogs from refuges (37%), breeding kennels (32.7%), veterinary clinics (15.6%) and exercise areas (5.3%). Protozoa, in particular Giardia, were detected more frequently (22.1%) than helminth parasites. After adjusting for other factors with multiple logistic regression, puppies less than 6 months of age, dogs living in households with more than one dog, and dogs from refuges were significantly more likely to be parasitized. The prevalence of Giardia was found to be directly associated with the number of doses of anthelmintics given in a year, increasing 1.2 times for each dose administered. The majority of owners were aware of the potential risk to human health from canine helminths, however only one third were aware of the means of transmission to humans. It is concluded that veterinarians can play an important role in increasing the level of awareness of canine zoonotic parasites.     

Prevalence and molecular characterization of Cryptosporidium and Giardia species and genotypes in sheep in Maryland

In the United Kingdom and Australia sheep have been implicated as sources of Cryptosporidium and Giardia that infect humans, but no such studies have been conducted in North America. Therefore, a study was undertaken to investigate the prevalence of these parasites in sheep on a farm in Maryland. Feces were collected from 32 pregnant ewes 1, 2, and 3 days after parturition and from each of their lambs 7, 14, and 21 days after birth. The presence of Cryptosporidium oocysts and Giardia cysts was determined by both immunofluorescence microscopy and PCR/gene sequence analysis. PCR was consistently more sensitive than microscopy. The prevalence, by PCR, of Cryptosporidium in ewes and lambs was 25 and 77.4%, respectively. Three species/genotypes of Cryptosporidium were identified: C. parvum, a novel C. bovis-like genotype, and Cryptosporidium cervine genotype. Cryptosporidium parvum and the cervine genotype have been reported worldwide in human infections. The novel C. bovis-like genotype is reported here for the first time. The prevalence of Giardia in ewes and lambs was 12 and 4%, respectively. Most infections were Assemblage E which is not zoonotic; however, one ewe was infected with zoonotic Assemblage A. The identification of only two lambs infected with C. parvum and one ewe infected with G. duodenalis Assemblage A suggests a low prevalence of these zoonoses. However, the high prevalence of the zoonotic cervine genotype indicates that sheep should be considered a potential environmental source of this human pathogen.     

Prevalence of Giardia duodenalis assemblages among dairy herds in the New York City Watershed

A longitudinal herd-level study was carried out to determine the cumulative incidence of Giardia duodenalis infections in dairy cattle in the New York City Watershed. We also sought to assess the changes in infection pattern of animals diagnosed as shedding Giardia over time, determine risk factors that may be associated with G. duodenalis infections, and identify potentially zoonotic infections. A total of 2109 fecal samples were randomly collected from dairy cattle at 34 farms in the New York City Watershed on a seasonal basis. A total of 504 Giardia-positive samples were identified by zinc sulfate flotation. The overall cumulative incidence of G. duodenalis based on flotation results was 23.9% with 73.8% of all infections occurring in animals under 180 days of age (372/504). The intensity of infection ranged from 2 to 563,200 cysts/gram of feces. Cattle shedding Cryptosporidium spp. oocysts were twice as likely to shed G. duodenalis cysts in comparison to the animals that did not shed oocysts (1.81 95% CI 1.26-2.60 p=0.0012). In the multivariate analysis, only the age of the animal and the presence of dogs on the farm were significantly associated with the likelihood of shedding G. duodenalis. DNA was extracted from positive samples and analyzed by polymerase chain reaction (PCR) of the beta-giardin and triosephosphate isomerase genes of Giardia spp. 304 samples were analyzed by PCR of which 131 were sequenced. 22.1% of sequenced samples were identified as assemblage A and 77.9% were identified as assemblage E. Interestingly, 100% of specimens identified as assemblage A were from calves under 84 days of age indicating that younger cattle are important reservoirs for potentially zoonotic assemblages of G. duodenalis.     

Enteric parasites of free-roaming,owned, and rural cats in prairie regions of Canada

The objective of this study was to determine prevalence, intensity, and zoonotic potential of gastrointestinal parasites in free-roaming and pet cats in urban areas of Saskatchewan (SK) and a rural region in southwestern Alberta (AB). Fecal samples were analyzed using a modified double centrifugation sucrose flotation to detect helminth eggs and coccidian oocysts, and an immunofluorescence assay to detect Giardia and Cryptosporidium. Endoparasite prevalence was higher in samples from rural AB cats (41% of 27) and free-roaming SK cats (32% of 161) than client-owned SK cats (6% of 31). Parasites identified using morphological and molecular techniques included Toxocara cati, Toxascaris leonina, Baylisascaris-type eggs, Eucoleus aerophilus, Taenia taeniaeformis, Isospora spp., Cryptosporidium spp., and zoonotic genotype A of Giardia duodenalis. This study demonstrates significant differences in endoparasite prevalence in feline populations, and the value of molecular techniques in fecal-based surveys to identify and determine parasite zoonotic potential.     

Prevalence and risk factors of Giardia duodenalis in dogs from Romania

The protozoan Giardia duodenalis is a mammalian-infecting parasite that produces diarrhoea and malabsorption in its hosts. A survey to investigate canine infections with G. duodenalis in Romania was undertaken between June 2008 and December 2009. The objectives of the study were to (i) estimate the prevalence of infection in different dog populations (kennels, shelters, shepherd, household) using microscopy and a commercially available enzyme-linked immunosorbent assay (ELISA) test kit; (ii) to establish the level of agreement and characteristics of the tests; and (iii) to identify risk factors for infection by multivariate logistic regression models. Faecal samples were collected from 614 dogs aged from 1 month to 16 years (mean ± SD=2.88 ± 2.86 years). Each sample was tested for the presence of cysts using a flotation method with saturated sodium chloride solution and 416 out of 614 stool samples were further examined for the presence of G. duodenalis specific antigens using Giardia Microwell ELISA (SafePath? Laboratories). Giardia cysts were identified in 8.5% of total dogs (52/614) and statistical significantly more frequently in dogs living in communities. The cysts prevalence according with dog populations was as follows: 7.2%(9/125) in kennel dogs; 16.5%(27/164 in shelter dogs; 4.3%(2/46) in shepherd dogs; 4.8%(4/84) in household dogs from urban areas; and 5.1%(10/195) in household dogs from rural areas. The overall prevalence of Giardia infection by ELISA was 34.6% (144/416). The prevalence was significantly higher in kennel dogs (50%; 13/26), shelter dogs (47.7%; 74/155) and shepherd dogs (40.5%; 17/42) than in household dogs from urban areas (34.1%; 15/44) and household dogs from rural areas (16.8%; 25/149). It was noticed poor agreement between microscopy and ELISA (k=0.19). The microscopy performed best, with an Youden Index of 0.74, a Se of 73.68% and a Sp of 100%. ELISA had 100% Sp, but only 19.44% Se. Young dogs (up to 12 months age) and living in communities were identified as risk factors for infection by multivariate logistic regression analysis. 71.2% (37/52) Giardia cysts positive dogs presented co-infections with other intestinal parasites: Toxocara canis (14/52; 26.9%), Isospora ohioensis (12/52; 23.1%), Ancylostoma caninum (9/52; 17.3%), Uncinaria stenocephala (7/52; 13.5%), Trichocephalus vulpis (6/52; 11.5%), Hammondia heydorni/Neospora caninum (5/52; 9.6%), Sarcocystis spp. (5/52; 9.6%), Isospora canis (4/52; 7.7%), Capillaria aerophila (3/52; 5.8%), Strongyloides stercoralis (2/52; 93.8%), Dipylidium caninum (1/52; 1.9%) and Toxascaris leonina (1/52; 1.9%).     

Prevalence of Giardia duodenalis and Cryptosporidium spp. in dogs from different living conditions in Uberlândia, Brazil

Infection rates with Giardia duodenalis and Cryptosporidium spp. were compared among dogs living under different conditions. Stool samples (n = 433) collected from dogs of different ages, gender, living conditions and origin were analyzed using three techniques, i.e., centrifugal flotation in zinc sulfate solution, centrifugal flotation in sucrose solution, and methylene blue gram safranin staining. Eighty-nine of the samples were from stay dogs living in shelters run by animal protection societies, 199 were from kennels and 122 from households. A total of 119 (29.0%) had G. duodenalis cysts and six (1.4%) were positive for Cryptosporidium spp. oocysts. Dogs from kennels were most frequently affected by G. duodenalis (49.7%) while those from shelters showed a higher prevalence of Cryptosporidium spp. (2.2%). A significant difference (p < or = 0.05) was observed between immature dogs and adults only with respect to Giardia infection. There was no significant difference between the gender with regard to the presence of either protozoan.     

Determining the zoonotic significance of Giardia and Cryptosporidium in Australian dogs and cats

In a recent study of intestinal parasites in dogs and cats in Australia, Giardia was found to be the most prevalent parasite in dogs. The aim of the current study through the use of molecular tools was to determine the zoonotic significance of the Giardia and Cryptosporidium isolates recovered from dogs and cats during the Australian study. Of the isolates successfully amplified all but one of the Giardia from dogs was either Assemblage C and/or D, with one Assemblage A. Of the cat samples amplified all but one were Assemblage F, with one Assemblage D. We hypothesize that the lack of zoonotic Giardia Assemblages recovered is a result of their being a low prevalence of Giardia in the human population. The Cryptosporidium recovered from dogs and cats was determined to be C. canis and C. felis, respectively, a finding which supports growing evidence that Cryptosporidiumin companion animals is of limited public health significance to healthy people.     

High prevalence of intestinal zoonotic parasites in dogs from Belgrade, Serbia--short communication

To identify areas of risk for canine-related zoonoses in Serbia, the aim of this study was to provide baseline knowledge about intestinal parasites in 151 dogs (65 household pets, 75 stray and 11 military working dogs) from Belgrade. The following parasites, with their respective prevalences, were detected: Giardia duodenalis (14.6%), Ancylostomatidae (24.5%), Toxocara canis (30.5%), Trichuris vulpis (47.0%) and Taenia-type helminths (6.6%). Of all examined dogs, 75.5% (114/151) were found to harbour at least one parasite species. Of these, mixed infections with up to four species per dog occurred in 44.7% (51/114). Infections with all detected species were significantly higher (p < 0.05) in military working (100%) and stray dogs (93.3%) versus household pets (50.8%). Among all parasites, agents with zoonotic potential including Giardia, Ancylostomatidae and Toxocara were detected in 58.3% (88/151) of all examined dogs with a significant difference (p < 0.05) among the subgroups (100%, 62.7% and 46.2% for military working dogs, stray dogs and household pets, respectively). The high prevalence of zoonotic parasites registered in the dog population from a highly urban area in south-eastern Europe indicates a potential risk to human health. Thus, veterinarians should play an important role in helping to prevent or minimise zoonotic transmission.