Genotyping of Giardia duodenalis from Southern Brown Howler Monkeys (Alouatta clamitans) from Brazil

Giardia duodenalis is a widespread intestinal protozoan that can infect humans and animals, both domestic and wild. Independent of host, infections present with the same symptoms. However, based on host specificity, Giardia isolates have been grouped into genotypes A to G. Parasites of assemblage A and B are known to infect humans, in addition to primates and a wide variety of mammals. In Brazil, hitherto Giardia genotypes were defined only for humans and domestic animals. To evaluate the genotypes of different Giardia present among other animals, fecal samples from 28 Southern Brown Howler Monkeys (Alouatta clamitans) kept in captivity from South Brazil were screened for G. duodenalis using parasitological methods. All of them were asymptomatic, but positive for Giardia. The genotype of the G. duodenalis circulating among these animals was ascertained by molecular typing, performed using amplification and sequencing of the beta-giardin gene. Sixteen of 28 samples were successfully amplified by PCR and sequencing of this gene s revealed that all of them were of the genotype A1. These findings suggest that A. clamitans represent a potential risk of environmental contamination of a G. duodenalis genotype that also infect humans, and therefore can be considered a potential reservoir for G. duodenalis of a genotype that can also infects humans. Therefore, these results highlight a potential public health problem due to the epidemiological and molecular evidence for anthropozoonotic transmission.     

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.     

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.     

Molecular characterization of Cryptosporidium and Giardia isolates from cattle from Portugal

Fecal samples from 291 calves and 176 adult cattle in Northern Portugal were screened for Cryptosporidium and Giardia using a formalin-ethyl acetate concentration method. Acid-fast staining techniques for Cryptosporidium oocyst identification and direct microscopic observation of fecal smears for Giardia cyst identification were performed so as immunofluorescence microscopy examination. Polymerase chain reaction methods were employed to determine the genotype of each isolate. Molecular characterization was performed using amplification and sequencing of the hsp70 and 18SrRNA genes of Cryptosporidium and beta-giardin gene and glutamate dehydrogenase for assemblage determination of Giardia duodenalis. Seventy-four out of 291 calves (25.4%) and 8 out of 176 adult bovines (4.5%) were positive for Cryptosporidium. Forty-one out of 291 calf samples (14.1%) and 1 out of 176 adults samples (0.57%) were positive for Giardia. From the Cryptosporidium positive samples we obtained 63 isolates from calves samples and 7 isolates from adult samples. Additionally, Giardia was isolated in 13 out of 41 positive samples from calves and it was also possible to isolate Giardia from the positive adult sample. Molecular characterization of the Cryptosporidium and Giardia isolates showed us that C. parvum and G. duodenalis assemblage E were the prevalent species. C. parvum may infect humans, representing a potential public health risk. On the other hand, the assemblages B and A2 of Giardia, previously described in humans, were here identified in cattle. Further studies will be needed for determine the importance of cattle as carrier of zoonotic assemblages of G. duodenalis.     

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).     

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.     

Prevalence and molecular typing of Giardia spp. in captive mammals at the zoo of Zagreb, Croatia

A total of 131 faecal samples from 57 mammalian species housed at the zoo of Zagreb, Croatia, were tested for the presence of Giardia spp. cysts using epifluorescence microscopy. The overall prevalence (29%) was high, yet all animals were asymptomatic at the time of sampling. Positive samples were characterized by PCR and sequence analysis of both conserved and variable loci, for the identification of Giardia species and G. duodenalis assemblages and genotypes. Assemblages A and C were identified in Artiodactyla, assemblage B in Primates, Rodentia and Hyracoidea, and assemblages A, B, C and D, as well as Giardia microti, in Carnivora. Genotyping at the ITS1-5.8S-ITS2 region, at the triose phosphate isomerase, glutamate dehydrogenase and beta-giardin genes revealed extensive polymorphisms, particularly among assemblage B isolates. A phylogenetic analysis of concatenated sequences showed that isolates from captive mammals housed at the zoo are genetically different from isolates of human and domestic animal origin. This is the first survey in a zoological garden to include a molecular characterization of the parasite, and provides novel sequence data of G. duodenalis from many previously uncharacterized hosts.     

Prevalence and genotyping of Giardia duodenalis from beef calves in Alberta,Canada

Giardia infections in domestic cattle has come under increasing scrutiny owing to the potential contamination of surface and ground waters through manure distribution on fields and pasture runoff. The objective of the study was to determine the prevalence and genotypes of Giardia duodenalis in beef calves in major beef cow calf farms in Alberta, Canada. Fecal samples were collected from beef calves aged 2-10 weeks at nine farms in Alberta. Samples were examined for the presence of G. duodenalis cysts by immunofluorescent staining. Giardia cysts were found in 168 of the 495 fecal samples examined, with prevalence ranging from 7 to 60% among farms. Genotypic analysis of positive isolates utilizing PCR and sequencing of a 292 bp fragment of the 16S-rRNA locus, revealed the hoofed livestock genotype in 41 of the 42 isolates. One isolate was identical to the Assemblage A genotype. The results of this study demonstrate that beef calves in this area are primarily infected with the livestock genotype which is thought to be specific to artiodactyl hosts and non-infective to humans. This suggests that the Giardia carried by beef cattle may be a minimal zoonotic threat.     

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.     

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.     

Molecular identification of Cryptosporidium parvum and Giardia duodenalis in the Italian water buffalo (Bubalus bubalis)

Livestock are commonly infected with protozoan parasites of the genera Cryptosporidium and Giardia, and some of the species and genotypes found in these animals have zoonotic significance. We characterized isolates of both parasites recovered from the Italian water buffalo (Bubalus bubalis), an economically important species whose milk is used for the production of "buffalo mozzarella" fresh cheese. Molecular analysis of the Cryptosporidium small subunit ribosomal DNA gene and of the Giardia beta-giardin gene shows the presence of both zoonotic parasites (Cryptosporidium parvum and Giardia duodenalis assemblage A) and host-specific parasites (G. duodenalis assemblage E), suggesting that water buffaloes can contribute to environmental contamination with oocysts and cysts potentially infectious to humans if their faeces are improperly disposed of. On the other hand, mozzarella cheese is probably a safe product, given that its production involves the treatment of cheese curd at 85-95 degrees C, which is likely to kill or inactivate the parasites.     

Infection of dogs by experimental inoculation with human isolates of Giardia duodenalis: clinical and laboratory manifestations

The susceptibility of dogs to experimental inoculation with trophozoites and cysts of human isolates of Giardia duodenalis and the clinical and laboratory profiles of infection of these animals were studied. Two groups (A and B), each comprising three dogs, were inoculated with G. duodenalis trophozoites and cysts, respectively. A third group of two dogs was not inoculated and remained as control. After inoculation feces were collected daily to determine the pre-patent period, by flotation in 33% zinc sulfate solution. Blood samples (5mL) were collected from animals at 15-day intervals during the 165 days of the experimental period and were used to carry out the hemogram and biochemical evaluation of the levels of total protein, albumin, alanine aminotransferase, gamma glutamyltransferase, urea and creatinine. A prepatent period was observed at 5-6 days post-inoculation (p.i.) in the inoculated dogs, with cysts eliminated for approximately 3 months. No alterations were seen in the clinical parameters evaluated. Anemia was observed at 15 p.i. in the inoculated dogs. The mean eosinophil count of inoculated groups was higher than that of the control (p< or =0.05) but none of the biochemical parameters analyzed presented significant differences. The results of this study show that G. duodenalis from human isolates is able to infect dogs with minimal systemic manifestations without producing clinical signs of giardiasis.     

Molecular epidemiology of cryptosporidium and giardia in humans on prince edward island, Canada: evidence of zoonotic transmission from cattle

To determine the zoonotic potential of Cryptosporidium and Giardia in Prince Edward Island (PEI), Canada, 658 human faecal specimens were screened that were submitted to the Queen Elizabeth Hospital diagnostic laboratory. Overall, 143 (22%) samples were Cryptosporidium positive, while three (0.5%) were positive for Giardia. Successful genotyping of 25 Cryptosporidium isolates by sequence analysis of the HSP70 gene revealed that 28 and 72% were C. hominis and C. parvum, respectively. Cryptosporidium isolates from humans and previously genotyped C. parvum from beef cattle were subtyped by sequence analysis of the GP60 gene. Subtyping identified three subtypes belonging to the family IIa. All three subtypes IIaA16G2RI (55%), IIaA16G3RI (22%) and IIaA15G2RI (22%) were found in the animal isolates, while two of the subtypes found in the animals, IIaA16G2RI (80%) and IIaA15G2RI (20%), were also identified in the human isolates. Cryptosporidium infection in humans peaked in April-June. Molecular epidemiological analysis of the human data showed a C. parvum peak in the spring and a relatively smaller peak for C. hominis in July-September. The majority (57%) of human Cryptosporidium isolates were found in children between 5 and 10 years of age. All three Giardia isolates were identified as G. duodenalis assemblage A. The overall Cryptosporidium prevalence in our human samples was high relative to other studies, but because the samples were submitted to a hospital diagnostic laboratory, the results may not be representative of the general population. Further, the presence of the same zoonotic C. parvum subtypes in cattle and human isolates implies that transmission is largely zoonotic and cattle may be a source of sporadic human infections on PEI. The presence of Giardia in people on PEI is rare, and the assemblage A found in humans might originate from humans, livestock or other domestic or wild animals.     

Occurrence of Giardia and Cryptosporidium in Norwegian red foxes (Vulpes vulpes)

Faecal samples from 269 Norwegian wild red foxes (Vulpes vulpes) shot during the hunting season (October-April) in 2002-2004 were examined for the presence of Giardia and Cryptosporidium. Cryptosporidium oocysts were detected in samples from 6 (2.2%) of the foxes, and Giardia cysts in 13 (4.8%) of the foxes. The prevalence of Giardia infection was significantly higher in juvenile male foxes than in adult male foxes, but no other significant differences between age and sex were found. No significant differences in prevalence related to geographical origin of animals were found. Insufficient nucleated Cryptosporidium oocysts were isolated for successful PCR, but genotyping of Giardia duodenalis isolates from seven foxes demonstrated a high degree of heterogeneity amongst them, with all isolates belonging to the zoonotic Assemblages A and B.     

Epidemiology of Cryptosporidium spp. and Giardia duodenalis on a dairy farm.

Prevalences of Cryptosporidium spp. and Giardia duodenalis in relation to age and season were investigated on a dairy farm in The Netherlands over the course of 1year. The whole herd was sampled five times, whereas calves younger than about 2 months were sampled every 2-3 weeks. Associations between diarrhoea and presence of one or more pathogens (Cryptosporidium spp., G. duodenalis, rotavirus) were investigated. Potential transmission routes of Cryptosporidium spp. were evaluated and positive samples of Cryptosporidium spp. and G. duodenalis were identified to genotype level by PCR microsatellite identification and fingerprinting. Shedding of Cryptosporidium spp. was found in all age categories but peaked in calves 1-3 weeks old (39.1%). Herd prevalence of shedding for Cryptosporidium spp. varied from 2.4% in June to 22.2% in December. Shedding of G. duodenalis was found in all age categories but peaked in animals 4-5 months old (54.5%). Herd prevalence of shedding for G. duodenalis varied from 0.8% in June to 15.5% in February. Cryptosporidium spp. and rotavirus appeared to be significantly associated with diarrhoea in calves. Microsatellite analysis showed two different subtypes (C3 and C1) of Cryptosporidium parvum calf strains. Two genotypes of G. duodenalis were found, one positive by A lineage specific PCR and thus closely related to human genotypes and one genotype, which was negative by A and B lineage specific PCR. The results indicate that cow-to-calf and indirect calf-to-calf transmission both are important routes for acquiring infection with Cryptosporidium spp.     

Presence and molecular characterisation of Giardia and Cryptosporidium in alpacas (Vicugna pacos) from Peru

The presence of Giardia and Cryptosporidium was investigated in 274 faecal samples of alpacas (Vicugna pacos) from 12 herds from Peru by immunofluorescence microscopy and PCR amplification and sequencing of fragments of the ssu-rRNA and β-giardin genes from Giardia spp., as well as the ssu-rRNA gene from Cryptosporidium spp. A total of 137 samples (50.0%) were positive for Giardia spp., and 12 samples (4.4%) for Cryptosporidium spp. In ten samples (3.6%), co-infection by both pathogens was found. Herd prevalence was found to be 91.7% (11/12 herds) for Giardia and 58.3% (7/12 herds) for Cryptosporidium. Regarding the age of the animals, although Giardia was detected in animals as young as 1 week, the prevalence increased with age, reaching 80% by 8 weeks. Similarly, the highest percentage of Cryptosporidium detection (20%) was also found in the 8 week-old group. By PCR, 92 of the 274 analysed samples were positive for Giardia. Sequencing of the amplicons showed the existence of Giardia duodenalis assemblage A in 67 samples; G. duodenalis assemblage E in 24 samples; and inconsistent results between the two molecular markers used in a further sample. Cryptosporidium was only detected by PCR in 3 of the 274 samples; Cryptosporidium parvum was identified in two samples and Cryptosporidium ubiquitum in one sample. This study is the first performing molecular characterisation of both parasites in Peruvian alpacas, and the first report of C. ubiquitum in this host. The identification of G. duodenalis assemblage A, C. parvum and C. ubiquitum, suggests that zoonotic transmission of these enteropathogens between alpacas and humans is possible.     

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.     

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.     

Giardia duodenalis trophozoites isolated from a parrot (Cacatua galerita) colonize the small intestinal tracts of domestic kittens and lambs

This study examines the ability of Giardia duodenalis trophozoites, isolated from a wild bird, to colonize the intestinal tracts of companion animals (kittens) and domestic ruminants (lambs). Trophozoites colonized the intestinal tracts of intraduodenally inoculated animals as demonstrated by increasing parasite burdens within the duodenum and jejunum and by fecal passage of cysts within 4 days post-inoculation. The pathogenesis of the trophozoites was further investigated in kittens. In these animals, infection significantly reduced jejunal brush border microvillous length and density, which resulted in a loss of overall epithelial brush border surface area. This injury was associated with the production of diarrhea in four of five infected kittens. These findings indicate that some bird species may carry G. duodenalis that represent a possible health threat to companion animals and livestock. Our results describe the first successful colonization of avian-derived G. duodenalis trophozoites in the small intestines of domestic kittens and lambs.     

Giardia and Cryptosporidium species and genotypes in coyotes (Canis latrans).

Feces and duodenal scrapings were collected from 22 coyotes (Canis latrans) killed in managed hunts in northeastern Pennsylvania. Polymerase chain reaction (PCR) methods were used to detect Giardia and Cryptosporidium spp. PCR-amplified fragments of Giardia and Cryptosporidium spp. SSU-rRNA genes were subjected to DNA sequence analysis for species/genotype determination. Seven coyotes (32%) were positive for G. duodenalis: three assemblage C, three assemblage D, and one assemblage B. Six coyotes (27%) were positive for Cryptosporidium spp. One isolate shared 99.7% homology with C. muris, whereas five others (23%) shared 100% homology with C. canis, coyote genotype. This is the first report on multiple genotypes of Giardia spp. in coyotes and on the prevalence of Cryptosporidium spp. genotypes in coyotes.