Molecular prevalence of trichomonad species from pet shop puppies and kittens in Japan.

Pentatrichomonas hominis and Tritrichomonas foetus (cat genotype) have been commonly identified as intestinal trichomonads in both dogs and cats. Although P. hominis is considered as non-pathogenic protozoa in many kinds of mammals, it has the potential for zoonotic transmission. T. foetus has been recognized as the emerging causative agent of diarrhea in cats without the risk of zoonotic transmission. As pet shops are the major source of young companion animals, the present study discusses the molecular prevalence of P. hominis and T. foetus from 544 pet shop puppies and 409 kittens. The results suggest that the prevalence of P. hominis (puppies: 7.0%; kittens: 0.5%) and T. foetus (puppies: 0%; kittens: 2.4%) in pet shop young animals are low. In addition, the infections of P. hominis and T. foetus are not always associated with the clinical signs (soft or diarrhea feces).

Molecular prevalence of trichomonad species from pet shop puppies and kittens in Japan / Molecular prevalência de espécies trichomonas em filhotes de cães e gatos de pet shop no Japão

Abstract Pentatrichomonas hominis and Tritrichomonas foetus (cat genotype) have been commonly identified as intestinal trichomonads in both dogs and cats. Although P. hominis is considered as non-pathogenic protozoa in many kinds of mammals, it has the potential for zoonotic transmission. T. foetus has been recognized as the emerging causative agent of diarrhea in cats without the risk of zoonotic transmission. As pet shops are the major source of young companion animals, the present study discusses the molecular prevalence of P. hominis and T. foetus from 544 pet shop puppies and 409 kittens. The results suggest that the prevalence of P. hominis (puppies: 7.0%; kittens: 0.5%) and T. foetus (puppies: 0%; kittens: 2.4%) in pet shop young animals are low. In addition, the infections of P. hominis and T. foetus are not always associated with the clinical signs (soft or diarrhea feces).

Resumo Pentatrichomonas hominis e Tritrichomonas foetus (genótipo de gato) têm sido comumente identificados como trichomonas intestinais em cães e gatos. Apesar de P. hominis ser considerado como protozoário não patogênico em muitos tipos de mamíferos, tem potencial para transmissão zoonótica. Enquanto o T. fetus foi reconhecido como o agente causador emergente de diarreia em gatos sem o risco de transmissão zoonótica. Devido às lojas de animais serem as principais fontes de filhotes de animais domésticos, o presente estudo discute a prevalência molecular e/ou o potencial zoonótico de P. hominis e T. foetus em 544 filhotes de cachorro e 409 gatos de "pet shop". Os resultados sugerem que a prevalência de P. hominis (cães: 7,0%; gatos: 0,5%) e T. foetus (cães: 0%; gatos: 2,4%) em animais jovens de "pet shop" é baixa. Além disso, as infecções de P. hominis e T. foetus nem sempre estão associadas aos sinais clínicos (fezes moles ou diarreia).

Molecular Prevalence of Cryptosporidium spp. in Breeding Kennel Dogs.

Cryptosporidium is a common intestinal protozoan that can lead to diarrhea in humans and dogs. The predominant species of infection are C. hominis and C. parvum in humans, and C. canis in dogs. However, C. canis can infect immunocompromised humans. Considering the close contact with humans, dogs have the potential to be reservoirs for human cryptosporidiosis. Breeding kennels are the major supply source of puppies for pet shops. The present study is to determine the molecular prevalence and characteristics of Cryptosporidium spp. found in breeding kennel dogs. A total of 314 fecal samples were collected from young and adult dogs kept in 5 breeding kennels. A polymerase chain reaction targeting the small subunit rRNA gene was employed for the detection of Cryptosporidium spp. To determine the species, the DNA sequences were compared to GenBank data. Overall, 21.0% of the fecal samples were positive for Cryptosporidium spp. infection. Cryptosporidium spp. was detected in all 5 facilities. A sequencing analysis demonstrated that all isolates shared 99-100% similarity with C. canis. The results suggest that Cryptosporidium spp. infection is present at a high-level in breeding kennel dogs. However, because dominant species in this survey was C. canis, the importance of breeding kennel dogs as reservoirs for Cryptosporidium spp. transmission to humans is likely to be low in Japan.

Molecular detection and genotyping of Enterocytozoon bieneusi in family pet dogs obtained from different routes in Japan.

The present study determined the prevalence of Enterocytozoon (E.) bieneusi infection and genotyped the isolates in family pet dogs obtained from three different routes in Japan. Fresh fecal samples were collected from 597 family pet dogs. A nested polymerase chain reaction (PCR) assay targeting the internal transcribed spacer (ITS) region of ribosomal DNA was employed for detection of E. bieneusi, and the obtained positive PCR amplicons were sequenced for genotyping. The overall prevalence of E. bieneusi in family pet dogs was 4.4%. Significant differences in prevalence were found among the categories of age (<1 year: 8.3% vs. ≥1 year: 3.4%), obtained route (from private owner: 3.2% vs. from pet shop: 3.9% vs. from breeding kennel: 14.3%), and living regions (ranged 0-10.3%). However, no significant differences were observed in the prevalence of living condition (indoor: 4.6% vs. outdoor: 2.0%), and fecal condition (formed: 4.5% vs. soft: 4.7% vs. diarrhea: 0%). Of the 26 sequenced samples, all were identified as genotype PtEb IX, which belongs to the dog-specific genotype. In conclusion, although E. bieneusi infections are relatively common in family pet dogs in Japan, the risk of zoonotic transmission from dogs to humans is likely to be low.

Molecular Prevalence of Cryptosporidium spp. among Companion Birds Kept in Pet Shops in Japan.

Cryptosporidium is the most common protozoan that can infect a wide range of animals, including mammals and birds. Avian Cryptosporidium spp. can cause enteric and respiratory diseases which can be fatal in birds and some species are zoonotic. Companion birds have the potential as reservoir due to their close contact with humans. Pet shops are the major source of companion birds. However, few reports are available regarding Cryptosporidium spp. infection among companion birds kept in pet shops. The present study reports the prevalence and molecular characteristics of Cryptosporidium spp. among companion birds kept in pet shops in Japan. A total of 265 fresh fecal samples were obtained from birds kept in 4 pet shops; these birds belonged to 41 species in 3 bird orders. A nested polymerase chain reaction (PCR) assay targeting the small subunit rRNA gene was employed for the detection of Cryptosporidium spp. A total of 24 samples (9.1%) were positive, and Cryptosporidium spp. were detected from all pet shops. The prevalence of Cryptosporidium spp. in each of the bird orders was 6.5% (10/153) in Psittaciformes, 14.4% (13/90) in Passeriformes, and 4.5% (1/22) in Galliformes. Based on sequence analysis, 13 (54.2%) isolates were classified to C. galli, 8 (33.3%) were avian genotype III, and the remaining 3 (12.5%) were C. baileyi. No infection with zoonotic C. meleagridis and no coinfection with multiple Cryptosporidium spp. and/or genotypes were observed. The zoonotic potential of Cryptosporidium spp. infecting companion birds kept in pet shops in Japan is likely to be low.

Multilocus genotyping of Giardia duodenalis isolates from household cats and pet shop kittens.

Giardia duodenalis is a pathogenic protozoan that causes diarrhea in mammalian hosts including humans and companion animals, and the host specificity differs depending on the genetic assemblages. The purpose of the present study is to perform multilocus genotyping at four loci of G. duodenalis isolates from household cats and pet shop kittens in Japan and evaluate the zoonotic potential. Fifty-seven fecal samples from cats (household cats: 13, pet shop kittens: 44), which were positive for Giardia-specific antigen as determined by an ELISA kit, were the subjects of our analysis. Nested or semi-nested PCRs targeting 18S ribosomal RNA (18S rRNA), glutamate dehydrogenase (GDH), beta-giardin (BG), and triosephosphate isomerase (TPI) were performed on all samples. All DNA amplicons were sequenced for genotyping. Forty-four isolates (household cats: 11, pet shop kittens: 33) were positive for at least one of the PCRs. Assemblage F was the most frequently detected (75%; 33/44), followed by assemblage A (13.6%; 6/44) and assemblage B (2.3%; 1/44). Four isolates (9.1%) showed the results as mixed assemblages F and A. Sub-genotyping of assemblage A isolates based on three loci (GDH, BG, and TPI) identified all of them as assemblage AI. DNA sequences of zoonotic G. duodenalis assemblages were detected from 25% of the genotyped specimens, and these were found at several shops and hospitals in Japan, which suggests that zoonotic G. duodenalis is prevalent among domestic cats in Japan, and that the considerable risk of zoonotic transmission of G. duodenalis from household cats or pet shop kittens to humans exists.

Multilocus genotyping of Giardia duodenalis isolates from breeding cattery cats in Japan.

OBJECTIVES: The present study reports the multilocus genotyping of Giardia duodenalis isolates from cats maintained in breeding catteries in Japan and discusses their potential for zoonotic transmission. METHODS: A total of 41 faecal samples positive for Giardia-specific antigen were procured from cats maintained in five breeding catteries and subjected to PCR to amplify four gene loci, namely small subunit ribosomal RNA (SSU rRNA), glutamate dehydrogenase (gdh), beta-giardin (bg) and triose phosphate isomerase (tpi ). The PCR-amplified DNA fragments were sequenced to determine the G duodenalis genotypes (synonym for assemblages). RESULTS: The most commonly occurring single assemblage was assemblage F (68.3%; n = 28/41), followed by assemblage A (12.2%; n = 5/41) and assemblage C (2.4%; n = 1/41). The mixed assemblages were identified as follows: assemblages F and A (9.8%; n = 4/41), assemblages F and C (4.9%; n = 2/41) and assemblages C and D (2.4%; n = 1/41). Additional sub-genotyping of assemblage A isolates based on three of the sequenced loci (gdh, bg and tpi ) revealed that all eight isolates were identified as sub-assemblage AI and/or AII. CONCLUSIONS AND RELEVANCE: The present study is the first to report the detection of dog-adapted assemblages C and D in feline isolates from Japan. In addition, zoonotic sub-assemblage AI and human-adapted sub-assemblage AII were also identified. Thus, we concluded that the risk of transmission of G duodenalis from breeding cattery cats to humans is considerable and cannot be ignored.

Molecular prevalence of Cryptosporidium species among household cats and pet shop kittens in Japan.

OBJECTIVES: To address the lack of up-to-date published data, the present study evaluates the PCR-based prevalence of Cryptosporidium species infection and molecular characteristics of isolates among household cats and pet shop kittens in Japan. METHODS: A total of 357 and 329 fresh faecal samples were collected from household cats and pet shop kittens, respectively, with or without clinical signs of infection. A nested PCR assay targeting the 18S rRNA gene was employed for the detection of Cryptosporidium species. After specific DNA fragments (approximately 826 base pairs) were confirmed, the amplicons were sequenced to determine species. RESULTS: Seven (2.0%) household cats and one (0.3%) pet shop kitten tested positive for the presence of Cryptosporidium species. In household cats, there was a significant difference in prevalence between cats aged <1 year (4.6%) and those aged ⩾1 year (0.4%). No significantly different prevalence was observed with regard to faecal condition in either household cats or pet shop kittens. A total of eight Cryptosporidium species isolates, seven from household cats and one from a pet shop kitten, were identified as Cryptosporidium felis. CONCLUSIONS AND RELEVANCE: The present study demonstrates the risk of zoonotic transmission of Cryptosporidium species from household cats and pet shop kittens to humans is low in Japan.