Molecular prevalence and characterization of Cryptosporidium in domestic free-range poultry in Anhui Province, China.

Free-range chickens might mediate the spread of Cryptosporidium oocysts to humans and other animals. Few studies have evaluated the prevalence of Cryptosporidium species in domestic free-range poultry in China. Here, we characterized the prevalence and distribution of species and genotypes of Cryptosporidium in domestic free-range chickens, ducks, and geese in Anhui Province, China. A total of 1910 fresh fecal samples from three poultry species were examined from 18 free-range poultry farms by nested PCR and analysis of the Cryptosporidium SSU rRNA gene. The overall prevalence of Cryptosporidium species was 2.9% (55/1910), with infection rates of 1.3% (11/829) in chickens, 7.3% (36/487) in ducks, and 1.4% (8/594) in geese. C. baileyi (0.6%), C. meleagridis (0.2%), C. galli (0.2%), and C. xiaoi-like genotype (0.2%) were identified in chickens, and only C. baileyi was identified in ducks and geese, with infection rates of 7.4% and 1.3%, respectively. C. baileyi was the most prevalent species. Sequencing of the GP60 gene revealed that the C. meleagridis isolates belonged to the IIIbA26G1R1b subtype. This is the first study to document C. galli and C. xiaoi-like genotype in domestic free-range chickens in China. These findings expand the range of avian hosts known for Cryptosporidium and highlight the need for additional studies to characterize the diversity of Cryptosporidium in avian species.

Molecular study and genotyping of Cryptosporidium baileyi and Cryptosporidium parvum from free-range and commercial broiler chickens in Guilan province, Iran.

Cryptosporidiosis acutely impacts the digestive and/or respiratory tract of the birds in many species of various orders. More importantly, it is also well known as a significant zoonotic disease, which can lead to diarrhea in humans and livestock. Regarding increasing demand for free-range products and increasing the number of free-range poultry farms, the present paper evaluated histopathological and molecular detection of Cryptosporidium baileyi and Cryptosporidium parvum in free-range and commercial broiler chickens in the north part of Iran. For this purpose, 100 fecal and tissue samples of the chickens in Guilan province were collected. After microscopic examination using Ziehl-Neelsen staining, molecular analyses of the fecal samples were processed by Nested-PCR targeting the 18S rRNA gene followed by sequencing of the amplicons and phylogenetic analyses. Eventually, the tissue samples were studied for histological lesions. Findings demonstrated the presence of Cryptosporidium baileyi and Cryptosporidium parvum in 6 % and 2 % of fecal samples, respectively. This is the first identification of C.parvum in avian hosts in Iran, and for the first time, C.baileyi and C.parvum are shown in native free-range chickens in Iran. All of the PCR positive birds with clinical symptoms showed gross lesions of respiratory infections. There was no significant difference between infection rate in free-range and commercial broiler chickens; however, the infection rate was significantly higher in chickens <25 days old. To conclude, we present here a notable Cryptosporidium infection rate in the free-range chicks in Iran, which notify the role of this host as a reservoir and should be more noted due to the economic and zoonotic importance.

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.

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.

Cryptosporidium infections of ring-necked pheasants (Phasianus colchicus) from an intensive artificial breeding programme in the Czech Republic.

From July to November 2012, preliminary coprological examinations were carried out on 85 pooled faecal samples of different aged ring-necked pheasants (Phasianus colchicus) (hatches from May until July) from an intensive artificial breeding programme in the Czech Republic. Cryptosporidium oocysts were detected in 12 samples (14.1 %) of ages >12 weeks (August-September). These results were supported by findings of Cryptosporidium baileyi and Cryptosporidium meleagridis oocysts in intestinal, or cloacal contents, and/or the bursa of Fabricius in 9 from 36 examined dead pheasants (prevalence 25 %). We describe in detail the various age groups of pheasants after hatching and present graphically the overall results of coprological examinations, showing pathways of infection of C. baileyi and C. meleagridis during the full rearing seasons of 2013 and 2014. We found very similar mean proportions of Cryptosporidium-positive samples over the entire 2013 period in pheasantry (173 pooled samples tested, 25 positive, 14.5 %) and 2014 (238 samples tested, 43 positive, 18.1 %). All tests were verified as being Cryptosporidium positive in 9 from 219 (prevalence 4.1 %) and 4 from 168 (prevalence 2.4 %) post-mortem examinations. Significantly, C. baileyi was found more frequently in faeces, with positivities ranging from 11.1 to 100 % (4->16-week-old pheasants). Oocysts of C. meleagridis were detected at ages 6->15 weeks ranging from 7.1 to 100 % in faeces during the rearing seasons. The burdens of C. baileyi (7 of 14 and 10 of 16) and C. meleagridis (5 of 14 and 7 of 16) for each year, in monitored brooder houses, flight pens and spread across all open areas were recorded. Oocysts of C. baileyi and C. meleagridis obtained from this study, and Cryptosporidium galli (obtained in another aviary from 36-week-old pheasants), were sequenced, and we characterized the highly variable 60-kDa glycoprotein gene of C. meleagridis. These results highlight the real risk of transmission of Cryptosporidium to susceptible wild birds and other potential hosts after termination of rearing and release.

First finding of spontaneous infections with Cryptosporidium baileyi and C. meleagridis in the red-legged partridge Alectoris rufa from an aviary in the Czech Republic.

This paper represents the first report of spontaneous infection with Cryptosporidium baileyi and Cryptosporidium meleagridis in the red-legged partridge (Alectoris rufa), as well as the percentage of positive samples and age-associated dynamics of cryptosporidial infections in an aviary in the Czech Republic. The entire infection process was monitored over two semesters (July-December 2012 and 2013) until release of birds for hunting purposes. Coprological examination of 663 pooled fecal samples and 89 post-mortem examinations of red-legged partridges were carried out. Our results indicated that infections with C. baileyi only occurred in 5-7 week-old birds during 2013 (percentage of positivity, 1%) and those with C. meleagridis in 18-22 week (17%) and 17-21 week-old birds (24%) during 2012 and 2013, respectively. Molecular characterization of isolates of C. baileyi and C. meleagridis heat shock protein 70 and actin genes were analyzed in order to support our coprological results. DNA sequence analysis of the 60kDa glycoprotein gene was used to subtype C. meleagridis. Our findings extend the host range for C. baileyi.