In this paper we report the occurrence of zoonotic species of Cryptosporidium (C. ubiquitum and C. parvum subtype IIaA15G2R1) and the host-specific Giardia duodenalis genotype E in a sheep farm from Brazil. Fecal specimens were collected from 100 animals and screened for the presence of G. duodenalis and Cryptosporidium spp. by enzyme immunoassay (EIA) and polymerase chain reaction (PCR). Restriction fragment length polymorphism (RFLP) and sequence analysis of selected samples was used to confirm the species and to characterize the isolates. Of 100 specimens, 34 were positive for Giardia (34%) and 25 were positive for Cryptosporidium (25%). The prevalence of Giardia and Cryptosporidium was higher in lambs than in ewes. Co-infection was observed in 14% of animals. All G. duodenalis isolates belonged to genotype E. The genetic characterization performed in 22 Cryptosporidium-positive samples revealed the presence of zoonotic C. ubiquitum and C. parvum in 76% (19/25) and 12% (3/25) of infected animals, respectively. The presence of zoonotic Cryptosporidium subtypes in lambs was of epidemiological and public health relevance.
In this study, we identified Cryptosporidium species and genotypes present in dairy cattle in the central region of São Paulo state, Brazil. Fecal specimens were collected from 200 animals (100 calves and 100 cows) in ten dairy farms. Fecal samples were examined using microscopic examination (ME), enzyme immunoassay (EIA) and polymerase chain reaction (PCR). Cryptosporidium species and genotypes were determined by restriction fragment length polymorphism (RFLP) or DNA sequencing analysis of the SSU-rRNA and GP60 genes. The occurrence of Cryptosporidium spp. infection was 14% (28/200). The occurrence in calves (26%) was significantly higher than in cows (2%). Of the 27 Cryptosporidium-positive specimens submitted to genotyping, C. andersoni was identified in 23 (85.1%), C. bovis in three (11.1%), and the zoonotic C. parvum subtype IIaA15G2R1 in one (3.7%). The study demonstrates that Cryptosporidium spp. infection was common and widespread in dairy cattle in this region and that calves have a high prevalence of C. andersoni. Furthermore, the presence of C. parvum subtype IIaA15G2R1 indicates that dairy calves from this region should be considered a potential source of zoonotic Cryptosporidium oocysts.Keywords: Cryptosporidium spp., cattle, PCR-RFLP, sequencing, genotyping. ResumoNo presente estudo foram identificadas espécies e genótipos de Cryptosporidium originadas de bovinos leiteiros na região central do estado de São Paulo, Brasil. Amostras fecais foram coletadas de 200 animais (100 bezerros e 100 vacas) em 10 propriedades leiteiras. As amostras foram examinadas utilizando os métodos de microscopia óptica (MO), ensaio imunoenzimático (EI) e reação em cadeia da polimerase (PCR). As espécies e genótipos de Cryptosporidium foram determinados pelo método de polimorfismo no tamanho dos fragmentos de restrição (RFLP) ou sequenciamento dos genes SSU-rRNA e GP60. A infecção por Cryptosporidium spp. teve ocorrência de 14% (28/200). A ocorrência em bezerros (26%) foi significativamente maior do que em vacas (2%). Do total de 27 amostras positivas submetidas à caracterização genética, C. andersoni foi identificado em 23 (85.1%), C. bovis em três (11.1%) e C. parvum subtipo IIaA15G2R1 em uma (3.7%). O presente estudo demonstrou que a infecção por Cyptosporidium é comum e difundida em bovinos leiteiros nessa região e que bezerros possuem uma alta prevalência de C. andersoni. A presença de C. parvum subtipo IIaA15G2R1 indica que bezerros leiteiros dessa região devem ser considerados uma fonte de oocistos de Cryptosporidium com potencial zoonótico.
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