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  • Title: Molecular characterization of Cryptosporidium in ruminants and observation of natural infection by Cryptosporidium andersoni in sheep from Paraná, Brazil.
    Author: Holsback L, Marquez ES, Silva MAD, Porto PP, Garcia JL, Martins FDC, Seixas M.
    Journal: Rev Bras Parasitol Vet; 2023; 32(4):e010023. PubMed ID: 38055435.
    Abstract:
    The aim of this study was to identify Cryptosporidium species found in cattle and sheep in Paraná, southern region of Brazil. Individual fecal samples from 458 bovines and 101 sheep were submitted for molecular analysis by PCR and nested PCR using specific primers for sequences of the 18S ribosomal unit (rRNA). Positive samples were analyzed using restriction fragment length polymorphism (RFLP), followed by genetic sequencing for species confirmation. The occurrence of Cryptosporidium was 11.27% (63/559). The highest occurrence was detected in lambs (12/59, 20.33%). From the 63 positive samples, it was possible to identify the species in 58 of them by RFLP and genetic sequencing. Five species of Cryptosporidium were identified: Cryptosporidium andersoni, Cryptosporidium bovis, Cryptosporidium ryanae, Cryptosporidium xiaoi, and Cryptosporidium parvum. The most prevalent species was C. andersoni (41.38%) and the least predominant was C. parvum (10.34%). The most abundant species of Cryptosporidium in dairy calves were C. andersoni (11/25) and C. ryanae (6/25). Of the 17 positive sheep, nine (52.94%) were infected with C. andersoni. This finding is the first report on the occurrence of C. andersoni in naturally infected sheep in Brazil and the first observation of a high absolute occurrence of this Cryptosporidium species in sheep. O objetivo deste estudo foi identificar espécies de Cryptosporidium em bovinos e ovinos do Paraná, região sul do Brasil. Amostras de fezes de 458 bovinos e 101 ovinos foram individualmente submetidas à análise molecular por PCR e nested PCR, utilizando-se iniciadores específicos para sequências da unidade ribossomal 18S (rRNA). As amostras positivas foram analisadas pelo polimorfismo de comprimento de fragmento de restrição (RFLP), seguido de sequenciamento genético para confirmação da espécie. A ocorrência de Cryptosporidium foi de 11,27% (63/559). Observou-se maior ocorrência em cordeiros (20,33%). Das 63 amostras positivas, foi possível identificar as espécies em 58 amostras por RFLP e sequenciamento genético. Foram identificadas cinco espécies de Cryptosporidium: Cryptosporidium andersoni, Cryptosporidium bovis, Cryptosporidium ryanae, Cryptosporidium xiaoi e Cryptosporidium parvum. A espécie mais predominantemente encontrada foi C. andersoni (41,38%) e a menos foi C. parvum (10,34%). As espécies mais abundantes de Cryptosporidium, em bezerros leiteiros, foram C. andersoni (11/25) e C. ryanae (6/25). Dos 17 ovinos positivos, nove (52,94%) estavam infectados com C. andersoni. Este achado é o primeiro relato sobre a ocorrência de C. andersoni em ovinos naturalmente infectados no Brasil e a primeira observação de alta ocorrência absoluta desta espécie de Cryptosporidium em ovinos.
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