Detecção molecular e subtipagem de Cryptosporidium spp. em caprinos, ovinos, bovinos, leitões e eqüinos jovens
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2011-08-25
Autores
Coelho, Willian Marinho Dourado [UNESP]
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Universidade Estadual Paulista (Unesp)
Resumo
A criptosporidiose é uma doença entérica grave, economicamente significante, caracterizada principalmente por desordens intestinais, podendo ocasionar manifestações clínicas variadas e eventual mortalidade, principalmente em animais jovens. Este estudo objetivou detectar molecularmente genótipos e subgenótipos de Cryptosporidium spp. em fezes de cabritos provenientes dos Estados de Goiás, Mato Grosso do Sul, Minas Gerais e São Paulo, Brasil. Amostras fecais foram colhidas diretamente do reto de 192 cabritos de diferentes raças, machos e fêmeas, com até doze meses de idade. Adicionalmente, foram analisadas amostras fecais de ovinos, bovinos, suínos e eqüinos jovens. A eliminação de oocistos de Cryptosporidium spp. foi observada por meio das técnicas de Sheather e Kinyoun, seguindo-se a micrometragem dos oocistos com ocular de campo amplo micrométrica 10x (Bioval®) em aumento microscópico de 400 e 1000x. A reação em cadeia da polimerase (PCR) foi realizada para amplificação dos fragmentos dos genes da subunidade 18S do rRNA e da glicoproteína GP60. Presença de oocistos de Cryptosporidium spp. foram observados pela análise microscópica em 11,45% (22/192) das amostras analisadas. Amplificação gênica positiva para Cryptosporidium foi demonstrada em 16,66% (32/192) destas amostras. Com o sequenciamento dos produtos da PCR do gene 18S rRNA, todas as amostras foram identificadas como Cryptosporidium parvum. Por meio da subgenotipagem com o sequenciamento do gene GP60, foi encontrado exclusivamente o subgenótipo de C. parvum IIaA15G2R1. Através dos resultados obtidos, pode-se inferir que a infecção por C. parvum está presente em rebanhos caprinos de diferentes Estados brasileiros podendo, esta espécie animal, atuar como uma importante fonte de infecção do subtipo zoonótico de Cryptosporidium para outras espécies animais, em especial para o ser humano
Cryptosporidiosis is a serious enteric disease, economically significant, mainly characterized by intestinal disorders, may cause various clinical manifestations and eventual mortality, especially in young animals. This study aimed to detect and molecularly genotypes and subgenotypes of Cryptosporidium spp. in feces of goat kids from the states of Goiás, Mato Grosso do Sul, Minas Gerais and São Paulo, Brazil. Fecal samples were collected directly from the rectum of 192 goat kids of different breeds, males and females, with up to twelve months old. Additionally, were analyzed fecal samples from cattle, sheep, pigs and young horses. The elimination of oocysts of Cryptosporidium spp. was observed using the Sheather and Kinyoun techniques, followed by micrometric analysis with ocular micrometer wide-field 10x (Bioval ®) in increase from 400 and 1000x. The polymerase chain reaction (PCR) was performed to amplify fragments of genes of the subunit 18S rRNA and glycoprotein GP60. Presence of Cryptosporidium spp. was observed by microscopic examination in 11.45% (22/192) of the samples. Gene amplification for Cryptosporidium was demonstrated in 16.66% (32/192) of these samples. With the sequencing of the PCR products of 18S rRNA gene, all samples were identified as Cryptosporidium parvum. Through of subgenotyping with sequencing of GP60 gene was found exclusively the subtype of C. parvum IIaA15G2R1. By the results obtained, it can be inferred that infection with C. parvum is present in goat kids in different brazilian states may, this animal species act as an important source of infection with zoonotic subtype of Cryptosporidium to other animal species, especially for humans
Cryptosporidiosis is a serious enteric disease, economically significant, mainly characterized by intestinal disorders, may cause various clinical manifestations and eventual mortality, especially in young animals. This study aimed to detect and molecularly genotypes and subgenotypes of Cryptosporidium spp. in feces of goat kids from the states of Goiás, Mato Grosso do Sul, Minas Gerais and São Paulo, Brazil. Fecal samples were collected directly from the rectum of 192 goat kids of different breeds, males and females, with up to twelve months old. Additionally, were analyzed fecal samples from cattle, sheep, pigs and young horses. The elimination of oocysts of Cryptosporidium spp. was observed using the Sheather and Kinyoun techniques, followed by micrometric analysis with ocular micrometer wide-field 10x (Bioval ®) in increase from 400 and 1000x. The polymerase chain reaction (PCR) was performed to amplify fragments of genes of the subunit 18S rRNA and glycoprotein GP60. Presence of Cryptosporidium spp. was observed by microscopic examination in 11.45% (22/192) of the samples. Gene amplification for Cryptosporidium was demonstrated in 16.66% (32/192) of these samples. With the sequencing of the PCR products of 18S rRNA gene, all samples were identified as Cryptosporidium parvum. Through of subgenotyping with sequencing of GP60 gene was found exclusively the subtype of C. parvum IIaA15G2R1. By the results obtained, it can be inferred that infection with C. parvum is present in goat kids in different brazilian states may, this animal species act as an important source of infection with zoonotic subtype of Cryptosporidium to other animal species, especially for humans
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Animais - Doenças entéricas, Animals - Enteric diseases
Como citar
COELHO, Willian Marinho Dourado. Detecção molecular e subtipagem de Cryptosporidium spp. em caprinos, ovinos, bovinos, leitões e eqüinos jovens. 2011. xi, 84 f. Tese (doutorado) - Universidade Estadual Paulista, Faculdade de Ciências Agrárias e Veterinárias, 2011.