RESUMO
Although Brazil is the world's fourth largest producer and exporter of pork, there is no information on Enterocytozoon bieneusi in pigs. This study was undertaken to determine the presence of E. bieneusi in pigs in the State of Rio de Janeiro, Brazil. Fecal samples were collected from 91 pigs (1- to 12-month-old) in 10 properties and examined by molecular methods. The presence of E. bieneusi was determined by PCR and all PCR positive specimens were sequenced to determine the genotype by nucleotide sequence analysis of the internal transcribed spacer of the rRNA gene. E. bieneusi was found in pigs in all farms. Fifty four (59.3%) samples were E. bieneusi-positive. A wide genetic diversity was found with 21 genotypes identified, 4 previously reported (O, EbpA, CS-1, and H) and 17 novel genotypes named PigEb1-PigEb17. All 17 novel genotypes identified in this study clustered within the previously designated zoonotic Group 1. The most prevalent genotypes were novel genotypes PigEb2 and PigEb4 (16/91, 17.6%, each). Mixed infections with 2 or 3 genotypes were detected in 13 pigs (24.1%). The high prevalence in pigs observed in this study, the description of two known zoonotic genotypes (EbpA and O), and the report of 17 new genotypes of E. bieneusi, represent an important advancement in the study of the wide genetic diversity of this organism, emphasizing the importance of further research, especially in geographical areas where little or no research has been conducted. The zoonotic risk of these novel genotypes and their importance to other animal species is still unknown, but needs to be further evaluated.
Assuntos
Enterocytozoon/isolamento & purificação , Fezes/microbiologia , Gado/microbiologia , Microsporidiose/veterinária , Sus scrofa/microbiologia , Doenças dos Suínos/microbiologia , Animais , Sequência de Bases , Brasil/epidemiologia , Coinfecção/microbiologia , Coinfecção/veterinária , DNA Fúngico/genética , DNA Espaçador Ribossômico/genética , Enterocytozoon/genética , Variação Genética , Genótipo , Humanos , Microsporidiose/epidemiologia , Microsporidiose/microbiologia , Dados de Sequência Molecular , Filogenia , Reação em Cadeia da Polimerase , Prevalência , Análise de Sequência de DNA , Suínos , Doenças dos Suínos/epidemiologiaRESUMO
Feces were collected from 68 dairy cattle, 1 to 12 mo of age, on 12 farms in the municipality of Campos dos Goytacazes, Rio de Janeiro, Brazil, and examined for the presence of Cryptosporidium sp. All samples were subjected to molecular analysis by polymerase chain reaction (nested PCR) of the 18S rRNA. Four positive samples (4.54%) were sequenced and identified as Cryptosporidium andersoni. This species represents a risk for Brazilian cattle because infection can affect cattle productivity. Moreover, C. andersoni is considered a zoonotic species.
Assuntos
Doenças dos Bovinos/epidemiologia , Criptosporidiose/veterinária , Cryptosporidium/isolamento & purificação , Animais , Brasil/epidemiologia , Bovinos , Doenças dos Bovinos/parasitologia , Doenças dos Bovinos/transmissão , Criptosporidiose/epidemiologia , Criptosporidiose/parasitologia , Criptosporidiose/transmissão , Cryptosporidium/classificação , Cryptosporidium/genética , DNA de Protozoário/química , DNA de Protozoário/isolamento & purificação , Indústria de Laticínios , Fezes/parasitologia , Humanos , Dados de Sequência Molecular , Reação em Cadeia da Polimerase/veterinária , RNA Ribossômico 18S/genética , Alinhamento de Sequência , Zoonoses/parasitologia , Zoonoses/transmissãoRESUMO
Pigs may represent a source of Cryptosporidium sp. infection to humans. The objective of this study was to identify the Cryptosporidium species present in pigs from the State of Rio de Janeiro, Brazil, and verify what risks pigs represent in the transmission of human cryptosporidiosis, because there is no such information to date in Brazil. Ninety-one samples of pig feces were collected from 10 piggeries in 2 municipalities located in the north and northwest regions of the State of Rio de Janeiro, Brazil. A nested polymerase chain reaction (PCR) protocol to amplify an 830-bp fragment of the small subunit rDNA (SSU rRNA) gene was followed by sequencing of all positive PCR samples. Two samples (2.2%) were Cryptosporidium sp. positive and were identified as pig genotype type II (PGII). This genotype has been observed in an immunocompetent person, in cattle without pigs nearby, and from a potential human source. Its potential for zoonotic transmission is little known and should be rigorously studied.
Assuntos
Criptosporidiose/veterinária , Cryptosporidium/classificação , Doenças dos Suínos/parasitologia , Animais , Brasil/epidemiologia , Criptosporidiose/epidemiologia , Criptosporidiose/parasitologia , Criptosporidiose/transmissão , Cryptosporidium/genética , DNA de Protozoário/química , DNA de Protozoário/isolamento & purificação , Fezes/parasitologia , Genótipo , Dados de Sequência Molecular , Reação em Cadeia da Polimerase/veterinária , Fatores de Risco , Alinhamento de Sequência/veterinária , Suínos , Doenças dos Suínos/epidemiologia , Doenças dos Suínos/transmissãoRESUMO
Cryptosporidiosis has recently attracted attention as an emerging waterborne and foodborne disease as well as an opportunistic infection in HIV infected individuals. The lack of genetic information, however, has resulted in confusion in the taxonomy of Cryptosporidium parasites and in the development of molecular tools for the identification and typing of oocysts in environmental samples. Phylogenetic analysis of the small subunit ribosomal RNA (SSU rRNA) gene has shown that the genus Cryptosporidium comprises several distinct species. Our data show the presence of at least four species: C. parvum, C. muris, C. baileyi and C. serpentis (C. meleagridis, C. nasorum and C. felis were not studied). Within each species, there is some sequence variation. Thus, various genotypes (genotype 1, genotype 2, guinea pig genotype, monkey genotype and koala genotype, etc.) of C. parvum differ from each other in six regions of the SSU rRNA gene. Information on polymorphism in Cryptosporidium parasites has been used in the development of species and strain-specific diagnostic tools. Use of these tools in the characterization of oocysts in various samples indicates that C. parvum genotype 1 is the strain responsible for most human Cryptosporidium infections. In contrast, genotype 2 is probably one of the major sources for environmental contamination, and has been found in most oysters examined from Chesapeake Bay that may serve as biologic monitors of estuarine waters.
Assuntos
Cryptosporidium/genética , Filogenia , RNA Ribossômico/análise , Água/parasitologia , Animais , Cryptosporidium/classificação , Cryptosporidium/isolamento & purificação , Genes de RNAr/genética , Genótipo , Humanos , Polimorfismo Genético/genéticaRESUMO
A cluster-sampling, cross-sectional study was conducted for assessing the prevalence of Cryptosporidium infection in children less than 16 years of age from three villages, Dondian, Linshan, and Fuziyin, in rural Anhui in eastern China. Among 320 apparently healthy children less than 10 years of age from Dondian who had stool specimens collected, cryptosporidial oocysts were found in stools of three children from Dondian, and no positive specimens were found in 239 children studied from Linshan. In addition, a total of 610 serum samples from children in these three villages were tested for specific IgG antibody to Cryptosporidium with an enzyme-linked immunosorbent assay (ELISA) and the prevalence rates were 42.3%, 51.7%, and 57.5%, respectively, in Dondian, Linshan, and Fuziyin. Seroprevalence increased progressively with age. No detectable antibody was found in infants between two and six months of age, and seropositivity steadily increased after one year of age. Among 36 sera from adults 15-60 years of age without diarrheal illness in Huanglu villages of rural Chaohu, 50% (18 of 36) were positive. As expected, a good correlation was found in the specific IgG antibody between the paired serum specimens from 30 matched mother-neonates who showed transplacental transfer of IgG. However, little or no IgM antibody was seen in the neonates even though several mothers had a positive anticryptosporidial IgM enzyme-linked immunoassay result. Forty randomly selected serum samples from children less than four years of age in a similarly impoverished semiurban community in Fortaleza, Brazil, where the majority of households also have pit toilets and shared community water supplies and 172 serum samples from patients one month to 29 years of age admitted to the University of Virginia Hospital without diarrhea were also examined. In Fortaleza, almost all children acquired antibody by their second year of life, demonstrating the high prevalence of this infection. In rural Anhui, only about half the children were infected by 5-7 years of age. The overall prevalence rate (16.9%) of seropositivity among children and young adults in Virginia was much lower than in China and Brazil. These results indicate that cryptosporidial infection is ubiquitous, and is highly endemic in these impoverished communities. The difference between China and Brazil may reflect earlier weaning, hygiene practices, poorer water or sanitation, multiple siblings in family and geographic environment in Brazil.