Edwards Frazão-Teixeira

Multi-locus DNA sequencing of Toxoplasma gondii isolated from Brazilian pigs identifies genetically divergent strains

Five Toxoplasma gondii isolates (TgPgBr1-5) were isolated from hearts and brains of pigs freshly purchased at the market of Campos dos Goytacazes, Northern Rio de Janeiro State, Brazil. Four of the five isolates were highly pathogenic in mice. Four genotypes were identified. Multi-locus PCR-DNA sequencing showed that each strain possessed a unique combination of archetypal and novel alleles not previously described in South America. The data suggest that different strains circulate in pigs destined for human consumption from those previously isolated from cats and chickens in Brazil. Further, multi-locus PCR-RFLP analyses failed to accurately genotype the Brazilian isolates due to the high presence of atypical alleles. This is the first report of multi-locus DNA sequencing of T. gondii isolates in pigs from Brazil.

Molecular characterization of Cryptosporidium in Brazilian sheep

Feces were collected from 125 sheep between January and December 2007, on ten farms in the State of Rio de Janeiro, Brazil, and examined for the presence of Cryptosporidium. Ninety samples were collected from lambs 2 to 6 months of age, and 35 were from sheep over 12 months of age. All samples were subjected to molecular analysis by polymerase chain reaction (nested PCR) in two steps of the SSU rRNA. Two samples (1.6%) from the lambs were positive, and after sequencing were identified as Cryptosporidium ubiquitum. This species has been reported worldwide and it is considered a zoonotic pathogen since it has been found and in several animal species and humans. However, because of the low frequency of C. ubiquitum found, the risk for public health in this region may not be high.

Immunohistochemical identification of Toxoplasma gondii in tissues from Modified Agglutination Test positive sheep

Toxoplasma gondii is a zoonotic agent of great importance in veterinary and public health. The aim of this study was to identify T. gondii by IHC (immunohistochemistry) in different sheep tissues and to determine if an association exists between the results obtained by this method and those obtained by the Modified Agglutination Test (MAT). Tissue specimens of twenty-six sheep seroreactive for T. gondii were selected for histopathological evaluation. The presence of T. gondii was investigated in brain, liver and heart samples by IHC and a possible anti-T. gondii antibody cross reactions with other parasites. McNemars, Chi-square and Fishers Exact Tests were applied for the statistical analysis of the results. The analysed tissues showed at least one of the following histopathological changes: mild-to-moderate congestion, focal polymorphonuclear inflammatory infiltrate and multifocal or focal mononuclear inflammatory infiltrate. Sarcocystis spp. were identified in the histological sections from both the heart and diaphragm tissues of 88.5% (23/26) of the animals. A total of 46.2% (12/26) of the T. gondii seroreactive sheep was also positive for T. gondii by IHC in at least one organ (brain, liver or heart). The liver IHC-positivity for T. gondii was statistically equivalent to the global individual IHC-positivity, according to McNemars test. In addition, IHC allowed the detection of T. gondii in infected animals regardless of the titration observed in the MAT. The statistical difference observed between the three organs when comparing the low titration group, suggested that the heart might be the most suitable organ to detect T. gondii infection by IHC. The IHC results in this study revealed that almost half of MAT positive animals could serve as potential sources of infection for humans because bradyzoites were identified in different tissues, regardless of the MAT titration.

Anti–Toxoplasma gondii Antibodies in Cattle and Pigs in a Highly Endemic Area for Human Toxoplasmosis in Brazil

Abstract Sera from 77 cattle and 61 pigs from herds of Campos dos Goytacazes and neighborhood, north of Rio de Janeiro state, Brazil, were analyzed for the presence of antibodies anti–Toxoplasma gondii via ELISA. The seropositivity was confirmed at 49.4% (38 of 77) for cattle and at 11.5% (7 of 61) for pigs. Pigs bred under free-ranging systems presented with 20.6% (7 of 34) seropositivity and no pigs bred under indoor systems were positive (n  =  27). The seropositivity verified for cattle and pigs confirmed the extensive distribution of the parasite in this region. In the face of these data, we discuss the potential risk for human infection by consumption of raw or undercooked meat in this geographic region of Brazil. This is the first report of seropositivity for anti–T. gondii antibodies in cattle in the north of Rio de Janeiro State, Brazil.

Cryptosporidium Pig Genotype II Diagnosed in Pigs From the State of Rio De Janeiro, Brazil

Abstract 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.

Characterization of the zoonotic potential of Toxoplasma gondii in horses from Rio de Janeiro State

The aim of this study was to perform a survey on the prevalence of anti-Toxoplasma gondii antibodies in horses from Rio de Janeiro State, Brazil. From 2012 to 2013, a total of 624 blood samples were collected from horses from the eight regions comprising Rio de Janeiro State (Baixadas Litorâneas, Serrana, Norte Fluminense, Noroeste Fluminense, Centro-Sul, Metropolitana, Médio Paraíba, and Costa Verde). All sera samples were tested for anti-T. gondii antibodies by performing the modified agglutination test with a cut-off of 1:25. Positive serology for T. gondii was detected in 22.8% (142/624) of the horses studied. Seropositivity was detected in all regions sampled; furthermore, statistical significance was observed when all locations were compared at once. The Médio Paraíba region had the highest number of positive animals 54.76% (23/42) in the Bonferroni correction among Costa Verde, Centro Sul, Metropolitana, and Serrana (p<0.001). Seropositivity was 39.58% (38/96) in Norte Fluminense, which was the second most prevalent region. The results indicated that the T. gondii parasite is widely distributed in horses in Rio de Janeiro State and represents a risk to public and animal health. These findings emphasize the need to increase control and prevention of this disease in horses.

Occurrence of Cryptosporidium andersoni in Brazilian Cattle

abstract:  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.

Compromised nutrition in gerbils infected by Cystoisospora felis detected through an animal performance analysis tool

The impact of Cystoisospora felis infection on the nutritional efficiency of gerbils was studied. The variables weight gain and feed intake were measured during four weeks in 28 laboratory gerbils, of which 14 were inoculated with 3.5 × 10(5) sporulated oocysts of C. felis and the remaining 14 were controls. The animals from both groups were weighted, killed, eviscerated and had their carcasses and tissues weighted and compared. A modern tool designed for measuring nutritional performance of farm animals was applied. The results showed compromised nutritional efficiency of the infected animals within the first week after infection. The consequences of these results are discussed here, including the potential impact of infection on farm animals performance.