The aim of this study was to report on detection of Toxoplasma gondii DNA in oysters (Crassostrea sp.) in the state of Maranhão. To conduct this study, 200 farmed oysters were acquired in the municipality of Raposa and 100 in Paço do Lumiar; and a further 100 oysters were taken from the natural stock in the municipality of Primeira Cruz. This total of 400 specimens sampled was divided into 80 pools composed of five animals each. The gills and visceral mass of each oyster were removed for DNA extraction (per pool of oysters), using a commercial kit. The nested PCR technique (with the primer SAG-1) was then used to investigate any presence of protozoa. This molecular technique demonstrated the presence of DNA of T. gondii in 2.5% of the pools of oysters (n = 2/80): these oysters were exclusively from farms. The results from this study allow the conclusion that oysters of the genus Crassostrea that are farmed in the state of Maranhão are capable of filtering oocysts of T. gondii and maintaining them in their tissues. They are therefore potential sources of contamination for humans and other animals.
A serological, molecular and histopathological study was carried out in order to investigate occurrences of Toxoplasma gondii in pigs slaughtered with and without inspection service. Serum samples were collected from 60 pigs to detect anti-T. gondii antibody by indirect fluorescent antibody (IFAT). Tongue, masseter and diaphragm fragments were also collected for parasite DNA detection by means of the polymerase chain reaction (PCR) and histopathological analysis. The serological results showed that 77% (44/60) of the pigs were positive. Regarding PCR, 66.67% (40/60) were positive for T. gondii. Among the tissues evaluated, the diaphragm was the one with the highest frequency of positivity (40%; 24/60), followed by the masseter (38.33%; 23/60) and tongue (33.3%; 20/60). Histopathological changes were only observed in the diaphragm, which presented inflammatory infiltrates of lymphohistiocytic and neutrophilic types. These results not only show the potential threat of T. gondii to human health, but also demonstrate the dynamic epidemiological situation of toxoplasmosis in pigs in the city of São Luís, providing support for food security regarding pigs and for T. gondii control programs in Brazil.
Enzootic bovine leukosis (EBL) is a chronic viral disease of wide distribution in cattle herds and may take several years for the first manifestation of clinical signs. Most animals do not present clinical signs. However, the economic losses are underestimated due to this disease. Thus, this work aimed to detect and characterize BLV in dairy cattle in the Maranhão state, northeastern Brazil. Blood samples were collected from 176 animals from 8 municipalities in the southeastern state of Maranhão. Bovine blood samples were subjected to DNA extraction and molecular diagnosis using nested PCR assays for BLV, targeting gp51 gene. Positive samples were then sequenced and then subjected to phylogenetic inferences. BLV DNA was detected in 16 cattle (16/176, 9.09%) in 4 municipalities. Phylogenetic analyzes showed that the sequence obtained clustered in a clade containing BLV sequences classified as genotype 6, with a high degree of support. Our data shows BLV occurrence in the Northeast of Brazil and the identification of genotype 6 in this region. These findings contribute to the molecular epidemiology of this agent in Brazil.
Histopathological and spermatogenesis classification by Johnsen is widely used in the germinal epithelium maturation analysis, besides identifying pathological alterations able to cause subfertility and even infertility. The aim of this study was to analyze cell-differentiation histopathological data and to correlate them with expression of PRM-1, TNP-2, 17β-HSD3, LHR, generic MHC-I, MIC-B, NC1 and NC3 genes, involved in bovine spermatogenesis using qRT-PCR from testicular parenchyma. Based on Johnsen’s criteria, the results showed normal spermatogenic activity in these animals, classified at 6, 7 and 8 scores. The qRT-PCR analysis expression showed that MHC-I (generic) gene was less expressed than all the other genes in evaluated scores (p < 0.05) and, PRM-1 and TNP-2 were the most expressed genes (p < 0.05). The PRM-1 gene expression was significantly higher than TNP-2 (p < 0.05). Comparing scores, 17β-HSD3 gene expression was lower (p < 0.05) in score 6 when compared to scores 7 and 8 animals. It was also observed that PRM-1 expression was lower in score 6 when compared to 7, as well as TNP-2 gene was less expressed in the score 6 (p < 0.05) when compared to 7 and 8 scores. Our results demonstrated that MHC I (generic), MIC-B, NC1, NC3, and LHR genes are poorly expressed in bovine testis, suggesting their marginal action on spermatogenesis. Instead, PRM-1, TNP-2, and 17β-HSD3 expression were higher, supporting the notion that these genes can act directly on the germ cells differential development during bovine spermatogenesis.
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