The aim of the present study was to evaluate a species-specific nested PCR based on a previously described species-specific PCR for detection of B. ovis in semen and urine samples of experimentally infected rams. The performance of the species-specific nested PCR was compared with the results of a genus-specific PCR. Fourteen rams were experimentally infected with the Brucella ovis REO 198 strain and samples of semen and urine were collected every week up to 180 days post infection. Out of 83 semen samples collected, 42 (50.6%) were positive for the species-specific nested PCR, and 23 (27.7%) were positive for the genus-specific PCR. Out of 75 urine samples, 49 (65.3%) were positive for the species-specific nested PCR, whereas 11 (14.6%) were genus-specific PCR positive. Species-specific nested PCR was significantly more sensitive (P<0.001) than the genus-specific PCR in semen and urine from experimentally infected rams. In conclusion, the species-specific nested PCR developed in this study may be used as a diagnostic tool for the detection of B. ovis in semen and urine samples from suspected rams.Keywords: Brucella ovis, species-specific, nested PCR, semen, urine de sêmen, 42 (50,6%) foram positivas pela nested e 23 (27,7%) foram positivas pela 49 (65,3%) foram positivas pela nested enquanto 11 (14,6%) PCR, sêmen, urina
RESUMO
O presente estudo objetivou avaliar uma técnica de nested PCR espécie-específica delineada a partir de PCR espécie-específica descrita anteriormente para detecção de B. ovis em sêmen e urina de carneiros infectados experimentalmente. O desempenho da nested PCR espécie-específica foi comparado com os resultados de uma PCR gênero-específica. Quatorze carneiros foram infectados experimentalmente com Brucella ovis REO 198 e amostras de sêmen e de urina foram colhidas semanalmente até 180 dias após a infecção. De 83 amostras
The objective of this study was to evaluate the testicular changes and detect the presence of Leishmania sp. in the testicles and semen of dogs with Visceral Leishmaniasis (VL). The animals were obtained from the Zoonoses Control Department of Teresina, PI, and taken to the kennel of the Agricultural Sciences Center of the Federal University of Piauí, where they remained were maintained for two months and subsequently euthanized for testicles removal. Semen samples were collected from 12 dogs, 06 positive and 06 negative for VL. The following diagnostic techniques readouts were assessed for the sampled animals: testosterone dosage, immunohistochemistry (IMH), histopathology of the slides containing the testicular material, and seminal evaluation by polymerase chain reaction (PCR). Testosterone values remained within the normal range for the canine specie and did not differ statistically among the experimental groups but displayed lower serum concentrations than those of the control group. All the testicular and semen samples from the dogs were negative for VL as determined by techniques IMH and PCR, respectively. The results of testicle histopathology revealed the presence of several lesions with statistical difference among the experimental groups. Parasitized dogs with VL have testicular lesions that may compromise the reproductive efficiency of these animals.
Communication [Comunicação] Adaptation and evaluation of polymerase chain reaction for Brucella ovis detection in semen, urine and organs of rams experimentally infected[Adaptação e avaliação da reação em cadeia da polimerase para detecção de Brucella ovis em sêmen, urina e órgãos de carneiros infectados experimentalmente]
The aim of this study was to detect Brucella spp. in artisanal cheese commercialized in Parnaíba city, Piauí state, Brazil. For this study, 30 samples of curd cheese (500g) were randomly collected from different points in the commercialization process. In the laboratory, 25g aliquots of the samples were suspended in Brucella broth; after this procedure, 10µL aliquots of this suspension were sown in Thayer Martin agar plates that were supplemented with 10% defibrinated sheep blood and VCNT antimicrobials. After inoculation, the samples were incubated at 37°C in microaerophilic conditions for 14 days. The suspected morphological colonies were identified and confirmed by polymerase chain reaction (PCR). From thirty samples of microbiologically analyzed cheeses, six samples (20%) were confirmed by PCR as bacteria from the Brucella genus. The detection of Brucella spp. was confirmed in cheese commercialized in markets or a public square (3.33%), bakery (3.33%) and small market (13.33%). Brucella spp. was detected in artisanal cheese commercialized in different points from Parnaíba city. Guidelines for the establishment of good practices for the production of artisanal cheese should be determined by the competent authorities. However, for the better control of human brucellosis it is necessary to control or eradicate bovine brucellosis in the herds.
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