RESUMO
Detection of Salmonella sp. is important for the broiler chicken production chain because it is one microorganisms involved in food-borne diseases. Thus, this study performed the optimization of a technique of Loop-mediated isothermal DNA amplification (LAMP) through the 3MTM Molecular Detection Assay 2: Salmonella (MDS®), in accordance with Ordinance number 126 of the Ministry of Agriculture, for the detection of Salmonella sp. in drag swab. The methodology followed ISO 16140-2: 2016, with the analysis naturally contaminated drag swab samples collected from broiler aviaries and artificially contaminated with salmonella ATCCs. Of the 300 samples processed in protocol A (pre-enrichment tetrathionate broth (TT)), 45 were positive for Salmonella sp., 242 negative, one false-positive, and 12 false-negative, while of the 300 samples analyzed in protocol B (pre-enrichment brain-heart infusion broth (BHI)), 40 were positive, 256 negative, one false-positive, and three false-negative. The result for protocol A was a sensitivity of 79%, specificity of 99.6%, Positive Predictive Value (PPV) of 98%, and Negative Predictive Value (NPV) of 95%; and for protocol B, 93% sensitivity, 99.6% specificity, 98% PPV, and 99% NPV. Both protocols were associated with the reference method (p>0.05), concluding that the MDS® can be used for the qualitative detection of Salmonella sp.
Assuntos
Galinhas , Técnicas de Amplificação de Ácido Nucleico , Salmonella , Sensibilidade e Especificidade , Salmonella/isolamento & purificação , Salmonella/genética , Animais , Técnicas de Amplificação de Ácido Nucleico/métodos , Galinhas/microbiologia , Técnicas de Diagnóstico Molecular/métodos , Salmonelose Animal/diagnóstico , Salmonelose Animal/microbiologia , DNA Bacteriano/análise , Reprodutibilidade dos TestesRESUMO
This study was designed to characterize the dynamics of infection of Mycoplasma hyopneumoniae in naïve replacement gilts after introduction to positive systems. Ninety-eight naïve gilts were monitored in three positive commercial farms (A, B, and C). The näive gilts were housed for 21 days in pens adjacently located to older gilt cohorts (named seeders), which have been naturally exposed to the positive farms. The infection dynamics was evaluated by PCR and ELISA, from laryngeal swabs and serum samples, respectively. Samples were collected at 150 (arrival), 165, 180, 210, 240, 270, 300 days of age (doa), and pre-farrowing. Infection occurred rapidly on farms A and B, taking 25.2 and 23.9 days for 95% of gilts to be PCR positive, respectively. There was no influence on the number of seeders at the time of exposure, but their absence (farm C) could explain the extended period it took for gilts to get infected (69.4 days). On average, it took 162.2 days after the first PCR detection for 85% of gilts to stop shedding the bacterium. The serology results were consistent with the herd infection curve. At pre-farrowing, 100% of gilts seroconverted and 36.7% remained PCR positive. A total of 1.33% of piglets were positive at weaning. Fifteen variants were detected among the three farms by MLVA. The acclimation protocol was efficient and easy to perform, and the presence of seeders was likely critical for early acclimation for M. hyopneumoniae.
Assuntos
Mycoplasma hyopneumoniae , Pneumonia Suína Micoplasmática , Doenças dos Suínos , Suínos , Animais , Feminino , Pneumonia Suína Micoplasmática/epidemiologia , Pneumonia Suína Micoplasmática/microbiologia , Mycoplasma hyopneumoniae/genética , Fazendas , Sus scrofa , Reação em Cadeia da Polimerase/veterináriaRESUMO
Chronic pleuritis is the main reason for sending pig carcasses to the Department of Final Inspection (DIF), condemnation and led to economic losses to industries and producers. Most pleura lesions detected after slaughter are sequelae from bacterial infections by agents that do not pose risks to pork consumers. The objective of the present study was to generate science-based information for decision making in the evaluation and destination of chronic pleuritis by the Federal Inspection Service (SIF). Therefore, 200 carcasses, with and without pleurisy, from a swine slaughterhouse with SIF were assessed following the visual classification of the inspection agent. The study was carried out in two stages. In stage 1, 50 carcasses with pneumonic lesions adjacent to chronic pleuritis and 50 carcasses with only chronic pleuritis lesions were evaluated, through macroscopy, histopathology, and bacterial culture. In stage 2, 50 swine carcasses with chronic pleuritis and 50 without this lesion were sampled in the parietal pleura region to bacterial culture and PCR. The economic impact of not exporting these carcasses with chronic pleuritis was also assessed. Considering the stages of evolution of the lesions, the macroscopic examination showed high correlation with the histological examination. There was no bacterial isolation through pleural swabs, regardless of the presence or not of adjacent pulmonary lesions. Isolation was restricted to the adjacent pulmonary lesions of 70% samples, with Pasteurella multocida type A found in 48% of them, followed by P. multocida type D and Streptococcus suis in 12%, and Actinobacillus pleuropneumoniae in 3%. Only Streptococcus suis DNA was detected in 5/100 samples, with no correspondence to the isolation of viable bacteria. The reliability demonstrated in the macroscopic evaluation carried out during inspection, the absence of viable bacteria in the chronic pleural lesions, and the negative economic impact suggest that carcasses with chronic pleuritis can be submitted to pleura removal, with no need of sending to DIF.
Pleurite crônica é a principal causa do desvio de carcaças de suínos para o Departamento de Inspeção Final (DIF), podendo causar condenação e prejuízos econômicos às indústrias e produtores. A maioria das lesões de pleura detectadas após o abate são sequelas de infecções bacterianas por agentes que não oferecem riscos aos consumidores de carne suína. O objetivo do presente estudo foi gerar informações científicas para a tomada de decisão na avaliação e destino da pleurite crônica pelo Serviço de Inspeção Federal (SIF). Para tanto, 200 carcaças, com e sem pleurisia, provenientes de um frigorífico de suínos com SIF foram avaliadas seguindo a classificação visual do agente fiscalizador. O estudo foi realizado em duas etapas. No estágio 1, 50 carcaças com lesões pneumônicas adjacentes à pleurite crônica e 50 carcaças apresentando somente lesões de pleurite crônica foram avaliadas macroscopicamente, por histopatologia e cultura bacteriana. No estágio 2, 50 carcaças suínas com pleurite crônica e 50 sem esta lesão foram amostradas na região da pleura parietal para cultura bacteriana e PCR. O impacto econômico de não exportar essas carcaças com pleurite crônica também foi avaliado. Considerando os estágios de evolução das lesões, o exame macroscópico apresentou alta correlação com o exame histológico. Não houve isolamento bacteriano por meio de swabs pleurais, independentemente da presença ou não de lesões pulmonares adjacentes. O isolamento foi restrito às lesões pulmonares adjacentes de 70% das amostras, sendo Pasteurella multocida tipo A encontrado em 48% delas, seguido por P. multocida tipo D e Streptococcus suis em 12%, e Actinobacillus pleuropneumoniae em 3%. Apenas DNA de Streptococcus suis foi detectado em 5/100 amostras, sem correspondência com o isolamento de bactérias viáveis. A confiabilidade demonstrada na avaliação macroscópica realizada durante a inspeção, a ausência de bactérias viáveis nas lesões pleurais crônicas e o impacto econômico negativo sugerem que carcaças com pleurite crônica podem ser submetidas à remoção da pleura, sem necessidade de envio para DIF.
Assuntos
Animais , Pleurisia/economia , Pleurisia/patologia , Pleurisia/veterinária , Doenças dos Suínos/economia , Doenças dos Suínos/microbiologia , Sus scrofa , Brasil , Matadouros , Fiscalização SanitáriaRESUMO
On-farm culturing is a technology booming in Brazil. It is based on the microbiological diagnosis of milk samples collected from clinical cases of bovine mastitis, on the farm where the animals are handled, quickly, simply, and at an affordable cost. With this resource, after 24 h of incubation, the isolated microorganisms are classified as gram positive or gram negative, helping to make decisions regarding the therapy of the animals. Considering the relevance of the Midwestern region of Santa Catarina State in the national dairy production scenario, the objectives of the study were: 1) to promote the technical training of the farmers in three dairy farms located in Treze Tílias-SC, 2) to assist in the implementation and evaluation of on-farm culturing as technological resource viewing the reduced use of antimicrobials in herds. In addition to the rapid acceptance by the farmers and their efficient training, with the use of the technology, there was a 45-50% reduction in the use of antimicrobials in dairy herds. This result demonstrated a significant effect on animal health, and determines an important cost reduction for farmers, as presented in detail in this manuscript.
A cultura de leite na fazenda (do inglês On farm culturing) é uma tecnologia em franca expansão no Brasil. Fundamenta-se no diagnóstico microbiológico de amostras de leite colhidas de casos clínicos de mastite bovina, na própria fazenda onde são manejados os animais, de forma rápida, simples e com custo acessível. Com este recurso, após 24 horas de incubação, os micro-organismos isolados são classificados em Gram-positivos ou Gram-negativos, auxiliando na tomada de decisões quanto à terapia dos animais. Considerando a relevância da região meio oeste catarinense no cenário nacional de produção leiteira, o objetivo do presente estudo foi promover a capacitação técnica dos produtores em três propriedades localizadas em Treze Tílias-SC, e auxiliar na implementação e avaliação da cultura de leite na fazenda como recurso tecnológico voltado ao uso reduzido de antimicrobianos nos rebanhos. Além da rápida aceitação pelos produtores e eficiente capacitação dos mesmos, verificou-se, com o emprego da tecnologia, redução de 45% a 50% do uso de antimicrobianos nos rebanhos. Este resultado, além de demonstrar significativo impacto em saúde animal, determinou importante redução de custos aos produtores, como apresentado detalhadamente no presente artigo.
Assuntos
Feminino , Animais , Bovinos , Doenças dos Bovinos/tratamento farmacológico , Fortalecimento Institucional/economia , Leite/microbiologia , Mastite Bovina/diagnóstico , Mastite Bovina/prevenção & controle , Agroindústria/economiaRESUMO
On-farm culturing is a technology booming in Brazil. It is based on the microbiological diagnosis of milk samples collected from clinical cases of bovine mastitis, on the farm where the animals are handled, quickly, simply, and at an affordable cost. With this resource, after 24 h of incubation, the isolated microorganisms are classified as gram positive or gram negative, helping to make decisions regarding the therapy of the animals. Considering the relevance of the Midwestern region of Santa Catarina State in the national dairy production scenario, the objectives of the study were: 1) to promote the technical training of the farmers in three dairy farms located in Treze Tílias-SC, 2) to assist in the implementation and evaluation of on-farm culturing as technological resource viewing the reduced use of antimicrobials in herds. In addition to the rapid acceptance by the farmers and their efficient training, with the use of the technology, there was a 45-50% reduction in the use of antimicrobials in dairy herds. This result demonstrated a significant effect on animal health, and determines an important cost reduction for farmers, as presented in detail in this manuscript.(AU)
A cultura de leite na fazenda (do inglês On farm culturing) é uma tecnologia em franca expansão no Brasil. Fundamenta-se no diagnóstico microbiológico de amostras de leite colhidas de casos clínicos de mastite bovina, na própria fazenda onde são manejados os animais, de forma rápida, simples e com custo acessível. Com este recurso, após 24 horas de incubação, os micro-organismos isolados são classificados em Gram-positivos ou Gram-negativos, auxiliando na tomada de decisões quanto à terapia dos animais. Considerando a relevância da região meio oeste catarinense no cenário nacional de produção leiteira, o objetivo do presente estudo foi promover a capacitação técnica dos produtores em três propriedades localizadas em Treze Tílias-SC, e auxiliar na implementação e avaliação da cultura de leite na fazenda como recurso tecnológico voltado ao uso reduzido de antimicrobianos nos rebanhos. Além da rápida aceitação pelos produtores e eficiente capacitação dos mesmos, verificou-se, com o emprego da tecnologia, redução de 45% a 50% do uso de antimicrobianos nos rebanhos. Este resultado, além de demonstrar significativo impacto em saúde animal, determinou importante redução de custos aos produtores, como apresentado detalhadamente no presente artigo.(AU)
Assuntos
Animais , Feminino , Bovinos , Doenças dos Bovinos/tratamento farmacológico , Mastite Bovina/diagnóstico , Mastite Bovina/prevenção & controle , Leite/microbiologia , Fortalecimento Institucional/economia , Agroindústria/economiaRESUMO
Dairy farming is an important social and economic activity in the Midwest region of Santa Catarina State, Brazil. However, despite technological advances in the productive chain, mastitis is still considered the main disease of dairy herds, due its high economic impact. Bacterial pathogens are the main cause of this disease and failures in the treatment of clinical cases are attributed in part to the antimicrobial resistance of these pathogens. As an alternative, in vitro antimicrobial susceptibility tests of strains isolated from bovine mastitis cases have been carried out; the information generated contributes to strategic treatment measures, and also to the control of microorganism resistance. Therefore, the aims of the present study were to evaluate the etiology and in vitro antimicrobial susceptibility profiles of bacterial strains isolated from cases of bovine mastitis in dairy herds from the Midwest region of Santa Catarina State, Brazil. For this purpose, 345 milk samples were collected from 70 herds located in 16 different municipalities. The main pathogens isolated were: staphylococci (55.9%), streptococci (34.8%), Corynebacterium spp. (7.8%) and Escherichia coli (1.4%). The resistance rates of staphylococcus strains to ampicillin and to gentamicin were lower (24.9%) and higher (33%), respectively, than those obtained in other Brazilian studies. The resistance profiles of streptococcus strains were similar to those described in the literature, while Corynebacterium spp., showed high resistance rates to tetracycline (85.2%). Moreover, a considerable prevalence of staphylococcus (27.5%) and streptococcus (11.7%) multiresistant strains was verified. These data reinforce the need for regional characterization of microorganisms.
Assuntos
Animais , Bovinos , Farmacorresistência Bacteriana , Mastite Bovina/etiologia , Mastite Bovina/patologia , Brasil , Corynebacterium , Staphylococcus , StreptococcusRESUMO
Dairy farming is an important social and economic activity in the Midwest region of Santa Catarina State, Brazil. However, despite technological advances in the productive chain, mastitis is still considered the main disease of dairy herds, due its high economic impact. Bacterial pathogens are the main cause of this disease and failures in the treatment of clinical cases are attributed in part to the antimicrobial resistance of these pathogens. As an alternative, in vitro antimicrobial susceptibility tests of strains isolated from bovine mastitis cases have been carried out; the information generated contributes to strategic treatment measures, and also to the control of microorganism resistance. Therefore, the aims of the present study were to evaluate the etiology and in vitro antimicrobial susceptibility profiles of bacterial strains isolated from cases of bovine mastitis in dairy herds from the Midwest region of Santa Catarina State, Brazil. For this purpose, 345 milk samples were collected from 70 herds located in 16 different municipalities. The main pathogens isolated were: staphylococci (55.9%), streptococci (34.8%), Corynebacterium spp. (7.8%) and Escherichia coli (1.4%). The resistance rates of staphylococcus strains to ampicillin and to gentamicin were lower (24.9%) and higher (33%), respectively, than those obtained in other Brazilian studies. The resistance profiles of streptococcus strains were similar to those described in the literature, while Corynebacterium spp., showed high resistance rates to tetracycline (85.2%). Moreover, a considerable prevalence of staphylococcus (27.5%) and streptococcus (11.7%) multiresistant strains was verified. These data reinforce the need for regional characterization of microorganisms.(AU)
Assuntos
Animais , Bovinos , Mastite Bovina/etiologia , Mastite Bovina/patologia , Farmacorresistência Bacteriana , Corynebacterium , Staphylococcus , Streptococcus , BrasilRESUMO
Protection against experimental fasciolosis in rats immunized with recombinant myosin regulatory light chain (MRLC) in TiterMax Gold® adjuvant was assessed. The experimental trial consisted of four groups of 15 animals; group 1 was unimmunized and infected, group 2 was immunized with MRLC in adjuvant and infected, group 3 was infected and immunized with adjuvant only and group 4 was unimmunized and uninfected. Immunization with MRLC in TiterMax Gold® adjuvant (group 2) induced a reduction in fluke burdens of 51.0% (p<0.001) when compared with the adjuvant control group, and 61.5% (p<0.001) when compared with the unimmunized infected controls. There was a reduction in fecal egg output in group 2 of 44.8% and 37.3% compared with group 1 and group 3, respectively; although this difference was not statistically significant. Measurement of cytokine levels revealed higher levels of TNF-alpha and IL-2 as well as lower levels of IL-4 in group 2 during the chronic stage of infection (p<0.05), along with higher levels of IFN-gamma during early stages of infection (p<0.05). These results suggest a mixed Th1/Th2 phenotype immune response; however predominance of Th1 cytokines was observed. Levels of anti-MRLC serum IgG in group 2 were significantly higher than controls at the time of euthanasia (p<0.05). This is the first report of immunization with recombinant MRLC in rats, demonstrating that this antigen significantly reduces fluke burdens, increases the Th1 immune response and encourages further studies to improve the vaccine's efficacy.
Assuntos
Adjuvantes Imunológicos/administração & dosagem , Antígenos de Helmintos/imunologia , Fasciolíase/prevenção & controle , Cadeias Leves de Miosina/imunologia , Poloxaleno/administração & dosagem , Vacinas Sintéticas/imunologia , Animais , Anticorpos Anti-Helmínticos/sangue , Antígenos de Helmintos/genética , Citocinas/metabolismo , Modelos Animais de Doenças , Fezes/parasitologia , Imunoglobulina G/sangue , Masculino , Cadeias Leves de Miosina/genética , Contagem de Ovos de Parasitas , Carga Parasitária , Ratos Wistar , Proteínas Recombinantes/genética , Proteínas Recombinantes/imunologia , Células Th1/imunologia , Células Th2/imunologia , Vacinas Sintéticas/administração & dosagem , Vacinas Sintéticas/genéticaRESUMO
Genomic fragments of the HN and L genes from Brazilian bovine parainfluenza 3 virus (bPIV-3) isolated as contaminants from cell cultures and clinical specimens were amplified by reverse transcription-polymerase chain reaction (RT-PCR), sequenced using specific degenerate primers and analyzed by phylogenetic comparison with reference strains of bPI3V. The Brazilian isolates revealed a high degree of genomic when compared to SF4/32 prototype strain, within the recently proposed genotype A of bPIV-3.
Assuntos
Animais , Bovinos , Sequência de Bases , Técnicas In Vitro , Filogenia , Infecções por Respirovirus , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Transcrição Reversa , /isolamento & purificação , /patogenicidade , Genótipo , Métodos , Métodos , Medicina VeterináriaRESUMO
A 2.4-kb phi29 polymerase amplification product from serum of a diseased chicken was cloned and sequenced. The 2383-nucleotide sequence showed about 40% identity to a representative genome of chicken anemia virus (CAV), the only member of the genus Gyrovirus, family Circoviridae. The new genome had an organization similar to that of CAV: a putative 5' untranscribed region of about 400 nt followed by three partially overlapping open reading frames encoding VP1, VP2 and VP3 homologs. The amino acid identities between these homologs and those of CAV were 38.8%, 40.3%, and 32.2%, respectively. Based on these limited similarities, it is proposed that the newly identified virus is a member of a new species in the genus Gyrovirus. For this new species, the name Avian gyrovirus 2 (AGV2) is proposed.