Leptospira spp. em bovinos leiteiros do estado do Maranhão, Brasil: frequência, fatores de risco e mapeamento de rebanhos reagentes / Leptospira spp. in dairy cattle of Maranhão state, Brazil: frequency, risk factors and mapping of reagent herds

O presente estudo teve como objetivo determinar a frequência, os fatores de risco e o mapeamento de focos em rebanhos leiteiros do estado do Maranhão, Brasil, associados à Leptospira spp. Foram colhidas 420 amostras de soros sanguíneos de fêmeas com idade ≥24 meses selecionadas de forma aleatória simples, distribuídas em 70 rebanhos provenientes de 14 municípios. Foi aplicado um questionário epidemiológico para investigar os principais fatores de riscos associados à infecção. As amostras foram submetidas à prova de soroaglutinação microscópica (SAM), sendo utilizados 24 sorovares de Leptospira spp. Das amostras analisadas, 70 (100%) para rebanhos e 420 (100%) para animais foram reagentes a pelo menos um dos sorovares de Leptospira spp., com títulos variáveis entre 100 e 800. Os sorovares mais prevalentes foram Patoc (410/420; 97%), Castellonis (351/420; 84%), Hardjo (347/420; 83%), Hebdomadis (335/420; 80%), Sentot (328/420; 78%), Wolffi (330/420; 79%), Icterohaemorrhagiae (300/420; 71%) e Pomona (286/420; 69%). As variáveis não realização de inseminação artificial, criação conjunta de caprinos, ovinos, equinos e caninos e maior taxa de nascimento de bezerros no período seco apresentaram associação estatística significativa (p<0,05) à ocorrência de animais sororreagentes para Leptospira interrogans e foram considerados fatores de risco. Os resultados revelaram elevados títulos aglutinantes para diversos sorovares de Leptospira no rebanho bovino leiteiro do estado do Maranhão. Esses resultados demonstram a necessidade de implementação de estratégias que consistem na realização de vacinações, preferencialmente com sorovares regionais, exames sorológicos, aquisição de animais de propriedades idôneas, efetivação de quarentena ao ingresso de novos animais e notificação da doença na Agência de Defesa Agropecuária do Estado.(AU)

The present study's aim was to determine frequency, risk factors and mapping outbreaks in dairy cattle of Maranhão state, Brazil, associated to Leptospira spp. Four hundred and twenty samples of blood serum from females were analyzed, age ≥24 months, selected in a random way, distributed in 70 herds from 14 townships. An epidemiological questionnaire was applied to investigate possible factors that could have been associated to the infection. Serum samples were submitted to the serum agglutination test (SAT). It was used 24 serovars from the Leptospira spp. complex. From the analyzed samples, 70 (100%) for herds and 420 (100%) for animals were reagent to at least one of the Leptospira spp. serovar, reaching titles between 100 and 800. The most prevalent serovars were Patoc (410/420; 97%), Castellonis (351/420; 84%), Hardjo (347/420; 83%), Hebdomadis (335/420; 80%), Sentot (328/420; 78%), Wolffi (330/420; 79%), Icterohaemorrhagiae (300/420; 71%) and Pomona (286/420; 69%). From the variables considered as risk factors, the not utilization of artificial insemination, mixed herds of goats, sheeps, equines and canines in the properties, and the birth of calves in the non-rainy period showed significant statistical association (p<0.05) to the risk of infection with Leptospira interrogans . The results show elevated levels of leptospirosis frequency in dairy cattle herd from Maranhão state. Therefore, there is a necessity to implement strategies consisting of vaccinations with region serovares and serological exams, establishment of quarantine for arrival of new animals, monitoring existent herds in the properties and notification of the disease to the State Animal Health Agency.(AU)

Application of DetectingVibrio Cholerae Combined with Serum Agglutination and Gene Sequencing / 现代检验医学杂志

Objective To avoid false positive detection ofVibrio cholerae and improve the detection correct rate.Methods 1~7 months of 2013 were randomly selected,the national various provinces and cities CDC to China cholera CDC positive screening 14 strains.LB nutrient agar 12 hours,take single colony to Vibrio cholera serum agglutination,extraction of strain DNA at the same time boiled template method.For Vibrio 16SrDNA sequence and design primers for PCR detection of Vib-rio,16SrDNA,electrophoresis were used to observe the 16SrDNA products,16SrDNA positive products sent to sequencing company sequencing,sequencing results were Blast comparison on the NCBI website for the analysis and comparison of ser-um agglutination and Blast alignment.Results 12 strains was positive for agglutination and 2 strains of non agglutination in 14 strains.The Vibrio 16SrDNA amplification,electrophoresis were used to observe the 14 isolates that were amplified frag-ment corresponding,that the selected strains were vibrio.The 16SrDNA positive products were 14 strains,and the sequen-cing the Blast results:2 strains of bacteria were Vibrio harveyi for non agglutination,in 12 positive strains of serum aggluti-nation;1 strains was Vibrio natriegen and 11 strains wereVibrio cholerae .Conclusion Detection ofVibrio cholerae cholerae combined with serum agglutination and gene sequencing can avoid false positive result ofVibrio cholerae ,and improve cor-rect rate of the detection.