FIRST ISOLATION OF Leptospira kirschneri SEROVAR CANICOLA AND Leptospira interrogans SEROVAR PYROGENES IN URINE SAMPLES FROM SLAUGHTERED CATTLE IN MIDWEST REGION OF SÃO PAULO STATE, BRAZIL

A leptospirose é uma zoonose de grande importância em saúde pública. Cento e quatro (104) amostras de urina de bovinos abatidos na região centro-oeste do Estado de São Paulo, Brasil, foram avaliadas para detectar Leptospira spp. As amostras foram mantidas em PBS e em culturas de EMJH e Fletcher. Observamos crescimento de microrganismos em 13 amostras do meio Fletcher, embora não tenha sido observado no meio EMJH. Por PCR, 25 animais (amostras) foram positivos com os primers LEP1 / LEP2, sendo que 16 amostras foram mantidas em PBS, seis em meio Fletcher e três em meio EMJH. As 25 amostras foram submetidas a PCR com o gene LipL32 para verificação da patogenicidade. Uma amostra amplificada e por sequenciamento resultou em 99,77% de similaridade com Leptospira kirschneri sorovar Canicola e Leptospira interrogans sorovar Pyrogenes. Este é o primeiro relato de isolamento de Leptospira kirschneri sorovar Canicola e Leptospira interrogans sorovar Pyrogenes em amostra de urina bovina na Região Centro-Oeste do estado de São Paulo, Brasil. Concluímos a importância da leptospirose no rebanho avaliado e a atenção para o risco ocupacional, devido à possibilidade de urina contaminada nas pastagens e disseminação da infecção para outros animais e pessoas contactantes.Leptospirosis is a zoonosis of great importance in public health. One hundred and four urine samples from slaughtered cattle in Midwest region of São Paulo State, Brazil were evaluated to detect Leptospira spp. The samples were kept in PBS and in cultures of EMJH and Fletcher. We observed growth of microorganisms in 13 samples of Fletcher medium, although it was not observed in EMJH medium. By PCR, 25 animals (samples) were positive with LEP1/LEP2 primers, which of them 16 samples were kept in PBS, six samples in Fletcher medium and three in EMJH medium. The 25 samples were submitted to PCR with the LipL32 gene to verify the pathogenicity. One sample amplified and by sequencing resulted in 99.77% of similarity to Leptospira kirschneri serovar Canicola and Leptospira interrogans serovar Pyrogenes. This is the first report of isolation of Leptospira kirschneri serovar Canicola and Leptospira interrogans serovar Pyrogenes from bovine urine sample in the Midwest Region of São Paulo state, Brazil. We concluded the importance of leptospirosis in the herd evaluated and the attention for the ocupacional risk, because of the possibility of contaminated urine in the pastures and the spread of infection to other animals and contactants persons

Epidemiologia molecular aplicada ao estudo de estirpes de Staphylococcus aureus na produção de queijo tipo Minas Frescal

Foi realizado o estudo epidemiológico molecular de estirpes de Staphylococcus aureus potencialmente toxigênicas isoladas no processo de produção do Queijo Tipo Minas Frescal em uma micro-usina no Estado de São Paulo. Para tanto, foram realizadas seis amostragens durante o período de junho de 2008 a julho de 2009, de modo a perfazer um total de 140 amostras, as quais foram colhidas do leite cru e pasteurizado, mãos do manipulador, queijo embalado para consumo e diversos pontos do fluxograma de seu processamento. A confirmação molecular da espécie dos isolados e a presença de genes codificadores das enterotoxinas SEA, SEB, SEC, SED e da toxina TSST-1, foram realizadas a partir da amplificação dos fragmentos de DNA genômico específico. Entre as 41 (29,3,4%) das estirpes confirmadas como sendo S. aureus, 25 (61%) mostraram-se positivas na pesquisa de genes codificadores de toxinas estafilocócicas. Os pontos de maior frequência de estirpes de S. aureus toxigênico foram as mãos do manipulador (16,0%), o leite cru do tanque de recepção (12,0%), o leite pasteurizado (12,0%) e o Queijo Tipo Minas Frescal pronto para consumo (12,0%). O gene codificador de enterotoxina de maior frequência identificada foi o sea. Na determinação do potencial toxigênico das estirpes com genes codificadores de enterotoxinas por imunodifusão, apenas uma estirpe isolada da superfície interna do tanque de estocagem do leite cru foi produtora da enterotoxina SEC. Houve variabilidade genética entre as 41 estirpes de S. aureus isoladas, uma vez que foram identificados 22 pulsotipos diferentes pelo PFGE. Os resultados obtidos permitem a adoção de medidas para a melhoria do processamento do Queijo Tipo Minas Frescal de modo a reduzir o risco potencial que estas toxinas podem representar para a saúde da população consumidora deste produtoWe performed molecular epidemiological study of strains of Staphylococcus aureus isolated toxigenic potential in the process of producing the Frescal Minas Type Cheese in a micro-mill in State of São Paulo. For this, six samples were taken during the period June 2008 to July 2009 in order to make a total of 140 samples, which were collected raw and pasteurized milk, the handler's hands, cheese packaged for consumption and different points flowchart of processing. Confirmation of the molecular species of the isolates and the presence of genes encoding enterotoxins SEA, SEB, SEC, SED and TSST-1 toxin, were made from the amplification of specific fragments of genomic DNA. Among the 41 (29.3%) of strains confirmed as S. aureus, 25 (61%) were positive in the research of genes encoding staphylococcal toxins. Points higher frequency of toxigenic strains of S. aureus were the hands of the handler (16.0%), raw milk receiving tank (12.0%), pasteurized milk (12.0%) and Frescal Minas Type Cheese ready for consumption (12.0%). The gene encoding enterotoxin most frequently identified was the sea. In determining the potential toxigenic strains with genes encoding enterotoxins by immunodiffusion, only one strain isolated from the inside surface of the storage tank of raw milk was the producer of enterotoxin SEC. There was genetic variability among the 41 strains of S. aureus isolated, since they were identified by 22 different PFGE pulsotypes. The results obtained allow the adoption of measures to improve the processing of Frescal Minas Type Cheese to reduce the potential risk that these toxins can pose to the health of consumers of this productCoordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES

Associação deagentes patogênicos isolados emanálise microbiológica da água, coma presença de mastite clínica ou subclínica, em vacas de propriedades leiteiras da região de Cerqueira César - SP

Objetivou-se, neste trabalho, pesquisar a relação entre os microrganismos patogênicos isolados e identificados em água utilizada na ordenha, com o isolamento e identificação dos mesmos em amostras de leite, de quartos mamários apresentando mastite clínica ou subclínica nas mesmas propriedades. Foram utilizadas 16 propriedades rurais leiteiras, escolhidas aleatoriamente, na região de Cerqueira César - SP, que utilizavam ordenha mecânica. A água utilizada na ordenha foi classificada em relação à presença de coliformes totais e fecais, como dentro dos padrões ou fora dos padrões de potabilidade humana. Nos resultados obtidos, 94% das amostras foram classificadas como fora dos padrões em relação a coliformes totais e fecais. Os microrganismos identificados foram: Escherichia coli (51%), Enterobacter spp. (25%), Enterobacter cloacae (8%) Edwardsiella tarda (8%) e Klebsiella oxytoca (8%). em relação ao leite, foram analisadas 373 amostras provenientes de vacas em lactação, com mastite clínica (n=19; 5%) e subclínica (n=354; 95%). Os animais com mastite subclínica foram identificados pela contagem de células somáticas (CCS), utilizando-se o aparelho eletrônico (Somacount 300, Bentley), onde a média observada foi de 1.631 x 10³ células/mL. Os principais microrganismos identificados foram: Staphylococcus aureus (30%), Corynebacterium bovis (23%) e Staphylococcus spp. (15%). Conforme os dados obtidos, os agentes coliformes encontrados na água, utilizada na ordenha, não estavam presentes nas análises das amostras de leite dos quartos mamários com mastite clínica ou subclínica das respectivas propriedades, demonstrando não haver associação entre a qualidade da água e a ocorrência de mastite.The aim of the present study was to research the relation between the isolated and identified pathogenic microorganisms in the water used in the milking, with the isolation and identification of the same in milk samples of teats showing clinical or subclinical mastitis. Sixteen dairy farms were randomly chosen, in Cerqueira Cesar town - SP, which used mechanical milking. Water of the farms was classified in relation to the presence of total and fecal coliforms such as in the standard or out of the standard of human potability. In the results obtained, 94% of the samples were classified as being out of the standards in relation to total and fecal coliforms. The identified microorganisms were Escherichia coli (51%), Enterobacter spp. (25%), Enterobacter cloacae (8%), Edwardsiella tarda (8%) and Klebsiella oxytoca (8%). Regarding the milk samples, 373 samples from suckling cows were analyzed, presenting clinical mastitis (n=19; 5%) and subclinical mastitis (n=354; 95%).Animals presenting subclinical mastitis were identified by somatic cell count (SCC), utilizing electronic equipment (Somacount 300 - Bentley), where the mean found was 1,631 x 10³ cells/mL. The main identified microorganisms were Staphylococcus aureus (30%), Corynebacterium bovis (23%) and Staphylococcus spp. (15%). According to the results obtained, coliform agents found in the water used in the milking were not present in the analysis of the milk samples of quarters presenting clinical or subclinical mastitis from the respective farms, showing that there was not an association between water quality and mastitis occurrence

Degradabilidade in situ do híbrido de sorgo e do capim-tanzania em vacas suplementadas no outono

O experimento foi conduzido na Agência Paulista de Tecnologia dos Agronegócios, em Ribeirão Preto, com objetivo de avaliar o efeito da suplementação concentrada na degradabilidade in situ dos colmos e das folhas do híbrido de sorgo e do capim-tanzânia colhidos no outono com e sem suplementação de concentrado em vacas. Amostras de folhas e colmos das gramíneas colhidas no início do outono foram incubadas em quatro vacas mestiças canuladas no rúmen, avaliando-se a fração solúvel, fração potencialmente degradável, taxa de degradação (kd) e a degradabilidade efetiva (DE), utilizando-se o delineamento em quadrado latino (4x4). Houve diferença (P<0,001) da fração solúvel do colmo entre as gramíneas, e o híbrido de sorgo apresentou maior resultado. Não houve efeito dos tratamentos (P=0,52) na taxa de degradação (kd), mas houve diferença (P<0,0001) entre os tratamentos para as degradações efetivas a 2%/h, 5%/h, e 8%/h, sendo as degradabilidades efetivas do colmo do capim-tanzânia menores que do híbrido de sorgo. Os resultados observados para fração solúvel, fração potencialmente degradável e DE com taxa de passagem de 2%/h para o hibrido de sorgo foram em média 25,12%; 48,78% e 52,5%/h para colmo e 18,6%; 64,54% e 56,9%/h para as folhas, respectivamente. Para o capim-tanzânia as médias obtidas foram de 12,27%; 43,33% e 36,6%/h para colmo e 14,96%; 60,95% e 54,3%/h para as folhas, respectivamente. A fração solúvel do colmo e das folhas é maior no híbrido de sorgo, e a degradabilidade efetiva é maior apenas no colmo do sorgo. O uso de suplementação concentrada não interfere na degradabilidade ruminal quando os animais consomem folhas e colmos de híbrido de sorgo ou capim-tanzânia em sistema de pastejo no outono

Teor de ácido cianídrico e taxa de crescimento do híbrido de sorgo x capim sudão durante o outono

In central Brazil after soybean or other annual agricultural species is harvested, sorghum hybrids are planted in the fall in order to establish pastures for grazing animals. This study conducted for two consecutive years aimed at quantifying the contents of hydrocyanic acid in the leaves and determining plant height, forage dry matter yield and the leaf/stem ratio for sorghum hybrid 1P400 at different ages. Statistical analysis was performed by regression analysis based on plant age. Leaf HCN content decreased with plant growth, ranging from 205.0 and 230.3 mg HCN/100 g leaf DM at two weeks old to 5.9 and 6.1 mg HCN/100 g leaf DM at five weeks old in the first and second year, respectively. The average heights measured varied from 60 and 56 cm, in the 4th week, to 117 and 151 cm, in the 8th week, during the first and second experimental year, respectively. Forage mass increased linearly with age and displayed average of 1.411 and 1.637 kg DM/ha in the first year and, 2.905 and 3.640 kg DM/ha in the second year, during the 7th and 8th week, respectively. Leaf proportion decreased while stem increased linearly with plant age. The leaf/stem ratio decreased with plant growth, elongation and increasing stem weight. The sorghum hybrid should be grazed only after five weeks or when the plant height is above 80 cm, in order to avoid the risk of cyanide poisoning.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP

Performance and rumen parameters of crossbred dairy cows fed two sugarcane varieties combined or not with soybean hulls

Two sugarcane varieties (IAC86-2480 and IAC91-2195) combined or not with soybean hulls (SH), partially replacing corn in concentrate, were included in the diet of crossbred dairy cows to evaluate the effects on feed intake, milk production, milk composition and rumen parameters of crossbred cows. The sugarcane varieties (IAC86-2480 and IAC91-2195) and concentrate compositions (with and without SH) were analyzed in a 2 × 2 factorial arrangement. Treatments were assigned to blocks by lactation period to evaluate feed intake, milk production and milk composition. A Latin square design was used to analyze rumen parameters. Feed intake, milk production and composition were similar among treatments. Average daily feed intake was 19.93 kg DM in cows fed IAC86-2480 sugarcane and 17.98 kg DM in those fed IAC91-2195, and their average milk production was 19.27 kg and 18.94 kg, respectively. The treatments with IAC86-2480 and IAC91-2195 had different effects on ruminal pH (6.47 and 6.61, respectively) and ammoniacal nitrogen (12.88 and 16.57 mg/dL, respectively). Sugarcane variety and concentrate composition had an interaction effect on volatile fatty acid levels and acetic acid/propionic acid ratio (93.62 mM and 2.54 for IAC86-2480 with SH; 106.70 mM and 2.41 for IAC86-2480 without SH; 115.70 mM and 3.30 for IAC91-2195 with SH; and 93.21 mM and 1.81 for IAC91-2195 without SH, respectively). Sugarcane variety and soybean hull inclusion in feed concentrate do not affect feed intake, milk production or composition in crossbred cows, although these variables change fatty acid production and ruminal pH

EPIDEMIOLOGIA MOLECULAR APLICADA AO MONITORAMENTO DE ESTIRPES DE Staphylococcus aureus NA PRODUÇÃO DE QUEIJO MINAS FRESCAL

We studied the molecular epidemiology of Staphylococcus aureus strains potentially toxigenic, isolated from the production process of Minas frescal cheese in a small dairy plant in the state of São Paulo. For this, samples were taken during the period from June 2008 to July 2009. Samples were collected from the surface of the receiving and storage tanks of raw milk, the surface of the balance tank of pasteurized milk, the water supply system, the pipes and equipments, the hands of the handler and from the packaged cheese, totaling 140 samples. The colonies isolated on Baird-Parker Agar confirmed as Gram positive and positive for catalase, coagulase and acetoin production, were submitted to extraction of bacterial DNA using the Invitek - Uniscience® kit. Confirmation of the isolated species and enterotoxins SEA, SEB, SEC, SED and TSST-1 toxin was carried out through the amplification of specific fragments of chromosomal DNA. Among the 74 strains of isolated coagulase-positive staphylococci, only 41 (55.4%) strains were confirmed as Staphylococcus aureus, of which 25 (61.0%) were positive to the presence of staphylococcal toxins. The most frequently identified enterotoxin was SEA. The toxigenic strains of Staphylococcus aureus were more frequently isolated from hands of the handler (16.0%), raw milk receiving tank (12.0%), pasteurized milk for cheese making (12.0%) and fresh white cheese ready for consumption (12.0%)