Lymph node metastases are an independent prognosis factor in gastric carcinoma (GC) patients. Radical lymphadenectomy can improve survival but it can also increase surgical morbidity. As a principle, sentinel node (SN) navigation surgery can avoid unnecessary lymphadenectomy without compromising prognosis. In this pilot study, 24 patients with untreated GC were initially screened for SN navigation surgery, of which 12 were eligible. Five patients had T2 tumors, 5 had T3 tumors and 2 had T1 tumors. In 33% of cases, tumor diameter was greater than 5.0 cm. Three hundred and eighty-seven lymph nodes were excised with a median of 32.3 per patient. The SN navigation surgery was feasible in all patients, with a median of 4.5 SNs per patient. The detection success rate was 100%. All the SNs were located in N1 and N2 nodal level. In 70.9% of cases, the SNs were located at lymphatic chains 6 and 7. The SN sensitivity for nodal staging was 91.6%, with 8.3% of false negative. In 4 patients who were initially staged as N0, the SNs were submitted to multisection analyses and immunohistochemistry, confirming the N0 stage, without micrometastases. In one case initially staged as negative for nodal metastases based on SN analyses, metastases in lymph nodes other than SN were found, resulting in a 20% skip metastases incidence. This surgery is a reproducible procedure with 100% detection rate of SN. Tumor size, GC location and obesity were factors that imposed some limitations regarding SN identification. Results from nodal multisection histology and immunohistochemistry analysis did not change initial nodal staging.

Diarrhoea among growing and finishing pigs is an important problem in many herds. The prevalence of L. intracellularis, B. pilosicoli, B. hyodysenteriae, Salmonella spp., enterotoxigenic E. coli, Trichuris suis and the occurrence of mixed infection were investigated. Fecal samples for forty-six herds with diarrhea or a history of diarrhea were randomly collected in Minas Gerais state, Brazil. The enteric pathogens were detected by culture (E. coli and Salmonella sp.), PCR (L. intracellularis and Brachyspira spp.) and eggs counts (T. suis). The overall herd prevalence of L. intracellularis, Salmonella enterica serotype Typhimurium and enterotoxigenic E. coli were 19.56%, 6.52%, 10.86% respectively. Mixed infection was diagnosed in 30.43% of herds, and L. intracellularis and Salmonella enterica serotype Typhimurium are main pathogens association (10.87%). B. pilosicoli was diagnosed only in two herds, always associated with mixed infections. B. hyodysenteriae and T. suis were not demonstrated in any sample. These pathogens have been reported world-wide but studies regarding epidemiology in Brazil are few. This study contributes to establish of prevention programs for the control enteropathogens in grower finish herds in Brazil.

O objetivo deste trabalho foi determinar a prevalência de anticorpos contra linfadenite caseosa (LC) em rebanhos ovinos comerciais do Distrito Federal (DF). Foram coletadas 1.028 amostras de soro entre março e junho de 2004, de todas as propriedades (32) do Distrito Federal com pelo menos 20 fêmeas adultas no rebanho. A soroprevalência da linfadenite caseosa foi determinada por ELISA com proteínas secretadas de Corynebacterium pseudotuberculosis. Cinquenta por cento das 32 propriedades apresentaram pelo menos um animal soropositivo para o LC e a prevalência real para animais foi de 44,0% (IC 95: 41,0; 47,0), portanto, esses dados sugerem que a LC está presente em rebanhos ovinos comerciais no Distrito Federal.

The objective of this study was to determinate the seroprevalence of caseous lymphadenitis (CL) in commercial sheep herds in the Federal District, Brazil. One thousand and twenty-eight serum samples were collected, between March and June 2004, from all the properties (32) with at least 20 adult females in the flock. The seroprevalence of caseous lymphadenitis was determined by ELISA with secreted proteins of Corynebacterium pseudotuberculosis. The real prevalence value for the animals was 44.0% (CI 95: 41.0; 47.0), and 50.0% of the 32 herds had at least one animal seropositive for CL. Therefore, these data show that CL is fairly widespread in commercial flocks of sheep in the Federal District.

A infecção por Brucella ovis é considerada uma das principais causas de epididimite e infertilidade em carneiros, resultando em falhas reprodutivas e perdas econômicas significativas em rebanhos ovinos ao redor do mundo. O estudo teve o objetivo de avaliar três testes sorológicos disponíveis para o diagnóstico da brucelose ovina por B. ovis, utilizando 181 soros ovinos. Amostras de soro provenientes de carneiros experimentalmente infectados foram coletadas ao longo de 192 dias pós-infecção (n=117) e durante o período pré-infecção (n=9). Adicionalmente, amostras de soro foram obtidas de ovinos provenientes de um rebanho livre para B. ovis (n=55). As técnicas de imunodifusão em gel de agar (IDGA), utilizando dois antígenos disponíveis comercialmente, e de fixação de complemento foram comparadas (FC). Foram obtidos resultados de sensibilidade especificidade semelhantes para ambos os métodos de IDGA e ainda, a técnica de IDGA foi mais eficiente do que a da FC para o diagnóstico sorológico da infecção por B. ovis.

A presente atualização trata de duas das mais importantes doenças sexualmente transmitidas de bovinos, a campilobacteriose genital bovina e a tricomonose genital bovina. São abordados aspectos relacionados à epidemiologia destas doenças, principalmente em relação a sua distribuição no Brasil. Também são revisados aspectos importantes de diagnóstico, incluindo as técnicas e interpretação dos resultados, além de medidas de controle para ambas as doenças.

The present update deals with two of the most important sexually transmitted diseases of cattle: bovine genital campylobacteriosis and bovine genital trichomonosis. Epidemiological aspects, mainly their distribution in Brazil, alongside with their diagnosis in cattle are presented and commented. The main points in their diagnoses, including the description of the techniques and the interpretation of the results are also reviewed. Finally the control and prevention of both diseases are discussed.

Foi padronizado um ensaio imunoenzimático do tipo indireto para detecção de imunoglobulina A (ELISA IgA) anti- Campylobacter fetus subp. venerealis em muco cérvico- vaginal bovino utilizando um extrato protéico de Campylobacter fetus subsp. venerealis produzido pelo método de extração ácida pelo tampão de glicina (0,2M; pH2,2). A média dos valores de densidade ótica (DO450) foi de 0,143±0,09. As bandas protéicas dos antígenos de Campylobacter fetus subsp. venerealis e de Campylobacter fetus subsp. fetus melhor reconhecidas pela IgA do muco cérvico- vaginal migraram em 42,6 kDa mas a proteina evidenciada em 93 kDa foi reconhecida exclusivamente pelo Campylobacter fetus subsp. venerealis. Os anticorpos presentes na amostra de muco vaginal testada no "immunoblotting" que apresentou resultado positivo no ELISA IgA, reconheceu antígenos de C. jejuni subsp. jejuni e C. fetus subsp. fetus.

An indirect enzyme-linked immunosorbent assay was developed to detect antigen-specific secretory IgA antibodies to Campylobacter fetus subsp. venerealis in bovine vaginal mucus with a protein extract of the Campylobacter fetus subsp. venerealis by the acid glycine extraction method. Mean optical density measurement (λ=450 nm) was 0.143±0.9. The most immunoreactive protein bands of the Campylobacter fetus subsp. venerealis or Campylobacter fetus subsp. fetus recognized by IgA in immunoblotting, using bovine vaginal mucus samples, migrate at 42.6 kDa. The protein that migrates at 93 kDa was recognized exclusively for C. fetus subsp. venerealis. A positive vaginal mucus sample of a cow from negative herd recognized antigens of C. jejuni subsp. jejuni e C. fetus subsp. fetus.

As estratégias de combate à brucelose bovina são bastante conhecidas e, até o momento, os resultados são divergentes. No Brasil, o Ministério da Agricultura, Pecuária e Abastecimento (MAPA) implementou, em 2001, o Programa Nacional de Controle e Erradicação da Brucelose e Tuberculose. Conhecer a situação epidemiológica da brucelose no início de um programa de controle permite: 1 - escolher as melhores estratégias de controle em função da frequência e padrão de distribuição da doença na população e 2 - acompanhar o programa com a finalidade de promover correções e evitar o desperdício de tempo e de recursos. Em razão disso, é necessário realizar estudos para dar suporte à escolha das melhores estratégias para os vários estados e regiões brasileiras e criar um mecanismo racional de verificação da efetividade das ações implementadas. Para tanto, o MAPA estabeleceu um Termo de Cooperação Técnica com a Faculdade de Medicina Veterinária da Universidade de São Paulo e envolveu também a Faculdade de Agronomia e Veterinária da Universidade de Brasília. Até o momento, foram concluídos os estudos de 15 unidades federativas, cujos resultados são apresentados nos artigos subsequentes. Além disso, há um 16º artigo que explora o impacto da vacinação de bezerras com a B19 na redução da prevalência da brucelose.

Although strategies for controlling and eradicating of bovine brucellosis are well known, the achievements vary widely. In 2001, the Brazilian Ministry of Agriculture, Livestock, and Food Supply (MAPA) started a new National Program for the Control and Eradication of Brucellosis and Tuberculosis (NPCEBT). In the beginning of a brucellosis control program, it is of utmost importance to gain insights into the epidemiological status of the disease in order to: (1) determine the sanitary measures according to the frequency and distribution patterns of the disease in the population; (2) monitor the development of the program and decide when to make adjustments, avoiding the waist of time and resources. As the epidemiological status of bovine brucellosis is not adequately known in Brazil, there was a case for undertaking cross-sectional studies targeted at identifying the best strategies for each state and region, as well as providing a basis for future appraisal of the NPCEBT. For this purpose, MAPA established a collaborative project with FMVZ-Universidade de São Paulo, which also included FAV-Universidade de Brasília. So far, 15 states completed the survey, whose results are described in the following papers. In addition, it is presented a paper that deals with modeling the impact of vaccination of heifers with strain 19 upon the reduction of brucellosis prevalence.

Avaliou-se a freqüência dos genes de fímbrias (K88, K99, 987P, F18 e F41) e toxinas (LT, Stb, StaP e Stx2e) de cepas de E. coli isoladas de leitões com diarréia usando a técnica de PCR multiplex com primers específicos para esses genes, e estudou-se o padrão de sensibilidade das cepas patogênicas pelo método de difusão em disco ao florfenicol, ceftiofur sódico, colistina, fosfomicina, neomicina, norfloxacina, sulfa + trimetoprim, doxiciclina, tetraciclina e lincomicina. Foram utilizadas 144 amostras de E.coli isoladas de leitões com diarréia, provenientes de granjas localizadas no estado de Minas Gerais. Dessas, 42 (29,2%) foram positivas para pelo menos um dos fatores de virulência testados. Dentre essas 42 amostras, 23 (54,8%) apresentaram genes de fímbria e toxina, sete (16,6%) apresentaram somente genes de toxinas e 12 (28,6%) amostras somente genes de fímbria. O resultado do teste de sensibilidade aos antimicrobianos demonstrou que o florfenicol (89,5 %) e o ceftiofur sódico (84,2%) foram as drogas de melhor eficácia in vitro sobre cepas de E. coli com fatores de virulência.

The frequency of virulence determinants genes for fimbrial adhesions (K88, K99, 987P, F18 and F41) and toxins (LT, Stb, StaP and Stx2e) in E. coli strains isolated from diarrheic piglets using the multiplex polymerase chain reaction assay with specific primers for these genes was studied. The antimicrobial sensitivity pattern of pathogenic isolates for florfenicol, sodium ceftiofur, colistin, fosfomycin, neomycin, norfloxacin, sulfa + trimetoprim, doxycycline, tetracycline and lincomycin was also tested using the disk diffusion method. E. coli were isolated from 144 diarrheic piglets from farms in the state of Minas Gerais. Forty-two out of 144 studied samples (29.2%) were positive for at least one tested virulence factor. Out of these 42, 23 samples (54.8%) contained fimbria and toxin genes, seven (16.6%) samples had genes for toxins only and 12 (28.6%) samples just fimbria genes. Disk diffusion in vitro antimicrobial sensitivity test demonstrated the best results for florfenicol (89.5%) and sodium ceftiofur (84.2%) against virulent E. coli strains.

RESUMO Foi realizada a estimativa de freqüência de infecção por Leptospira sp. em bovinos do Estado da Paraíba, Brasil. Seis municípios foram escolhidos ao acaso em cada uma das 3 ecorregiões do Estado: Agreste/Litoral, Cariri/Curimatau e Sertão. Estes 18 municípios foram divididos em quadrantes e uma propriedade foi selecionada por quadrante. As amostras de sangue foram coletadas aleatoriamente de, pelo menos, 32 animais por propriedade. Todos os municípios incluídos no estudo (100%) e 87,75% das propriedades apresentaram no mínimo uma amostra positiva para pelo menos uma das 15 diferentes sorovariedades de Leptospira sp. (16 amostras) testadas. Dos 2.343 animais examinados, 759 reagiram positivamente para pelo menos uma das 16 amostras de Leptospira sp. testadas. Quatrocentos e setenta animais reagiram positivamente para múltiplas amostras. A sorovariedade Hardjo, do sorogrupo Sejroe, foi a mais freqüente com 16,05% (95% IC – 12,69% a 19,41%) dos resultados positivos (376 reações positivas). Houve diferenças na freqüência dos resultados positivos para as amostras da sorovariedade Hardjo testadas: amostra Norma (15,62% 95% IC – 12,38% a 18,86%), isolada no Brasil, e a amostra de referência Hardjoprajitino (2,43% 95% IC – 1,12% a 3,75%). As demais sorovariedades apresentaram freqüências inferiores a 2,8%. Os resultados obtidos no presente estudo confirmam que Leptospira interrogans serovar Hardjo é amplamente distribuída no Estado da Paraíba, Brasil.

ABSTRACT A survey was conducted to estimate the frequency of Leptospira sp. infection in cattle in the State of Paraíba, Brazil. A random sample of 6 municipalities was chosen from each of 3 ecoregions of the state: the Agreste/Litoral, the Cariri/Curimatau and the Sertão. These 18 municipalities were broken down into quadrants and one property was selected at random from each quadrant. Random samples were collected from at least eight animals at each property, in four-age categories, summing at least 32 animals per property. All municipalities that were included in the study (100%) and 87.75% of the properties had at least one positive sample for one of the 15 different Leptospira sp. serovars (16 strains) studied. From 2,343 samples, 759 showed positive reaction to, at least, one of the 16 strains tested. Four hundred and seventy animals were positive to multiple strains. The serovar Hardjo, of the Sejroe serogroup, was the most frequent and accounted for 16.05% (95% CI 12.69% to 19.41%) of the positive results (376 positive reactions). There were differences in the frequency of positive results according to the two strains of serovar Hardjo used: strain Norma (15.62% 95% CI 12.38% to 18.86%), isolated in Brazil, and reference strain Hardjoprajitino (2.43% 95% CI 1.12% to 3.75%). The remaining serovars tested had frequencies lower than 2.8%. Results obtained in the present study confirmed that Leptospira interrogans serovar Hardjo is widely spread in the State of Paraíba, Brazil.

Foi adotado o tratamento intermitente da tuberculose com isoniazida em 240 bovinos de um rebanho naturalmente infectado pelo M. bovis. No início do tratamento 36,6% dos animais foram reagentes positivos e 2,9% inconclusivos. Todos os animais do rebanho receberam isoniazida por via oral em doses de 25mg/kg de peso vivo, administrada três vezes por semana, às segundas, quartas e sextas-feiras, durante 10 meses, perfazendo 120 doses. A cura de 98,9% dos animais tratados foi verificada por meio da dessensibilização alérgica realizada pela tuberculinização e controle bacteriológico de 39 animais abatidos. O tratamento neste rebanho não resultou em seleção de cepas resistentes a isoniazida, comprovada pelo teste de sensibilidade a essa droga, realizado em cultura de M. bovis isolada de um animal não curado.

The intermittent treatment with isoniazid was adopted in 240 bovines from a naturally infected herd. At the beginning of the treatment, 36.6% of the animals were positive reactors and 2.9% inconclusive reactors. The drug was administered orally at a dose of 25mg/kg of live weight, three times a week (on Mondays, Wednesdays and Fridays) during 10 months in a total of 120 doses. The cure of 98.9% of the treated animals was verified by the allergic desensitization demonstrated through the tuberculin test and the bacteriological control of 39 slaughtered animals. The treatment in this herd did not cause selection of isoniazid-resistant strains as demonstrated by the isoniazid sensitivity test carried out in M. bovis culture isolated from a non-cured animal.

Produziu-se uma bacterina de Escherichia coli enterotoxigênica (ETEC) contendo as fímbrias K99 e F41 e avaliaram-se a capacidade de indução de anticorpos anti-K99 e anti F-41 no colostro de vacas vacinadas e no soro de bezerros e a persistência dos anticorpos nos neonatos. Três experimentos foram realizados em duas fazendas comerciais. Os animais foram aleatoriamente alocados em blocos, de duas fêmeas prenhes (uma vacinada e outra controle) e seus respectivos bezerros. No experimento A (fazenda 1), com 18 blocos, os animais receberam uma dose da vacina, 30 dias antes do parto. No experimento B (fazenda 1), com 26 blocos, os animais receberam duas doses de vacina, aos 60 e 30 dias antes do parto. No experimento C (fazenda 2), com 22 blocos, os animais receberam o mesmo esquema de vacinação do experimento B. Nos experimentos A e B foram coletados colostro das parturientes e soro dos bezerros entre 24 e 36 horas de vida. No experimento C, foram usadas novilhas receptoras de embriões e coletados colostro e soro dos bezerros aos 7, 14, 28 e 42 dias de idade. Anticorpos anti-K99 e anti-F41 foram detectados por ELISA utilizando antígenos K99 e F41 purificados. No experimento A não foi observada diferença entre o grupo vacinado e o controle quanto à detecção de anticorpos. No experimento B foi observada diferença (P<0,001) entre o colostro de fêmeas vacinadas e o soro de seus bezerros. No C houve diferença entre o grupo vacinado e o controle para o colostro e o soro dos bezerros aos 7, 14, 28 (P<0,001) e 42 dias de idade (P= 0,003). A bacterina utilizada foi eficiente para a indução de resposta imune humoral detectável.

A bacterin from enterotoxigenic Escherichia coli (ETEC), containing fimbriae K99 and F41, was produced and its capacity to induce anti-K99 and anti-F41 antibodies in colostrum of vaccinated cows and in calf serum, and the persistence of these antibodies in neonates were determined. Three experiments were performed on two commercial farms. In all experiments animals were allotted randomly to the blocks, each block consisting of two pregnant females (a vaccinated one and a control one) and their respective calves. In experiment A (farm 1), comprised of 18 blocks, the animals received a vaccine dose 30 days before delivery. In experiment B (farm 1), consisted of 26 blocks, the animals received two vaccine doses (60 and 30 days before delivery). In experiment C (farm 2), consisted of 22 blocks, the animals received two vaccine doses (60 and 30 days before delivery). In experiments A and B pregnant cows and heifers were used and colostrum and serum from 24- to 36-hour-old calves were collected. In experiment C, pregnant embryo-recipient heifers were used and colostrum and sera from calves at 7, 14, 28 and 42 days of age were collected. Anti-K99 and anti-F41 antibodies were detected by ELISA using purified K99 and F41 fimbrial antigens. In experiment A no difference between treated and control groups was observed for the concentration of anti-K99 and anti-F41 antibodies in colostrum and calf serum. In experiment B a difference (P<0.001) was observed for colostrum of vaccinated females and for serum of their calves. In experiment C, difference between vaccinated and control animals was observed for colostrum and calf serum at 7, 14, 28 (P<0.001 in all cases) and 42 days of age (P= 0.003). The results showed the efficiency of the bacterin to induce detectable humoral immune response.

The frequency of bovine genital campylobacteriosis (BGC) in 157 cows from nine herds from the microregion of Varginha - Minas Gerais - Brazil was evaluated. Farmers use either artificial insemination and natural breeding after two insemination procedures or natural breeding. The diagnosis of BGC was performed by the direct fluorescent antibody test (DFAT) using vaginal mucus. All studied herds showed C. fetus infected animals and, of 157 animals, 40 (25.5%) were positive in DFAT: 7 (26.9%) from herds with natural breeding and 33 (25.1%) from herds with both artificial insemination and natural breeding after the second-to-third insemination. The high frequency of BGC found in this study shows that this disease is present among herds which have reproductive problems and the use of natural breeding after the second-to-third unsuccessful insemination could be a risk factor for the disease.

A comparative tuberculinization test was performed in 1303 animals from seven farms and 266 showed positive to the test. Mycobacterium bovis was isolated from a pool of lymphatic nodes from 14 reagents animals using Stonebrink media and confirmed by biochemical tests.

This report describes the isolation of Mycobacterium bovis from a dog with a history of co-habitation with bufallos infected with Mycobacterium bovis. After necropsy, the microrganism was isolated from a mesenteric lymphatic node in Stonebrink media and bacterial identification was confirmed by biochemical tests.

Cromatografia líqüida de alta eficiência (HPLC) foi utilizada na identificação de isoniazida (hidrazida do ácido isonicotínico) no leite de bovinos tratados com a dose de 25 mg/kg/dia em dias alternados. Também foi verificado o efeito da pasteurização sobre a concentração residual de isoniazida no leite. A excreção da droga no leite apresentou variação cíclica com picos no primeiro dia após a administração da droga, média de 1104,48µg/l, e queda dois dias após, média de 104,12µg/l. Quatro dias após a última administração da droga não foi mais possível a identificação de resíduos de isoniazida no leite. Observou-se influência do peso corporal e da produção de leite sobre a quantidade da droga excretada. A pasteurização diminuiu, em média 47,07%, a concentração de resíduo de isoniazida no leite.

The high pressure liquid chromatography (HPLC) was used for the identification of isoniazid (isonicotinic acid hydrazide) in the milk of cattle treated with a dose of 25 mg/kg/day in alternated days. The effect of milk pasteurization on the isoniazid residue concentration was also studied. The drug excretion presented a cyclic variation, with higher levels in the first day after administration (aa), a mean of 1104.48µg/l, and a decrease two days aa, with a mean of 104.12µg/l. Four days after the last administration of the drug it was not possible to identify residues of isoniazid in the milk of treated animals. Body weight and milk yield influenced the amount of the excreted drug, and pasteurization decreased (mean 47.07%) the concentration of isoniazid residue in milk.