Antimicrobial susceptibility profile of Streptococcus agalactiaestrains isolated from bovine mastitis / Perfil de susceptibilidade a antimicrobianos em Streptococcus agalactiae isolados de mastite bovina

Bovine mastitis is the most important disease of dairy herds worldwide. Its main etiologic agents are bacteria, including Streptococcus agalactiae. The importance of this agent in bovine mastitis is because it is highly contagious and has a high impact on the occurrence of clinical mastitis cases and in the increase of the bulk milk somatic cell counts. The dry cow therapy and the treatment of the clinical mastitis cases stand out among the measures to control intramammary infections in cows. However, these strategies require knowledge about the antimicrobial susceptibility of the causal microorganisms. Thus, this study aimed to evaluate the antimicrobial susceptibility of 89 S. agalactiae strains isolated from bovine mastitis between the years 2004 and 2008 in dairy herds from Campo das Vertentes region, Minas Gerais State, Brazil. The disc diffusion technique was used and the antimicrobials currently used in mastitis therapy were tested. The isolates tested showed 100% susceptibility to chloramphenicol, ceftiofur, cefotaxime, enrofloxacin, and cefquinome. High frequencies of susceptibility (>95%) were also observed for the beta-lactams (penicillin G, ampicillin, and oxacillin), cephalosporins (cephalotin, ceftiofur, cefotaxime, cefoperazone, and cefquinome), florfenicol, gentamicin, lincomycin, nitrofurantoin, and sulfamethoprim. The strains showed high frequencies of resistance to neomycin (15.74%), and tetracycline (21.35%). Multidrug resistance was detected in 2.25% of the tested isolates. The results pointed to variations in the antimicrobial susceptibility profiles of the studied strains and the importance of the use of the susceptibility tests to determine the correct antimicrobial to be applied in the treatment of bovine mastitis caused by S. agalactiae. The high frequencies of resistance observed to some antimicrobials, such as neomycin and tetracycline, commonly used in the treatment of mastitis and other pathologies, highlighted the need for more judicious use of antimicrobials on dairy farms.(AU)

A mastite é a principal doença de bovinos leiteiros em todo o mundo e tem como principais agentes as bactérias, entre as quais Streptococcus agalactiae. Esse agente se destaca por ser altamente contagioso e pelos reflexos que causa na incidência de casos clínicos e no incremento da contagem de células somáticas do leite do tanque. Para o controle desta enfermidade, destacam-se a terapia de vacas secas e o tratamento de casos clínicos, medidas que requerem o conhecimento do perfil de sensibilidade dos agentes causais aos antimicrobianos. Este estudo teve como objetivo avaliar os perfis de suscetibilidade a antimicrobianos em 89 amostras de S. agalactiae isoladas de casos de mastite bovina em rebanhos da região de Campo das Vertentes, Minas Gerais, no período de 2004 a 2008. A técnica de difusão em discos foi utilizada e os antimicrobianos correntemente empregados na terapia da mastite foram testados. Os testes de suscetibilidade antimicrobiana apontaram 100% de susceptibilidade para cloranfenicol, ceftiofur, cefotaxima, enrofloxacina e cefquimona. Níveis elevados de susceptibilidade (>95%) foram observados para os betalactâmicos, florfenicol, gentamicina, lincomicina, nitrofurantoína e sulfametoprim. Altas frequências de resistência foram observadas para neomicina (15,74%) e tetraciclina (21,35%). Dois isolados multirresistentes (2,25%) foram encontrados. Os resultados apontaram variações nos perfis se suscetibilidade aos antimicrobianos na população analisada, indicando a importância do uso do antibiograma para a escolha mais criteriosa dos antibacterianos a serem utilizados para o tratamento da mastite bovina causada por S. agalactiae. As altas frequências de resistência detectadas para alguns dos antimicrobianos comumente utilizados para o tratamento de mastite e outras patologias, tais como a neomicina e a tetraciclina, salientam a necessidade de monitoramento permanente do perfil de suscetibilidade aos antimicrobianos e do uso mais criterioso dos mesmos nos rebanhos leiteiros.(AU)

Antimicrobial susceptibility profile of Streptococcus agalactiaestrains isolated from bovine mastitis / Perfil de susceptibilidade a antimicrobianos em Streptococcus agalactiae isolados de mastite bovina

Bovine mastitis is the most important disease of dairy herds worldwide. Its main etiologic agents are bacteria, including Streptococcus agalactiae. The importance of this agent in bovine mastitis is because it is highly contagious and has a high impact on the occurrence of clinical mastitis cases and in the increase of the bulk milk somatic cell counts. The dry cow therapy and the treatment of the clinical mastitis cases stand out among the measures to control intramammary infections in cows. However, these strategies require knowledge about the antimicrobial susceptibility of the causal microorganisms. Thus, this study aimed to evaluate the antimicrobial susceptibility of 89 S. agalactiae strains isolated from bovine mastitis between the years 2004 and 2008 in dairy herds from Campo das Vertentes region, Minas Gerais State, Brazil. The disc diffusion technique was used and the antimicrobials currently used in mastitis therapy were tested. The isolates tested showed 100% susceptibility to chloramphenicol, ceftiofur, cefotaxime, enrofloxacin, and cefquinome. High frequencies of susceptibility (>95%) were also observed for the beta-lactams (penicillin G, ampicillin, and oxacillin), cephalosporins (cephalotin, ceftiofur, cefotaxime, cefoperazone, and cefquinome), florfenicol, gentamicin, lincomycin, nitrofurantoin, and sulfamethoprim. The strains showed high frequencies of resistance to neomycin (15.74%), and tetracycline (21.35%). Multidrug resistance was detected in 2.25% of the tested isolates. The results pointed to variations in the antimicrobial susceptibility profiles of the studied strains and the importance of the use of the susceptibility tests to determine the correct antimicrobial to be applied in the treatment of bovine mastitis caused by S. agalactiae. The high frequencies of resistance observed to some antimicrobials, such as neomycin and tetracycline, commonly used in the treatment of mastitis and other pathologies, highlighted the need for more judicious use of antimicrobials on dairy farms.(AU)

A mastite é a principal doença de bovinos leiteiros em todo o mundo e tem como principais agentes as bactérias, entre as quais Streptococcus agalactiae. Esse agente se destaca por ser altamente contagioso e pelos reflexos que causa na incidência de casos clínicos e no incremento da contagem de células somáticas do leite do tanque. Para o controle desta enfermidade, destacam-se a terapia de vacas secas e o tratamento de casos clínicos, medidas que requerem o conhecimento do perfil de sensibilidade dos agentes causais aos antimicrobianos. Este estudo teve como objetivo avaliar os perfis de suscetibilidade a antimicrobianos em 89 amostras de S. agalactiae isoladas de casos de mastite bovina em rebanhos da região de Campo das Vertentes, Minas Gerais, no período de 2004 a 2008. A técnica de difusão em discos foi utilizada e os antimicrobianos correntemente empregados na terapia da mastite foram testados. Os testes de suscetibilidade antimicrobiana apontaram 100% de susceptibilidade para cloranfenicol, ceftiofur, cefotaxima, enrofloxacina e cefquimona. Níveis elevados de susceptibilidade (>95%) foram observados para os betalactâmicos, florfenicol, gentamicina, lincomicina, nitrofurantoína e sulfametoprim. Altas frequências de resistência foram observadas para neomicina (15,74%) e tetraciclina (21,35%). Dois isolados multirresistentes (2,25%) foram encontrados. Os resultados apontaram variações nos perfis se suscetibilidade aos antimicrobianos na população analisada, indicando a importância do uso do antibiograma para a escolha mais criteriosa dos antibacterianos a serem utilizados para o tratamento da mastite bovina causada por S. agalactiae. As altas frequências de resistência detectadas para alguns dos antimicrobianos comumente utilizados para o tratamento de mastite e outras patologias, tais como a neomicina e a tetraciclina, salientam a necessidade de monitoramento permanente do perfil de suscetibilidade aos antimicrobianos e do uso mais criterioso dos mesmos nos rebanhos leiteiros.(AU)

Antimicrobial susceptibility profile of Campylobacter spp. isolated from different animal species in Minas Gerais / Perfil de suscetibilidade a antimicrobianos de amostras Campylobacter spp isoladas de diferentes espécies animais em Minas Gerais

Susceptibility pattern of 45 Campylobacter spp.isolates ­ 16 C. jejuni, eight C. coli, and 21 C. fetus isolated from different animal species in Brazil ­ to twelve antimicrobial agents was determined. All Campylobacter spp. isolates were susceptible to gentamicin, sulfadiazine, and sulfamethoxazole. C. jejuni and C. coli were also sensitive to chloramphenicol, whereas all C. fetus strains were susceptible to kanamycin. Cefoperazone showed the highest percentage of resistance among C. jejuni (68.75%), followed by nalidixic acid (31.25%), ampicillin (37.50%), tetracycline (37.50%), erythromycin (12.50%), and kanamycin (6.25%). Likewise, cefoperazone exhibited the highest percentage of resistance among C. coli (75.00%), followed by nalidixic acid (50.00%), tetracycline (50.00%), erythromycin (37.50%), ampicillin (12.50%), and kanamycin (12.50%). Among C. fetus strains, nalidixic acid showed the highest resistance rate (85.71%), followed by cefoperazone (71.43%), tetracycline (42.86%), ampicillin (19.05%), chloramphenicol (9.52%), and erythromycin (4.76%). Therefore, it was found that the majority of Campylobacter spp. isolated from animals was sensitive to gentamycin, chloramphenicol, kanamacyn, and sulfonamides; however, a high proportion of the strains showed reduced susceptibility to nalidixic acid, ampicillin, cefoperazone, and tetracycline. Moreover, C. coli and C. fetus isolates showed a high percentage of multidrug resistant strains.(AU)

O padrão de sensibilidade de 45 amostras de Campylobacter spp, incluindo 16 amostras de C. jejuni, 8 de C. coli e 21 C. fetus, isoladas de diferentes espécies de animais do Brasil, foi determinado para doze antimicrobianos. Todas as amostras de Campylobacter spp foram sensíveis à gentamicina, sulfadiazina e sulfametoxazol. C. jejuni e C. coli foram também sensíveis ao cloranfenicol, enquanto todas as amostras de C. fetus foram sensíveis à canamicina. Cefoperazona mostrou o maior percentual de resistência entre C. jejuni (68,75%), seguido pelo ácido nalidíxico (31,25%), ampicilina (37,50%), tetraciclina (37,50%), eritromicina (12,50%) e canamicina (6,25%). Similarmente, cefoperazona também exibiu o maior percentual de resistência entre as amostras de C. coli (75,00%), seguido pelo ácido nalidíxico (50,00%), tetraciclina (50,00%), eritromicina (37,50%), ampicilina (12,50%) e canamicina (12,50%). Entre os isolados de C. fetus, ácido nalidíxico apresentou maior taxa de resistência (85,71%), seguido de cefoperazona (71,43%), tetraciclina (42,86%), ampicilina (19,05%), cloranfenicol (9,52%) e eritromicina (4,76%). Assim, os nossos resultados mostraram que a maioria das amostras de Campylobacter spp isolados de animais foram sensíveis à gentamicina, cloranfenicol, canamicina e sulfonamidas. No entanto, uma proporção elevada das amostras apresentou susceptibilidade reduzida ao ácido nalidíxico, ampicilina, cefoperazona e tetraciclina. Além disso, C. coli e C. fetus mostraram uma alta porcentagem de amostras resistentes a múltiplas drogas.(AU)

Antimicrobial susceptibility profile of Campylobacter spp. isolated from different animal species in Minas Gerais / Perfil de suscetibilidade a antimicrobianos de amostras Campylobacter spp isoladas de diferentes espécies animais em Minas Gerais

Susceptibility pattern of 45 Campylobacter spp.isolates ­ 16 C. jejuni, eight C. coli, and 21 C. fetus isolated from different animal species in Brazil ­ to twelve antimicrobial agents was determined. All Campylobacter spp. isolates were susceptible to gentamicin, sulfadiazine, and sulfamethoxazole. C. jejuni and C. coli were also sensitive to chloramphenicol, whereas all C. fetus strains were susceptible to kanamycin. Cefoperazone showed the highest percentage of resistance among C. jejuni (68.75%), followed by nalidixic acid (31.25%), ampicillin (37.50%), tetracycline (37.50%), erythromycin (12.50%), and kanamycin (6.25%). Likewise, cefoperazone exhibited the highest percentage of resistance among C. coli (75.00%), followed by nalidixic acid (50.00%), tetracycline (50.00%), erythromycin (37.50%), ampicillin (12.50%), and kanamycin (12.50%). Among C. fetus strains, nalidixic acid showed the highest resistance rate (85.71%), followed by cefoperazone (71.43%), tetracycline (42.86%), ampicillin (19.05%), chloramphenicol (9.52%), and erythromycin (4.76%). Therefore, it was found that the majority of Campylobacter spp. isolated from animals was sensitive to gentamycin, chloramphenicol, kanamacyn, and sulfonamides; however, a high proportion of the strains showed reduced susceptibility to nalidixic acid, ampicillin, cefoperazone, and tetracycline. Moreover, C. coli and C. fetus isolates showed a high percentage of multidrug resistant strains.(AU)

O padrão de sensibilidade de 45 amostras de Campylobacter spp, incluindo 16 amostras de C. jejuni, 8 de C. coli e 21 C. fetus, isoladas de diferentes espécies de animais do Brasil, foi determinado para doze antimicrobianos. Todas as amostras de Campylobacter spp foram sensíveis à gentamicina, sulfadiazina e sulfametoxazol. C. jejuni e C. coli foram também sensíveis ao cloranfenicol, enquanto todas as amostras de C. fetus foram sensíveis à canamicina. Cefoperazona mostrou o maior percentual de resistência entre C. jejuni (68,75%), seguido pelo ácido nalidíxico (31,25%), ampicilina (37,50%), tetraciclina (37,50%), eritromicina (12,50%) e canamicina (6,25%). Similarmente, cefoperazona também exibiu o maior percentual de resistência entre as amostras de C. coli (75,00%), seguido pelo ácido nalidíxico (50,00%), tetraciclina (50,00%), eritromicina (37,50%), ampicilina (12,50%) e canamicina (12,50%). Entre os isolados de C. fetus, ácido nalidíxico apresentou maior taxa de resistência (85,71%), seguido de cefoperazona (71,43%), tetraciclina (42,86%), ampicilina (19,05%), cloranfenicol (9,52%) e eritromicina (4,76%). Assim, os nossos resultados mostraram que a maioria das amostras de Campylobacter spp isolados de animais foram sensíveis à gentamicina, cloranfenicol, canamicina e sulfonamidas. No entanto, uma proporção elevada das amostras apresentou susceptibilidade reduzida ao ácido nalidíxico, ampicilina, cefoperazona e tetraciclina. Além disso, C. coli e C. fetus mostraram uma alta porcentagem de amostras resistentes a múltiplas drogas.(AU)

EFICÁCIA DA PREDNISOLONA ASSOCIADA À CEFOPERAZONA NO TRATAMENTO INTRAMAMÁRIO DE CASOS NATURAIS DE MASTITE CLÍNICA EM VACAS LEITEIRAS

O objetivo deste estudo foi avaliar a eficácia da associação entre cefoperazona e prednisolona no tratamento de casos naturais de mastite clínica (MC) em bovinos leiteiros durante a lactação. Animais de 14 rebanhos comerciais foram distribuídos aleatoriamente em dois grupos experimentais, denominados como: 1) grupo PCEF: receberam tratamento intramamário contendo 250 mg de cefoperazona e 4 mg de prednisolona a cada 48h, totalizando 2 aplicações (N=147) e 2) grupo CEF: tratados somente com cefoperazona, conforme tratamento do grupo PCEF (N=141). Foram coletadas amostras de leite para cada quarto mamário incluído no estudo no dia da detecção do caso (D0), e 14 (D14) e 21 (D21) dias após D0, para realização de diagnóstico microbiológico e contagem de células somáticas (CCS). Os desfechos estudados foram cura microbiológica, cura clínica, recorrência de MC, risco de novas IMI em D14 e D21, CCS em D14 e D21 e produção de leite e CCS no 1º, 2º e 3º exames mensais após MC. Um total de 288 casos de MC foram randomizados em PCEF (n = 147) ou CEF (n = 141). A cura clínica (PCEF = 81,63% e CEF = 82,27%) e o tempo mediano (1º - 3º quartil) até a cura clínica [PCEF = 3,0 (2,5-4,0 dias) e CEF = 3,50 (3,0-4,0) dias] não foram diferentes entre os tratamentos. O risco de cura clínica foi 1,86 (IC 95%: 1,38-2,51) vezes maior para os casos leves, em relação aos moderados. A cura microbiológica (PCEF = 68,87% e CEF = 54,21%) não diferiu entre os tratamentos. Os casos associados a patógenos Gram-negativos foram 5,40 (IC 95%: 1,84-15,91) vezes mais propensos a serem curados do que aqueles associados a patógenos Gram-positivos. A recorrência de MC (PCEF = 26,67% e CEF = 21,93%) e o tempo mediano até a recorrência (PCEF = 63,5 e CEF = 65 dias) não diferiram entre os tratamentos. Para os casos com cultura negativa em D0, a incidência de nova IMI em D14 (PCEF = 18,75% e CEF = 34,78%) e D21 (PCEF = 28,13% e CEF = 39,13%) não foi diferente entre os tratamentos. A CCS foi numericamente maior em CEF do que em PCEF em D14 e D21, embora as diferenças não tenham sido estatisticamente significantes. Da mesma forma, não foram encontradas diferenças significantes entre a CCS ou a produção de leite no 1º, 2º e 3º teste mensal após o caso de MC. Os resultados deste estudo indicam que o tratamento intramamário de casos naturais de MC com PCEF não foi superior ao tratamento com CEF, com base no risco de cura clínica e microbiológica, recorrência de MC, risco de nova IMI, produção de leite e CCS.

The objective of this study was to evaluate the efficacy of a combination of cefoperazone and prednisolone for treatment of natural cases of clinical mastitis (CM) in dairy cattle. Cows from 14 commercial herds were randomly assigned to two experimental groups, as follows: 1) PCEF: received intramammary treatment (IMM) containing 250 mg of cefoperazone and 4 mg of prednisolone every 48 hours, totaling 2 applications (N = 143) and 2) CEF: treated only with cefoperazone, as previously described (N = 130). Milk samples were collected for each mammary quarter included in the study on the day of detection of the case (D0), and 14 (D14) and 21 (D21) days after D0, to perform microbiological diagnosis and somatic cell count (SCC). The outcomes were microbiological cure, clinical cure, recurrence of CM, risk of new IMI at D14 and D21, SCC at D14 and D21, and cow-level milk production and SCC at the 1st, 2nd, and 3rd monthly tests after CM. A total of 288 CM cases were randomized into PCEF (n = 147) or CEF (n = 141). Clinical cure (PCEF = 81.63% and CEF = 82.27%) and the median (1st - 3rd quartile) time to clinical cure [PCEF = 3.0 (2.5-4.0 days) and CEF = 3.50 (3.0-4.0) days] were not different between the treatments. The risk of clinical cure was 1.86 (95% CI: 1.38-2.51) times higher for mild cases, as compared with moderate cases. Microbiological cure (PCEF = 68.87% and CEF = 54.21%) was not different between the treatments. Cases associated with Gram-negative pathogens were 5.40 (95% CI: 1.84-15.91) times more likely to cure than those associated with Gram-positive pathogens. Recurrence of CM (PCEF = 26.67% and CEF = 21.93%) and the median time to recurrence (PCEF = 63.5 and CEF = 65 days) were not different between the treatments. For culture-negative quarters at D0, the incidence of new IMI at D14 (PCEF = 18.75% and CEF = 34.78%) and D21 (PCEF = 28.13% and CEF = 39.13%) were not different between the treatments. Quarter milk SCC was numerically higher in CEF than in PCEF at D14 and D21, although the differences were not statistically significant. Likewise, no significant differences were found between cow-level SCC or milk production at the 1st, 2nd, and 3rd monthly DHI test after CM. The results of this study indicate that intramammary treatment of natural CM cases with PCEF was not superior to treatment with CEF, based on the risk of clinical and microbiological cure, CM recurrence, risk of new IMI, milk production, and SCC

Eficácia "in vivo" da cefoperazona sódica, gentamicina e ciprofloxacino no tratamento intramamário da mastite clínica bovina / In vivo efficacy of sodium cefoperazone, gentamicin and ciprofloxacin for the intramammary treatment of clinical mastitis in cows / Eficacia in vivo de la cefoperazona sódica, la gentamicina y el ciprofloxacino en el tratamiento intramamario de la mastitis clínica bovina

O presente estudo avaliou a eficácia "in vivo" do tratamento intramamário, convencional e estendido, utilizando a cefoperazona sódica (250 mg), gentamicina (250 mg) e ciprofloxacino (100 mg) em casos de mastite clínica bovina. Foram utilizadas 30 vacas com mastite clínica, não sistêmica, distribuídas em três grupos de dez animais. O tratamento convencional da mastite consistiu em três aplicações do anti-mastítico, a cada 12 horas, e o tratamento estendido foi realizado com seis aplicações do anti-mastítico, a cada 12 horas. As vacas receberam tratamento intramamário, a cada 12 horas, com seis aplicações por animal, constituindo os seguintes momentos (M): M1 (dia zero - diagnóstico da mastite e início do tratamento), M2 (12 horas após M1), M3 (24 horas após M1), M4 (36 horas após M1), M5 (48 horas após M1) e M6 (60 horas após M1). Foi considerado, no mesmo animal, o tratamento convencional encerrado na terceira aplicação do antimicrobiano (M3) e o tratamento estendido ao final da sexta aplicação (M6). Os principais micro-organismos isolados das vacas com mastite foram Staphylococcus aureus, Staphylococus spp., Corynebacterium sp., Streptocccus spp., Streptococcus agalactiae e Escherichia coli. A cura clínica do tratamento convencional foi de 50% para o grupo tratado com cefoperazona sódica, 70% para o grupo tratado com gentamicina e 50% para o grupo tratado com ciprofloxacino. Para os mesmos grupos, a cura clínica do tratamento estendido foi 90%, 100% e 80%, respectivamente. A cura bacteriológica foi de 100% para o grupo tratado com cefoperazona sódica e de 90% para os grupos utilizando a gentamicina e ciprofloxacino no tratamento convencional. Para os mesmos grupos, foram observados 100% de cura bacteriológica para a gentamicina e 90% para os grupos com cefoperazona sódica e ciprofloxacino no tratamento estendido. Não foram encontradas diferenças entre cura clínica e cura bacteriológica dentro de cada grupo e entre os grupos de antimicrobianos, tampouco entre os tratamentos convencional e estendido dentro do mesmo grupo ou entre os grupos. Os resultados do presente estudo indicam a importância da lactocultura e do teste de sensibilidade microbiana "in vitro" como respaldo para os tratamentos intramamários de vacas com mastite clínica. Inferiu-se também a boa eficácia na cura clínica e bacteriológica da cefoperazona sódica, gentamicina e ciprofloxacino no tratamento da mastite clínica em vacas na lactação, de forma convencional ou estendida, contanto que respaldado na sensibilidade microbiana "in vitro".

This study evaluated the in vitro and in vivo efficacy of sodium cefoperazone, gentamicin and ciprofloxacin for the intramammary treatment of clinical mastitis in cows. Thirty cows with non-systemic, clinical mastitis were divided into three groups of ten animals. One of the antibiotics was used in each group for conventional (i.e., three doses, given every 12 hours) and extended (i.e., six doses, every 12 hours) treatments of mastitis. The cows were treated every 12 hours using six doses per animal at the next moments (M): M1, day zero ­ diagnosis and beginning of treatment; M2, 12 hours from diagnosis and treatment; M3, 24 hours; M4, 36 hours; M5, 48 hours; and M6 60 hours. The most common microorganisms isolated from cows were Staphylococcus aureus, Staphylococcus spp., Corynebacterium sp., Streptococcus spp., Streptococcus agalactiae and Escherichia coli. The efficacy of clinical cure using conventional treatment was 50%, 70% and 50% for the groups treated with sodium cefoperazone, gentamicin and ciprofloxacin, respectively. Regarding the extended treatment, efficacy was 90%, 100% and 80%, for sodium cefoperazone, gentamicin and ciprofloxacin, respectively. Efficacy of bacteriological cure was 100% for sodium cefoperazone, and 90% for both gentamicin and ciprofloxacin using the conventional treatment. By using the extended treatment, efficacy of bacteriological cure was 100% for gentamicin and 90% for both cefoperazone and ciprofloxacin. Statistical differences were not found in clinical or in bacteriological cure into the same group or among groups. Similarly, statistical differences between the conventional and extended treatments were not found. The results from this study reinforce the importance of performing microbiological culture of milk and in vitro antimicrobial sensitivity tests for supporting the intramammary treatment of clinical mastitis in cows. The clinical and bacteriological efficacy of sodium cefoperazone, gentamicin and ciprofloxacin in lactating cows suffering from clinical mastitis using either conventional or extended treatments was also inferred, provided the use of sensitivity tests in vitro.

El presente estudio comparó la eficacia in vitro e in vivo de la cefoperazona sódica, la gentamicina y el ciprofloxacino en el tratamiento intramamario de la mastitis clínica bovina. Fueron utilizadas 30 vacas con mastitis clínica sin signos sistémicos, divididas en tres grupos de diez animales. En cada grupo fue utilizado uno de los antibióticos citados antes, para realizar tratamiento convencional (tres aplicaciones a cada 12 horas) y extendido (seis aplicaciones a cada 12 horas) contra mastitis. Las vacas recibieron tratamiento intramamario a cada 12 horas, consistente de seis aplicaciones por animal en los siguientes momentos (M): M1, día cero - diagnóstico e inicio del tratamiento; M2, doce horas después del M1; M3, 24 horas después del M1; M4, 36 horas después del M1; M5, 48 horas después del M1; y M6 60 horas después del M1. Los principales microorganismos aislados de vacas con mastitis fueron Staphylococcus aureus, Staphylococcus spp., Corynebacterium sp. Streptococcus agalactiae spp. y Escherichia coli. La eficacia de la cura clínica usando tratamiento convencional fue de 50% para el grupo de cefoperazona sódica, 70% para gentamicina y 50% para ciprofloxacino. Con el tratamiento extendido la eficacia de la cura clínica fue de 90%, 100% y 80%, respectivamente. La eficacia de la cura bacteriológica fue de 100% para el grupo cefoperazona sódica y de 90% para los grupos gentamicina y ciprofloxacino utilizando el tratamiento convencional. Así mismo, se observó 100% de cura bacteriológica para gentamicina y 90% para los grupos cefoperazona sódica y ciprofloxacino utilizando el tratamiento extendido. No fueron observadas diferencias estadísticas para la cura clínica y bacteriológica ni dentro ni entre los grupos de antimicrobianos. Tampoco se encontró diferencia entre los tratamientos convencional y extendido dentro del mismo grupo o entre los grupos. Los resultados del presente estudio refuerzan la importancia del cultivo de la leche y de las pruebas de sensibilidad microbiana in vitro como respaldo para los tratamientos intramamarios en vacas con mastitis clínica. Los resultados permiten inferir también que la cefaperazona sódica, la gentamicina y el ciprofloxacino son eficaces para la cura clínica y bacteriológica de la mastitis clínica en vacas lactantes, tanto en un esquema de tratamiento convencional como en el tratamiento extendido siempre que se utilicen en combinación con exámenes de sensibilidad microbiana in vitro.

Evaluation of the different rinse volumes for performing the direct technique for Campylobacter spp. enumeration in chilled broiler carcasses / Avaliação de diferentes alíquotas de enxaguadura para a contagem pelo método direto de Campylobacter spp. em carcaças de frango resfriadas

The direct method for detecting and enumerating Campylobacter spp. in chicken meat is easy to perform, but the volumes of 100μL and 400μL, as recommended in some methodologies, are often unable to achieve the colonies counts due to the occurrence of confluent growth or microbial contaminants. This study aimed at evaluating the different volumes for rinsing chicken meats in order to minimize the interference of microbial contaminants, without compromising the sensitivity of direct methodology. Rinse volumes of 5, 10, 50, 100 and 400μL were tested using the modified charcoal cefoperazone deoxycholate agar (mCCDA) and modified Bolton agar. The presence of Campylobacter was confirmed by phenotypic methods and PCR assay. However, the strategy of using less than 100 µL of rinse volumes did not improve the isolation and enumeration of Campylobacter colonies, because it reduces the sensitivity of the assay. Thus, a possible solution to minimize the interference of microbial contaminants would be in developing new selective media or incorporate them into other existing antimicrobial drugs.(AU)

O método direto de detecção e contagem de Campylobacter spp. em carne de frango é de fácil execução, porém os volumes de 100 µL e de 400 µL, preconizados em algumas metodologias, muitas vezes impossibilitam a contagem de colônias pela ocorrência de crescimento confluente ou de microbiota contaminante. O objetivo do presente estudo foi de avaliar os diferentes volumes de enxaguadura de carne de frango com a finalidade de minimizar a interferência da microbiota contaminante, sem comprometer a sensibilidade do método. Os volumes de enxaguadura de 5, 10, 50, 100 e 400 µL foram testados utilizando-se os meios seletivo diferenciais ágar carvão cefoperazona desoxicolato modificado (mCCDA) e ágar Bolton modificado. A presença de Campylobacter spp. foi confirmada por métodos fenotípicos e por PCR. No entanto, a estratégia de utilização de volumes menores do que 100 µL de enxaguadura não melhoraram o isolamento e a contagem de colônias de Campylobacter, porque houve diminuição da sensibilidade do ensaio. A provável solução para minimizar a interferência da microbiota contaminante seria desenvolver novos meios seletivos ou incorporá-los aos antimicrobianos já existentes.(AU)

Prevalence and etiology of buffalo mastitis and milk somatic cell count in dry and rainy seasons in a buffalo herd from Analândia, São Paulo State, Brazil / Prevalência e etiologia da mastite bubalina e contagem de células somáticas no leite, nas estações seca e chuvosa, em um rebanho bubalino do município de Analândia, Estado de São Paulo, Brasil

The aim of this study was to evaluate somatic cell count (SCC), prevalence and etiology of mastitis in a dairy buffalo herd from Analândia, São Paulo State, Brazil, in the dry and rainy seasons. Additionally, antimicrobial susceptibility profile of microorganisms isolated from milk samples was also evaluated. 1,042 milk samples from female Murrah buffaloes in a dairy farm located in Analândia, São Paulo State, Brazil, collected between May 2011 and November 2012 were analyzed. After the mammary gland physical examination, strip cup test and California Mastitis Test (CMT) were performed. Afterwards, 50mL of milk samples from each mammary quarter were collected aseptically for SCC in automatic equipment and microbiological examination. The antimicrobial sensitivity profile to ampicillin, cefoperazone, ceftiofur, enrofloxacin, gentamicin, neomycin, oxacillin, penicillin, and sulfamethoxazole/trimethoprim was evaluated by disk diffusion method. The monthly average temperature and pluviometric index were obtained from "Centro Integrado de Informações Agrometeorológicas" (CIIAGRO) of "Instituto Agronômico de Campinas" (IAC). Milk samples with positive results in the microbiological test showed average SCC of 137,720 cells/mL in the dry period and 190,309 cells/mL in the rainy period. Although a higher number of isolated microorganisms was observed in buffalo milk samples during the rainy period (69/600) compared to the dry period (50/442), the season had no significant effect on the frequency of isolation of microorganisms. The main genera of microorganisms isolated were coagulase-negative Staphylococcus (38.4%), Streptococcus agalactiae (28.8%), and Bacillus spp. (7.56%) during the dry season and Corynebacterium sp. (23.5%), Streptococcus spp. (32.3%), and Streptococcus agalactiae (9.24%) during the rainy period. Multidrug resistance was observed in 30.1% of the isolated microorganisms.(AU)

O objetivo do presente estudo foi avaliar a contagem de células somáticas, a prevalência e a etiologia da mastite bubalina nas estações seca e chuvosa em um rebanho de bubalinos do município de Analândia, estado de São Paulo, Brasil. Adicionalmente, verificou-se o perfil de sensibilidade antimicrobiana dos micro-organismos isolados nas amostras de leite das búfalas. Foram avaliadas 1.042 amostras de leite de búfalas da raça Murrah pertencentes a uma propriedade rural localizada no município de Analândia-SP, obtidas no período de maio de 2011 e novembro de 2012. Após o exame físico da glândula mamária, foram realizados o teste da caneca de fundo escuro e o California Mastits Test (CMT); em seguida, foram colhidas, de forma asséptica, amostras de 50mL de leite de cada quarto mamário, para a contagem de células somáticas (CCS) em aparelho automático e exame microbiológico. Também, foi avaliado o perfil de sensibilidade antimicrobiana a ampicilina, cefoperazona, ceftiofur, enrofloxacina, gentamicina, neomicina, oxacilina, penicilina e sulfametoxazol/trimetoprim, pelo método de difusão em disco. A temperatura média e o índice pluviométrico mensais foram obtidos no Centro Integrado de Informações Agrometeorológicas (CIIAGRO) do Instituto Agronômico de Campinas (IAC). Notou-se que as amostras de leite com resultado positivo no exame microbiológico apresentaram CCS média de 137.720 células/mL, no período seco, e 190.309 células/mL, no período chuvoso. Embora tenha se constatado maior índice de isolamentos de micro-organismos nas amostras de leite obtidas no período chuvoso (69/600) do que no período seco (50/442), a frequência de isolamentos não foi influenciada significativamente pela estação do ano. Os principais gêneros de micro-organismos isolados durante o período seco foram Staphylococcus coagulase negativa (38,4%), Streptococcus agalactiae (28,8%) e Bacillus spp. (7,56%), e no período chuvoso Corynebacterium sp. (23,5%), Streptococcus spp. (32,3%) e Streptococcus agalactiae (9,24%). Verificou-se resistência a três ou mais antimicrobianos em 30,1% dos micro-organismos isolados.(AU)

Resistência a antimicrobianos em Staphylococcus aureus isolados de mastite em bovinos leiteiros de Minas Gerais, Brasil

Staphylococcus aureus (S. aureus) stands out as one of the most common etiological agents of bovine mastitis, currently causing major economic losses to dairy farming. In order to evaluate the sensitivity of this micro-organism to antimicrobials, 352 isolates from 35 dairy herds located in the state of Minas Gerais were submitted to antibiogram tests, using the disc diffusion technique. Nitrofurantoin and the Associations of neomycin, bacitracin and tetracycline (NBT), as well as penicillin, nafcillin and dihydrostreptomycin (PND), showed 100% efficiency in vitro. There were low resistance rates in the group of cephalosporins, with values of 0, 0.28 and 0.40% for cefquimona, cephalothin, and ceftiofur, respectively. Among the aminoglycosides, there was 1.69% resistance to gentamicin and 3.35% for neomycin. The highest resistance was observed for polymyxin B (82%), followed by betalactams, ampicillin and penicillin, with resistance rates of 80.92 and 80.45%, respectively. Moderate levels of resistance were observed for tetracycline, lincomycin, cefoperazone and sulfazotrim. Among the tested isolates, 65 (18.15%), coming from 24 of the 35 herds studied, showed multidrug resistance (MAR index 0.2). The results pointed out to high variation in antimicrobial resistance profiles and the occcurrence of multidrug resistance among some studied strains, highlighting the importance of antibiogram tests for the choice of an appropriate drug to be used for the treatment or prevention of mastitis caused by S. aureus.

Staphylococcus aureus (S. aureus) destaca-se como um dos agentes etiológicos mais frequentes da mastite bovina, que causa os maiores prejuízos econômicos à pecuária leiteira brasileira. Visando avaliar o perfil de sensibilidade deste agente aos antimicrobianos, 352 isolados provenientes de 35 rebanhos leiteiros localizados em Minas Gerais foram submetidos a testes de antibiograma, utilizando-se a técnica de difusão em disco. Nitrofurantoína, bem como as associações de neomicina, bacitracina e tetraciclina (NBT) e penicilina, nafcilina e dihidroestreptomicina (PND), apresentaram 100% de eficiência in vitro. Foram verificados baixos índices de resistência no grupo das cefalosporinas, com valores de 0, 0,28 e 0,40% para cefquimona, cefalotina e ceftiofur, respectivamente. Entre os aminoglicosídeos, observou-se 1,69% de resistência para gentamicina e 3,35% para a neomicina. O maior índice de resistência foi observado para polimixina B (82%), seguido pelos betalactâmicos, ampicilina e penicilina, com índices de resistência de 80,92 e 80,45%, respectivamente. Níveis intermediários de resistência foram observados para tetraciclina, lincomicina, cefoperazona e sulfazotrim. Entre os isolados testados, 65 (18,15%), oriundos de 24 dentre os 35 rebanhos estudados, apresentaram multirresistência (índice MAR 0,2). Os resultados apontaram grande variação nos perfis de resistência aos antimicrobianos, assim como a ocorrência de múltipla resistência entre algumas cepas estudadas, salientando a necessidade de testes de antibiograma para a escolha dos antimicrobianos mais adequados para o tratamento ou prevenção de mastite causada por S. aureus.

Antimicrobial resistance in Staphylococcus aureus isolated from mastitis in dairy herds from the state of Minas Gerais, Brazil / Resistência a antimicrobianos em Staphylococcus aureus isolados de mastite em bovinos leiteiros de Minas Gerais, Brasil

Staphylococcus aureus (S. aureus) stands out as one of the most common etiological agents of bovine mastitis, currently causing major economic losses to dairy farming. In order to evaluate the sensitivity of this micro-organism to antimicrobials, 352 isolates from 35 dairy herds located in the state of Minas Gerais were submitted to antibiogram tests, using the disc diffusion technique. Nitrofurantoin and the Associations of neomycin, bacitracin and tetracycline (NBT), as well as penicillin, nafcillin and dihydrostreptomycin (PND), showed 100% efficiency in vitro. There were low resistance rates in the group of cephalosporins, with values of 0, 0.28 and 0.40% for cefquimona, cephalothin, and ceftiofur, respectively. Among the aminoglycosides, there was 1.69% resistance to gentamicin and 3.35% for neomycin. The highest resistance was observed for polymyxin B (82%), followed by betalactams, ampicillin and penicillin, with resistance rates of 80.92 and 80.45%, respectively. Moderate levels of resistance were observed for tetracycline, lincomycin, cefoperazone and sulfazotrim. Among the tested isolates, 65 (18.15%), coming from 24 of the 35 herds studied, showed multidrug resistance (MAR index 5; 0.2). The results pointed out to high variation in antimicrobial resistance profiles and the occcurrence of multidrug resistance among some studied strains, highlighting the importance of [...]

Staphylococcus aureus (S. aureus) destaca-se como um dos agentes etiológicos mais frequentes da mastite bovina, que causa os maiores prejuízos econômicos à pecuária leiteira brasileira. Visando avaliar o perfil de sensibilidade deste agente aos antimicrobianos, 352 isolados provenientes de 35 rebanhos leiteiros localizados em Minas Gerais foram submetidos a testes de antibiograma, utilizando-se a técnica de difusão em disco. Nitrofurantoína, bem como as associações de neomicina, bacitracina e tetraciclina (NBT) e penicilina, nafcilina e dihidroestreptomicina (PND), apresentaram 100% de eficiência in vitro. Foram verificados baixos índices de resistência no grupo das cefalosporinas, com valores de 0, 0,28 e 0,40% para cefquimona, cefalotina e ceftiofur, respectivamente. Entre os aminoglicosídeos, observou-se 1,69% de resistência para gentamicina e 3,35% para a neomicina. O maior índice de resistência foi observado para polimixina B (82%), seguido pelos betalactâmicos, ampicilina e penicilina, com índices de resistência de 80,92 e 80,45%, respectivamente. Níveis intermediários de resistência foram observados para tetraciclina, lincomicina, cefoperazona e sulfazotrim. Entre os isolados testados, 65 (18,15%), oriundos de 24 dentre os 35 rebanhos estudados, apresentaram multirresistência (índice MAR 5; 0,2). Os resultados apontaram grande variação nos perfis de resistência aos [...]

Antimicrobial resistance in Staphylococcus aureus isolated from mastitis in dairy herds from the state of Minas Gerais, Brazil / Resistência a antimicrobianos em Staphylococcus aureus isolados de mastite em bovinos leiteiros de Minas Gerais, Brasil

Staphylococcus aureus (S. aureus) stands out as one of the most common etiological agents of bovine mastitis, currently causing major economic losses to dairy farming. In order to evaluate the sensitivity of this micro-organism to antimicrobials, 352 isolates from 35 dairy herds located in the state of Minas Gerais were submitted to antibiogram tests, using the disc diffusion technique. Nitrofurantoin and the Associations of neomycin, bacitracin and tetracycline (NBT), as well as penicillin, nafcillin and dihydrostreptomycin (PND), showed 100% efficiency in vitro. There were low resistance rates in the group of cephalosporins, with values of 0, 0.28 and 0.40% for cefquimona, cephalothin, and ceftiofur, respectively. Among the aminoglycosides, there was 1.69% resistance to gentamicin and 3.35% for neomycin. The highest resistance was observed for polymyxin B (82%), followed by betalactams, ampicillin and penicillin, with resistance rates of 80.92 and 80.45%, respectively. Moderate levels of resistance were observed for tetracycline, lincomycin, cefoperazone and sulfazotrim. Among the tested isolates, 65 (18.15%), coming from 24 of the 35 herds studied, showed multidrug resistance (MAR index 5; 0.2). The results pointed out to high variation in antimicrobial resistance profiles and the occcurrence of multidrug resistance among some studied strains, highlighting the importance of [...](AU)

Staphylococcus aureus (S. aureus) destaca-se como um dos agentes etiológicos mais frequentes da mastite bovina, que causa os maiores prejuízos econômicos à pecuária leiteira brasileira. Visando avaliar o perfil de sensibilidade deste agente aos antimicrobianos, 352 isolados provenientes de 35 rebanhos leiteiros localizados em Minas Gerais foram submetidos a testes de antibiograma, utilizando-se a técnica de difusão em disco. Nitrofurantoína, bem como as associações de neomicina, bacitracina e tetraciclina (NBT) e penicilina, nafcilina e dihidroestreptomicina (PND), apresentaram 100% de eficiência in vitro. Foram verificados baixos índices de resistência no grupo das cefalosporinas, com valores de 0, 0,28 e 0,40% para cefquimona, cefalotina e ceftiofur, respectivamente. Entre os aminoglicosídeos, observou-se 1,69% de resistência para gentamicina e 3,35% para a neomicina. O maior índice de resistência foi observado para polimixina B (82%), seguido pelos betalactâmicos, ampicilina e penicilina, com índices de resistência de 80,92 e 80,45%, respectivamente. Níveis intermediários de resistência foram observados para tetraciclina, lincomicina, cefoperazona e sulfazotrim. Entre os isolados testados, 65 (18,15%), oriundos de 24 dentre os 35 rebanhos estudados, apresentaram multirresistência (índice MAR 5; 0,2). Os resultados apontaram grande variação nos perfis de resistência aos [...](AU)

Mastite caprina: aspectos epidemiológicos e moleculares das infecções persistentes por Staphylococcus aureus e Escherichia coli

A mastite é uma enfermide de grande importância nos ruminantes, principalmente nos animais com aptidão leiteira, causando prejuízos econômicos diretos e indiretos além de riscos à saúde dos consumidores de leite e derivados, pois esses alimentos podem veicular microrganismos patogênicos e/ou toxinas. Este estudo objetivou revisar quanto aos tratamentos para mastite caprina, identificar e caracterizar as bacterias isoladas de leite de cabras com diferentes mastites, determinar o perfil das propriedades e os fatores de risco para mastite, identificar alterações da infecção intramamária persistente por Staphylococcus aureus após o tratamento com enrofloxacina e avaliar a diversidade de Esherichia coli isolados de cabras leiteiras com mastite clínica, subclínica e persistente. Um total de 539 cabras em lactação foram examinados e 268 amostras individuais de leite (uma/teto) foram coletadas de animais positivos no teste da caneca de fundo escuro e/ou Califórnia Mastitis teste (CMT). A prevalência de mastite subclínica foi de 28,0% e da clínica, 2,8%. A multiplicação bacteriana foi obtida em 62,0% das amostras. Um total de cento e oitenta e sete bactérias foram identificadas (fenotípica e genotípica). As espécies mais isoladas foram S. aureus (60,4%), Staphylococcus epidermidis (9.1%,), Escherichia coli (6.9%), Staphylococcus saprophyticus (5.9%) e Staphylococcus caprae (4,3%). No gênero Staphylococcus predominou perfil de resistência à classe de beta-lactâmicos: penicilina, ampicilina e oxacilina, além da tetraciclina. Doze por cento dos isolados apresentaram resistência múltipla a antimicrobianos (MDR). Entre as bactérias com maior prevalência de MDR, 38,5% E. coli e 10,6% S. aureus. Os produtores são tecnificados, trabalham com raças leiteiras especializadas e possuem bom manejo. Os fatores de risco: ordenha manual, utilização de cama, baixa frequencia de limpeza dos bebedouros, intervalo entre partos menor de 12 meses. S. aureus isolado de mastite persistente após o tratamento com enrofloxacina foi encontrado somente um complexo clonal, (133) e apresentaram pulsotipos idênticos ou estreitamente relacionados, confirmando a persistencia. Nos isolados após o tratamento com enrofloxacina aumentaram a concentração inibitória mínima para enrofloxacina, ampicilina e gentamicina. Houve pouca variação na presença dos genes de resistência, entretanto para genes de sistema de efluxo multidrogas variaram consideravelmente. A presença do gene norA aumentou em 50% nos isolados obtidos após o tratamento. A resistência antimicrobiana é o fator predominante na persistência mastite caprina. Foram encontrados 15 E. coli, sendo: nove (60,0 %) de mastite subclínica, quatro (26,7 %) de clínica e dois (13,3 %) de persistente. Houve maior número de isolados apresentando resistência frente a cefatiofur, tetraciclina e cefalexina e o índice de resistência multipla ocorreu em 33,3 %, variando de 0,2 a 0, 5. E. coli obtidos de animais com mastite clínica, a CIM foi maior para tetraciclina, ampicilina e gentamicina, os obtidos de mastite persistente apresentaram valores maiores da MIC frente a tetraciclina, ampicilina, cefoperazona e ciprofloxacina, e os isolados de animais com mastite subclínica observou-se maiores MIC para tetraciclina e cefoperazona. No perfil clonal não houve associação entre o cluster e o tipo de mastite. E. coli isolados de cabras com mastite persistente possui maior capacidade de adesão e invadem as células epiteliais mamária bovina (MAC-T). São necessários mais estudos para validar a eficácia e segurança dos antimicrobianos para tratamento da mastite caprina. As propriedades da Zona da Mata de Minas Gerais os Produtores são bem instruidos e tecnificados, criam raças especializadas na produção de leite, e não possuem assistência veterinária. Staphylococcus aureus foi a bactéria mais isolada com perfil de resistência principalmente aos beta-lactâmicos. Esherichia coli foi a bacteria Gram negativa mais prevalente. Foram observadas alterações nos perfis de resistência nos isolados de S. aureus obtidos após o tratamento com enrofloxacina. Não há uma relação entre os perfis estudados de E. coli e o tipo de mastite, entretanto E. coli isolados de cabras com mastite persistente apresentou maior capacidade de adesão e invasão.

The mastitis is one disease of great importance ruminants, mainly animals with dairy fitness, causing direct and indirect economic losses, risks to the health of consumers of milk and milk products, as these foods may carry pathogenic microorganisms and / or toxins This study objected revise how many treatments for goats mastitis, identify and characterize the bacteria isolated from goat leite with different mastites, determine or profile the farms of the mastitis, identify alterations of persistent intramammary infection by Staphylococcus aureus after or enrofloxacin treatment and to evaluate the diversity of Esherichia coli isolates of dairy goats with clinical mastitis, subclinical and persistent. A total of 539 goats in lactation foram examined in 268 individual samples of milk (one / teat) was coleted of positive animals strip cup testo r Califórnia Mastitis test (CMT). A prevalence of subclinical mastitis was 28.0% and clinic, 2.8%. A bacterial multiplication was obtained in 62.0% of the days. A total of seven cents and seven identified bacteria (phenotypic and genotypic). As well as more isolates were S. aureus (60.4%), Staphylococcus epidermidis (9.1%), Escherichia coli (6.9%), Staphylococcus saprophyticus (5.9%) and Staphylococcus caprae (4.3%). Genus Staphylococcus predominated resistance profile à class of betalactamics: penicillin, ampicillin and oxacillin, besides the tetracycline. Twelve percente of isolates presented multiple antimicrobial resistance (MDR). Among the bacterias by greater prevalence of MDR, 38.5% E. coli and 10.6% S. aureus. The producers are technified, work with specialized dairy breeds and have good management. Risk factors: manual milking, utilization of bed, low frequency of cleaning two drinkers, interval between parturition less than 12 months. S. aureus isolated of persistent mastitis after treatment with enrofloxacin was found only one clonal complex (133) and apresented identical pulsotypes or closely related, confirming persistence. The isolates afther treatment of enrofloxacin increased concentration inhibitory minimal for enrofloxacin, ampicillin and gentamicin. There was no variation for resistenfe genes, the genes of multidrug pump efflux varied. The presence of norA gene increased in 50% isolates obtained após or treatament. Antimicrobial resistance is factor predominant of persistence goat mastitis. Foram found 15 E. coli: Nine (60.0%) of subclinical mastitis, four (26.7%) of clinic and two (13.3%) of persistente. There was number of isolates exhibiting resistance to cefatiofur, tetracycline and cephalexin and resistance index multipla in 33.3%, varying from 0.2 to 0, 5. E. coli obtained from animais with clinical mastitis, to MIC incresed. For tetracycline, ampicillin and gentamicin, obtained of persistent mastitis presented higher values of MIC against tetracycline, ampicillin, cefoperazone and ciprofloxacin, the isolates of animals with subclinical mastitis observed MICs for tetracycline and cefoperazone. The clonal profile there was no association between or cluster and o type of mastite. E. coli isolates of goats with persistent mastitis have incresed fitness of adhesion and invasion of mammary bovine epithelial cells (MAC-T). There are more studies to validate the effectiveness and safety of two antimicrobials for goat mastitis treatment. As farms of Zona da Mata of Minas Gerais, farmers are trained and technified, specialized breeds of goat milk and not have veterinary assistance. Staphylococcus aureus is bacteria with a resistance profile mainly beta-lactams. Esherichia coli was more prevalent Gram negative bacteria. Foram observed alterações nos perfis de resistência we isolados of S. aureus obtidos após or tratamento com enrofloxacina. There is not relationship between E. coli and type of mastitis. E. coli isolated from goats with persistent mastitis apresented greater capacity of adhesion and invasion.

Study of thermophilic campylobacter contamination of a broiler batch at slaughter / Study of thermophilic campylobacter contamination of a broiler batch at slaughter

Background: Broilers are a reservoir of Campylobacter (C.), an important causal agent of gastroenteritis mostly associated to handling and consumption of broiler meat. The majority of broiler fl ocks are colonized by thermophilic Campylobacter at the slaughter age, and carcasses might be contaminated throughout the processing line. Since surveillance is crucial to evaluate and improve approaches to reduce Campylobacter spread during broiler processing, a cross-sectional study was carried out to detect the level of Campylobacter contamination in a broiler at slaughterMaterials, Methods & Results: Cloacal swabs, caeca and whole carcasses were taken from a broiler fl ock slaughtered in Southern Brazil. Samples were individually inoculated in Bolton Broth (BB) and incubated at 41.5C in a microaerobic atmosphere for 44 h, when the enriched culture was inoculated onto modifi ed Charcoal Cefoperazone Deoxycholate Agar (mCCDA) and Campy-Cefex Agar (CCA) plates. All plates were incubated at 41.5C in the microaerobic atmosphere for 44 h. Aliquots of each enriched BB were collected and submitted to polymerase chain reaction (PCR), while the genetic relatedness of isolates was analyzed by pulsed-fi eld gel electrophoresis (PFGE). A total of 3 (9.4%) cloacal swabs were positive for C. jejuni. No Campylobacter was isolated from any of the caecal contents or broiler carcasses analyzed.

Background: Broilers are a reservoir of Campylobacter (C.), an important causal agent of gastroenteritis mostly associated to handling and consumption of broiler meat. The majority of broiler fl ocks are colonized by thermophilic Campylobacter at the slaughter age, and carcasses might be contaminated throughout the processing line. Since surveillance is crucial to evaluate and improve approaches to reduce Campylobacter spread during broiler processing, a cross-sectional study was carried out to detect the level of Campylobacter contamination in a broiler at slaughter.Materials, Methods & Results: Cloacal swabs, caeca and whole carcasses were taken from a broiler fl ock slaughtered in Southern Brazil. Samples were individually inoculated in Bolton Broth (BB) and incubated at 41.5C in a microaerobic atmosphere for 44 h, when the enriched culture was inoculated onto modifi ed Charcoal Cefoperazone Deoxycholate Agar (mCCDA) and Campy-Cefex Agar (CCA) plates. All plates were incubated at 41.5C in the microaerobic atmosphere for 44 h. Aliquots of each enriched BB were collected and submitted to polymerase chain reaction (PCR), while the genetic relatedness of isolates was analyzed by pulsed-fi eld gel electrophoresis (PFGE). A total of 3 (9.4%) cloacal swabs were positive for C. jejuni. No Campylobacter was isolated from any of the caecal contents or broiler carcasses analyzed.

Study of thermophilic Campylobacter contamination of a broiler batch at slaughter

Background: Broilers are a reservoir of Campylobacter (C.), an important causal agent of gastroenteritis mostly associated to handling and consumption of broiler meat. The majority of broiler flocks are colonized by thermophilic Campylobacter at the slaughter age, and carcasses might be contaminated throughout the processing line. Since surveillance is crucial to evaluate and improve approaches to reduce Campylobacter spread during broiler processing, a cross-sectional study was carried out to detect the level of Campylobacter contamination in a broiler at slaughter. Materials, Methods & Results: Cloacal swabs, caeca and whole carcasses were taken from a broiler flock slaughtered in Southern Brazil. Samples were individually inoculated in Bolton Broth (BB) and incubated at 41.5°C in a microaerobic atmosphere for 44 h, when the enriched culture was inoculated onto modified Charcoal Cefoperazone Deoxycholate Agar (mCCDA) and Campy-Cefex Agar (CCA) plates. All plates were incubated at 41.5°C in the microaerobic atmosphere for 44 h. Aliquots of each enriched BB were collected and submitted to polymerase chain reaction (PCR), while the genetic relatedness of isolates was analyzed by pulsed-field gel electrophoresis (PFGE). A total of 3 (9.4%) cloacal swabs were positive for C. jejuni. No Campylobacter was isolated from any of the caecal contents or broiler carcasses analyzed. In addition, negative mCCDA and CCA plates showed an abundant growth of contaminant cells. The PCR assay detected all thermophilic Campylobacter reference strains tested and also the Arcobacter species. No amplified product was obtained from the non-related bacterial species analyzed. It was possible to identify 29 (90.6%) cloacal swabs, 32 (97.0%) caecal contents and 31 (100%) broiler carcasses Campylobacter-positive by PCR analysis. PFGE typing of the C. jejuni isolated resulted in two clearly distinguished genotypes which were grouped into different clusters. Discussion: The detection of C. jejuni in only few cloacal swabs sampled contrasts with higher frequencies of Campylobacter previously described in broilers. However, the enrichment culture of fecal samples might be compromised by the many competing non-target bacteria present, which may have prevented the detection of Campylobacter-positive samples. In addition, the BB and selective media containing cefoperazone might have allowed the growth of cefoperazone-resistant contaminant cells from fecal and carcasses samples, which masked Campylobacter cells onto mCCDA and CCA. To improve the detection of Campylobacter in broiler samples, alternative antimicrobial supplements or reduction of the time of enrichment has already been suggested. PCR showed a higher number of positive samples, which might reflect the increased ability of the PCR assay to detect either injured cells in conventional enrichment culture or Campylobacter that were masked by the proliferation of competing cells onto selective media used. The PCR assay was able to detect all the reference strains of thermophilic Campylobacter, but also the related Arcobacter species. However, the temperature of incubation of the enriched cultures associated to the selective pressure of the antimicrobials present in the BB restricts the growth of Arcobacter and the false-positive results observed using PCR. The subtypes of the C. jejuni strains isolated showed that the target broiler flock was simultaneously colonized by more than one C. jejuni strain which might be the result of introduction of Campylobacter from different sources at farm. PCR analysis showed high Campylobacter contamination level of the target flock at slaughter, pointing to the need for additional studies to investigate Campylobacter sources at broiler processing.

Utilização de caldo Bolton no enriquecimento seletivo em comparação ao plaqueamento direto na pesquisa de Campylobacter spp. em carcaças resfriadas de frango / Use of Bolton broth for selective enrichment and comparative analysis of its performance with direct plating methodology for isolating Campylobacter spp. from chilled chicken carcasses

O objetivo deste estudo foi comparar o enriquecimento seletivo e o plaqueamento direto no isolamento das espécies termofílicas de Campylobacter spp. em 30 amostras de carcaças de frango resfriadas adquiridas em supermercados, feiras livres e abatedouros no município do Rio de Janeiro, no período de julho de 2009 a julho de 2010. Foi realizada a enxaguadura da carcaça com água peptonada tamponada a 1% para recuperação das bactérias. Para o plaqueamento direto, foram utilizados ágar carvão cefoperazone desoxicolato modificado e ágar campy-cefex. Para o enriquecimento seletivo, foi empregado o caldo Bolton em concentrações simples e dupla. Foi detectada a presença de Campylobacter spp. em 21 amostras (70%), sendo 6 (28,6%) de abatedouros (3 com Serviço de Inspeção Estadual e 3 sem inspeção), 8 (38,1%) de supermercados e 7 (33,3%) de feiras livres. Não houve diferença estatisticamente significativa entre os resultados obtidos nesses estabelecimentos. As 21 amostras positivas foram isoladas do plaqueamento direto, 2 (9,5%) foram isoladas também no caldo Bolton simples e nenhuma no caldo Bolton duplo. O plaqueamento direto foi considerado o método mais rápido e eficiente, e apresenta menor custo que o enriquecimento seletivo na recuperação de Campylobacter spp. em carcaças resfriadas de frango.(AU)

This study aimed at comparing the use of selective enrichment and direct plating for isolating the thermophilic species of Campylobacter spp. from 30 samples of chilled chicken carcasses purchased in supermarkets, marketplaces and slaughterhouses in the Rio de Janeiro city, from July 2009 to July 2010. Chicken carcasses were rinsed out with 400 mL of 1% buffered peptone water for bacteria recovering. Charcoal Cefoperazone Deoxycholate agar and Campy Cefex agar were used for direct plating. Selective enrichment was performed employing Bolton broth in single and double concentrations. The presence of Campylobacter spp. was detected in 21 samples (70%), being 6 (28.6%) from slaughterhouses (3 with State inspection service and 3 without inspection), 8 (38.1%) from supermarkets and 7 (33.3%) from marketplaces. No statistically significant differences were found among the results obtained from different establishments. Of 21 positive samples, 2 (9.5%) were isolated from single Bolton broth and none of them was positive in double Bolton broth. Therefore, the direct plating was considered easier, faster and more cost-effective than the selective enrichment methodology for recovering Campylobacter spp. from chilled chicken.(AU)

Multiple drug resistance patterns in various phylogenetic groups of uropathogenic E.coli isolated from Faisalabad region of Pakistan

The objective of this work was the phylogenetic characterization of local clinical isolates of uropathogenic E. coli with respect to drug resistance. A total of 59 uropathogenic E. coli responsible for community acquired urinary tract infections were included in this study. A triplex PCR was employed to segregate each isolate into four different phylogenetic groups (A, B1, B2 and D). Drug resistance was evaluated by disc diffusion method. The drugs used were ampicillin, aztreonam, cefixime, cefoperazone, ceftriaxone, cephradine among -lactam group; amikacin, gentamicin, and streptomycin among aminoglycosides; nalidixic acid and ciprofloxacin from quinolones; trimethoprim-sulfomethoxazole, and tetracycline. Among 59 uropathogenic E. coli isolates majority belonged to phylogenetic group B2 (50%) where as 19% each belonged to groups A and B1, and 12% to group D. All the isolates were multiple drug resistant (MDR). Most effective drugs against Group A, B1, and B2 were gentamicin, amikacin and cefixime; ceftriaxone and quinolones; and ceftriaxone and amikacin, respectively. Group D isolates were found to be highly resistant to all drugs. Our results have shown emergence of MDR isolates among uropathogenic E. coli with dominance of phylogenetic group B2. However, it was found that group D isolates were though less frequent, more drug resistant as compared with group B2. Groups A and B1 were relatively uncommon. Amikacin, ceftriaxone and gentamicin were the most effective drugs in general.

Thermophilic Campylobacter survey in chilled and frozen poultry meatat retail in Concórdia, Santa Catarina

Background: Human campylobacteriosis has become a major foodborne disease, although little information is available about the role it plays in the scenario of foodborne diseases in Brazil. Since thermophilic Campylobacter (C.) species are often found in the intestinal tract of broiler chickens, consumption of contaminated poultry meat has been considered a risk factor for Campylobacter human infection. Campylobacter has been described as extremely susceptible to a variety of environmental stresses; hence the difficulty to establish cultures of the microorganism in the laboratory. In addition, it has been shown to decline in refrigerated and frozen foods. Currently there is a need for data on Campylobacter contamination level in Brazilian poultry meat. This work describes a survey performed in chilled and frozen poultry meat obtained from different retailers in Concórdia, Santa Catarina, Brazil. Materials, Methods & Results: This study analyzed 24 samples of fresh (chilled or frozen) poultry meat portions (thighs and drumsticks) produced by three different Brazilian broiler chicken processors, which were purchased from three different retailers in Concórdia, Santa Catarina. Campylobacter isolation was performed according to ISO 10272-1:2006. Individual samples (25g) were enriched in 225 mL of Bolton Broth in microaerobic atmosphere at 37°C for 4h to 6h, then at 41.5°C for 44h (+/- 4h). Aliquots were streaked in Modif ed Charcoal Cefoperazone Deoxycholate Agar (mCCDA) and Campy-Cefex Agar at 41.5°C in a microaerobic atmosphere for 44h (+/- 4h). Suspected colonies were subcultured in Blood Agar no 2 plates for confirmation by morphology, Gram staining, tests for catalase, oxidase and hippurate and indoxyl acetate hydrolysis test. Enriched Bolton Broth aliquots were also analyzed by Polymerase Chain Reaction (PCR) to amplify a 287 bp sequence of the 16S rRNA gene from thermophilic Campylobacter. Although C. jejuni positive control was isolated and confirmed by the morphological and biochemical tests described, all samples analyzed were negative for the presence of thermophilic Campylobacter. However, either mCCDA or Campy-Cefex Agar showed an abundant growth of non-Campylobacter cells. Moreover, all samples analyzed were negative in PCR analysis, although the C. jejuni reference strain used as PCR positive control showed the expected DNA fragment amplified. Discussion: Higher levels of Campylobacter contamination in poultry meat have been found by other studies, which used different isolation protocols in comparison to the present work. This study shows that Campylobacter isolation procedure according to ISO 10272-1:2006 allowed the growth of contaminants in both selective media used. It might be result of enrichment for 48 h or proliferation of ESBL producing Escherichia coli, able to hydrolyze cefoperazone in the selective medium used, which could underestimate the presence of Campylobacter in samples. In this sense, the PCR assay was essential to corroborate the negative result in Campylobacter isolation. Because poultry meat analyzed was negative for the presence of thermophilic Campylobacter, it was not possible to assess any difference between chilled or frozen storage on the bacteria survival. The present study might reflect low rates of Campylobacter contamination in poultry meat available at retail in Concórdia, Santa Catarina, Brazil, although it cannot replace good hygienic practices as well as consumer education.

Thermophilic Campylobacter survey in chilled and frozen poultry meatat retail in Concórdia, Santa Catarina

Background: Human campylobacteriosis has become a major foodborne disease, although little information is available about the role it plays in the scenario of foodborne diseases in Brazil. Since thermophilic Campylobacter (C.) species are often found in the intestinal tract of broiler chickens, consumption of contaminated poultry meat has been considered a risk factor for Campylobacter human infection. Campylobacter has been described as extremely susceptible to a variety of environmental stresses; hence the difficulty to establish cultures of the microorganism in the laboratory. In addition, it has been shown to decline in refrigerated and frozen foods. Currently there is a need for data on Campylobacter contamination level in Brazilian poultry meat. This work describes a survey performed in chilled and frozen poultry meat obtained from different retailers in Concórdia, Santa Catarina, Brazil. Materials, Methods & Results: This study analyzed 24 samples of fresh (chilled or frozen) poultry meat portions (thighs and drumsticks) produced by three different Brazilian broiler chicken processors, which were purchased from three different retailers in Concórdia, Santa Catarina. Campylobacter isolation was performed according to ISO 10272-1:2006. Individual samples (25g) were enriched in 225 mL of Bolton Broth in microaerobic atmosphere at 37°C for 4h to 6h, then at 41.5°C for 44h (+/- 4h). Aliquots were streaked in Modif ed Charcoal Cefoperazone Deoxycholate Agar (mCCDA) and Campy-Cefex Agar at 41.5°C in a microaerobic atmosphere for 44h (+/- 4h). Suspected colonies were subcultured in Blood Agar no 2 plates for confirmation by morphology, Gram staining, tests for catalase, oxidase and hippurate and indoxyl acetate hydrolysis test. Enriched Bolton Broth aliquots were also analyzed by Polymerase Chain Reaction (PCR) to amplify a 287 bp sequence of the 16S rRNA gene from thermophilic Campylobacter. Although C. jejuni positive control was isolated and confirmed by the morphological and biochemical tests described, all samples analyzed were negative for the presence of thermophilic Campylobacter. However, either mCCDA or Campy-Cefex Agar showed an abundant growth of non-Campylobacter cells. Moreover, all samples analyzed were negative in PCR analysis, although the C. jejuni reference strain used as PCR positive control showed the expected DNA fragment amplified. Discussion: Higher levels of Campylobacter contamination in poultry meat have been found by other studies, which used different isolation protocols in comparison to the present work. This study shows that Campylobacter isolation procedure according to ISO 10272-1:2006 allowed the growth of contaminants in both selective media used. It might be result of enrichment for 48 h or proliferation of ESBL producing Escherichia coli, able to hydrolyze cefoperazone in the selective medium used, which could underestimate the presence of Campylobacter in samples. In this sense, the PCR assay was essential to corroborate the negative result in Campylobacter isolation. Because poultry meat analyzed was negative for the presence of thermophilic Campylobacter, it was not possible to assess any difference between chilled or frozen storage on the bacteria survival. The present study might reflect low rates of Campylobacter contamination in poultry meat available at retail in Concórdia, Santa Catarina, Brazil, although it cannot replace good hygienic practices as well as consumer education.(AU)

Microrganismos patogênicos, celularidade e resíduos de antimicrobianos no leite bovino produzido no sistema orgânico

In last years increase the importance of milk quality and conditions of bovine milking. Simultaneously, increase the interest about organic milk and derivates. The aim of present study was investigate the milk pathogens, sensitivity and multiple drug resistance of isolates, somatic cell count and residues of drugs in milk, from cattle with and without mastitis, come from four little organic dairy farms in State of São Paulo, Brazil. Were used 148 cattle on the middle period of lactation. From these, two showed clinical mastitis, 72 subclinical mastitis and 74 without signs of mammary inflammation (controls). Staphylococcusaureus (25.7%), Streptococcus spp. (21.4%), Corynebacterium bovis (12.9%), Streptococcus agalactiae (4.3%) and Staphylococcus spp. (4.3%) were the more-frequent microorganisms isolated from animals with mastitis. Aspergillus spp. was isolated from one animal. Ceftiofur (95.2%), oxacillin (84.2%), gentamicin (76.3%) and cefoperazone (70.3%) were the more effective drugs. High resistance of isolates were found to penicillin (53.5%), ampicillin (41.6%) and neomycin (38.6%). Multiple drug resistance to three or more drugs was observed in 40 (39.6%) isolates. Media of somatic cell count encountered in animals with mastitis and controls were 175,742.67cs/mL and 58,227.6 cs/mL, respectively. Antimicrobials residues in milk were detected in four (2.7%) animals. The present findings showed the low somatic cell count of animals, indicative of good quality of milk. However, pointed the need of control measures for contagious pathogens of bovine mastitis and more attention for prohibition of antimicrobial use in organic dairy farms.

Nos últimos anos cresce a preocupação dos consumidores quanto à qualidade do leite e às condições de produção e bem-estar dos animais. Simultaneamente, aumenta o interesse e o consumo de produtos e subprodutos de origem animal produzidos no sistema orgânico, com destaque para o leite e derivados. O presente estudo investigou a presença de microrganismos patogênicos, a sensibilidade e a multi-resistência dos isolados aos antimicrobianos, a celularidade e a presença de resíduos de drogas no leite de vacas, com e sem mastite, produzido no sistema orgânico. Foram amostradas 148 vacas no período médio de lactação, das quais duas com mastite clínica, 72 com mastite subclínica e 74 sem mastite (controles), provenientes de quatro pequenas propriedades do interior do Estado de São Paulo, certificadas como orgânicas. Staphylococcusaureus (25,7%), Streptococcus spp. (21,4%), Corynebacterium bovis (12,9%), Streptococcus agalactiae (4,3%) e Staphylococcus spp. (4,3%) foram os microrganismos mais frequentemente isolados nos animais com mastite. Aspergillus spp. foi isolado de um animal. Ceftiofur (95,2%), oxacilina (84,2%), gentamicina (76,3%) e cefoperazona (70,3%) foram os antimicrobianos mais efetivos frente aos isolados. As maiores taxas de resistência das linhagens foram constatadas para penicilina (53,5%), ampicilina (41,6%) e neomicina (38,6%). Multi-resistência a três ou mais fármacos foi encontrada em 40 (39,6%) linhagens. A média da contagem de células somáticas das propriedades para animais com mastite foi 175.742,67 céls/mL, enquanto que para os animais controles foi 58.227,6 céls/mL. A presença de resíduos de antimicrobianos foi observada no leite de quatro (2,7%) animais. Os resultados revelaram baixa celularidade média nos animais, indicativo de boa qualidade do leite. Entretanto, apontam para a necessidade da adoção de medidas de controle para microrganismos contagiosos e maior rigor na proibição do uso de antimicrobianos em fazendas de leite orgânico.

Microrganismos patogênicos, celularidade e resíduos de antimicrobianos no leite bovino produzido no sistema orgânico

In last years increase the importance of milk quality and conditions of bovine milking. Simultaneously, increase the interest about organic milk and derivates. The aim of present study was investigate the milk pathogens, sensitivity and multiple drug resistance of isolates, somatic cell count and residues of drugs in milk, from cattle with and without mastitis, come from four little organic dairy farms in State of São Paulo, Brazil. Were used 148 cattle on the middle period of lactation. From these, two showed clinical mastitis, 72 subclinical mastitis and 74 without signs of mammary inflammation (controls). Staphylococcusaureus (25.7%), Streptococcus spp. (21.4%), Corynebacterium bovis (12.9%), Streptococcus agalactiae (4.3%) and Staphylococcus spp. (4.3%) were the more-frequent microorganisms isolated from animals with mastitis. Aspergillus spp. was isolated from one animal. Ceftiofur (95.2%), oxacillin (84.2%), gentamicin (76.3%) and cefoperazone (70.3%) were the more effective drugs. High resistance of isolates were found to penicillin (53.5%), ampicillin (41.6%) and neomycin (38.6%). Multiple drug resistance to three or more drugs was observed in 40 (39.6%) isolates. Media of somatic cell count encountered in animals with mastitis and controls were 175,742.67cs/mL and 58,227.6 cs/mL, respectively. Antimicrobials residues in milk were detected in four (2.7%) animals. The present findings showed the low somatic cell count of animals, indicative of good quality of milk. However, pointed the need of control measures for contagious pathogens of bovine mastitis and more attention for prohibition of antimicrobial use in organic dairy farms.

Nos últimos anos cresce a preocupação dos consumidores quanto à qualidade do leite e às condições de produção e bem-estar dos animais. Simultaneamente, aumenta o interesse e o consumo de produtos e subprodutos de origem animal produzidos no sistema orgânico, com destaque para o leite e derivados. O presente estudo investigou a presença de microrganismos patogênicos, a sensibilidade e a multi-resistência dos isolados aos antimicrobianos, a celularidade e a presença de resíduos de drogas no leite de vacas, com e sem mastite, produzido no sistema orgânico. Foram amostradas 148 vacas no período médio de lactação, das quais duas com mastite clínica, 72 com mastite subclínica e 74 sem mastite (controles), provenientes de quatro pequenas propriedades do interior do Estado de São Paulo, certificadas como orgânicas. Staphylococcusaureus (25,7%), Streptococcus spp. (21,4%), Corynebacterium bovis (12,9%), Streptococcus agalactiae (4,3%) e Staphylococcus spp. (4,3%) foram os microrganismos mais frequentemente isolados nos animais com mastite. Aspergillus spp. foi isolado de um animal. Ceftiofur (95,2%), oxacilina (84,2%), gentamicina (76,3%) e cefoperazona (70,3%) foram os antimicrobianos mais efetivos frente aos isolados. As maiores taxas de resistência das linhagens foram constatadas para penicilina (53,5%), ampicilina (41,6%) e neomicina (38,6%). Multi-resistência a três ou mais fármacos foi encontrada em 40 (39,6%) linhagens. A média da contagem de células somáticas das propriedades para animais com mastite foi 175.742,67 céls/mL, enquanto que para os animais controles foi 58.227,6 céls/mL. A presença de resíduos de antimicrobianos foi observada no leite de quatro (2,7%) animais. Os resultados revelaram baixa celularidade média nos animais, indicativo de boa qualidade do leite. Entretanto, apontam para a necessidade da adoção de medidas de controle para microrganismos contagiosos e maior rigor na proibição do uso de antimicrobianos em fazendas de leite orgânico.