Resumo
This study was undertaken to compare selenite-cystine-novobiocin (SCN) broth, tetrationate-novobiocin (TN) broth and Rappaport-Vassiliadisnovobiocin (RVN) broth as enrichment and MacConkey (MC) agar, brilliant green (BG) agar, Hektoen (HE) agar, Salmonella-Shigella (SS) agar and xylose Iysine desoxycholate (XLD) agar as plating media for isolating Salmonella from broiler carcasses and chicken feces. The carcasses were washed with 300mL of Buffer Peptone Water 0.1% (BPW). The BPW solation was incubated 6h at ambient temperature and up to 24 h/43ºC. From this, 2 mL were inoculate in 20 mL of enrichment broth, incubated at 43ºC/24h. The enrichment broth was plated on the five tested selective agars, also incubated at 43ºC/24h. Up to five colonies from each plate were inoculated in TSI agar and LIA agar, incubated at 37ºC/24h. The genus of Salmonella was confirmed by slide agglutination test using poly serum anti-somatic Salmonella antigens (O) and poly serum anti-fiagellar (H) Salmonella antigens. The feces were experimentally contaminated with eight Salmonella serotypos (Agona, Anatum, Enteritidis, Havana, Intantis, Owakam, Schwazengrundand Typhimurium). Each serotype was tested five times. Salmonella serotypes were added in the sample of feces individually so that the final concentration of the bacterium was 1,2 x iO³ ufclg. Two grams of each sample were inoculated in 20 mL of the enrichment broth. The proceduring from this point wes the same adopted to carcasses examination. To broiler carcasses there was no statistic difference (p>0, 001) among the enrichment broth and among plating media. However the use of two enrichment broth (SCN and TN) and two agars between XLD agar, SS agar and HE agar gave the bost results. The assay done with feces showed that the TN broth was much better than the others and, in this case it can be plated either HE agar, or VB agar or SS agar The results from the broiler carcasses examination and the feces examination showed that it is not easy to determine the better bacteriologic routine to search for Salmonella and suggest that the association of two eorichments broth and two plating media may improve the test.
Este trabalho foi desenvolvido para avaliar, comparativamente, os caldos de enriquecimento Rapapport-novobiocina (RVN), selenito-cistinanovobiocina (SCN) e tetrationato-novobiocina (TN) e os meios para plaqueamento ágar Hektoen (HE), ágar MacConkey (MC), ágar Salmonella-Shigella (SS), ágar verde brilhante (VB) e ágar xilose lisina desoxicolato (XLD), utilizados no isolamento de Salmonella em carcaças de frango e fezes de aves. O procedimento bacteriológico consistiu das etapas de pré-enriquecimento, enriquecimento em caldos seletivos, plaqueamento, testes bioquímicos presuntivos e confirmação sorológica com soros polivalentes anti antígenos somáticos e flagelares de Salmonella.. As fezes foram experimentalmente contaminadas com 8 sorotipos de Salmonella (Agona, Anatum, Enteritidis, Havana, Infantis, Owakam, Schwazengrund e Typhimurium) e a concentração final foi aproximadamente de 1,2 x 10² ufc/g. As fezes foram inoculadas nos caldos enriquecedores e a partir daí, seguiu-se o mesmo procedimento utilizado para as carcaças. Os resultados referentes às carcaças de frango não foram estatisticamente diferentes (p>0,01) para os caldos e as placas. Todavia, verificou-se superioridade numérica em relação aos caldos SCN e TN sobre o caldo RVN, e em relação ao ágar XLD sobre os demais. Verificou-se também que com o emprego de dois caldos de enriquecimento e de dois meios para plaqueamento pode-se obter maior positividade. Quanto ao exame das fezes, o caldo TN mostrou-se superior aos demais (p>0,01), não havendo diferença (p>0,01) de resultados para os meios de plaqueamento. Os resultados sugerem que a utilização de mais de um meio de enriquecimento e de plaqueamento poderia aumentar as chances de isolamento de Salmonella.
Resumo
This study was undertaken to compare selenite-cystine-novobiocin (SCN) broth, tetrationate-novobiocin (TN) broth and Rappaport-Vassiliadisnovobiocin (RVN) broth as enrichment and MacConkey (MC) agar, brilliant green (BG) agar, Hektoen (HE) agar, Salmonella-Shigella (SS) agar and xylose Iysine desoxycholate (XLD) agar as plating media for isolating Salmonella from broiler carcasses and chicken feces. The carcasses were washed with 300mL of Buffer Peptone Water 0.1% (BPW). The BPW solation was incubated 6h at ambient temperature and up to 24 h/43ºC. From this, 2 mL were inoculate in 20 mL of enrichment broth, incubated at 43ºC/24h. The enrichment broth was plated on the five tested selective agars, also incubated at 43ºC/24h. Up to five colonies from each plate were inoculated in TSI agar and LIA agar, incubated at 37ºC/24h. The genus of Salmonella was confirmed by slide agglutination test using poly serum anti-somatic Salmonella antigens (O) and poly serum anti-fiagellar (H) Salmonella antigens. The feces were experimentally contaminated with eight Salmonella serotypos (Agona, Anatum, Enteritidis, Havana, Intantis, Owakam, Schwazengrundand Typhimurium). Each serotype was tested five times. Salmonella serotypes were added in the sample of feces individually so that the final concentration of the bacterium was 1,2 x iO³ ufclg. Two grams of each sample were inoculated in 20 mL of the enrichment broth. The proceduring from this point wes the same adopted to carcasses examination. To broiler carcasses there was no statistic difference (p>0, 001) among the enrichment broth and among plating media. However the use of two enrichment broth (SCN and TN) and two agars between XLD agar, SS agar and HE agar gave the bost results. The assay done with feces showed that the TN broth was much better than the others and, in this case it can be plated either HE agar, or VB agar or SS agar The results from the broiler carcasses examination and the feces examination showed that it is not easy to determine the better bacteriologic routine to search for Salmonella and suggest that the association of two eorichments broth and two plating media may improve the test.
Este trabalho foi desenvolvido para avaliar, comparativamente, os caldos de enriquecimento Rapapport-novobiocina (RVN), selenito-cistinanovobiocina (SCN) e tetrationato-novobiocina (TN) e os meios para plaqueamento ágar Hektoen (HE), ágar MacConkey (MC), ágar Salmonella-Shigella (SS), ágar verde brilhante (VB) e ágar xilose lisina desoxicolato (XLD), utilizados no isolamento de Salmonella em carcaças de frango e fezes de aves. O procedimento bacteriológico consistiu das etapas de pré-enriquecimento, enriquecimento em caldos seletivos, plaqueamento, testes bioquímicos presuntivos e confirmação sorológica com soros polivalentes anti antígenos somáticos e flagelares de Salmonella.. As fezes foram experimentalmente contaminadas com 8 sorotipos de Salmonella (Agona, Anatum, Enteritidis, Havana, Infantis, Owakam, Schwazengrund e Typhimurium) e a concentração final foi aproximadamente de 1,2 x 10² ufc/g. As fezes foram inoculadas nos caldos enriquecedores e a partir daí, seguiu-se o mesmo procedimento utilizado para as carcaças. Os resultados referentes às carcaças de frango não foram estatisticamente diferentes (p>0,01) para os caldos e as placas. Todavia, verificou-se superioridade numérica em relação aos caldos SCN e TN sobre o caldo RVN, e em relação ao ágar XLD sobre os demais. Verificou-se também que com o emprego de dois caldos de enriquecimento e de dois meios para plaqueamento pode-se obter maior positividade. Quanto ao exame das fezes, o caldo TN mostrou-se superior aos demais (p>0,01), não havendo diferença (p>0,01) de resultados para os meios de plaqueamento. Os resultados sugerem que a utilização de mais de um meio de enriquecimento e de plaqueamento poderia aumentar as chances de isolamento de Salmonella.