Enterococcus Spp. Survival Through the Use of Standard Protocol for Clostridium Sp. Isolation

The genera Clostridium and Enterococcus are very different from each other, both morphologically and physiologically. Due to the high resistance by the sporulation capacity of Clostridium species, the thermal shock is a characteristic tool used for the isolation and identification of these microorganisms, this way, it would eliminate any other bacteria that did not present spores. The objective of this work is to show that Enterococcus sp. resist the temperature treatment and grow in culture media used for the isolation of Clostridium sp. For this, the present study initially attempted to identify reducing sulfite clostridia in poultry products, through the use of specific culture media and heat shock treatment. However, the PCR did not detect the presence of Clostridium sp. Then, sequencing of the 16S rDNA region was performed, which showed that the reducing sulfite colonies that were being isolated were, actually, Enterococcus spp. With this, some tests were carried out using different temperature and time combinations in the thermal shock, as well as the use of five different selective and differential culture media, in an attempt to eliminate any contaminants, but all without success, because these bacteria resisted to all modification. Therefore, the standard protocol for the isolation of bacteria of the genus Clostridium does not eliminate Enterococcus, which can lead to failures in the quantification and qualification of sulfite reducing microorganisms, a fact that can significantly affect food safety and animal health.(AU)

Enterococcus Spp. Survival Through the Use of Standard Protocol for Clostridium Sp. Isolation

The genera Clostridium and Enterococcus are very different from each other, both morphologically and physiologically. Due to the high resistance by the sporulation capacity of Clostridium species, the thermal shock is a characteristic tool used for the isolation and identification of these microorganisms, this way, it would eliminate any other bacteria that did not present spores. The objective of this work is to show that Enterococcus sp. resist the temperature treatment and grow in culture media used for the isolation of Clostridium sp. For this, the present study initially attempted to identify reducing sulfite clostridia in poultry products, through the use of specific culture media and heat shock treatment. However, the PCR did not detect the presence of Clostridium sp. Then, sequencing of the 16S rDNA region was performed, which showed that the reducing sulfite colonies that were being isolated were, actually, Enterococcus spp. With this, some tests were carried out using different temperature and time combinations in the thermal shock, as well as the use of five different selective and differential culture media, in an attempt to eliminate any contaminants, but all without success, because these bacteria resisted to all modification. Therefore, the standard protocol for the isolation of bacteria of the genus Clostridium does not eliminate Enterococcus, which can lead to failures in the quantification and qualification of sulfite reducing microorganisms, a fact that can significantly affect food safety and animal health.

Antisepsys and profilatic antimicrobial therapy in prevention of surgical site infection of horses / [Antissepsia e terapia antimicrobiana profilática no controle infeccioso do foco cirúrgico em equinos

The efficacy of an antisepsis protocol comprising chlorhexidine gluconate and ethyl alcohol in combination with prophylactic antimicrobial therapy in controlling surgical site infection in horses was studied. To that purpose, seven mixed breed horses received potassium penicillin and gentamicin at least 30 minutes prior to surgery. The surgical site was scrubbed with chlorhexidine gluconate and rinsed with ethyl alcohol. Samples were collected at four time points: (A) - before and (B) - immediately following shaving of the hair coat, (C) - at the end of antisepsis procedures, and (D) - at the end of the surgical procedure. Duration of surgery was recorded. Samples were cultured in three different culture mediums: Mitis Salivarus (Streptococcus sp.), Staphylococcus 110 (Staphylococcus sp.), and Mac Conkey (Enterobacteria). A high level of bacterial growth was observed in all culture mediums at (A) and (B), with no bacterial growth in (C). Staphylococcus sp. growth was observed in (D) in a single patient whose surgical procedure lasted for 120 minutes. Shaving of the hair coat reduced microbial flora on the surface of the skin. Antisepsis in combination with prophylactic antimicrobial therapy was effective in controlling surgical site infection in elective procedures with an average duration of 90 minutes.(AU)

Objetivou-se averiguar a eficácia do protocolo de antissepsia com clorexidina degermante e álcool etílico hidratado 70%, em associação com terapia antimicrobiana profilática, no controle microbiano do foco cirúrgico de equinos submetidos a procedimentos cirúrgicos. Foram utilizados 07 cavalos adultos de raças variadas, onde ambos receberam o mesmo tratamento (terapia antimicrobiana profilática e antissepsia com clorexidina degermante 2% e álcool etílico hidratado 70%), coletando-se amostras em quatro tempos distintos [(A - antes da tricotomia), (B - imediatamente após tricotomia), (C - ao término da antissepsia), (D - ao término do procedimento cirúrgico)]. O tempo de cada procedimento cirúrgico foi contabilizado. Foram utilizados três meios de cultura diferentes, cada um com especificidade para um tipo de crescimento bacteriano. Constatou-se alta incidência de crescimento bacteriano nos três meios utilizados nos tempos de coleta A e B. Para o tempo C, não foi observado crescimento bacteriano. No tempo D averiguou-se crescimento bacteriano do tipo Staphylococcus sp. em um único paciente, cujo tempo cirúrgico foi de 120 minutos de duração. Desta forma, a tricotomia reduziu a carga microbiana na superfície da pele. A antissepsia associada à terapia antimicrobiana profilática mostrou-se eficaz no controle microbiano do foco cirúrgico em procedimentos eletivos, com duração média de 90 minutos.(AU)

Antisepsys and profilatic antimicrobial therapy in prevention of surgical site infection of horses / Antissepsia e terapia antimicrobiana profilática no controle infeccioso do foco cirúrgico em equinos

The efficacy of an antisepsis protocol comprising chlorhexidine gluconate and ethyl alcohol in combination with prophylactic antimicrobial therapy in controlling surgical site infection in horses was studied. To that purpose, seven mixed breed horses received potassium penicillin and gentamicin at least 30 minutes prior to surgery. The surgical site was scrubbed with chlorhexidine gluconate and rinsed with ethyl alcohol. Samples were collected at four time points: (A) - before and (B) - immediately following shaving of the hair coat, (C) - at the end of antisepsis procedures, and (D) - at the end of the surgical procedure. Duration of surgery was recorded. Samples were cultured in three different culture mediums: Mitis Salivarus (Streptococcus sp.), Staphylococcus 110 (Staphylococcus sp.), and Mac Conkey (Enterobacteria). A high level of bacterial growth was observed in all culture mediums at (A) and (B), with no bacterial growth in (C). Staphylococcus sp. growth was observed in (D) in a single patient whose surgical procedure lasted for 120 minutes. Shaving of the hair coat reduced microbial flora on the surface of the skin. Antisepsis in combination with prophylactic antimicrobial therapy was effective in controlling surgical site infection in elective procedures with an average duration of 90 minutes.(AU)

Objetivou-se averiguar a eficácia do protocolo de antissepsia com clorexidina degermante e álcool etílico hidratado 70%, em associação com terapia antimicrobiana profilática, no controle microbiano do foco cirúrgico de equinos submetidos a procedimentos cirúrgicos. Foram utilizados 07 cavalos adultos de raças variadas, onde ambos receberam o mesmo tratamento (terapia antimicrobiana profilática e antissepsia com clorexidina degermante 2% e álcool etílico hidratado 70%), coletando-se amostras em quatro tempos distintos [(A - antes da tricotomia), (B - imediatamente após tricotomia), (C - ao término da antissepsia), (D - ao término do procedimento cirúrgico)]. O tempo de cada procedimento cirúrgico foi contabilizado. Foram utilizados três meios de cultura diferentes, cada um com especificidade para um tipo de crescimento bacteriano. Constatou-se alta incidência de crescimento bacteriano nos três meios utilizados nos tempos de coleta A e B. Para o tempo C, não foi observado crescimento bacteriano. No tempo D averiguou-se crescimento bacteriano do tipo Staphylococcus sp. em um único paciente, cujo tempo cirúrgico foi de 120 minutos de duração. Desta forma, a tricotomia reduziu a carga microbiana na superfície da pele. A antissepsia associada à terapia antimicrobiana profilática mostrou-se eficaz no controle microbiano do foco cirúrgico em procedimentos eletivos, com duração média de 90 minutos.(AU)

Highly Pathogenic and Multidrug Resistant Avian Pathogenic Escherichia Coli in Free-Range Chickens from Brazil

Avian pathogenic Escherichiacoli (APEC) virulence mechanism has been continuously studied and it is believed to be multifactorial and because of this, this work aimed to characterize potentially APEC strains isolated from free-range hens. Isolates were submitted to PCR for the detection of virulence genes, which were of high prevalence. In vivo inoculation of day-old chicks revealed that 49 of these strains were of high and intermediate pathogenicity. In addition, isolates were submitted to antimicrobials susceptibility test with the majority of the strains presenting multiresistance. Phylogenetic analysis showed a greater presence of potentially APEC isolates in-group B2. In addition, high heterogeneity was detected among the isolates byXbaI enzyme. Fifteen serogroups were identified, being the O8 the most frequent. These results strengthen the fact that a combination of diverse factors are associated with the pathogenicity APEC strains, as well as to highlight its importance to public health and that free-range hens can act as a reservoirs of potentially zoonoticbacteria.(AU)

Highly Pathogenic and Multidrug Resistant Avian Pathogenic Escherichia Coli in Free-Range Chickens from Brazil

Avian pathogenic Escherichiacoli (APEC) virulence mechanism has been continuously studied and it is believed to be multifactorial and because of this, this work aimed to characterize potentially APEC strains isolated from free-range hens. Isolates were submitted to PCR for the detection of virulence genes, which were of high prevalence. In vivo inoculation of day-old chicks revealed that 49 of these strains were of high and intermediate pathogenicity. In addition, isolates were submitted to antimicrobials susceptibility test with the majority of the strains presenting multiresistance. Phylogenetic analysis showed a greater presence of potentially APEC isolates in-group B2. In addition, high heterogeneity was detected among the isolates byXbaI enzyme. Fifteen serogroups were identified, being the O8 the most frequent. These results strengthen the fact that a combination of diverse factors are associated with the pathogenicity APEC strains, as well as to highlight its importance to public health and that free-range hens can act as a reservoirs of potentially zoonoticbacteria.

Quantification of enterobacteria and clostridium spp. In poultry slaughterhouses sanitary conveyors belts / Quantificação de enterobactérias e clostridium spp. Em esteiras condutoras de cortes de frango em frigoríficos

The increase in production and consumption of chicken meat has occurred due to modernization in this area. Such increase caused the concern about the transmission of pathogens to humans; however, with proper hygiene this transmission can be controlled. Thus, this study aimed to evaluate the pre-operational and operational hygiene insanitary conveyors belts of chicken cuts in slaughterhouses through Clostridium spp. and Enterobacteria quantification.Statistical data analysis for Clostridium spp. colony count showed a difference between the studied slaughterhouses and the types of cleaning performed on sanitary conveyors belts (p<0,0001). Already statistical analysis for Enterobacteriaceae colony count showed significant differences only between the visited slaughterhouses (p<0,0001),with no difference between the types of conveyors belts cleaning (p=0,4057). The results showed that there was avariation in bacterial count among the slaughterhouses and the hygiene process in sanitary conveyors belts were deficient because they presented counts higher than the values recommended by the international organizations.

O aumento da produção e do consumo per capita de carne de frango ocorreu devido a modernização neste setor. Tal aumento gerou preocupação com a transmissão de patógenos para o ser humano, porém com uma higienização adequada essa transmissão pode ser controlada. Assim, o objetivo deste estudo foi avaliar a higiene pré-operacional e operacional das esteiras condutoras de cortes de frangos através da quantificação de Clostridium spp. e Enterobactérias. As análises estatísticas da contagem de Clostridium spp. mostraram uma diferença entre os frigoríficos visitados e entre os tipos de limpeza realizados nas esteiras (p<0,0001). Já as análises estatísticas para a contagem de Enterobactérias mostraram diferenças significativas somente entre os frigoríficos visitados (p<0,0001), não havendo diferença entre os tipos de limpeza das esteiras (p=0,4057). Os resultados demonstraram que houve uma variação na contagem bacteriana entre os frigoríficos e que a higienização das esteiras foram deficientes pois apresentaram contagens superiores aos valores recomendado pelas organizações internacionais.

Quantification of enterobacteria and clostridium spp. In poultry slaughterhouses sanitary conveyors belts / Quantificação de enterobactérias e clostridium spp. Em esteiras condutoras de cortes de frango em frigoríficos

The increase in production and consumption of chicken meat has occurred due to modernization in this area. Such increase caused the concern about the transmission of pathogens to humans; however, with proper hygiene this transmission can be controlled. Thus, this study aimed to evaluate the pre-operational and operational hygiene insanitary conveyors belts of chicken cuts in slaughterhouses through Clostridium spp. and Enterobacteria quantification.Statistical data analysis for Clostridium spp. colony count showed a difference between the studied slaughterhouses and the types of cleaning performed on sanitary conveyors belts (p<0,0001). Already statistical analysis for Enterobacteriaceae colony count showed significant differences only between the visited slaughterhouses (p<0,0001),with no difference between the types of conveyors belts cleaning (p=0,4057). The results showed that there was avariation in bacterial count among the slaughterhouses and the hygiene process in sanitary conveyors belts were deficient because they presented counts higher than the values recommended by the international organizations.(AU)

O aumento da produção e do consumo per capita de carne de frango ocorreu devido a modernização neste setor. Tal aumento gerou preocupação com a transmissão de patógenos para o ser humano, porém com uma higienização adequada essa transmissão pode ser controlada. Assim, o objetivo deste estudo foi avaliar a higiene pré-operacional e operacional das esteiras condutoras de cortes de frangos através da quantificação de Clostridium spp. e Enterobactérias. As análises estatísticas da contagem de Clostridium spp. mostraram uma diferença entre os frigoríficos visitados e entre os tipos de limpeza realizados nas esteiras (p<0,0001). Já as análises estatísticas para a contagem de Enterobactérias mostraram diferenças significativas somente entre os frigoríficos visitados (p<0,0001), não havendo diferença entre os tipos de limpeza das esteiras (p=0,4057). Os resultados demonstraram que houve uma variação na contagem bacteriana entre os frigoríficos e que a higienização das esteiras foram deficientes pois apresentaram contagens superiores aos valores recomendado pelas organizações internacionais.(AU)

Efeito da utilização de próbióticos em dietas para bovinos nelore terminados em confinamento / Effect of probiotics inclusion in diets for nellore cattle finished in feedlot

O objetivo do presente trabalho foi avaliar se o probiótico DBR® pode substituir a monensina sódica na alimentação de bovinos em confinamento. Foram utilizados 18 bovinos da raça Nelore, com média de idade de 18 meses e 362±30kg de peso vivo, no início do experimento. Os animais foram distribuídos em baias individuais cobertas, com acesso a sombra e água, em um delineamento inteiramente casualizado. A alimentação foi fornecida ad libitum, duas vezes ao dia, às 8h e 16h, com uma dieta a base de bagaço de cana-de-açúcar como fonte de volumoso (18%) e concentrado (82%), que apresentava 72,4% de NDT e 14,2% de PB. Além da dieta citada, os animais receberam diariamente 100g de uma mistura contendo milho moído, pela qual foram fornecidas quantidades específicas de monensina sódica e DBR® Probiótico conforme os seguintes tratamentos: DBR (milho moído + 2,0 g DBR® Probiótico); MON (milho moído + 275 mg de monensina sódica) e DBR + MON (milho moído + 1,0g de DBR® Probiótico + 138mg de monensina sódica). Não foram verificados efeitos dos diferentes aditivos sobre o consumo de matéria seca, ganho de peso, eficiência, conversão alimentar e características da carcaça dos bovinos submetidos aos diferentes tratamentos dietéticos (P>0,05). Conclui-se que o DBR® probiótico pode substituir total ou parcialmente a monensina sódica em dietas com alta proporção de concentrados para bovinos Nelore em confinamento.

The aim of this study was to evaluate if the DBR ® probiotic can replace monensin in cattle feed in confinement. Eighteen animals of Nellore breed were used, with an average age of 18 months and 362 ± 30 kg live weight at the beginning of the experiment. The animals were distributed into individual pen, with access to shade and water, in a completely randomized design. Feed was provided ad libitum, twice daily at 8am and 16h, with a diet of crushed cane sugar as a source of roughage (18%) and concentrate (82%), which had 72.4 % TDN and 14.2% CP). Besides the aforementioned diet, the animals received daily 100 g of a mixture containing corn, which were provided by specific amounts of sodium monensin and probiotic DBR® according to the following treatments: DBR (corn meal + 2.0 g of probiotic DBR® ); MON (corn meal + 275 mg monensin sodium) and MON + DBR (corn meal + 1.0 g probiotic DBR® + 138mg of sodium monensin). The aim of this work was evaluate whether Probiotic DRB could replace the monesin. Data were subjected to analysis of variance using the SAS statistical software. There were no effects of different additives on the dry matter intake, weight gain, efficiency, feed conversion and carcass characteristics of cattle subjected to different dietary treatments (P> 0.05). We conclude that the probiotic DBR® can replace all or part of sodium monensin in diets with high levels of concentrates in feedlot for Nellore cattle.

Efeito da utilização de próbióticos em dietas para bovinos nelore terminados em confinamento / Effect of probiotics inclusion in diets for nellore cattle finished in feedlot

O objetivo do presente trabalho foi avaliar se o probiótico DBR® pode substituir a monensina sódica na alimentação de bovinos em confinamento. Foram utilizados 18 bovinos da raça Nelore, com média de idade de 18 meses e 362±30kg de peso vivo, no início do experimento. Os animais foram distribuídos em baias individuais cobertas, com acesso a sombra e água, em um delineamento inteiramente casualizado. A alimentação foi fornecida ad libitum, duas vezes ao dia, às 8h e 16h, com uma dieta a base de bagaço de cana-de-açúcar como fonte de volumoso (18%) e concentrado (82%), que apresentava 72,4% de NDT e 14,2% de PB. Além da dieta citada, os animais receberam diariamente 100g de uma mistura contendo milho moído, pela qual foram fornecidas quantidades específicas de monensina sódica e DBR® Probiótico conforme os seguintes tratamentos: DBR (milho moído + 2,0 g DBR® Probiótico); MON (milho moído + 275 mg de monensina sódica) e DBR + MON (milho moído + 1,0g de DBR® Probiótico + 138mg de monensina sódica). Não foram verificados efeitos dos diferentes aditivos sobre o consumo de matéria seca, ganho de peso, eficiência, conversão alimentar e características da carcaça dos bovinos submetidos aos diferentes tratamentos dietéticos (P>0,05). Conclui-se que o DBR® probiótico pode substituir total ou parcialmente a monensina sódica em dietas com alta proporção de concentrados para bovinos Nelore em confinamento.(AU)

The aim of this study was to evaluate if the DBR ® probiotic can replace monensin in cattle feed in confinement. Eighteen animals of Nellore breed were used, with an average age of 18 months and 362 ± 30 kg live weight at the beginning of the experiment. The animals were distributed into individual pen, with access to shade and water, in a completely randomized design. Feed was provided ad libitum, twice daily at 8am and 16h, with a diet of crushed cane sugar as a source of roughage (18%) and concentrate (82%), which had 72.4 % TDN and 14.2% CP). Besides the aforementioned diet, the animals received daily 100 g of a mixture containing corn, which were provided by specific amounts of sodium monensin and probiotic DBR® according to the following treatments: DBR (corn meal + 2.0 g of probiotic DBR® ); MON (corn meal + 275 mg monensin sodium) and MON + DBR (corn meal + 1.0 g probiotic DBR® + 138mg of sodium monensin). The aim of this work was evaluate whether Probiotic DRB could replace the monesin. Data were subjected to analysis of variance using the SAS statistical software. There were no effects of different additives on the dry matter intake, weight gain, efficiency, feed conversion and carcass characteristics of cattle subjected to different dietary treatments (P> 0.05). We conclude that the probiotic DBR® can replace all or part of sodium monensin in diets with high levels of concentrates in feedlot for Nellore cattle.(AU)