Biofilm-forming and antimicrobial resistance traits of staphylococci isolated from goat dairy plants.

INTRODUCTION: Biofilm-associated antimicrobial resistance is of increasing importance to the maintenance and spread of foodborne pathogens in the food industry. This study aimed to investigate the ability to form biofilm and the antimicrobial resistance of staphylococci contaminating small-scale goat milk dairy plants. METHODOLOGY: Sixty isolates were tested for antimicrobial resistance against 20 drugs by the microdilution method. Biofilm-forming traits were assessed by the microtiter plate method (MtP), Congo red agar method (CRA), and icaD gene detection by polymerase chain reaction (PCR). RESULTS: High antimicrobial resistance to ampicillin (60/60; 100%), penicillin G (21/60; 35%), and erythromycin (15/60; 25%) was observed, but all isolates were susceptible to amoxicillin/K-clavulanate, ceftriaxone, ciprofloxacin, gentamicin, levofloxacin, linezolid, and moxifloxacin. No resistance to oxacillin or vancomycin was seen among Staphylococcus aureus. Twenty-seven isolates (27/60; 45%) were considered to form biofilm according to MtP, and similar biofilm-producing frequencies were observed in coagulase-negative staphylococci (CoNS) (20/44; 45.4%) and S. aureus (7/16; 43.7%). The icaD gene was observed only in S. aureus isolates. There was no association between biofilm production and antimicrobial resistance. A higher frequency of biofilm-producing staphylococci was found in isolates from bulk tank milk and hand swabs. On the other hand, isolates from pasteurized milk showed lower frequency of biofilm formation. CONCLUSIONS: Staphylococci contaminating goat dairy plants are potential biofilm producers. The results suggest no association between the ability to form biofilm and antimicrobial resistance.

Use of a bacteriophage cocktail to control Salmonella in food and the food industry.

The use of lytic bacteriophages for the biocontrol of food-borne pathogens in food and in the food industry is gaining increasing acceptance. In this study, the effectiveness of a bacteriophage cocktail composed of three different lytic bacteriophages (UAB_Phi 20, UAB_Phi78, and UAB_Phi87) was determined in four different food matrices (pig skin, chicken breasts, fresh eggs, and packaged lettuce) experimentally contaminated with Salmonella enterica serovar Typhimurium and S. enterica serovar Enteritidis. A significant bacterial reduction (>4 and 2 log/cm(2) for S. Typhimurium and S. Enteritidis, respectively; p≤0.005) was obtained in pig skin sprayed with the bacteriophage cocktail and then incubated at 33 °C for 6h. Significant decreases in the concentration of S. Typhimurium and S. Enteritidis were also measured in chicken breasts dipped for 5 min in a solution containing the bacteriophage cocktail and then refrigerated at 4 °C for 7 days (2.2 and 0.9 log10 cfu/g, respectively; p≤0.0001) as well as in lettuce similarly treated for 60 min at room temperature (3.9 and 2.2 log10 cfu/g, respectively; p≤0.005). However, only a minor reduction of the bacterial concentration (0.9 log10 cfu/cm(2) of S. Enteritidis and S. Typhimurium; p≤0.005) was achieved in fresh eggs sprayed with the bacteriophage cocktail and then incubated at 25 °C for 2 h. These results show the potential effectiveness of this bacteriophage cocktail as a biocontrol agent of Salmonella in several food matrices under conditions similar to those used in their production.

Molecular characterization of Rhodococcus equi isolates of horse breeding farms from an endemic region in South of Brazil by multiplex PCR

Rhodococcus equi is a gram-positive coco-bacillus and an intracellular opportunistic pathogen which causes pneumonia in foals. It is widely detected in environment and has been isolated from several sources, as soil, feces and gut from health and sick foals. The goal of this study was to characterize the epidemiological status (endemic, sporadic or no infection) of horse breeding farms from Bage County in South of Brazil, using a multiplex PCR. One hundred and eighteen R. equi isolates were identified by biochemical tests and submitted to a specie-specific and vapA multiplex PCR. These isolates were obtained from: three farms where the R. equi infection has been noticed, two farms where the disease has been not reported and one farm where the disease is frequent. All clinical isolates from horse breeding farms where the disease is endemic and/or sporadic were vapA-positive. None environmental isolates were vapA-positive. In three horse breeding farms with sporadic R. equi infection, 11.54 percent of the isolates from adult horse feces were vapA-positive. The multiplex PCR technique has proven to be effective for the molecular and epidemiological characterization of the R. equi isolates in horse breeding farms. An important finding in this study was the isolation of vapA-positive R. equi from adult horse feces, which is an evidence for other routes of dissemination of this pathogen in the farms.

Rhodococcus equi é um coco-bacilo gram positivo que causa pneumonia em potros. Trata-se de um patógeno oportunista amplamente detectado no ambiente e isolado de várias fontes, como solo, fezes e intestino de potros doentes e sadios. O presente estudo visa caracterizar a situação epidemiológica de criatórios eqüinos da região de Bagé, RS, Brasil, pela técnica de PCR multiplex. Cento e dezoito isolados de R. equi foram identificados por testes bioquímicos e, posteriormente, submetidos a um PCR multiplex para caracterização da espécie e da presença do gene vapA. Estes isolados eram provenientes de três haras com histórico da doença, dois haras onde não havia casos da doença e uma propriedade onde a infecção por R. equi é relatada frequentemente. Todos os isolados clínicos provenientes de haras onde a doença é endêmica e/ou esporádica foram vapA positivos. Nenhum isolado ambiental foi vapA positivo. Nos três haras onde a doença é esporádica, 11,54 por cento dos isolados de fezes de eqüinos adultos foram positivos para o gene vapA. A técnica de PCR multiplex mostrou-se efetiva para caracterização epidemiológica e molecular dos criatórios equinos, estando de acordo com o histórico da propriedade. Um fato relevante demonstrado pela aplicação desta técnica foi a detecção de R. equi vapA positivo nas fezes de eqüinos adultos. Esta observação pode pressupor que haja outras vias de disseminação da bactéria dentro de uma propriedade.

Molecular characterization of Rhodococcus equi Isolates of horse breeding farms from an endemic region in South of Brazil by multiplex PCR.

Rhodococcus equi is a gram-positive coco-bacillus and an intracellular opportunistic pathogen which causes pneumonia in foals. It is widely detected in environment and has been isolated from several sources, as soil, feces and gut from health and sick foals. The goal of this study was to characterize the epidemiological status (endemic, sporadic or no infection) of horse breeding farms from Bage County in South of Brazil, using a multiplex PCR. One hundred and eighteen R. equi isolates were identified by biochemical tests and submitted to a specie-specific and vapA multiplex PCR. These isolates were obtained from: three farms where the R. equi infection has been noticed, two farms where the disease has been not reported and one farm where the disease is frequent. All clinical isolates from horse breeding farms where the disease is endemic and/or sporadic were vapA-positive. None environmental isolates were vapA-positive. In three horse breeding farms with sporadic R. equi infection, 11.54% of the isolates from adult horse feces were vapA-positive. The multiplex PCR technique has proven to be effective for the molecular and epidemiological characterization of the R. equi isolates in horse breeding farms. An important finding in this study was the isolation of vapApositive R. equi from adult horse feces, which is an evidence for other routes of dissemination of this pathogen in the farms.

Caracterização epidemiológica, molecular e perfil de resistência aos antimicrobianos de Escherichia coli isoladas de criatórios suínos do sul do Brasil / Epidemiology, molecular characterization and resistance to antimicrobials of Escherichia coli isolated from South-Brazilian pig herds

A colibacilose é a enfermidade entérica de maior impacto na suinocultura, sendo ocasionada por cepas enterotoxigênicas de Escherichia coli. Quarenta isolados clínicos de suínos com diarréia e 13 isolados ambientais foram analisados quanto ao perfil genotípico, relação genética e resistência antimicrobiana. O gene que codifica para a toxina Stb foi identificado em 50 por cento dos isolados clínicos, seguido por Sta e Lt, com 35 por cento. Dentre os fatores de adesinas pesquisados, a F18 foi encontrada em 27,5 por cento das amostras. A técnica de ERIC-PCR utilizada para caracterização epidemiológica dos isolados, não demonstrou poder discriminatório esperado, e apesar de permitir a separação dos isolados em grupos, estes não evidenciaram grupos relacionados aos fatores de virulência. No teste de susceptibilidade antimicrobiana a maior resistência foi observada à tetraciclina, em 88,6 por cento. O índice de resistência múltipla aos antimicrobianos (IRMA), variou entre 0 a 0,69.