Identificação por "Multiplex PCR" do sorotipo monofásico e atípico Salmonella enterica subsp. enterica sorotipo 1,4,[5],12:i:-, no Estado de São Paulo, Brasil: freqüência e resistência antimicrobiana

Salmonella spp. are the etiologic agents of salmonellosis, a worldwide spread zoonoses causing foodborne outbreaks and clinical diseases. By serological identification, Salmonella enterica subsp. enterica serotype 1,4,[5],12:i:- accounted for 8.8% of human and 1.6% of nonhuman Salmonella strains isolated in São Paulo State, during 1991-2000. A total of 28.6% of them amplified a fragment corresponding to H:1,2 (flagellar phase two) through PCR analysis and were further assigned as S. Typhimurium. Antimicrobial resistance was detected in 36.3% of the 369 PCR-negative strains tested, including the multiresistance to ampicillin, chloramphenicol, sulfonamides, tetracycline, and streptomycin.Salmonella spp. é o agente etiológico da salmonelose, zoonose mundialmente distribuída e responsável por surtos de doenças transmitidas por alimentos e doenças clínicas. Sorologicamente, Salmonella enterica subsp. enterica sorotipo 1,4,[5],12:i:- correspondeu a 8,8% e 1,6% das cepas de Salmonella de origem humana e não-humana, respectivamente, isoladas no Estado de São Paulo, no decênio 1991-2000. Aproximadamente 28,6% destas cepas amplificaram o fragmento correspondente a H:1,2 (fase flagelar dois) em testes de PCR e foram, então, identificadas como S. Typhimurium. Das 369 cepas negativas em PCR, 36,3% apresentou resistência antimicrobiana, incluindo multirresistência a ampicilina, cloranfenicol, sulfonamidas, tetraciclina e estreptomicina

Stool processing-methods for Salmonella enterica isolation and PCR detection

The aim of this study was to compare the efficiency of three protocols for bacteriological isolation of Salmonella enterica and detection by PCR in swine feces samples. Pool of feces (n=62) were processed by three different methods. Method 1: samples (10g) were pre-enriched in BPW (1:10) and enriched in Rappaport-Vassiliadis broth (1:100). Method 2: samples (1g) were first enriched in GN-Hajna broth (1:10) and secondly enriched in Muller-Kaufmann tetrathionate broth (1`:10). Method 3: Single step enrichment of feces (1g) in selenite-cystine broth (1:10). PCR was performed using DNA extracted from the last enrichment broth of each bacteriological method. Salmonella enterica was cultured from 13 out of 62 samples (20.9%) and seven different serotypes were isolated. The methods 1, 2 and 3 resulted in 9 (14.5%), 6 (9.6%) and 2 (3.2%) positive samples, respectively. PCR was significantly superior than conventional bacteriology for Salmonella detection only when Rappaport-Vassiliadis was used for DNA-template preparation

Salmonella Typhimurium CARRIAGE at slaughter AFTER an enterocolitis outbreak in a swine herd

The aim of this study was to estimate the prevalence of slaughter pigs carrying Salmonella Typhimurium after a entericolitis outbreak in a commercial pig farm. A cross-sectional study was done during the slaughter of a batch of 86 animals. Mesenteric lymph nodes from 43 pigs were collected and pre-enriched in buffered peptone water (1:10) overnight at 37∞C Afterwards, aliquots of 1mL and 0.1mL were transferred to selenite-cystine and Rappaport-Vassiliadis broth, respectively. A loopful of each sample was streaked onto XLT4 and brilliant green agar plates, which were incubated at 37∞C for 24 hours. Salmonella was cultured in 23 out of 43 collected samples (53.48%). Salmonella Typhimurim (13 strains) and Salmonella enterica subs. enterica 1,4,5,12:i:- (10 strains) were isolated. These results indicated that the slaughter of pigs from batches previously affected by enteric salmonellosis may represent a high risk for pork contamination, since there is an positive association between infected pigs before slaughter and carcass contamination

Sorovares de Salmonella isolados de humanos no Estado de São Paulo, Brasil, 1996-2003

A salmonelose permanece uma importante causa de doença diarréica em humanos no Estado de São Paulo, Brasil. Nesse estudo foram identificados 3554 isolados de Salmonella de infecções humanas, durante o período 1996-2003. Entre os 68 diferentes sorovares determinados, S. Enteritidis foi predominante em infecções gastrointestinais e extra-intestinais, responsável por 67.4% de todos os isolados. S. Typhimurium e S. enterica subsp. enterica (4,5,12:i:-) foram mais freqüentemente isolados de crianças com idade de < 1 a 4 anos, em contraste, infecções por S. Enteritidis foram predominantes em pessoas entre 20 e 50 anos. A contínua vigilância laboratorial das salmoneloses, incluindo a sorotipagem, tem evidenciado os sorovares de Salmonella causando infecções humanas em nossa área geográfica ao longo do tempo.Salmonellosis remains an important cause of diarrheal illness in humans in São Paulo State, Brazil. In this study were identified 3554 Salmonella isolates from human infections, during the period 1996-2003. Among 68 different serovars determined, S. Enteritidis was the most frequent one in gastrointestinal and extra-intestinal infections accounting for 67.4% of all isolates. S. Typhimurium and S. enterica subsp. enterica (4,5,12:i:-) were most frequently isolated from children aged < 1-4 year-old, in contrast, people with S. Enteritidis infections were most likely to be 20-50 year-old. In our geographic area the continued laboratorial surveillance of salmonellosis, including serotyping, has showed the trends in Salmonella serovars causing infections in humans throughout the time

Antimicrobial resistance patterns of Salmonella isolated from finishing pigs and environmental samples

The aim of this study was to determine the antimicrobial resistance patterns of Salmonella isolates from pigs and environmental samples collected from modern swine facilities in Brazil. Sixteen samples from a total of 1,026 were positive to Salmonella and six serotypes were identified: Salmonella Typhimurium (1), Salmonella Agona (5), Salmonella Sandiego (4), Salmonella Rissen (1), Salmonella Senftenberg ( 4) and Salmonella Javiana (I). Resistance patterns were different among serotypes, but different isolates from a single serotype had the same antimicrobial pattern. The highest percentage of resistance was to tetracycline (100%), streptomycin (100%), nalidixic acid (100%), cefotaxime (12.5) and tobramycin ( 12.5). All serotypes were I 00% susceptible to ceftriaxone, norfloxacin, ciprofloxacin, ampicillin, gentamycin and chloramphenicol. Intermediate resistance to neomycin (93.5%), amikacin (12.5%) and trimethoprim (12.5%) was seen. The high resistance to tetracycline and streptomycin may be due to its extensive use in pig production

Epidemiology of Salmonella in two different finishing swine barns in Brazil

Many finishing barns in Brazil have lamina d\u27agua , a continuous water flow at the back of solid-floored adjacent pens. Prevalence of Salmonella shedding and environmental contamination in finishing barns with lamina d\u27agua and without it were assessed through a cross sectional study conducted in 6 farms. No difference was found between the two systems. Sixteen Salmonella strains were isolated from 4 farms, comprising 6 serotypes: S. Agona, S. Typhimurium, S. Senftenberg, S. Sandiego, S. Rissen and S. Javiana. Serotypes varied among farms and differed from those recently identified in Brazil. Epidemiology of salmonella in swine farms is complex and might vary between farms or even between barns in a same farm

"Multiplex PCR" identification of the atypical and monophasic Salmonella enterica subsp. enterica serotype 1,4,[5],12:i:- in São Paulo State, Brazil: frequency and antibiotic resistance patterns Identificação por "Multiplex PCR" do sorotipo monofásico e atípico Salmonella enterica subsp. enterica sorotipo 1,4,[5],12:i:-, no Estado de São Paulo, Brasil: freqüência e resistência antimicrobiana

Salmonella spp. are the etiologic agents of salmonellosis, a worldwide spread zoonoses causing foodborne outbreaks and clinical diseases. By serological identification, Salmonella enterica subsp. enterica serotype 1,4,[5],12:i:- accounted for 8.8% of human and 1.6% of nonhuman Salmonella strains isolated in São Paulo State, during 1991-2000. A total of 28.6% of them amplified a fragment corresponding to H:1,2 (flagellar phase two) through PCR analysis and were further assigned as S. Typhimurium. Antimicrobial resistance was detected in 36.3% of the 369 PCR-negative strains tested, including the multiresistance to ampicillin, chloramphenicol, sulfonamides, tetracycline, and streptomycin.<br>Salmonella spp. é o agente etiológico da salmonelose, zoonose mundialmente distribuída e responsável por surtos de doenças transmitidas por alimentos e doenças clínicas. Sorologicamente, Salmonella enterica subsp. enterica sorotipo 1,4,[5],12:i:- correspondeu a 8,8% e 1,6% das cepas de Salmonella de origem humana e não-humana, respectivamente, isoladas no Estado de São Paulo, no decênio 1991-2000. Aproximadamente 28,6% destas cepas amplificaram o fragmento correspondente a H:1,2 (fase flagelar dois) em testes de PCR e foram, então, identificadas como S. Typhimurium. Das 369 cepas negativas em PCR, 36,3% apresentou resistência antimicrobiana, incluindo multirresistência a ampicilina, cloranfenicol, sulfonamidas, tetraciclina e estreptomicina