Norma S. Lázaro

Salmonella spp. in healthy swine and in abattoir environments in Brazil

Tonsils and inguinal, mesenteric, and prescapular lymph node samples collected from 115 swine carcasses from two abattoirs and a family-run operation in the State of Rio de Janeiro, Brazil, were cultured for the presence of Salmonella species. Salmonella spp. were detected in 40 (34.8%) of the swine samples with the following distribution; tonsils (31/40, 77.5%), mesenteric lymph nodes (16/40, 40.0%), inguinal lymph nodes, (9/40, 22.5%), and prescapular lymph nodes (7/40, 17.5%), Scalding tank water and environmental swabs collected from the abattoirs were also analyzed. Salmonella spp. were recovered from 13 of 51 (22.5%) of the environmental samples from one of the two abattoirs, none from those from the other abattoir. Salmonella spp. were recovered from the evisceration tables (5/11, 45.5%), the killing room (3/10, 30.0%), the holding pen (2/10, 20.0%), the butchering saw (2/10, 20.0%), and the scalding tank (1/10, 10.0%). The most frequently detected serovar was Salmonella Muenster. The results show the necessity of adopting more effective hygienic measures in the abattoirs as well as in the areas where swine are raised in order to reduce the role of abattoirs and storage facilities in the spread of Salmonella contamination.

Antimicrobial susceptibility and serovars of salmonella circulating in commercial poultry carcasses and poultry products in Brazil.

Antimicrobial resistance was evaluated for 12,582 strains of Salmonella isolated by public and private laboratories from commercial poultry carcasses and poultry products from different regions in Brazil between 2007 and 2011. These isolates were submitted unsolicited to the National Reference Laboratory for Bacterial Enteroinfections, Oswaldo Cruz Institute, Oswaldo Cruz Foundation, Rio de Janeiro, Brazil, for conclusive antigenic characterization. The prevalent Salmonella enterica serogroups were O:4 (B) (28.9%), O:9 (D1) (22.3%), O:21 (L) (15.9%), and O:7 (C1) (14.3%); the most prevalent serovars were Enteritidis, Minnesota, Typhimurium, Schwarzengrund, and Mbandaka. The antimicrobial susceptibility profiles of 1,234 strains indicated that 54.5% were resistant to up to eight antimicrobial drugs. Multidrug resistance (three or more classes) was observed in 16.4% of the strains, with 190 distinct patterns. The results showed increased resistance to ampicillin (12.4 to 18.9%), tetracycline (≈ 15.2 to ≈ 18.9%), and gentamicin (f7.0 to ≈ 9.6%) during the study period. Decreased resistance to nitrofurans (61.9 to 9.2%), quinolones (44.4 to 15.5%), and folate inhibitors (11.7 to 7.2%) was observed. Resistance to third-generation cephalosporins was detected in 5.5% of the strains throughout the study period; resistance to fluoroquinolones has been observed in 0.3% of the strains since 2009. These findings highlight the importance of surveillance along the food chain for detection of altered patterns among foodborne zoonotic bacteria that are important to public health.

Effects of gamma irradiation on the viability and phenotypic characteristics of Salmonella Enteritidis inoculated into specific-pathogen-free eggs.

The goal of this study was to determine the effects of various levels of gamma irradiation on the phenotypic characteristics of 20 strains of Salmonella Enteritidis inoculated separately into specific-pathogen-free shell eggs. Bacterial strains were inoculated into egg yolks and exposed to (60)Co radiation at doses of 0.49 to 5.0 kGy. The eggs were maintained at 25°C and analyzed for the presence of Salmonella on days 1, 2, 4, and 7, and the recovered Salmonella isolates were characterized biochemically. All strains were resistant to doses of 0.49, 0.54, 0.59, 0.8, and 1 kGy; colony counts were ≥10(5) CFU/ml of egg yolk except for one strain, which was detected at 96 h and at 7 days after irradiation at 1 kGy, with a population reduction of 2 log CFU/ml. For the other evaluated doses, 12 strains (60.0%) were resistant at 1.5 kGy and 7 strains (35.0%) were resistant at 3.0 kGy. Among all analyzed strains, 5.0 kGy was more effective for reducing and/or eliminating the inoculated bacteria; only two (10%) strains were resistant to this level of irradiation. Salmonella colony counts were significantly reduced (P < 0.01) with increasing doses from the day 1 to 7 of observation, when microbial growth peaked. Loss of mobility, lactose fermentation, citrate utilization, and hydrogen sulfide production occurred in some strains after irradiation independent of dose and postirradiation storage time. Increases in antibiotic susceptibility also occurred: seven strains became sensitive to β-lactams, two strains became sensitive to antifolates, and one strain each became sensitive to fluoroquinolone, phenicol, nitrofurans, tetracyclines, and aminoglycosides. The results indicate that up to 5.0 kGy of radiation applied to shell eggs inoculated with Salmonella Enteritidis at 4 log CFU per egg is not sufficient for complete elimination of this pathogen from this food matrix.

Antimicrobial resistance and R-plasmid in Salmonella spp from swine and abattoir environments

Salmonella serovars isolated from swine are of particular interest not only because of the pathogenic potential for this animal species, but also due to its relevance with regard to public health. On basis of the profile of resistance to antimicrobials, 13 Salmonella strains were selected which belonged to the serovars Muenster (7), Derby (4), Typhimurium (1), and Braenderup (1). They were isolated from healthy swine as well as from the abattoir environment in the state of Rio de Janeiro. All strains of Salmonella were subjected to bacterial conjugation, and the E. coli K12 Nalr Lac+ F standard strain was used as receptor, with the purpose to verify the ability to transfer the resistance marks. Gene transfer phenomenon was detected in seven strains, and except SalmonellaTyphimurium which transconjugated to Sm, Tc and Su, the remaining ones were characterized by transferring mark Su only. By plasmidial analysis of strains used and their respective transconjugants, 63 Kb plasmid was found, which was probably related to S. Typhimurium resistance.

Enteropatógenos bacterianos em peixes criados em uma estação de reciclagem de nutrientes e no ecossistema relacionado

The presence of bacterial enteropathogens from fishes of a nutrient recycle system from a Experimental Station in Petropolis, RJ, was evaluated in 72 samples from april 2000 to july 2001 Simultaneously was collected the mud used as organic manure and poultry beds localized next to the tanks. The isolation procedures included preenrichment in Peptone Water followed by enrichment with alcaline Peptone Water (pH 8.4-8.6), and streaked onto GSP Agar for the isolation of Aeromonas spp. and Plesiomonas shigelloides. For enteropathogenic bacteria of the Enterobacteriaceae family, 1ml samples were transferred for enrichment in Rappaport- Vassiladis broth and Kauffmann Tetrathionate Broth followed by streak onto Hektoen Enteric Agar and Salmonella-Shigella Agar. Simultaneously at each visit samples of water from fish and macrophyte tanks were collected for monitoring faecal coliforms (MPN) using A1 medium. Among the 116 isolates, Aeromonas spp. were the most frequent (67.2%) with 9 species (A. veronii, biogroup sobria, A. hidrophila, A. sobria, A. trota, A. eucrenophila, A. veronii biog. veronii, A. media, A. cavia and A. jandaei), followed by Edwardsiella tarda (16.4%), Plesiomonas shigelloides (12.9%) and Salmonella spp. (3.4%). The NMP of fecal coliforms showed higher values in the fish tanks (>1800/100ml).

Perfil de susceptibilidade a antimicrobianos em amostras de cocos Gram-positivos, catalase negativos, isoladas de mastite subclínica bubalina

The susceptibility of antimicrobials was studied in Gram positive and catalase negative cocci (21 samples of Lactococcus garvieae and 6 Enterococcus gallinarum), isolated from the milk of cows with subclinical mastitis, belonging to six buffalo herds in the State of Rio de Janeiro. The test used was diffusion of disks in agar Muller Hinton, according to recommendations of the National Committee for Clinical Laboratory Standards - NCCLS. There were tested disks with ampicillin (10mg), cefalotin (30mg), cefotaxime (30mg), cefoxitin (30mg), cloranfenicol (30mg), eritromycin (15mg), gentamycin (10mg), nitrofurantoin (300mg), norfloxacin (10mg), penicillin (10 IU), tetracyclin (30mg) and vancomycin (30mg). The results showed that with Lactococcus garvieae, the most efficient antimicrobial was nitrofurantoin, revealing 85.71% sensibility, followed by cefotaxime (61.90%), vancomycin (52.38%), norfloxacin (47.62&) and cefalotin (47.62%). The highest resistance was developed against penicillin and ampicillin, with 95.24% resistance for the two antimicrobials. The susceptibility profile developed by the strains of Enterococcus gallinarum showed low sensibility against the tested antimicrobials; the highest resistance observed was against eritromycin and gentamycin, with 33.34% sensibility for both. The antimicrobial evaluation showed 100% resistance against vancomycin and tetracyclin, followed by cloranfenicol, penicillin, ampicillin, cefoxitin, cefotaxim, norfloxacin and nitrofurantoin; all of them showed a resistance of 83.33% with the samples tested.

ANTI-Yersinia enterocolitica SEROTYPE 3 AGGLUTININS IN SWINE SERA FROM RIO DE JANEIRO

The slow serum agglutination test was applied to 119 healthy pigs for the determination of the possible presence of anti-Yersinia enterocolitica 0:3 agglutinins. Of the 63.9% reactive animals (s1:20), 8.4% presented positive titers (s1:80), suggesting the presence of this pathogen among swine and consequently an additional public health problem.

Padrão de susceptibilidade a antimicrobianos e perfil plasmidial em Salmonella Muenster isoladas de suínos e do ambiente de abatedouros

Thirty-eight strains of Salmonella Muenster, isolated from swine and the abattoir environment, in the State of Rio de Janeiro, Brazil, from March 1991 to February 1992, were analyzed for the presence of plasmids. The strains were selected according to their profile regarding the antimicrobials: streptomycin, tetracycline, sulphonamide and sulfametoxazole-trimethoprim. Thirteen strains were resistant to one or several antimicrobials, 18 with intermediate degree and seven were sensitive. Plasmids varying in size from 1.2 Kb to 42 Kb were detected in 37 (97.36%) of the 38 samples, corresponding to 11 different profiles (P1- P11), varying from 1 to 6 plasmids per model. The number and plasmids diversity was greater than the resistance marks for strains. The plasmid of 2.85 Kb was the most frequent, being present in 83.78% of the 37 strains; only the one of 7.5 Kb was detected at the two slaughterhouses. There was no parallelism between resistance pattern and plasmidial profile, and a same antibiotype was found in several plasmidial profiles. The results of the present investigation, allowed us to conclude that the plasmid characterization is an useful and simple tool for the epidemiological typing of this sorovar.