Gabriela Isabel Favier

Genotypic and phenotypic characteristics of Yersinia enterocolitica isolated from the surface of chicken eggshells obtained in Argentina.

The prevalence of Yersinia enterocolitica on chicken eggshell surfaces in San Luis, Argentina, was investigated. The pathogenic potential of recovered isolates was assessed by means of phenotypic virulence tests and the presence of the 72-kb pYV plasmid. Antimicrobial susceptibility was determined by the agar diffusion method. DNA digested with XbaI was analyzed by pulsed-field gel electrophoresis (PFGE), and relationships between genomic DNA profiles were established. Eight Y. enterocolitica B2 O:9 strains were recovered after enrichment, for a prevalence of 2.27%. All strains harbored the virulence pYV plasmid, bound Congo red, grew in a low-calcium medium, and autoagglutinated at 37 degrees C. They lacked pyrazinamidase activity and did not hydrolyze esculin. These Y. enterocolitica strains were susceptible to amikacin, ciprofloxacin, chloramphenicol, and trimethoprim-sulfamethoxazole and were resistant to rifampin. According to the genomic DNA patterns obtained by PFGE, the isolates clustered into two groups, I and II. The highest similarity coefficient observed between Y. enterocolitica strains was 0.947. Microbiological controls on production stages of eggs and good culinary practices are necessary to reduce the risk of Y. enterocolitica infection for consumers.

Effect of Chlorine, Sodium Chloride, Trisodium Phosphate, and Ultraviolet Radiation on the Reduction of Yersinia enterocolitica and Mesophilic Aerobic Bacteria from Eggshell Surface

Eggshell sanitizing practices are necessary to improve microbiological safety of fresh hen eggs and their products. In this work, the effects of 100 mg/liter free chlorine (chl), 3% sodium chloride (NaCl), 1, 5, and 12% trisodium phosphate (TSP) in wash solutions, and UVR (ultraviolet radiation; 4.573 microW/cm2) were studied at different times on uninoculated and Yersinia enterocolitica-inoculated eggs. On uninoculated eggs, the best results were obtained with 100 mg/liter chlorine and UV exposure for >25 min, with reductions of 1.28 and 1.60 log cycles, respectively, compared to the average bacterial count (4.55 log CFU/egg) on the control (untreated eggs). On Y. enterocolitica-inoculated eggs, highest reductions of the average bacterial count (7.35 log CFU/egg) were obtained with 5 and 12% TSP and 100 mg/liter chl. The decrease obtained with 12% TSP (3.74-log reduction) was significantly higher (P < 0.05) than those obtained with the remaining treatments. Y. enterocolitica was more resistant to UVR than the eggshell natural mesophilic aerobic microflora, except when low inoculum (4.39 log CFU/egg) was assayed. Changes in eggshell microstructure were measured by the blue lake staining method. The presence of Yersinia and Salmonella in eggshell natural flora was also investigated.

Survival of Yersinia enterocolitica and Mesophilic Aerobic Bacteria on Eggshell after Washing with Hypochlorite and Organic Acid Solutions

Populations of Yersinia enterocolitica 0:9 and mesophilic aerobic bacteria on the shell of fresh chicken eggs were assessed prior and after washing with 0.75%, 1%, and 3% acetic and lactic acids, 50, 100, and 200 mg/liter (ppm) of chlorine, and water. Highest reductions of mesophilic aerobic bacterial populations (normal flora) on trypticase soy agar were 1.28 and 2.15 log10 cycles with 100 and 200 mg/liter of chlorine, 0.28 and 0.36 log10 cycles with 1% and 3% acetic acid, and 0.70 and 0.71 log10 cycles with 1% and 3% lactic acid, respectively, as compared to the control group. No Salmonella or Yersinia were detected among the natural flora of the eggs. On Y. enterocolitica O:9-inoculated eggs, reductions of 2.66, 2.77, and 2.92 log10 cycles by 50, 100, and 200 mg/liter of chlorine, of 2.47, 2.48, and 2.49 log10 cycles by 0.75%, 1%, and 3% of acetic acid, and of 2.48 and 2.72 log10 cycles with 1% and 3% of lactic acid, respectively, were observed with respect to the control. Organic acids at 3% caused detachment of the surface cuticle of the eggshell. Y. enterocolitica was more sensitive to the wash treatments than the natural microflora. The absence of potentially pathogenic Y. enterocolitica, observed for other fresh foods, should be a norm for fresh eggs sold in retail stores.

Pulsed field, PCR ribotyping and multiplex PCR analysis of Yersinia enterocolitica strains isolated from meat food in San Luis Argentina

The characterization of phenotypic and genotypic virulence markers of Yersinia enterocolitica strains belonging to biotypes (B) 1A, 2 and 3, mostly isolated from food in San Luis, Argentina, and the assessment of their genotypic diversity using PFGE and PCR ribotyping, were performed in our laboratory for the first time. Thirty five Y. enterocolitica strains, two reference strains and 33 strains isolated in our laboratory were studied. The presence of virF, ail, ystA, and myfA genes was investigated by multiplex PCR. The pathogenic potential of B1A strains, the most predominant biotype of Y. enterocolitica strains isolated from meat in our region, was investigated by simple PCR. Four B1A strains were positive for ystB gene. Four Y. enterocolitica 2/O:9 (bio/serotype) and two 3/O:5 strains isolated in our laboratory showed virulence-related results in the phenotypic tests and multiplex PCR. A good correlation between the expression of virulence markers and their corresponding genotypes was observed for most strains. Sixteen genomic types (GT) and 9 different intergenic spacer region (SR) groups were generated by PFGE and PCR ribotyping, respectively. In both cases the Y. enterocolitica 2/O:9 strains were separately clustered from 1A and 3/O:5 strains. Meat foods might be vehicles of transmission of pathogenic Y. enterocolitica strains in our region.

Detection and Characterization of Shiga Toxin Producing Escherichia coli, Salmonella spp., and Yersinia Strains from Human, Animal, and Food Samples in San Luis, Argentina

Shiga toxin producing Escherichia coli (STEC), Salmonella spp., and Yersinia species was investigated in humans, animals, and foods in San Luis, Argentina. A total of 453 samples were analyzed by culture and PCR. The antimicrobial susceptibility of all the strains was studied, the genomic relationships among isolates of the same species were determined by PFGE, and the potencial virulence of Y. enterocolitica strains was analyzed. Yersinia species showed higher prevalence (9/453, 2.0%, 95% CI, 0.7–3.3%) than STEC (4/453, 0.9%, 95% CI, 0–1.8%) and Salmonella spp. (3/453, 0.7%, 95% CI, 0–1.5%). Y. enterocolitica and Y. intermedia were isolated from chicken carcasses (6/80, 7.5%, 95% CI, 1.5–13.5%) and porcine skin and bones (3/10, 30%, 95% CI, 0–65%). One STEC strain was recovered from human feces (1/70, 1.4%, 95% CI, 0–4.2%) and STEC stx1/stx2 genes were detected in bovine stools (3/129, 2.3%, 95% CI, 0–5.0%). S. Typhimurium was isolated from human feces (1/70, 1.4%, 95% CI, 0–4.2%) while one S. Newport and two S. Gaminara strains were recovered from one wild boar (1/3, 33%, 95% CI, 0–99%). The knowledge of prevalence and characteristics of these enteropathogens in our region would allow public health services to take adequate preventive measures.

Inhibitory effects of sucrose fatty acid esters on survival of Yersinia enterocolitica on eggshell surface and use of blue lake as indicator of bacterial penetration into eggs.

Aims:  To determine the effectiveness of sucrose monolaurate (SML) and sucrose monocaprate (SMC), alone and in combination with ethylenediaminetetraacetic acid (EDTA), propionic acid (PA) or citric acid (CA) in reducing mesophilic aerobic bacteria (MAB) and Yersinia enterocolitica O:9 populations on eggshells and their damage potential on the microstructure of shell cuticle.

Evaluation of the pathogenic potential, antimicrobial susceptibility, and genomic relations of Yersinia enterocolitica strains from food and human origin

Yersinia enterocolitica is a food-borne pathogen that causes gastroenteritis with occasional postinfection sequels. This study was aimed to determinate the pathogenic potential, antimicrobial susceptibility, and genomic relationships of Y. enterocolitica strains of different bioserotypes (B/O) isolated from foods and human samples in San Luis, Argentina. Strains obtained by culture were bioserotyped and characterized by phenotypic and genotypic virulence markers, antimicrobial susceptibility, and pulsed-field gel electrophoresis (PFGE). Yersinia enterocolitica was detected in 9.2% of 380 samples, with a distribution of 10.6% (30/284) for food products and 5.2% (5/96) for human samples. Regarding the pathogenic potential, B1A strains of different serotypes were virF(-) ail(-), of which 72.0% (13/18) were ystB(+) with virulence-related phenotypic characteristics. Among B2/O:9 isolates, 75.0% (9/12) exhibited the genotype virF(+) ail(+) ystB(-) along with phenotypic traits associated with virulence; the same genotype was observed in 80.0% (4/5) of B3/O:3 and B3/O:5 strains. By PFGE, it was possible to separate Y. enterocolitica biotypes into 4 clonal groups (A to D) with 23 genomic types, generating a discriminatory index of 0.96. All isolates were susceptible to antimicrobials used for clinical treatment. This study highlights the presence of pathogenic bioserotypes and the high genomic diversity of the Y. enterocolitica strains isolated in our region.