An overview of Yersinia enterocolitica and related species in samples of different origin from San Luis, Argentina.

This study is aimed at offering an overview of the prevalence of Yersinia enterocolitica and related species in San Luis, Argentina, from samples of diverse origin received in our laboratory between 1984 and 2014, and providing an analysis of the distribution of Yersinia isolates according to their isolation sources, highlighting bioserotypes and potential reservoirs and vehicles of transmission to humans. From a total of 4572 samples of human, animal, food and environmental origins analyzed by traditional culture methods and molecular techniques, 229 (5%) samples were Yersinia positive. The highest frequency of Yersinia isolates was observed in environmental specimens (14.3%), followed by animal (9.2%), food (5%) and human (0.6%) samples. A total of 255 Yersinia isolates were characterized, including 183 Y. enterocolitica and 72 isolates of other Yersinia species. Biotype 1A associated to several serotypes was identified in Y. enterocolitica isolates from environment (100%), animals (95.5%), foods (71.7%) and human samples (40%); bioserotype 2/O:9 was identified in isolates from foods (25.5%), and biotype 3 was associated with strains from humans (60%), animals (4.5%) and foods (2.8%). This biotype included three strains O:3 and six strains O:5. The data highlight animals and foods as the main Y. enterocolitica sources in our region.

Dysprosium Removal from Water Using Active Carbons Obtained from Spent Coffee Ground.

This paper describes the physicochemical study of the adsorption of dysprosium (Dy3+) in aqueous solution onto two types of activated carbons synthesized from spent coffee ground. Potassium hydroxide (KOH)-activated carbon is a microporous material with a specific Brunauer-Emmett-Teller (BET) surface area of 2330 m2·g-1 and pores with a diameter of 3.2 nm. Carbon activated with water vapor and N2 is a solid mesoporous, with pores of 5.7 nm in diameter and a specific surface of 982 m2·g-1. A significant dependence of the adsorption capacity on the solution pH was found, but it does not significantly depend on the dysprosium concentration nor on the temperature. A maximum adsorption capacity of 31.26 mg·g-1 and 33.52 mg·g-1 for the chemically and physically activated carbons, respectively, were found. In both cases, the results obtained from adsorption isotherms and kinetic study were better a fit to the Langmuir model and pseudo-second-order kinetics. In addition, thermodynamic results indicate that dysprosium adsorption onto both activated carbons is an exothermic, spontaneous, and favorable process.

Design of an internal amplification control for a duplex PCR used in the detection of Shiga toxin producing Escherichia coli in pediatric feces.

A conventional PCR targeted directly to the detection of Shiga toxin-producing Escherichia coli (STEC) in diarrheal stools of symptomatic patients may require the introduction of internal controls to detect false negative results. In the present study, we designed a competitive internal amplification control (IAC) to be included in a well-known PCR protocol used to amplify the stx1and stx2 genes from STEC isolates. The IAC was introduced in the PCR reaction and amplified when E. coli O157:H7 cultures and contaminated pediatric feces were assayed. When STEC concentration was 10(3) CFU ml(-1) in pure culture and 10(4) CFU g(-1) in contaminated stools, the IAC at concentration of 0.143 pg µl(-1) in the PCR reaction mixture was co-amplified with the stx2 sequence, producing bands of 279 and 349 bp, respectively. These STEC values were considered the detection limits of the duplex PCR. The specific detection of STEC by duplex PCR including IAC might be achieved directly on pediatric feces when the pathogen load reaches concentrations of at least 10(4) CFU g(-1).

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.

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.

Detection of Yersinia spp. in meat products by enrichment culture, immunomagnetic separation and nested PCR.

The prevalence of Yersinia enterocolitica in meat products was assessed by four methods: cold enrichment in trypticase soy broth (A), enrichment in modified Rappaport broth at 25 °C (B), concentration by immunomagnetic separation (C) and yadA nested PCR (D). Furthermore, the pathogenic potentials of the isolates were established by phenotypic and genotypic tests, and their genomic relationships were determined by pulsed-field gel electrophoresis (PFGE). A total of 238 samples were collected at retail level in the city of San Luis, Argentina, during the period 2007-2008. The highest Yersinia prevalence in meat products was observed by method D (92 positive samples), followed by methods A (13 positive samples) and C (5 positive samples); however, no isolation was obtained by method B. Fourteen Y. enterocolitica and 4 Yersinia intermedia strains were recovered by culture. All Y. enterocolitica 2/O:9 strains gave results related to virulence by phenotypic tests and exhibited the genotype virF(+)myfA(+)ail(+)ystA(+). Two biotype 1A strains showed a genotype virF(-)myfA(-)ail(+)ystA(+)ystB(+). The 14 Y. enterocolitica strains isolated during this work plus one reference strain were separated into 11 genomic types by PFGE. This genomic heterogeneity of the isolates shows the diversity of Y. enterocolitica strains in our region. It is the first time that IMS was used to search Y. enterocolitica strains from naturally contaminated meat products.

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.