ESßL E. coli isolated in pig's chain: Genetic analysis associated to the phenotype and biofilm synthesis evaluation.

Resistance to new generation cephalosporins is an important public health problem globally, in terms of economic and social costs, morbidity and mortality. Βeta-lactamase enzymes are mainly responsible for the antibiotic resistance of Gram negative bacteria and extended-spectrum-ß-lactamases (ESßLs) are one of the major determinants of resistance against oxymino-cephalosporins in Enterobacteriaceae. Food-producing animals represent one of the sources of antibiotic resistant bacteria, including pigs. Here we analysed the presence of E. coli resistant to III generation cephalosporins isolated from different matrices collected from intensively bred pigs. A total of 498 E. coli were isolated from faeces and carcasses of pigs at slaughterhouse as well as from pork meat and sausages. Among these, 73 were phenotypically confirmed to be ESßL producers. Genetic analysis revealed that all except two harboured at least one of the three selected genes: blaCTX-M, blaTEM, and blaSHV. Furthermore, six of the E. coli ESßL isolated from faeces and carcasses swabs, were also able to produce biofilm, highlighting the virulence potential of these strains. The presence of Multi-Drug-Resistance patterns (MDR) recorded by the 73 ESßL E. coli was significant (60% of the strains were resistant to more than six antibiotics in MIC test). Results from the present study show that the transmission of resistant bacteria is possible along the food chain, including production of pork, one the most highly consumed meats around the world. Transmission is possible through the ingestion of raw meat products, and following cross-contamination between raw and cooked foods during preparation. The potential risk for human health demonstrated here, associated with the consumption of pork contaminated with bacterial strains characterized by multidrug resistance patterns, and the ability to produce ESßL and biofilm, is cause for concern. It is imperative to study future control strategies to avoid or limit as much as possible the transmission of these highly pathogenic strains through food consumption and/or contact with the environment.

Toxoplasma gondii in the Cornigliese sheep breed in Italy: Meat juice serology, in vitro isolation and genotyping.

Toxoplasma gondii is considered one of the most important food-borne parasitic zoonoses globally and sheep are important intermediate hosts of the parasite. Meat and milk from infected sheep are considered an important source of infection for humans. Here, the authors evaluated T. gondii infection in the Italian Cornigliese sheep breed using meat juice ELISA, and in vitro assay for followed by Real Time-PCR and PCR-RFLP. Twenty-one hearts were collected at slaughter. Meat juice serology was carried out on all samples, while eleven hearts with the highest antibody titres were subjected to acid-peptic digestion and seeding onto Vero cells. DNA was extracted at three different time points following seeding. PCR-positive samples were then genotyped by PCR-RFLP. All the meat juice samples were positive for IgG antibodies against p30 protein of T. gondii. Five of the 11 samples, seeded onto Vero cells, were positive in PCR made on DNA extracted after 21days of culture and the PCR-RFLP revealed a Type-II or Type II variant profile at 9/10 loci. Two out of five samples showed an increase in terms of parasite growth by comparing the Cq values at three different time points. To the authors' knowledge, this is the first report of in vitro cultivation of T. gondii from muscle tissue of naturally-infected sheep. In vitro assays may be a promising alternative to bioassays and further studies are necessary in order to improve assay performance and to identify possible early markers of parasite proliferation.