Phenotypic and Genetic Characterization of Klebsiella pneumoniae Isolates from Wild Animals in Central Italy.

Despite Klebsiella pneumoniae being widely recognized as a nosocomial pathogen, there is a critical lack in defining its reservoirs and sources of infections. Most studies on risk factors have focused on multidrug-resistant (MDR) isolates and clinically-oriented questions. Over a two-year period, we sampled 131 wild animals including mammal and bird species from three regions of Central Italy. All typical colonies isolated from the analytical portions were confirmed by real-time PCR and identified by MALDI-TOF mass spectrometry (MALDI-TOF MS). All confirmed K. pneumoniae isolates were tested for antimicrobial susceptibility to 29 antimicrobials and subjected to whole genome sequencing. Typical colonies were detected in 17 samples (13%), which were identified as K. pneumoniae (n = 16) and as K. quasipneumoniae (n = 1) by MALDI-TOF MS. The antimicrobial susceptibility profile showed that all the isolates were resistant to ß-lactams (ceftobiprole, cloxacillin, cefazolin) and tetracycline; resistance to ertapenem and trimethoprim was observed and nine out of 16 K. pneumoniae isolates (56.2%) were classified as MDR. Genomic characterization allowed the detection of fluoroquinolone resistance-associated efflux pumps, fosfomycin and ß-lactamase resistance genes, and virulence genes in the overall dataset. The cluster analysis of two isolates detected from wild boar with available clinical genomes showed the closest similarity. This study highlights the link between humans, domestic animals, and wildlife, showing that the current knowledge on this ecological context is lacking and that the potential health risks are underestimated.

Draft Genome Sequence of Serratia rubidaea, a Potential Opportunistic Pathogen Isolated from Food in Italy.

Serratia rubidaea has emerged in recent years as an opportunistic nosocomial pathogen. Here, we present the draft genome sequence of an isolate derived from an industrial meat food product purchased in a large-scale retail store that revealed fluoroquinolone, ß-lactam, and aminoglycoside resistance genes and two different host-unspecific prophages.

Campylobacter jejuni and Campylobacter coli: prevalence, contamination levels, genetic diversity and antibiotic resistance in Italy.

A research was carried out in Italy with the aim of assessing Campylobacter contamination in broilers from breeding to slaughter, of defining the genetic diversity of isolates and their antibiotic resistance. Sampling was carried out in a slaughterhouse, and in farms representative of the most common broiler production in Italy. At farm, the 78.8% (95% C.I.: 74.5%­82.5%) of cloacal samples tested positive for Campylobacter spp. C. jejuni showed higher prevalence in winter than in spring and summer (p < 0.00001, χ2 = 32.9), while C. coli showed an opposite trend (p < 0.00001, χ2= 41.1). At slaughterhouse, the 32.3% (95% C.I.: 30.2%­35.2%) and the 23.9% (95% C.I.: 21.7%­26.3%) of skin samples tested positive for C. jejuni for C. coli, respectively. C. coli showed higher prevalence than C. jejuni at washing (p < 0.05, χ2 = 11.11) and at chilling (p < 0.05, χ2 = 9.26). PFGE revealed high heterogeneity among isolates. Some clones were identified within the same farm in more than one season, suggesting environmental conditions able to support their persistence; other clones resulted to be spatially distant, suggestive of cross­contamination. Both Campylobacter species showed high resistance to nalidixic acid and ciprofloxacin, while resistance to erythromycin was more frequent in C. coli than C. jejuni (p < 0.05; χ2 test).

Prevalence of thermotolerant Campylobacter in broiler flocks and broiler carcasses in Italy.

In accordance with European Union regulations, from 5 February until 15 December 2008, sampling and analysis activities were conducted in Italy to assess the extent of contamination caused by thermotolerant Campylobacter in broiler chickens farmed nationwide. The survey involved 48 poultry slaughterhouses distributed across eleven regions of Italy, where the caeca and carcasses of 393 slaughter batches were sampled. A total of 284 batches (72.3%) gave positive results for Campylobacter spp. as follows: 52.1% were contaminated by C. jejuni, 55.6% by C. coli and 1.1% by C. lari. C. jejuni and C. coli were isolated together in 37 batches (13% of positive results). Campylobacter spp. was isolated only from the caeca in 251 slaughter batches (63.9%) including caecal isolates of C. jejuni (48.2%), C. coli (50.6%), and C. lari (1.2%). Carcasses from 182 batches (46.3%) were contaminated by C. jejuni in 40.7% of cases, C. coli in 57.7% and the absence of C. lari from all batches examined. The contamination level observed in the carcasses ranged between 10 and 1.6 × 10(7) cfu/g.

Thermotolerant Campylobacter in poultry meat marketed in the Abruzzo and Molise regions of Italy: prevalence and contamination levels.

The prevalence and level of contamination of Campylobacter were determined for poultry meat taken from small and large retailers in Abruzzo and Molise. Of a total of 392 samples analysed, 160 (40.8%) revealed low-level thermotolerant Campylobacter contamination (0.3-9.3 most probable number [MPN]/g), 17 samples (10.6%) showed a contamination level above 9.3 MPN/g and one sample (0.6%) had 110 MPN/g. Campylobacter jejuni and C. coli were isolated from 81.9% and 32.5% of the samples, respectively. More than one species of Campylobacter were isolated from 23.1% of the positive samples.

Prevalence of Salmonellae, Listeriae, and Yersiniae in the Slaughterhouse Environment and on Work Surfaces, Equipment, and Workers.

In 1995 and 1996 a nine-month study was carried out in 11 pig abattoirs located in the Molise region (Italy) to evaluate the degree of contamination of- the slaughterhouse environment, work surfaces, equipment, and personnel by Salmonella spp., Listeria spp., and Yersinia spp. A total of 219 samples were taken over three replications including slaughtering floor and wall, hooks, work-tables, chopping blocks, knives, cleavers, dehairing devices, hands of personnel, clothing, hand-wash basins, and cold room handles, floor, wall, and hooks. Overall, six abattoirs (54.5%) had one or more positive sites, while only 14 of the 219 sites (6.4%) tested were positive for any of considered microorganisms. Salmonella spp. were isolated from 1 of 9 cleavers (11.1 %), 1 of 16 worktables (6.25%), and 1 of 18 slaughtering floors (5.6%). Yersinia enterocolitica was found on 3 slaughtering floors (16.7%) and on 2 worktables (12.5%). Yersinia kristensenii was detected on 2 slaughtering floor swabs (11.1 %). Listeria monocytogenes was isolated from 2 of 20 cold room floor swabs (13.3%) and from 1 of 14 hand-wash basins (7.1%). Other species of Listeria were detected on slaughtering wall and floor swabs and on chopping blocks. Our study indicates that slaughtering floors, cold room floors, and worktables are important sites in abattoirs that may possibly harbor pathogens like Salmonella spp., Yersinia enterocolitica , and Listeria monocytogenes , and that cleaning and sanitizing of the slaughterhouse environment and equipment need a greater emphasis.