Sanitisation of fresh-cut celery and radicchio by gas plasma treatments in water medium

open7noNome progetto: STARTEC: “Decision Support Tools to ensure safe, tasty and nutritious Advanced Ready-to-eat foods for healthy and vulnerable Consumers”.The antimicrobial efficacy of dielectric barrier discharge atmospheric gas plasma (DBD) was tested against Listeria monocytogenes and shigatoxin-producing Escherichia coli serogroups O157 and O26. Challenge tests were carried out with samples of cut celery and radicchio leaves inoculated with a mix of five strains of L. monocytogenes or the two strains of E. coli immersed in deionised water. The treatment efficacy was also assessed considering only the contaminated deionised water. For deionised inoculated water alone, a treatment time-dependent strong effect was observed and a pathogens reduction higher than 6LogCFU/mL was obtained after 40min of treatment. With the vegetables presence in the liquid medium, the efficacy appeared reduced and related to the treatment time, microorganism, substrate and storage duration (reduction up to 2.5 and 3.7LogCFU/cm2 for L. monocytogenes and E. coli, respectively). No significant changes were observed on celery visual attributes, soluble solids content and textural parameters. A significant decrease of the chroma colour parameter during storage was noted in treated radicchio samples respect to control ones.openBerardinelli, Annachiara; Pasquali, Frederique; Cevoli, Chiara; Trevisani, Marcello; Ragni, Luigi; Mancusi, Rocco; Manfreda, GerardoBerardinelli, Annachiara; Pasquali, Frederique; Cevoli, Chiara; Trevisani, Marcello; Ragni, Luigi; Mancusi, Rocco; Manfreda, Gerard

Whole genome sequencing based typing and characterisation of Shiga-toxin producing Escherichia coli strains belonging to O157 and O26 serotypes and isolated in dairy farms

In the present study, the genetic relationships as well as the virulome and resistome of newly sequenced O26 and O157 Shiga-toxin producing E. coli (STEC) isolates, collected from dairy farms in Italy, were investigated in comparison to publicly available genomes collected worldwide. The whole genome of Italian isolates was sequenced on Illumina MiSeq Platform. Reads quality control, de novo draft genome assembly, species confirmation and the 7-loci Multi-Locus Sequence Type assignment were performed using INNUca pipeline. Reference-based SNPs calling was performed on O157 and O26 genomes, separately, mapping contigs to high-quality finished genomes. Virulence and antimicrobial resistance determinants were detected in silico using the tool ABRicate. Phylogenetic reconstructions revealed that genomes clustered mainly based on their 7-loci MLST type. The virulome of tested genomes included 190 determinants. O157 genomes carried chu genes associated to heme mediated iron uptake, whereas O26 genomes harboured genes ybt associated to siderophore mediated iron uptake. Resistome analysis showed the presence of tet(34) on all but one O157 genomes and on only one O26 genomes. Only 4 genomes carried genes associated to multiresistance. In the present study, the genes chu and ybt were identified as potential biomarker for the differentiation of O157 and O26 serotypes

Microbiota analysis and microbiological hazard assessment in poultry carcasses from conventional and antibiotic free farms

The aim of this study was to assess microbiota and microbiological hazards in poultry carcasses from animals reared in conventional (n=15) and antibiotic free (n=15) farms. An aliquot of neck and breast skin was obtained from each individual carcass at the end of the refrigeration tunnel and submitted to DNA extraction. Total DNA was sequenced in the 16S rRNA and reads analysed with MG-RAST to classify the colonising bacteria up to the genus level and compare each taxonomic group in terms of mean relative frequency of abundance in conventional and antibiotic free carcasses. Firmicutes displayed abundances always higher than 38% but did not show statistically significative differences between conventional and antibiotic free carcasses. On the contrary, Bacteroidetes and Actinobacteria were significantly higher in antibiotic free then conventional carcasses (21.57 vs 10.95%; 19.29 vs 12.05%), whereas Proteobacteria were higher in the latter (33.19 vs 19.52%). The genera significantly higher in antibiotic free than conventional carcasses were Chryseobacterium (10.07 vs 1.94%), Rothia (3.08 vs 0.77%) and Micrococcus (1.12 vs 0.16%), while Shewanella was significantly higher in conventional carcasses (1.38 vs 0.26%). Among Firmicutes, the genera significantly higher in conventional carcasses were Ureibacillus (1.45 vs 0.11%) and Bacillus (3.28 vs 0.56%). The higher abundance of Proteobacteria in conventional carcasses might suggest that hygienic conditions in conventional farms are worse than antibiotic free farms. However, from a food safety point of view, Salmonella was not detected in both kinds of carcasses and the Campylobacter mean relative frequency of abundance was always lower than 0.4%

Hospitalized Pets as a Source of Carbapenem-Resistance

The massive and irrational use of antibiotics in livestock productions has fostered the occurrence and spread of resistance to “old class antimicrobials.” To cope with that phenomenon, some regulations have been already enforced in the member states of the European Union. However, a role of livestock animals in the relatively recent alerts on the rapid worldwide increase of resistance to last-choice antimicrobials as carbapenems is very unlikely. Conversely, these antimicrobials are increasingly administered in veterinary hospitals whose role in spreading bacteria or mobile genetic elements has not adequately been addressed so far. A cross-sectional study was carried out on 105 hospitalized and 100 non-hospitalized pets with the aim of measuring the prevalence of carbapenem-resistant Gram-negative bacteria (GNB) colonizing dogs and cats, either hospitalized or not hospitalized and estimating the relative odds. Stool samples were inoculated on MacConkey agar plates containing 1 mg/L imipenem which were then incubated aerobically at 37°C ± 1 for 48 h. Isolated bacteria were identified first by Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and were confirmed by 16S rRNA sequencing. The genetic basis of resistance was investigated using PCR methods, gene or whole genome sequencing (WGS). The prevalence of pets harboring carbapenem-resistant bacteria was 11.4 and 1.0% in hospitalized and not-hospitalized animals, respectively, with an odds ratio of 12.8 (p < 0.01). One pet carried two diverse isolates. Overall, 14 gram-negative non-fermenting bacteria, specifically, one Acinetobacter radioresistens, five Acinetobacter baumannii, six Pseudomonas aeruginosa and two Stenotrophomonas maltophilia were isolated. The Acinetobacter species carried acquired carbapenemases genes encoded by blaNDM-1 and blaOXA-23. In contrast, Pseudomonas phenotypic resistance was associated with the presence of mutations in the oprD gene. Notably, inherent carbapenem-resistant isolates of S. maltophilia were also resistant to the first-line recommended chemotherapeutic trimethoprim/sulfamethoxazole. This study estimates the risk of colonization by carbapenem-resistant non-fermenting GNB in pets hospitalized in veterinary tertiary care centers and highlights their potential role in spreading resistance genes among the animal and human community. Public health authorities should consider extending surveillance systems and putting the release of critical antibiotics under more strict control in order to manage the infection/colonization of pets in veterinary settings

DMTs and Covid-19 severity in MS: a pooled analysis from Italy and France

We evaluated the effect of DMTs on Covid-19 severity in patients with MS, with a pooled-analysis of two large cohorts from Italy and France. The association of baseline characteristics and DMTs with Covid-19 severity was assessed by multivariate ordinal-logistic models and pooled by a fixed-effect meta-analysis. 1066 patients with MS from Italy and 721 from France were included. In the multivariate model, anti-CD20 therapies were significantly associated (OR = 2.05, 95%CI = 1.39–3.02, p < 0.001) with Covid-19 severity, whereas interferon indicated a decreased risk (OR = 0.42, 95%CI = 0.18–0.99, p = 0.047). This pooled-analysis confirms an increased risk of severe Covid-19 in patients on anti-CD20 therapies and supports the protective role of interferon

Modelling survival behaviour of Salmonella enterica ser. Enteritidis, Typhimurium and Tennessee on table eggs during storage at different temperatures

Quantitative risk assessment studies on the health risk of Salmonella due to consumption of contaminated table eggs are based on the assumption that Salmonella is inside the egg and that the pathogen belongs to serovar Enteritidis. However different serovars of Salmonella may contaminate the surface of table eggs and spread to other foods at consumer's kitchen due to improper food handling. In the present study the survival behaviour of one strain each from Salmonella enterica serovars Enteritidis, Typhimurium and Tennessee on table egg surface during storage at 4, 8, and 20\ub0C have been described. Besides, in those cases observed data were subjected for modelling; linear, log-linear tail and Weibull models were compared in terms of model fitting and model performance. Overall, in most cases, inactivation kinetics presented a linear trend on Salmonella behaviour so that Weibull and linear models adequately described observed data. Regarding log-linear tail models, though they presented a better fitting, their adequacy could not be assessed given the lack of data in the tail region. Regarding storage temperatures, 4\ub0C was predicted to be the most inhibitory temperature for table eggs externally contaminated by a strain of S.enterica serovar Enteritidis. After 28 days of storage, a reduction of 4 log10cfu/g of eggshell on the S.enterica ser. Enteritidis load was registered at 4\ub0C. S.enterica ser. Typhimurium and Tennessee showed higher survival rates at all tested temperatures. The results highlighted the importance of keeping constant the storage temperature of table eggs in order to reduce the risk of S.enterica ser. Enteritidis contaminating the surface of table eggs. However this temperature might not be the optimal one in view of S.enterica serovars other than Enteritidis

Microbiological profile of chicken carcasses: A comparative analysis using shotgun metagenomic sequencing

In the last few years metagenomic and 16S rRNA sequencing have completly changed the microbiological investigations of food products. In this preliminary study, the microbiological profile of chicken carcasses collected from animals fed with different diets were tested by using shotgun metagenomic sequencing. A total of 15 carcasses have been collected at the slaughetrhouse at the end of the refrigeration tunnel from chickens reared for 35 days and fed with a control diet (n=5), a diet supplemented with 1500 FTU/kg of commercial phytase (n=5) and a diet supplemented with 1500 FTU/kg of commercial phytase and 3g/kg of inositol (n=5). Ten grams of neck and breast skin were obtained from each carcass and submited to total DNA extraction by using the DNeasy Blood & Tissue Kit (Qiagen). Sequencing libraries have been prepared by using the Nextera XT DNA Library Preparation Kit (Illumina) and sequenced in a HiScanSQ (Illumina) at 100 bp in paired ends. A number of sequences ranging between 5 and 9 million was obtained for each sample. Sequence analysis showed that Proteobacteria and Firmicutes represented more than 98% of whole bacterial populations associated to carcass skin in all groups but their abundances were different between groups. Moraxellaceae and other degradative bacteria showed a significantly higher abundance in the control compared to the treated groups. Furthermore, Clostridium perfringens showed a relative frequency of abundance significantly higher in the group fed with phytase and Salmonella enterica in the group fed with phytase plus inositol. The results of this preliminary study showed that metagenome sequencing is suitable to investigate and monitor carcass microbiota in order to detect specific pathogenic and/or degradative populations