Vancomycin resistance and virulence genes evaluation in Enterococci isolated from pork and wild boar meat.

Enterococci are considered valuable sentinel Gram-positive bacteria for monitoring vancomycin antibiotic resistance due to their widespread presence and characteristics. The use of antimicrobials in farming animals has a role in the increasing of Antimicrobial Resistance (AMR) and the anthropogenic transformation of the landscape has forced wildlife into greater contact with humans and their livestock. The transmission of resistant bacteria by their meat products is a significant contributor to AMR development. The present study aimed to assess the prevalence of vancomycin resistant Enterococci spp. In antimicrobial-treated farmed pigs meat and in antimicrobial-free wild boars meat. A total of 341 Enterococci were isolated from 598 pork meat samples (57 %) and 173 Enterococci were isolated from 404 wild boar meat samples (42.8 %). Data found showed that low-resistance was detected more in wild boars meat Enterococci (52.6 %) than in pork meat once (48.4 %). However, the prevalence of resistance genes was at low level (33.9 % in pork meat Enterococci and 4.4 % in wild boar meat ones) and the only gene found was vanC1/C2, related to intrinsic AMR. Normally, Enterococci persist in the normal intestinal flora of animals including humans. However, the presence of resistance genes was frequently linked to the detection of pathogenic genes, mostly gelE in pork meat isolates and asa1 in wild boars meat isolates. Pathogenic bacteria can cause severe infections in human that can become more risky if associated to the presence of AMR. Pathogenic bacteria were characterized and a high presence of E. gallinarum and E. casseliflavus was found. Given the growing interest in wild game meat consumption the monitoring of AMR in these matrices is essential. Further surveillance studies are needed to fully evaluate the emergence and spread of vancomycin-resistant Enterococci (VRE) and pathogenic Enterococci from animal-derived food to humans, including the role of wildlife in this phenomenon. Giving the higher interest in wild animals meat consumption, it is important to better evaluate the spread of AMR phenomenon in the future and intensify hygienic control of wild animals derived food.

Emerging of Shiga toxin-producing Escherichia coli O177:H11 and O177:H25 from cattle at slaughter in Italy.

Shiga toxin-producing Escherichia coli (STEC) are zoonotic pathogens frequently carried by cattle, responsible in humans of mild to bloody diarrhoea, haemolytic uraemic syndrome (HUS) and even death. In 2023-2024, a study on STEC contamination of hide and carcasses of dairy cattle at slaughter was planned in Emilia-Romagna region (northern Italy). When the study was still in progress and 60 animals were sampled, the detection of STEC O177 isolates reached high rates and gained our attention. A total of five O177 STEC strains were detected, namely four from three carcasses (5.0 %) and one from a hide sample (1.7 %). The isolates were typed by WGS as following: 1) STEC O177:H11 sequence type (ST) 765 (stx2a+, eae+), detected from one carcass; 2) STEC O177:H25 ST659 (stx2c+, eae+) detected from three carcasses and one hide sample. One carcass was contaminated by both STEC serotypes. The isolates carried other virulence determinants often found in STEC strains associated with HUS, namely the exha, astA and espP genes, together with genes for adhesion to the epithelial cells of the gut (lpfA, fdeC, fimH) and non-Locus for Enterocyte Effacement (LEE) effector protein genes (nleA, nleB). The STEC O177:H11 isolate harboured antimicrobial resistance (AMR) genes to ß-lactams (blaTEM-1A), aminoglycosides (aadA1, aph(3″)-Ib, aph(6)-Id), trimethoprim (dfrA1), sulphonamides (sul1, sul2), tetracyclines (tetA), (tetB), streptothricin (sat2), and quaternary ammonium compounds (qacEdelta1). On the contrary, the STEC O177:H25 isolates carried no AMR genes. Persistent carriage of STEC O177:H25 ST659 (stx2c+, eae+) at farm level was assessed by testing animals of the same herd sent to slaughter. Interestingly, the colonies of STEC O177:H11 and STEC O177:H25 had different morphology on CHROMagar™ STEC plates, being mauve and colourless, respectively. Since mauve is the colour STEC colonies commonly have on the CHROMagar™ STEC medium, our findings can help microbiologists in the selection of uncommon serotypes. To the best of our knowledge, this is the first detection of STEC O177 from carcasses and hides of dairy cattle at slaughter. Noteworthy, the STEC-positive hide was classified as "very dirty" thus stressing the need of clean animals entering the slaughter chain, as required by Regulation (EC) No 853/2004. Since STEC O177 has been responsible of HUS in Europe, our data could add information on the source of uncommon serogroups in human infections.

Feasibility of Near-Infrared Spectroscopy in the Classification of Pig Lung Lesions.

Respiratory diseases significantly affect intensive pig farming, causing production losses and increased antimicrobial use. Accurate classification of lung lesions is crucial for effective diagnostics and disease management. The integration of non-destructive and rapid techniques would be beneficial to enhance overall efficiency in addressing these challenges. This study investigates the potential of near-infrared (NIR) spectroscopy in classifying pig lung tissues. The NIR spectra (908-1676 nm) of 101 lungs from weaned pigs were analyzed using a portable instrument and subjected to multivariate analysis. Two distinct discriminant models were developed to differentiate normal (N), congested (C), and pathological (P) lung tissues, as well as catarrhal bronchopneumonia (CBP), fibrinous pleuropneumonia (FPP), and interstitial pneumonia (IP) patterns. Overall, the model tailored for discriminating among pathological lesions demonstrated superior classification performances. Major challenges arose in categorizing C lungs, which exhibited a misclassification rate of 30% with N and P tissues, and FPP samples, with 30% incorrectly recognized as CBP samples. Conversely, IP and CBP lungs were all identified with accuracy, precision, and sensitivity higher than 90%. In conclusion, this study provides a promising proof of concept for using NIR spectroscopy to recognize and categorize pig lungs with different pathological lesions, offering prospects for efficient diagnostic strategies.

Transmission of ß-lactamases in the pork food chain: A public health concern.

Antimicrobial resistance (AMR) is a risk for public health that requires management in a One Health perspective, including humans, animals, and the environment. The food production chain has been identified as a possible route of transmission of AMR bacteria to humans. The most critical issue regards resistance to the Critically Important Antimicrobials (CIAs), such as ß-lactams antibiotics. Here, pigs were analysed along the entire food producing chain, including feces, carcasses and pork products (fresh meat, fermented and seasoned products) ensuring treaciability of all samples. Escherichia coli were isolated and their ability to produce ESBL and AmpC ß-lactamases was evaluated both phenotypically and genotypically. Strains with the same AMR profile from feces, carcasses, and meat products were selected for phylogenetic and comparative genomic analyses to evaluate the possible "farm-to-fork" transmission of ß-lactams resistant bacteria. Results showed that the percentage of ESBL strains in fecal E. coli was approximately 7% and increased slightly in the pork food chain: the 10% of ESBL E. coli isolated from carcasses and the 12.5% of isolates from fresh meat products. AmpC E. coli were found only in feces, carcasses, and fresh meat with a low prevalence. Results showed that of the 243 pigs followed along the entire food chain genetic similarities in E. coli isolated from farm-to-fork were found in only one pig (feces, carcasses and fresh meat). Frequent similarities were shown in resistant E. coli isolates from carcasses and fresh meat or fermented product (three pork food chain). Moreover, in one case, bacteria isolated from fresh meat and fermented product were genotypically similar. Concluding, direct transmission of ß-lactams resistance from farm-to-fork is possible but not frequent. Further studies are needed to improve risk communication to consumers and access to clear and reliable information and health concerns on food.

A Preliminary Study on the Relationship between Gastric Lesions and Anti-Inflammatory Drug Usage in Heavy Pigs.

Gastric lesions in pigs cause welfare and economic losses. Their prevalence in heavy pigs reared for premium products (e.g., Parma ham) requires further investigation. Stress, nutrition, and farm management are known risk factors, but the effects of steroidal and non-steroidal anti-inflammatory drugs (NSAIDs) are largely unknown. The aim of this study was to evaluate the prevalence of gastric lesions in Italian heavy pigs and their possible association with the use of anti-inflammatory drugs. A total of 9371 pig stomachs from 76 farms were evaluated. Among these, 20.3% showed no lesions, while 30.7%, 42.1%, and 6.8% were scored 1, 2 and 3, respectively. A tendency for an inverse relationship with farm size emerged. The use of steroids and NSAIDs was estimated by calculating a treatment incidence per 1000 (TI1000) in a subset of 36 farms. At least one prescription for NSAIDs and/or steroids was found in 80.6% of the farms (55.6% used NSAIDs and 63.9% used steroids). Median TI1000 was 0.07 (range: 0-30.1) and 0.18 (range: 0-6.2) for NSAIDs and steroids, respectively. Gastric scores were positively associated with NSAID use, but not with steroid use. Although the role of these drugs in gastric lesions needs to be further clarified, these findings suggest a cautious use of non-selective NSAIDs.

Comparing Visual-Only and Visual-Palpation Post-Mortem Lung Scoring Systems in Slaughtering Pigs.

Respiratory diseases continue to pose significant challenges in pig production, and the assessment of lung lesions at the abattoir can provide valuable data for epidemiological investigations and disease surveillance. The evaluation of lung lesions at slaughter is a relatively simple, fast, and straightforward process but variations arising from different abattoirs, observers, and scoring methods can introduce uncertainty; moreover, the presence of multiple scoring systems complicates the comparisons of different studies, and currently, there are limited studies that compare these systems among each other. The objective of this study was to compare validated, simplified, and standardized schemes for assessing surface-related lung lesions in slaughtered pigs and analyze their reliability under field conditions. This study was conducted in a high-throughput abattoir in Italy, where two different scoring methods (Madec and Blaha) were benchmarked using 637 plucks. Statistical analysis revealed a good agreement between the two methods when severe or medium lesions were observed; however, their ability to accurately identify healthy lungs and minor injuries diverged significantly. These findings demonstrate that the Blaha method is more suitable for routine surveillance of swine respiratory diseases, whereas the Madec method can give more detailed and reliable results for the respiratory and welfare status of the animals at the farm level.

Isotope Fingerprinting as a Backup for Modern Safety and Traceability Systems in the Animal-Derived Food Chain.

In recent years, due to the globalization of food trade and certified agro-food products, the authenticity and traceability of food have received increasing attention. As a result, opportunities for fraudulent practices arise, highlighting the need to protect consumers from economic and health damages. In this regard, specific analytical techniques have been optimized and implemented to support the integrity of the food chain, such as those targeting different isotopes and their ratios. This review article explores the scientific progress of the last decade in the study of the isotopic identity card of food of animal origin, provides the reader with an overview of its application, and focuses on whether the combination of isotopes with other markers increases confidence and robustness in food authenticity testing. To this purpose, a total of 135 studies analyzing fish and seafood, meat, eggs, milk, and dairy products, and aiming to examine the relation between isotopic ratios and the geographical provenance, feeding regime, production method, and seasonality were reviewed. Current trends and major research achievements in the field were discussed and commented on in detail, pointing out advantages and drawbacks typically associated with this analytical approach and arguing future improvements and changes that need to be made to recognize it as a standard and validated method for fraud mitigation and safety control in the sector of food of animal origin.

Filling gaps in animal welfare assessment through metabolomics.

Sustainability has become a central issue in Italian livestock systems driving food business operators to adopt high standards of production concerning animal husbandry conditions. Meat sector is largely involved in this ecological transition with the introduction of new label claims concerning the defense of animal welfare (AW). These new guarantees referred to AW provision require new tools for the purpose of authenticity and traceability to assure meat supply chain integrity. Over the years, European Union (EU) Regulations, national, and international initiatives proposed provisions and guidelines for assuring AW introducing requirements to be complied with and providing tools based on scoring systems for a proper animal status assessment. However, the comprehensive and objective assessment of the AW status remains challenging. In this regard, phenotypic insights at molecular level may be investigated by metabolomics, one of the most recent high-throughput omics techniques. Recent advances in analytical and bioinformatic technologies have led to the identification of relevant biomarkers involved in complex clinical phenotypes of diverse biological systems suggesting that metabolomics is a key tool for biomarker discovery. In the present review, the Five Domains model has been employed as a vademecum describing AW. Starting from the individual Domains-nutrition (I), environment (II), health (III), behavior (IV), and mental state (V)-applications and advances of metabolomics related to AW setting aimed at investigating phenotypic outcomes on molecular scale and elucidating the biological routes most perturbed from external solicitations, are reviewed. Strengths and weaknesses of the current state-of-art are highlighted, and new frontiers to be explored for AW assessment throughout the metabolomics approach are argued. Moreover, a detailed description of metabolomics workflow is provided to understand dos and don'ts at experimental level to pursue effective results. Combining the demand for new assessment tools and meat market trends, a new cross-strategy is proposed as the promising combo for the future of AW assessment.

Different Roles of Wild Boars and Livestock in Salmonella Transmission to Humans in Italy.

Wild boar (Sus scrofa) is the most widely distributed large wildlife mammal worldwide. To investigate the transmission of Salmonella enterica amongst wild boars (Sus scrofa), humans, and livestock, we compared via pulsed-field gel electrophoresis and whole genome sequences the isolates of S. enterica serovar Typhimurium (biphasic and monophasic variants) and Enteritidis collected from wild boars, food-producing animals, and human patients in Emilia-Romagna region (Northern Italy) between 2017 and 2020. Specifically, we analysed 2175 isolates originated from human (1832), swine (117), bovine (128), poultry (76), and wild boar (22). The genomic analyses showed that wild boars shared most of their lineages of biphasic Typhimurium with bovines and most of Enteritidis with poultry, whilst we did not find any lineage shared with swine. Moreover, almost 17% of human biphasic Typhimurium and Enteritidis belonged to genomic clusters including wild boar isolates, but the inclusion of bovine and poultry isolates in the same clusters and the peculiar spatial distribution of the isolates suggested that human cases (and wild boar infections) likely originated from bovines and poultry. Consequently, wild boars appear not to play a significant role in infecting humans with these serovars, but seem to get infected themselves from livestock, probably through the environment.

The Pork Food Chain as a Route of Transmission of Antimicrobial Resistant Escherichia coli: A Farm-to-Fork Perspective.

Antimicrobial resistance (AMR) is a public health risk that needs to be faced from a One Health perspective that includes humans, animals, and environmental health. The food production chain has been identified as a possible route of transmission of AMR bacteria to humans. The most critical phenomenon is related to Critically Important Antimicrobial (CIA) resistance. ß-lactams antibiotics (cephalosporin of 3rd, 4th generation, carbapenem, monobactams, and penicillins), quinolones, aminoglycosides, polymyxin, and glycylcyclines were the CIAs chosen in this study. Samples derived from all the stages of the pork food production chain were collected, including pig feces, carcasses, and pork food products (fresh meat, fermented, and seasoned). Escherichia coli were isolated, and AMR and MDR profiles were evaluated. Enterobacterial Repetitive Intragenic Consensus (ERIC-PCR) was used to evaluate phylogenetic similarities. Data showed that 50% of phenotypical AMR observed in the entire pork food chain were related phylogenetically. The contamination of fresh meat, in half of the cases, was not directly related to contamination from feces or carcasses. Despite this, some similarities were found between feces and carcasses. In group analysis, phylogenetic similarities were detected in a 3/36 cluster (8.3%). Nevertheless, further studies are needed to improve consumer risk communication and access to clear and reliable information and health concerns on food labels.

Antimicrobial Resistant E. coli in Pork and Wild Boar Meat: A Risk to Consumers.

Antimicrobial-resistant foodborne microorganisms may be transmitted from food producing animals to humans through the consumption of meat products. In this study, meat that was derived from farmed pigs and wild boars was analyzed and compared. Escherichia coli (E. coli) were isolated and tested phenotypically and genotypically for their resistance to quinolones, aminoglycosides and carbapenems. The co-presence of AMR-associated plasmid genes was also evaluated. A quinolone AMR phenotypic analysis showed 41.9% and 36.1% of resistant E. coli derived from pork and wild boars meat, respectively. A resistance to aminoglycosides was detected in the 6.6% of E. coli that was isolated from pork and in 1.8% of the wild boar meat isolates. No resistant profiles were detected for the carbapenems. The quinolone resistance genes were found in 58.3% of the phenotypically resistant pork E. coli and in 17.5% of the wild boar, thus showing low genotypic confirmation rates. The co-presence of the plasmid-related genes was observed only for the quinolones and aminoglycosides, but not for the carbapenems. Wild boar E. coli were the most capable to perform biofilm production when they were compared to pork E. coli. In conclusion, the contamination of pork and wild boar meat by AMR microorganisms could be a threat for consumers, especially if biofilm-producing strains colonize the surfaces and equipment that are used in the food industry.

Prevalence and antimicrobial resistance profile in Salmonella spp. isolates from swine food chain.

The aim of this survey was to examine the prevalence and the antimicrobial resistance (AMR) of Salmonella spp. isolated from swine food chain. A total of 435 samples were collected: 360 from slaughterhouse (150 carcasses, 30 cecal samples, 180 environmental samples) and 75 from Italian traditional pork dry sausages. Thirty-six Salmonella were isolated and identified by Polymerase Chain Reaction (PCR): 13,3% (4/30) in fecal samples, 5,5% (10/180) in environmental samples, 7,3% (11/150) in carcasses, and 14,6% (11/75) in Italian traditional dry sausages. Salmonella serotypes were: S. Typhimurium (44,4%), S. Typhimurium monophasic variant (8,3%), S. Typhi (2,8%), S. Enteritidis (22,2%), S. Rissen (16,6%) and S. Derby (5,5%). Phenotypic and genotypic characterization of AMR Salmonella spp. isolates was executed through automatic system (VITEK 2, bioMèrieux) and PCR assays. Salmonella spp. showed phonotypical and genotypical resistance to at least one or more classes of antibiotic. All Salmonella spp. were resistant to aminoglycoside (amikacin and tobramycin) and gentamicin, 86,1% strains were resistant to tetracycline, 55,5% strains were resistant to ampicillin and piperacillin, 25% strains to trimethoprim, 5,5% strains to chloramphenicol, 2,8% strains to amoxicillin/ clavulanic acid, and nitrofurantoin. Among Salmonella isolates, the most detected AMR genes were catA for chloramphenicol (94,4%), nitrofuran nfsA (77.7%), nfsB (86,1%) and, for fluoroquinolone par C (100%) and gyrA (94,4%). This study reported epidemiological data regarding Salmonella spp. and AMR's circulation in the swine food chain. This phenomenon (AMR) has critical repercussions on the final consumer health; therefore, it represents a crucial One-Health issue.

Predictivity of Antemortem Findings on Postmortem Inspection in Italian Heavy Pigs Slaughterhouses.

Pigs slaughtered in European abattoirs must be submitted to antemortem inspection (AMI) and postmortem inspection (PMI), as required by the current European legislation in the matter of official controls. AMI and PMI are equally essential to guarantee food safety and to monitor swine health and welfare. However, little is known about the ability of AMI to predict conditions that are possibly found during PMI. In this study, such a correlation was explored together with the assessment of conditions typically found during AMI and PMI in heavy pigs slaughtered in two Italian slaughterhouses. An assessment scheme containing 13 variables for AMI and 34 lesions for PMI was used for the scope. The herd size was also considered as a variable and included in the study. A total of 24,510 pigs and 30,961 pigs were assessed during AMI and PMI, respectively. The most common conditions found were manure on the body covering more than 30% of the body (dirt >30%) and pluck lesions ('pleurisy', 'pericarditis', and 'pneumonia') for AMI and PMI, respectively. A significant correlation (p < 0.05) between some antemortem (AM) findings and postmortem (PM) conditions was found. In particular, the AM conditions 'dirt >30%'and 'skin lesions' were positively related with PM conditions 'skin wounds' and 'dermatitis', while the complexes of respiratory and kidney lesions were predicted only by the condition 'dirt >30%'. The variable 'standardized herd size' was negatively associated with 'milk spot liver' and positively associated with 'arthritis/bursitis'. The results of this study show that findings reported during AMI can potentially be used to predict certain conditions found in pigs at PMI. These data can be useful for the competent authorities in characterizing swine farms using a risk-based approach and in developing systems and specific plans for official controls.

Longitudinal study on the sources of Listeria monocytogenes contamination in cold-smoked salmon and its processing environment in Italy.

The aim of the present study was to investigate the sources of Listeria monocytogenes contamination in a cold smoked salmon processing environment over a period of six years (2003-2008). A total of 170 samples of raw material, semi-processed, final product and processing surfaces at different production stages were tested for the presence of L. monocytogenes. The L. monocytogenes isolates were characterized by multiplex PCR for the analysis of virulence factors and for serogrouping. The routes of contamination over the six year period were traced by PFGE. L. monocytogenes was isolated from 24% of the raw salmon samples, 14% of the semi-processed products and 12% of the final products. Among the environmental samples, 16% were positive for L. monocytogenes. Serotyping yielded three serovars: 1/2a, 1/2b, 4b, with the majority belonging to serovars 1/2a (46%) and 1/2b (39%). PFGE yielded 14 profiles: two of them were repeatedly isolated in 2005-2006 and in 2007-2008 mainly from the processing environment and final products but also from raw materials. The results of this longitudinal study highlighted that contamination of smoked salmon occurs mainly during processing rather than originating from raw materials, even if raw fish can be a contamination source of the working environment. Molecular subtyping is critical for the identification of the contamination routes of L. monocytogenes and its niches into the production plant when control strategies must be implemented with the aim to reduce its prevalence during manufacturing.

Mineral composition of Italian salami and effect of NaCl partial replacement on compositional, physico-chemical and sensory parameters.

In 2008 the European Commission developed an EU framework for dietary sodium chloride (NaCl) reduction in order to achieve the World Health Organization recommendations for no more than 5 g/day/person. This initiative is based on four elements: investigate the national data available on NaCl consumption and current NaCl levels of foods, develop actions to raise public awareness, develop reformulation actions with industry/catering, monitor and evaluate actions and reformulations. The initiative is working towards a reduction in NaCl of 16% over 4 years against the 2008 levels and is concentrated on meat products, bread, cheese, and ready meals. In this context, NaCl content and mineral composition of commercial Italian salami were investigated to provide information on their current mineral levels. Moreover, a technological intervention based on NaCl partial replacement by other chloride salts was investigated on Cacciatore salami, a typical Italian dry fermented sausage, as a strategy to decrease the sodium (Na) content of cured meat products. The effect of NaCl partial replacement by KCl, MgCl2, and CaCl2 in some compositional, physicochemical, and sensory properties of Cacciatore salami was evaluated. A 50% reduction of NaCl used for salami manufacture and its replacement by a mixture of KCl, CaCl2, and MgCl2 allowed a 40% lowering of Na content with limited detrimental effects on sensory attributes. Although no effects were observed on pH, water activity, proximate, and free fatty acid composition in reduced sodium Cacciatore salami formulation compared to the traditional one, the NaCl partial replacement induced a significant increase of lipid oxidation.

Polymorphism of actA gene is not related to in vitro virulence of Listeria monocytogenes.

Listeria monocytogenes is a foodborne pathogen which is able to cause serious disease both in humans and in animals. Several studies have demonstrated variations in the levels of virulence among L. monocytogenes strains. Invasion and growth ability of L. monocytogenes into cultured cells have been used to evaluate its pathogenicity. In particular, invasiveness and growth ability have been typically investigated using HeLa cell line. This study aimed to provide further insights on the virulence potential as well as on the molecular and phenotypic characteristics of L. monocytogenes isolated both from food sources and food environments. Thirty-eight isolates were tested for cell invasion and intracellular growth. Among the latter, 15 strains exhibited a high invasion index (I.I.); 18 strains showed intermediate II and 5 isolates revealed a low II. Regarding intracellular growth, all tested isolates had a replication time between 2 and 6h. Furthermore, nine virulence-associated genes (hlyA, actA, inlA, inlB, iap, plcA, plcB, mpl, prfA) were investigated by the multiplex PCR assay. All tested virulence genes were detected in all strains. Interestingly, a polymorphism was observed in the actA gene. However, the polymorphism could not be related to a different level of invasion or intracellular growth. In conclusion, data presented in this study have revealed considerable differences in the ability of L. monocytogenes strains to invade host cells and suggest the presence of additional factors that may contribute to adhesion and invasion. Virulence of L. monocytogenes is still not fully understood in some respects. Further studies focused on the mechanisms of L. monocytogenes pathogenicity together with the development of more reliable and efficient methods for virulence determination in this species are still required.

Characterization of antimicrobial resistance of foodborne Listeria monocytogenes.

The objective of this study was to evaluate the susceptibility of 120 Listeria monocytogenes strains isolated from food and food-processing environments to 19 antibiotics currently used in veterinary and human therapy. Susceptibility tests were performed by using the automated VITEK2 system. Apart from penicillin, ampicillin and trimethoprim-sulfamethoxazole, for which clinical breakpoints for Listeria susceptibility testing are defined according to the Clinical and Laboratory Standard Institute (CLSI), in the present study the CLSI criteria for staphylococci were applied. Among the 120 tested strains, 14 (11.7%) displayed resistance to at least one antibiotic. In particular, resistance to one antibiotic was more common than multiple resistance, i.e., 10 (8.3%) isolates were resistant to one antibiotic, 3 (2.5%) to two antibiotics and one (0.8%) to five antibiotics. Resistance to clindamycin was the most common, followed by linezolid, ciprofloxacin, ampicillin and rifampicin, trimethoprim/sulphamethoxazole and, finally, vancomycin and tetracycline. This study shows that L. monocytogenes strains from food and food-processing environments are susceptible to the antibiotics commonly used in veterinary and human listeriosis treatment. Considering that L. monocytogenes is slowly becoming antibiotic resistant, a continued surveillance of emerging antimicrobial resistance of this pathogen is important to ensure effective treatment of human listeriosis. These data are useful in improving background data on antibiotic resistance of strains isolated from food and food environment.

Safety improvement and preservation of typical sensory qualities of traditional dry fermented sausages using autochthonous starter cultures.

Traditional dry fermented sausages are manufactured without addition of starter cultures in small-scale processing units, their fermentation relying on indigenous microflora. Characterisation and control of these specific bacteria are essential for the sensory quality and the safety of the sausages. The aim of this study was to develop an autochthonous starter culture that improves safety while preserving the typical sensory characteristics of traditional sausages. An autochthonous starter composed of Lactobacillus sakei, Staphylococcus equorum and Staphylococcus succinus isolated from a traditional fermented sausage was developed. These strains were tested for their susceptibility to antibiotics and their production of biogenic amines. This starter was evaluated in situ at the French traditional processing unit where the strains had been isolated. Effects of the autochthonous starter were assessed by analysing the microbial, physico-chemical, biochemical and sensory characteristics of the sausages. Inoculation with the chosen species was confirmed using known species-specific PCR assays for L. sakei and S. equorum and a species-specific PCR assay developed in this study for S. succinus. Strains were monitored by pulse-field gel electrophoresis typing. Addition of autochthonous microbial starter cultures improved safety compared with the traditional natural fermentation of sausages, by inhibiting the pathogen Listeria monocytogenes, decreasing the level of biogenic amines and by limiting fatty acid and cholesterol oxidation. Moreover, autochthonous starter did not affect the typical sensory quality of the traditional sausages. This is the first time to our knowledge that selection, development and validation in situ of autochthonous starter cultures have been carried out, and also the first time that S. equorum together with S. succinus have been used as starter cultures for meat fermentation. Use of autochthonous starter cultures is an effective tool for limiting the formation of unsafe compounds in traditional sausage while preserving their original and specific sensory quality.

Evaluation of 2-alkylcyclobutanones in irradiated cured pork products during vacuum-packed storage.

The 2-Alkylcyclobutanones (2-ACBs) content was determined in three Italian cured pork products (salame Milano, coppa, and pancetta) irradiated at different targeted irradiation doses (2, 5, and 8 kGy) during vacuum-packed storage. Among 2-ACBs, three different compounds were investigated, namely, 2-dodecylcyclobutanone, 2-tetradecylcyclobutanone, and 2-(tetradec-5'-enyl)cyclobutanone. 2-ACBs were absent from the nonirradiated samples, whereas their content increased with irradiation dose. Their presence was recorded occasionally at 2 kGy and constantly at higher irradiation doses (5 and 8 kGy). The plot of 2-ACBs content against targeted irradiation doses showed an exponential relationship. The effect of vacuum-packed storage time on the 2-ACBs content was dependent on the irradiation dose. During vacuum-packed storage for up to 60 days, the 2-ACBs content remained unchanged in the cured pork products irradiated at 2 and 5 kGy, whereas a significant increase was observed in the pork products irradiated at 8 kGy.

Rapid detection and identification of Brachyspira aalborgi from rectal biopsies and faeces of a patient.

This study reports for the first time the detection of Brachyspira aalborgi in faeces and rectal biopsies of a female suffering for 3-4 months of abdominal pain with long-standing mucosal diarrhoea, rectal bleeding and suspected carcinoma of the rectum. After pre-treatment of samples (faeces and biopsies) with a liquid medium (trypticase soy broth-TSB) containing foetal calf serum (FCS, 10%) and spectinomycin and rifampicin (TSB-SR) the first detection of B. aalborgi isolate HBS1 was observed after 48 h in the primary plates of selective blood agar modified medium (BAM) containing spectinomycin and rifampicin (BAM-SR), where growth zones were signalled by a small weakly beta-haemolytic halo. Attempts to subculture spirochaetes in agar media failed. The new HBS1 isolate was only propagated in TSB broth and at electron microscopy it showed 4 endoflagella inserted at each tapered end. The phenotypic characterization of HBS1 demonstrated absence of hippurate hydrolysis, indole production, alpha-galactosidase, alpha- and beta-glucosidase activities in accordance with the B. aalborgi type strain. Rapid identification of B. aalborgi isolate HBS1 was performed directly from faeces and rectal biopsies and subsequently from pure cultures by a genetic method based on 16S DNA restriction fragment length polymorphism (RFLP)-polymerase chain reaction (PCR). The sequence of 16S DNA amplicon of the isolate HBS1 was found 99.2% corresponding to that of the B. aalborgi type strain. Our results encourage further investigations for the development of a suitable selective agar medium for the isolating and cultivating B. aalborgi from human specimens.