An Integrated Analysis of Abattoir Lung Lesion Scores and Antimicrobial Use in Italian Heavy Pig Finishing Farms.

Respiratory diseases significantly affect intensive pig finishing farms, causing production losses and increased antimicrobial use (AMU). Lesion scoring at slaughter has been recognized as a beneficial practice to evaluate herd management. The integrated analysis of abattoir lesion scores and AMU data could improve decision-making by providing feedback to veterinarians and farmers on the effectiveness of antimicrobial treatments, thus rationalizing their use. This study compared lung and pleural lesion scores collected at Italian pig slaughterhouses with on-farm AMU, estimated through a treatment index per 100 days (TI100). Overall, 24,752 pig carcasses, belonging to 236 batches from 113 finishing farms, were inspected. Bronchopneumonia and chronic pleuritis were detected in 55% and 48% of the examined pigs, respectively. Antimicrobials were administered in 97% of the farms during the six months prior to slaughter (median TI100 = 5.2), notwithstanding compliance with the mandatory withdrawal period. EMA category B (critical) antimicrobials were administered in 15.2% of cases (median TI100 = 0.06). The lung score was not associated with the total AMU, but significant, positive associations were found with the past use of critical antimicrobials (p = 0.041) and macrolides (p = 0.044). This result highlights the potential of abattoir lung lesion monitoring to rationalize antimicrobial stewardship efforts, contributing to AMU reduction.

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.

Cadmium, lead, and mercury in two commercial squid species from the north Adriatic Sea (central Mediterranean): contamination levels and health risk assessment.

In this study, lead (Pb), cadmium (Cd), and total mercury (Hg) concentrations in European squids (Loligo vulgaris) and flying squids (Todarodes sagittatus) from the northern Adriatic Sea (Italy) were analyzed. The risk of the Italian population being exposed to potentially hazardous metal concentrations through the consumption of these products was also assessed. Compared to European squids, flying squids showed three times higher total Hg concentrations and one hundred times higher Cd concentrations to the point that more than 6 and 25% of the samples exceeded the maximum Hg and Cd limits established by the current legislation. From the evaluation of dietary exposure levels, it emerged that the consumption of flying squids was associated with the highest Pb intake by children and, consequently, with the lower margin of exposure values in relation to the risk of neurotoxicity (margin of exposure=33). Consumption of flying squids, especially by children, was also associated with higher intakes of Cd, inorganic, and methyl-Hg, which, respectively, accounted for 156, 113, and 23% of the tolerable weekly intakes established for these contaminants at European level. The obtained results raise concern and it may be necessary to provide specific dietary advice on the moderate dietary consumption of some cephalopod species, especially to the youngest and most vulnerable segment of the population. However, besides the highly conservative deterministic method adopted in this study, a refined consumer exposure assessment should be performed through the probabilistic methodology, which is more suitable to represent the real exposure scenario.

Elemental profiles of swine tissues as descriptors for the traceability of value-added Italian heavy pig production chains.

The increasing demand for reliable traceability tools in the meat supply chain has prompted the exploration of innovative approaches that meet stringent quality standards. In this work, 57 elements were quantified by inductively coupled plasma mass spectrometry and direct mercury analysis in 80 muscle and 80 liver samples of Italian heavy pigs to investigate the potential of new tools based on multi-elemental profiles in supporting value-added meat supply chains. Samples from three groups of animals belonging to the protected designation of origin (PDO) Parma Ham circuit (conventionally raised; raised with genetically modified organism (GMO)-free feeds; raised with GMO-free feeds plus the supplementation of omega-3 polyunsaturated fatty acids (n-3 PUFA)) and a fourth group of samples from animals not compliant with the PDO Parma Ham production process were analyzed. Hierarchical cluster analysis allowed for the identification of three macro-clusters of liver or muscle samples, highlighting some inhomogeneities among the target groups. Following SIMCA analysis, better classification models were obtained by using liver elemental profiles (95% correct classification rate), with the highest classification accuracy observed for GMO-free livers (100%). The elements contributing the most to the separation of livers by class membership were La, Ce, and Pb for conventional, Li, Cr, Fe, As, and Sr for GMO-free + n-3 PUFA, and Lu for non-PDO samples. Given these findings, the analysis of the elemental profiles of pig tissues can be regarded as a promising method to confirm the declared pig meat label attributes, deter potential complex fraud, and support meat traceability systems.

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.

1H NMR Metabolomics on Pigs' Liver Exposed to Antibiotics Administration: An Explorative Study.

An untargeted Nuclear Magnetic Resonance (NMR) spectroscopy-based metabolomics approach was applied as a first attempt to explore the metabolome of pigs treated with antibiotics. The final goal was to investigate the possibility of discriminating between antibiotic-treated (TX group) and untreated pigs (CTRL group), with the further perspective of identifying the authentication tools for antibiotic-free pork supply chains. In particular, 41 samples of pig liver were subjected to a biphasic extraction to recover both the polar and the non-polar metabolites, and the 1H NMR spectroscopy analysis was performed on the two separate extracts. Unsupervised (principal component analysis) and supervised (orthogonal partial least squares discriminant analysis) multivariate statistical analysis of 1H NMR spectra data in the range 0-9 ppm provided metabolomic fingerprinting useful for the discrimination of pig livers based on the antibiotic treatment to which they were exposed. Moreover, within the signature patterns, significant discriminating metabolites were identified among carbohydrates, choline and derivatives, amino acids and some lipid-class molecules. The encouraging findings of this exploratory study showed the feasibility of the untargeted metabolomic approach as a novel strategy in the authentication framework of pork supply chains and open a new horizon for a more in-depth investigation.

Microbial contamination, antimicrobial resistance and biofilm formation of bacteria isolated from a high-throughput pig abattoir.

The aim of this work was to assess the level of microbial contamination and resistance of bacteria isolated from a highthroughput heavy pig slaughterhouse (approx. 4600 pigs/day) towards antimicrobials considered as critical for human, veterinary or both chemotherapies. Samples, pre-operative and operative, were obtained in 4 different surveys. These comprised environmental sampling, i.e. air (ntotal = 192) and surfaces (ntotal = 32), in four different locations. Moreover, a total of 40 carcasses were sampled in two different moments of slaughtering following Reg. (CE) 2073/2005. Overall, 60 different colonies were randomly selected from VRBGA plates belonging to 20 species, 15 genera and 10 families being Enterobacteriaceae, Moraxellaceae and Pseudomonadaceae the most represented ones. Thirty-seven isolates presented resistance to at least one molecule and seventeen were classified as multi-drug resistant. Enterobacteriaceae, particularly E. coli, displayed high MIC values towards trimethoprim, ampicillin, tetracycline and sulphametoxazole with MICmax of 16, 32, 32 and 512 mg/L, respectively. Moreover, isolated Pseudomonas spp. showed high MIC values in critical antibiotics such as ampicillin and azithromycin with MICmax of 32 and 64 mg/L, respectively. Additionally, in vitro biofilm formation assays demonstrated that fifteen of these isolates can be classified as strong biofilm formers. Results demonstrated that a high diversity of bacteria containing antibiotic resistant and multiresistant species is present in the sampled abattoir. Considering these findings, it could be hypothesised that the processing environment could be a potential diffusion determinant of antibiotic resistant bacteria through the food chain and operators.

Occurrence of Toxic Metals and Metalloids in Muscle and Liver of Italian Heavy Pigs and Potential Health Risk Associated with Dietary Exposure.

Muscle and liver tissues from Italian heavy pigs were analyzed to investigate whether the chronic consumption of these products by local consumers could represent a health risk in relation to the contamination by some toxic metals and metalloids (TMMs). The concentrations of Al, As, Cd, Cr, Cu, Fe, Ni, Pb, Sn, U, and Zn were measured with an inductively coupled plasma-mass spectrometer, while Hg analysis was performed by using a mercury analyzer. Fe, Zn, and Cu were the most abundant elements in both tissues, while U was detected only at ultra-trace levels. As, Cd, Cu, Fe, Hg, Pb, U, and Zn showed significantly higher concentrations in livers compared to muscles (p ≤ 0.01), with Cd and Cu being 60- and 9-fold more concentrated in the hepatic tissue. Despite this, concentrations of all TMMs were found to be very low in all the samples to the point that the resulting estimated dietary intakes did not suggest any food safety concern. Indeed, intakes were all below the toxicological health-based guidance values or resulted in low margins of exposure. Nevertheless, in the calculation of the worst-case exposure scenario, the children's estimated intake of Cd, Fe, and Zn through the sole consumption of pig liver contributed to more than 23, 38, and 39% of the tolerable weekly intakes of these elements, while the combined consumption of pig liver and pig muscle to more than 24, 46, and 76%. These findings alert about the probability of exceeding the toxicological guidance values of Cd, Fe, and Zn though the whole diet, suggesting long-term negative health effects for the younger population.

Relationship between Bruises on Carcasses of Beef Cattle and Transport-Related Factors.

The assessment of bruises on carcasses at the slaughterhouse has been lately indicated as a valid method to evaluate cattle welfare. However, little is known about the prevalence and the causes of bruises of cattle slaughtered in Italy. The aim of this study was to collect information concerning the prevalence of bruises on the carcasses of beef cattle slaughtered in an Italian abattoir and to determine a relationship between fresh bruises and transport-related factors. In total, 1265 animals were included in this study, with 21.6% of them being positive for at least one bruise, either fresh or old. In most cases, the bruising was mild, with lesions exclusively located in one area of the carcass. Most of the bruised animals (63%) showed at least one red lesion. Occurrence of such red, fresh bruises varied significantly depending on the body parts (p < 0.0001), with the flank being the most affected area (39.5%), followed by the butt (36.0%) and the front (23.8%). The probability of fresh bruising varied significantly depending on the category of each animal (p < 0.0001), with steers showing fewer red bruises than both heifers and veal. Finally, animals transported in conditions of a high density had a lower probability of bruising (p = 0.0003). These findings support the use of a monitoring scheme based on the presence of bruises to assess cattle welfare at the abattoir level in order to provide feedback to farmers and to implement procedures carried out during transport.

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.

ESBL/AmpC-Producing Escherichia coli in Wild Boar: Epidemiology and Risk Factors.

The complex health problem of antimicrobial resistance (AMR) involves many host species, numerous bacteria and several routes of transmission. Extended-spectrum ß-lactamase and AmpC (ESBL/AmpC)-producing Escherichia coli are among the most important strains. Moreover, wildlife hosts are of interest as they are likely antibiotics free and are assumed as environmental indicators of AMR contamination. Particularly, wild boar (Sus scrofa) deserves attention because of its increased population densities, with consequent health risks at the wildlife-domestic-human interface, and the limited data available on AMR. Here, 1504 wild boar fecal samples were microbiologically and molecularly analyzed to investigate ESBL/AmpC-producing E. coli and, through generalized linear models, the effects of host-related factors and of human population density on their spread. A prevalence of 15.96% of ESBL/AmpC-producing E. coli, supported by blaCTX-M (12.3%), blaTEM (6.98%), blaCMY (0.86%) and blaSHV (0.47%) gene detection, emerged. Young animals were more colonized by ESBL/AmpC strains than older subjects, as observed in domestic animals. Increased human population density leads to increased blaTEM prevalence in wild boar, suggesting that spatial overlap may favor this transmission. Our results show a high level of AMR contamination in the study area that should be further investigated. However, a role of wild boar as a maintenance host of AMR strains emerged.

S. aureus Biofilm Protein Expression Linked to Antimicrobial Resistance: A Proteomic Study.

Antimicrobial resistance (AMR) represents one of the most critical challenges that humanity will face in the following years. In this context, a "One Health" approach with an integrated multidisciplinary effort involving humans, animals and their surrounding environment is needed to tackle the spread of AMR. One of the most common ways for bacteria to live is to adhere to surfaces and form biofilms. Staphylococcus aureus (S. aureus) can form biofilm on most surfaces and in a wide heterogeneity of environmental conditions. The biofilm guarantees the survival of the S. aureus in harsh environmental conditions and represents an issue for the food industry and animal production. The identification and characterization of biofilm-related proteins may provide interesting insights into biofilm formation mechanisms in S. aureus. In this regard, the aims of this study were: (i) to use proteomics to compare proteomes of S. aureus growing in planktonic and biofilm forms in order to investigate the common features of biofilm formation properties of different strains; (ii) to identify specific biofilm mechanisms that may be involved in AMR. The proteomic analysis showed 14 differentially expressed proteins among biofilm and planktonic forms of S. aureus. Moreover, three proteins, such as alcohol dehydrogenase, ATP-dependent 6-phosphofructokinase, and fructose-bisphosphate aldolase, were only differentially expressed in strains classified as high biofilm producers. Differentially regulated catabolites metabolisms and the switch to lower oxygen-related metabolisms were related to the sessile conformation analyzed.

Histamine Control in Raw and Processed Tuna: A Rapid Tool Based on NIR Spectroscopy.

The present study was designed to investigate whether near infrared (NIR) spectroscopy with minimal sample processing could be a suitable technique to rapidly measure histamine levels in raw and processed tuna fish. Calibration models based on orthogonal partial least square regression (OPLSR) were built to predict histamine in the range 10-1000 mg kg-1 using the 1000-2500 nm NIR spectra of artificially-contaminated fish. The two models were then validated using a new set of naturally contaminated samples in which histamine content was determined by conventional high-performance liquid chromatography (HPLC) analysis. As for calibration results, coefficient of determination (r2) > 0.98, root mean square of estimation (RMSEE) ≤ 5 mg kg-1 and root mean square of cross-validation (RMSECV) ≤ 6 mg kg-1 were achieved. Both models were optimal also in the validation stage, showing r2 values > 0.97, root mean square errors of prediction (RMSEP) ≤ 10 mg kg-1 and relative range error (RER) ≥ 25, with better results showed by the model for processed fish. The promising results achieved suggest NIR spectroscopy as an implemental analytical solution in fish industries and markets to effectively determine histamine amounts.

Advances in Troubleshooting Fish and Seafood Authentication by Inorganic Elemental Composition.

The demand for fish and seafood is growing worldwide. Meanwhile, problems related to the integrity and safety of the fishery sector are increasing, leading legislators, producers, and consumers to search for ways to effectively protect themselves from fraud and health hazards related to fish consumption. What is urgently required now is the availability of reliable, truthful, and reproducible methods assuring the correspondence between the real nature of the product and label declarations accompanying the same product during its market life. The evaluation of the inorganic composition of fish and seafood appears to be one of the most promising strategies to be exploited in the near future to assist routine and official monitoring operations along the supply chain. The present review article focuses on exploring the latest scientific achievements of using the multi-elemental composition of fish and seafood as an imprint of their authenticity and traceability, especially with regards to the geographical origin. The scientific literature of the last 10 years focusing on the analytical determination and statistical elaboration of elemental data (alone or in combination with methodologies targeting other compounds) to verify the identity of fishery products is summarized and discussed.

Abattoir-Based Measures to Assess Swine Welfare: Analysis of the Methods Adopted in European Slaughterhouses.

The assessment of swine welfare requires feasible, reliable, and reasonable indicators. On-farm evaluation of pig welfare can provide valuable information to veterinarians and farmers. However, such protocols can result expensive and time-consuming. With this regard, an interest in the appraisal of swine welfare at abattoir has grown over the recent years. In particular, the use of certain lesions collected directly from slaughtered animals to determine the welfare status of pigs has been evaluated by several authors. In the present review, the different methods developed to score lesions collected directly from the body and the viscera of animals slaughtered in European abattoirs ("abattoir-based measures") are presented. The text specifically focuses on the methods currently available in the literature for the scoring of body, pluck and gastric lesions during post-mortem activities. Moreover, the strengths and weaknesses of abattoir-based measures schemes are discussed. To conclude, the future perspectives of the assessment of pig welfare at the slaughterhouse are described, appealing for a benchmarking system that can be systematically used by veterinarians and other professional figures involved in the process.

The Use of Antimicrobials in Italian Heavy Pig Fattening Farms.

Data on antimicrobial use (AMU) in heavy pig production (>150 kg) are limited. The aim of this study was to investigate the AMU in this production. Data from 2015 were collected for 143 fattening farms. The AMU was estimated through a treatment index per 100 days (TI100) using the defined daily dose animal for Italy (DDDAit). When possible, a comparison with the European Medicines Agency's defined daily doses for animals (DDDvet) was performed. The median TI100 was 10.7 (range, 0.2-49.5). Group treatments represented 94.6% of overall consumption. The AMU calculated using DDDAit and DDDvet were strongly correlated (ρ = 0.976; p < 0.001). The AMU was negatively correlated with injectables use (ρ = -0.46, p < 0.001) and positively correlated with oral products (ρ = 0.21, p = 0.014), premixes (ρ = 0.26, p = 0.002), and mortality (ρ = 0.18; p = 0.027). Farm size was negatively correlated with AMU (ρ = -0.29, p < 0.001). Smaller farms were more frequently above the median TI100 (odds ratio = 2.3, 95% confidence interval = 1.2-4.7), suggesting that they may have lower biosecurity and management standards. The results of this study should provide useful insights for the development of an Italian monitoring system.

Molecular characterization of Methicillin-resistant Staphylococcus aureus isolated from the pig production chain in Northern Italy.

This study aimed to evaluate the molecular characteristics of methicillin resistant Staphylococcus aureus (MRSA) isolated in the swine chain in Northern Italy. A sample of 50 fattening units located in Lombardy was selected. Five cutaneous samples at slaughtering and three environmental samples at farm were collected from each unit giving a total of 250 and 150 samples, respectively. A total of 25 MRSA isolates were isolated from 400 samples, in 17 different fattening units. At farm, 12 out of 250 samples were positive for MRSA (4,8 %), and 13 out of 150 samples at slaughter were identified as MRSA (8,7 %), giving an overall incidence among samples of 6,25 % (n = 25). Molecular characterization was carried out using multi-locus sequence typing (MLST) and spa-typing. Outcomes showed that most of the isolates belonged to ST398, carrying spa-types t899, t011, t18498, t1939, t1200, and t304. Nonetheless, three isolates were identified as ST97 (t1730 and t4795), and one as ST30, showing spa-type t318. Furthermore, a novel ST was identified, namely 5422, showing spa-type t1730. Heterogeneity in genotypes within the same farm was also found in different fattening units, with concern for the possibility of the exchange of genetic determinants among different lineages. Genetic diversity among MRSA isolates in pig fattening units has been observed, highlighting the possibility that some isolates could be able to infect different hosts, including human.

Assessment of the Antibiotic Resistance Profile, Genetic Heterogeneity and Biofilm Production of Methicillin-Resistant Staphylococcus aureus (MRSA) Isolated from The Italian Swine Production Chain.

The main aim of the present study was to evaluate the level of antibiotic resistance, prevalence and virulence features of methicillin-resistant Staphylococcus aureus (MRSA) isolated from heavy swine at abattoir level and farming environments in Lombardy (Northern Italy). With this scope, 88 different heavy swine farms were surveyed, obtaining a total of n = 440 animal swabs and n = 150 environmental swabs. A total of n = 87 MRSA isolates were obtained, with an overall MRSA incidence of 17.50% (n = 77) among animal samples and a 6.67% (n = 10) among environmental. Molecular characterisation using multilocus sequence typing (MLST) plus spa-typing showed that sequence type ST398/t899 and ST398/t011 were the most commonly isolated genotypes, although other relevant sequence types such as ST1 or ST97 were also found. A lack of susceptibility to penicillins, tetracycline and ceftiofur was detected in >91.95, 85.05 and 48.28% of the isolates, respectively. Resistance to doxycycline (32.18%), enrofloxacin (27.59%) and gentamicin (25.29%) was also observed. Additionally, a remarkable level of antibiotic multiresistance (AMR) was observed representing a 77.01% (n = 67) of the obtained isolates. Genetic analysis revealed that 97.70% and 77.01% of the isolates harboured at least one antibiotic resistance or enterotoxin gene, respectively, pointing out a high isolate virulence potential. Lastly, 55.17% (n = 48) were able to produce measurable amounts of biofilm after 24 h. In spite of the current programmes for antibiotic reduction in intensively farming, a still on-going high level of AMR and virulence potential in MRSA was demonstrated, making this pathogen a serious risk in swine production chain, highlighting once more the need to develop efficient, pathogen-specific control strategies.