New Stable Gallium(III) and Indium(III) Complexes with Thiosemicarbazone Ligands: A Biological Evaluation.

The aim of this work is to explore a new library of coordination compounds for medicinal applications. Gallium is known for its various applications in this field. Presently, indium is not particularly important in medicine, but it shares a lot of chemical traits with its above-mentioned lighter companion, gallium, and is also used in radio imaging. These metals are combined with thiosemicarbazones, ligating compounds increasingly known for their biological and pharmaceutical applications. In particular, the few ligands chosen to interact with these hard metal ions share the ideal affinity for a high charge density. Therefore, in this work we describe the synthesis and the characterization of the resulting coordination compounds. The yields of the reactions vary from a minimum of 21% to a maximum of 82%, using a fast and easy procedure. Nuclear Magnetic Resonance (NMR) and Infra Red (IR) spectroscopy, mass spectrometry, elemental analysis, and X-ray Diffraction (XRD) confirm the formation of stable compounds in all cases and a ligand-to-metal 2:1 stoichiometry with both cations. In addition, we further investigated their chemical and biological characteristics, via UV-visible titrations, stability tests, and cytotoxicity and antibiotic assays. The results confirm a strong stability in all explored conditions, which suggests that these compounds are more suitable for radio imaging applications rather than for antitumoral or antimicrobic ones.

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.

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.

Bismuth complex of quinoline thiosemicarbazone restores carbapenem sensitivity in NDM-1-positive Klebsiella pneumoniae.

Resistant bacteria represent an urgent worldwide threat. NDM-1-producing strains are rendering the last line antibiotics less effective. Six bismuth complexes of general formula BiLCl2, where L is a thiosemicarbazone bearing a quinoline moiety, have been synthesized and fully characterized, including their X-ray crystal structures. The synergistic relationship between the compounds and meropenem have been tested in a combination therapy in carbapenem-resistant Klebsiella pneumoniae (NTCT14331) carrying the NDM-1 gene. Quinoline-2-carboxaldehyde-N4-phenyl-3-thiosemicarbazone bismuth dichloride and carbapenem showed synergism in a dose dependent manner with negligible antibacterial activity when used in a monotherapy and could restore antibiotic sensitivity in the strain producing NDM-1 enzyme. The minimum inhibitory concentration (MIC) of meropenem lowered down 128 folds up to 2 µgmL-1, a concentration lower to the sensitivity level. The IC50 of the compound against A549 human lung carcinoma cells and HuDe human epithelial tissue was 46.96 ± 16.66 µM and 54.26 ± 9.89 µM respectively. The cytotoxicity against human cells was higher than the effective concentration needed for the synergistic effect in bacterial cells, indicating that a structural optimization of the compounds is needed.

Evolution of ß-lactams, fluroquinolones and colistin resistance and genetic profiles in Salmonella isolates from pork in northern Italy.

The European Food Safety Authority and European Centre of Disease Prevention and Control antimicrobial resistance report published in 2021 shows increasing levels of antimicrobial resistance in Salmonella against antibiotics of choice for human salmonellosis (s-lactams and fluoroquinolones). The aim of the study was to follow the evolution of resistance against some Critical Important Antimicrobials in Salmonella isolates from fresh pork collected in Emilia-Romagna region, northern Italy, over two decades. Emilia-Romagna region is characterized by production of well-known pork derived products, as Parma Ham. The samples were collected in three different periods, ranging from 2000 to 2003, 2012 to 2016 and 2018 to 2021. After serotyping, the isolates were phenotypically tested for resistance to three classes of antibiotics: s-lactams, fluoroquinolones and polymyxins. End-point polymerase chain reaction (PCR) and PCRReal Time were used for genotypical analyses. The phenotypical resistance to s-lactams and fluoroquinolones were clearly increasing when comparing the results obtained from isolates collected in the first period (16.7% and 16.7%, respectively) with those of the third period (29.7% and 32.4%, respectively). On the contrary, the resistance to colistin decreased from 33.3% to 5.4%. Genotypically, the 71.4% and 83.3% of the strains harboured s-lactams and fluoroquinolones genes, respectively, while colistin resistance genes were not detected in the phenotypically resistant strains.

Gallium(III)-pyridoxal thiosemicarbazone derivatives as nontoxic agents against Gram-negative bacteria.

Many bacterial strains are developing mechanism of resistance to antibiotics, rendering last-resort antibiotics inactive. Therefore, new drugs are needed and in particular metal-based compounds represent a valid starting point to explore new antibiotic classes. In this study, we have chosen to investigate gallium(III) complexes for their potential antimicrobial activity against different strains of Klebsiella pneumoniae, Escherichia coli, and Pseudomonas aeruginosa which have developed different type of resistance mechanism, including the expression of ß-lactamases (NDM-1, ESßL, or AmpC) or the production of biofilm. We studied a series of thiosemicarbabazones derived from pyridoxal, their related Ga(III) complexes, and the speciation in solution of the Ga(III)/ligand systems as a function of the pH. Proton dissociation constants and conditional stability constants of Ga(III) complexes were evaluated by UV/Vis spectroscopy, and the most relevant species at physiological pH were identified. The compounds are active against resistant Gram-negative strain with minimal inhibitory concentration in the µM range, while no cytotoxicity was detected in eukaryotic cells.

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.

Survey on Carbapenem-Resistant Bacteria in Pigs at Slaughter and Comparison with Human Clinical Isolates in Italy.

This study is focused on resistance to carbapenems and third-generation cephalosporins in Gram-negative microorganisms isolated from swine, whose transmission to humans via pork consumption cannot be excluded. In addition, the common carriage of carbapenem-resistant (CR) bacteria between humans and pigs was evaluated. Sampling involved 300 faecal samples collected from slaughtered pigs and 300 urine samples collected from 187 hospitalised patients in Parma Province (Italy). In swine, MIC testing confirmed resistance to meropenem for isolates of Pseudomonas aeruginosa and Pseudomonas oryzihabitans and resistance to cefotaxime and ceftazidime for Escherichia coli, Ewingella americana, Enterobacter agglomerans, and Citrobacter freundii. For Acinetobacter lwoffii, Aeromonas hydrofila, Burkolderia cepacia, Corynebacterium indologenes, Flavobacterium odoratum, and Stenotrophomonas maltophilia, no EUCAST MIC breakpoints were available. However, ESBL genes (blaCTXM-1, blaCTX-M-2, blaTEM-1, and blaSHV) and AmpC genes (blaCIT, blaACC, and blaEBC) were found in 38 and 16 isolates, respectively. P. aeruginosa was the only CR species shared by pigs (4/300 pigs; 1.3%) and patients (2/187; 1.1%). P. aeruginosa ST938 carrying blaPAO and blaOXA396 was detected in one pig as well as an 83-year-old patient. Although no direct epidemiological link was demonstrable, SNP calling and cgMLST showed a genetic relationship of the isolates (86 SNPs and 661 allele difference), thus suggesting possible circulation of CR bacteria between swine and humans.

Prevalence of ESßL, AmpC and Colistin-Resistant E. coli in Meat: A Comparison between Pork and Wild Boar.

A global increase in Escherichia coli (E. coli) resistant to cephalosporins (extended-spectrum ß-lactamases (ESßLs) and AmpC ß-lactamases) has been recorded in the last 20 years. Similarly, several studies have reported the spread of colistin resistance in Enterobacteriaceae isolated from food and the environment. The aim of the present study was to evaluate the prevalence of ESßL, AmpC and colistin-resistant E. coli isolated from pork and wild boar meat products in the Emilia Romagna region (North Italy). The isolates were analysed phenotypically (considering both resistant and intermediate profiles) and genotypically. The prevalence of genotypically confirmed ESßL and AmpC E. coli was higher in pork meat products (ESßL = 11.1% vs. AmpC = 0.3%) compared to wild boar meat (ESßL = 6.5% vs. AmpC = 0%). Intermediate profiles for cefotaxime (CTX) and ceftazidime (CAZ) were genotypically confirmed as ESßL in pork meat isolates but not for wild boar. Four E. coli from wild boar meat were resistant to colistin but did not harbour the mcr-1 gene. E. coli isolated from wild boar meat seem to show aspecific antimicrobial resistance mechanisms for cephalosporins and colistin. The prevalence of resistant isolates found in wild boar is less alarming than in pork from farmed domestic pigs. However, the potential risk to consumers of these meat products will require further investigations.

Antimicrobial Resistance Profile and ExPEC Virulence Potential in Commensal Escherichia coli of Multiple Sources.

We recently described the genetic antimicrobial resistance and virulence profile of a collection of 279 commensal E. coli of food-producing animal (FPA), pet, wildlife and human origin. Phenotypic antimicrobial resistance (AMR) and the role of commensal E. coli as reservoir of extra-intestinal pathogenic Escherichia coli (ExPEC) virulence-associated genes (VAGs) or as potential ExPEC pathogens were evaluated. The most common phenotypic resistance was to tetracycline (76/279, 27.24%), sulfamethoxazole/trimethoprim (73/279, 26.16%), streptomycin and sulfisoxazole (71/279, 25.45% both) among the overall collection. Poultry and rabbit were the sources mostly associated to AMR, with a significant resistance rate (p > 0.01) to quinolones, streptomycin, sulphonamides, tetracycline and, only for poultry, to ampicillin and chloramphenicol. Finally, rabbit was the source mostly associated to colistin resistance. Different pandemic (ST69/69*, ST95, ST131) and emerging (ST10/ST10*, ST23, ST58, ST117, ST405, ST648) ExPEC sequence types (STs) were identified among the collection, especially in poultry source. Both ST groups carried high number of ExPEC VAGs (pandemic ExPEC STs, mean = 8.92; emerging ExPEC STs, mean = 6.43) and showed phenotypic resistance to different antimicrobials (pandemic ExPEC STs, mean = 2.23; emerging ExPEC STs, mean = 2.43), suggesting their role as potential ExPEC pathogens. Variable phenotypic resistance and ExPEC VAG distribution was also observed in uncommon ExPEC lineages, suggesting commensal flora as a potential reservoir of virulence (mean = 3.80) and antimicrobial resistance (mean = 1.69) determinants.

Escherichia coli Strains Isolated from Retail Meat Products: Evaluation of Biofilm Formation Ability, Antibiotic Resistance, and Phylogenetic Group Analysis.

ABSTRACT: Escherichia coli is a ubiquitous organism capable of forming a biofilm. This is an important virulence factor and is critical in certain diseases and in the development of antibiotic resistance, which is increased by biofilm synthesis. In the present study, the potential health risk associated with handling and consumption of foods of animal origin contaminated with E. coli-producing biofilm was evaluated. We analyzed the ability of 182 E. coli strains isolated from pork, poultry, and beef, purchased in three different supermarkets in the area of the "Italian Food Valley" (Parma, northern Italy), to form biofilms. Positive strains were also tested for the presence of 12 biofilm-associated genes. Moreover, the 182 E. coli were characterized for antibiotic resistance, presence of multidrug resistance, extended-spectrum ß-lactamase strains, and phylogenetic diversity through PCR. Twenty-five percent of the isolates produced biofilm. The majority showed weak adherence, five were moderate, and three were strong producers. E. coli with a strong adherence capability (three of three) harbored eight biofilm-associated genes, while weak and moderate producers harbored only five (frequencies ranging from 80 to 100%). Multidrug resistance was observed in 20 biofilm-producing E. coli, and 15 of these belonged to phylogenetic group D. Among nonbiofilm producers, the percentage of strains belonging to phylogenetic groups B2 and D was approximately 40%, highlighting a potential health risk for consumers and people handling contaminated products. The present study underlines the importance of monitoring the prevalence and characteristics of E. coli contaminating retail meat in relation to the potential virulence highlighted here.

Antibiotic Treatment Administered to Pigs and Antibiotic Resistance of Escherichia coli Isolated from Their Feces and Carcasses.

Antimicrobial resistance (AMR) in bacteria is a frequent and widespread phenomenon. The European Food Safety Authority (EFSA) reports that multidrug resistant (MDR) Escherichia coli is considered an important hazard to public health. The lack of data on the correlation between the administration of antibiotics to pigs and the diffusion of MDR E. coli necessitates an in-depth study. The aims of our study were first of all to determine the presence of MDR and/or extended spectrum ß-lactamase (ESßL) E. coli isolated from feces and carcasses of pigs; and second, to evaluate the correlation between antibiotic resistance and the antibiotic treatment administrated to the animals considered. The examined E. coli was isolated from 100 fecal swabs and 100 carcass sponges taken from farms and slaughterhouses located in Reggio Emilia province in Italy. The MDR isolates were tested following the protocol defined by EUCAST (2015). Subsequently, a real-time PCR and an endpoint-PCR were used for the genomic analysis. Data highlighted 76.5% of MDR E. coli with a marked presence of the ampicillin (AMP)-streptomycin (STRE)-tetracycline (TETRA) pattern. Moreover, 13 isolates were ESßL producers, and the blaCTXM gene was the most frequently observed in genomic analysis. Results confirm the complexity of the AMR phenomenon showing a partial correlation between the administration of antibiotics and the resistance observed. Pigs destined to the production of Protected Designation of Origin items are colonized by bacteria resistant to a wide range of antibiotic classes even if data are encouraging for colistin and third generation cephalosporin. Furthermore, in-depth study focused on food production could be useful in a view of high safety standards for consumers.

Antibacterial activity of metal complexes based on cinnamaldehyde thiosemicarbazone analogues.

The development of microbial antibiotic resistance has become one of the biggest threats to global health and the search for new molecules active against resistant pathogenic strains is a challenge that must be tackled. In many cases nosocomial infections are caused by bacteria characterized by multi-drug resistance patterns and by their ability to produce biofilms. These properties lead to the persistence of pathogens in the hospital environment. This paper reports the synthesis and characterization of three thiosemicarbazone derivatives based on a compound containing the cinnamaldehyde natural scaffold but possessing different logPow values. These molecules are then used as ligands to prepare complexes of the Cu(II) and Zn(II) ions. All these compounds, ligands and complexes, were screened in vitro on stains of Escherichia coli and Klebsiella pneumoniae for their antibacterial activity. Despite their molecular similarity they revealed variegated behaviors. Only two of them present interesting antimicrobial properties and have also been studied to verify their stability in solution. The compound with the lowest partition coefficient is the most promising. The minimal bactericidal concentration on K. pneumoniae and E. coli of these substances are very interesting and demonstrate that the use of metalloantibiotics is a promising device to fight antibiotic resistance.

Occurrence and Antimicrobial Profile of Bacterial Pathogens in Former Foodstuff Meat Products Used for Pet Diets.

European legislation stipulates that food no longer intended for human consumption, due to commercial reasons, manufacturing problems, or some defect, can be used in pet feed. However, the presence of former foodstuffs in pet diets could constitute a public health issue because pets can act as reservoirs of antimicrobial resistance genes. In this study, for the first time, biological hazards due to the presence of Escherichia coli and Salmonella in former foodstuff meat products were evaluated. Among the 112 samples of packaged fresh meat (poultry, pork, and beef) collected from cold storage warehouses of a mass market retailer, the overall prevalence of E. coli and Salmonella, the prevalence of strains with multidrug resistance, the phenotypic and genotypic characteristics of strains that produce extended-spectrum ß-lactamase, and the presence of biofilm producers were assessed. A high prevalence of E. coli was observed in former foodstuffs of poultry (100%), pork (100%), and beef (93.3%). Salmonella Derby and Salmonella Typhimurium were found in 11.5% of the poultry samples, and Salmonella Typhimurium was found in 13.3% of the pork samples. Salmonella was not recovered from the beef samples. Multidrug resistance was found in E. coli and Salmonella isolates from poultry, pork, and beef. Overall, 18.2% of E. coli isolates and 20% of Salmonella isolates were resistant to various types of antibiotics with various mechanisms of action. Salmonella isolates from pork are an important source of extended-spectrum ß-lactamase production. Both E. coli and Salmonella were carriers of antibiotic resistance marker genes ( blaTEM, blaSHV, and blaCTX-M) associated with ß-lactamase production in poultry and pork. Approximately 30% of the E. coli isolates from the various types of meat were phenotypically biofilm producers, and all carried the fimH gene. These findings indicate that the use of former foodstuff meat products in pet diets can represent a risk for public health.

Antimicrobial resistance, biofilm synthesis and virulence genes in Salmonella isolated from pigs bred on intensive farms.

Salmonella is the second cause of food-borne infection in humans in the USA and Europe. Pigs represent the second most important reservoir for the pathogen and the consumption of pork meat is a major risk factor for human salmonellosis. Here, we evaluated the virulence patterns of eleven Salmonella isolated from pigs (carcasses and faces) bred in intensive farms in the north of Italy. The two serotypes identified were S. Typhimurium and its monophasic variant 1,4,5,12:i:-. None of the isolates was an ESBL producer, as confirmed also by PCR. However, the presence of a multi-drug resistant pattern was evident, with all the isolates being resistant to at least to five antimicrobial agents belonging to various classes. Moreover, six out of eleven isolates showed important resistance profiles, such as resistance against colistin and ciprofloxacin, with nine to twelve recorded resistances. The isolates were negative for the biofilm synthesis test, while four different virulotypes were characterized. All the isolates showed the presence of invA, hilA, stn, ssrA, sipC. One sample also harbored ssaR and spvC genes. One strain was positive for all the virulence genes tested and was resistant to 12 antimicrobial agents. The present study contributes new data to the surveillance program for antibiotic resistance. Furthermore, the presence of eleven highly virulent isolates poses concern for human health in relation to their diffusion in the environment.

Novel Activity of a Synthetic Decapeptide Against Toxoplasma gondii Tachyzoites.

The killer peptide KP is a synthetic decapeptide derived from the sequence of the variable region of a recombinant yeast killer toxin-like microbicidal single-chain antibody. KP proved to exert significant activities against diverse microbial and viral pathogens through different mechanisms of action, but little is known of its effect on apicomplexan protozoa. The aim of the present study was to evaluate the in vitro activity of KP against Toxoplasma gondii, a globally widespread protozoan parasite of great medical interest. The effect of KP treatment and its potential mechanism of action on T. gondii were evaluated by various methods, including light microscopy, quantitative PCR, flow cytometry, confocal microscopy, and transmission electron microscopy. In the presence of KP, the number of T. gondii tachyzoites able to invade Vero cells and the parasite intracellular proliferation were significantly reduced. Morphological observation and analysis of apoptotic markers suggested that KP is able to trigger an apoptosis-like cell death in T. gondii. Overall, our results indicate that KP could be a promising candidate for the development of new anti-Toxoplasma drugs with a novel mechanism of action.

Toxoplasma Gondii and Pre-treatment Protocols for Polymerase Chain Reaction Analysis of Milk Samples: A Field Trial in Sheep from Southern Italy.

Toxoplasmosis is a zoonotic disease caused by the protozoan Toxoplasma gondii. Ingestion of raw milk has been suggested as a risk for transmission to humans. Here the authors evaluated pre-treatment protocols for DNA extraction on T. gondii tachyzoite-spiked sheep milk with the aim of identifying the method that resulted in the most rapid and reliable polymerase chain reaction (PCR) positivity. This protocol was then used to analyse milk samples from sheep of three different farms in Southern Italy, including real time PCR for DNA quantification and PCR-restriction fragment length polymorphism for genotyping. The pre-treatment protocol using ethylenediaminetetraacetic acid and Tris-HCl to remove casein gave the best results in the least amount of time compared to the others on spiked milk samples. One sample of 21 collected from sheep farms was positive on one-step PCR, real time PCR and resulted in a Type I genotype at one locus (SAG3). Milk usually contains a low number of tachyzoites and this could be a limiting factor for molecular identification. Our preliminary data has evaluated a rapid, cost-effective and sensitive protocol to treat milk before DNA extraction. The results of the present study also confirm the possibility of T. gondii transmission through consumption of raw milk and its unpasteurised derivatives.

Associations between animal welfare indicators and Campylobacter spp. in broiler chickens under commercial settings: A case study.

Few studies have previously investigated how poor animal welfare might be associated with infection of zoonotic pathogens in humans. This paper assesses the predictive value of the presence of Campylobacter spp. in broiler chicken flocks when animal-based measures related to footpad dermatitis, hock burns, body lesions and arthritis are identified under commercial conditions (high density). The study population included 32 flocks analysed on farm and at slaughter, slaughtered between April and August 2008 in six different slaughter plants in Brittany, France. Welfare and health indicators are those indicated by the European legislation and sampling was carried out in the framework of the European baseline survey on the prevalence of Campylobacter in broiler chicken. Caecal contents, sampled both on farm and at slaughter, and carcass skin samples from the neck and breast at slaughter, were investigated for the presence of Campylobacter spp. Logistic models/classification trees were used to estimate the probability of the presence (or absence) of a specific foodborne pathogen in a flock based on specific animal-based measures (or combinations of measures) in order to study the potential relationship between welfare indicators and foodborne pathogen prevalence/incidence levels. On farm, flocks with more than 25% animals with severe lesions on between 25 and 50% of the footpad are predicted to be Campylobacter-positive whereas flocks where less than 13 individuals have arthritis are predicted to be Campylobacter-negative. The error rate on farm and at slaughter was 10 and 4% respectively indicating good predicting abilities. A poor welfare environment may result in stress, which reduces chicken immunocompetence making them more susceptible to Campylobacter spp. An infection with Campylobacter spp may lead to impaired defence and susceptibility to other pathogens which may result in greater intestinal excretion. Poor welfare and high growing rate lead to digestive troubles that lead to litter humidity. Litter humidity that, among other things, causes footpad dermatitis may also influence the horizontal transmission of the Campylobacter spp. infection due to the normal coprophagic behaviour of poultry. Reducing welfare problems by a better management of rearing conditions would not only improve broiler welfare, but it would also decrease the risks of Campylobacter contamination, of carcass condemnations and of economic loss for the poultry industry.

Meat Juice Serology for Toxoplasma Gondii Infection in Chickens.

Toxoplasma gondii is an important foodborne zoonosis. Free-range chickens are at particularly high risk of infection and are also excellent indicators of soil contamination by oocysts. In the present study, hearts of 77 free-range chickens were collected at slaughter. T. gondii meat juice enzyme-linked immunosorbent assay was performed with a commercial kit, following validation with positive controls, from experimentally infected chickens, and negative ones. Out of 77 samples, only 66 gave sufficient meat juice for serology. Of these, 24 (36.4%) were positive for T. gondii considering the 5*standard deviation values (calculated on the optical density of negative controls), while all the samples were negative considering sample/positive% values. Parasite-specific polymerase chain reaction was carried out on all samples obtained from heart tissue and none were positive for the presence of T. gondii DNA. Results would suggest that further study on the use of meat juice with a validated serological test to detect T. gondii in chickens could lead to widespread epidemiological studies in this important intermediate host. However, sample collection and test specificity require further evaluation.

Salmonella Enterica Prevalence in Finishing Pigs at Slaughter Plants in Northern Italy.

Finishing pigs carrying Salmonella enterica are believed to be the main source of carcass contamination at the beginning of slaughtering. The aim of this study was to assess the S. enterica carrier status of finishing pigs at herd level by sampling pooled faeces on farm and mesenteric lymph nodes at slaughter in the North East of Italy. Environmental faecal samples belonging to 30 batches of pigs were collected on farm. At slaughter, mesenteric lymph nodes were collected from five randomly selected pigs per batch. S. enterica was isolated from 16 lymph nodes out of 150 (10.6%) and from seven out of 30 (23.3%) faecal samples. Four batches (13.3%) were positive to S. enterica both in lymph nodes and in faeces. The number of batches positive to S. enterica either in lymph nodes or in faeces was 13 out of 30 (43.3%). The most prevalent serovars from lymph nodes were S. Derby (25.0%) and S. Typhimurium monophasic variant 1, 4,[5],12:i:- (18.6%), which were also isolated from faecal material (14.3 and 42.8% respectively). Contaminated faecal material or lymph nodes could be a primary source of carcass contamination at slaughter during evisceration. S. enterica contamination is widespread on pig farms and carrier pigs pass undetected the inspection visits at slaughter, entering the food chain. Therefore, in order to control S. enterica in pigs, the need to quantify possible risk factors at slaughter and develop effective management strategies on farm is of paramount importance to ensure food safety.