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Milk has been consumed by humans for thousands of years for its nutritional properties. In recent years, raw milk demand has increased, valued for its authenticity and connection to local traditions. In Italy, the sale of raw milk is allowed exclusively through direct sale from the producing farm to the final consumer, either at the producing farm itself or through vending machines. However, the consumption of raw milk is not without risks. Among these, microbiological ones are relevant. These can lead to severe symptoms, particularly in vulnerable populations. For this reason, although consumers are advised to boil raw milk before consumption, producing farms in Italy are required to meet the microbiological criteria outlined in the Provision of 25 January 2007. In this retrospective study, the results of the analyses performed on 355 raw milk samples collected in Liguria between 2014 and 2023 for the detection of Campylobacter spp., Salmonella spp., Listeria monocytogenes , Staphylococcus aureus , and Escherichia coli O157 were analysed to better characterise the associated risk for consumers. The samples were collected during official controls by the local veterinary health services at vending machines of seven producing farms. Overall, six samples tested positive for C. jejuni , while only one sample tested positive for Salmonella enterica subsp. enterica , Serovar Veneziana. Listeria monocytogenes , S. aureus , and E. coli O157 were never responsible for non-compliances. Interestingly, three of the six samples positive for C. jejuni derived from the same producer. In farms where positive samples were detected, certain structural and/or operational non-compliances were identified. It can be concluded that, although the scenario in question does not present any cause for concern, it is nevertheless essential to implement a series of preventive measures in order to guarantee the safety of raw milk. These measures include the implementation of biosecurity practices, the maintenance of strict hygiene protocols during milking, and the adherence to the cold chain distribution protocol until the final stage of distribution.

The occurrence of antibiotic-resistant bacteria in foodstuff involves a human health risk. Edible insects are a precious resource; however, their consumption raises food safety issues. In this study, the occurrence of antibiotic resistant bacteria in laboratory-reared fresh mealworm larvae ( Tenebrio molitor L.) and frass was assessed. Antibiotics were not used during the rearing. Enterobacteriaceae and enterococci were isolated from 17 larvae and eight frass samples. In total, 62 and 69 isolates presumed to belong to Enterobacteriaceae and Enterococcus spp., respectively, were obtained and tested for antibiotic susceptibility via disk diffusion. Based on the results, isolates were grouped, and representative resistant isolates were identified at species level through 16S rRNA gene sequencing. For enterococci resistance, percentages higher than 15% were observed for vancomycin and quinupristin-dalfopristin, whereas Enterobacteriaceae resistance higher than 25% was found against cefoxitin, ampicillin, and amoxicillin-clavulanic acid. Based on the species identification, the observed resistances seemed to be intrinsic both for enterococci and Enterobacteriaceae , except for some β-lactams resistance in Shigella boydii (cefoxitin and aztreonam). These could be due to transferable genetic elements. This study suggests the need for further investigations to clarify the role of edible insects in the spreading of antibiotic resistance determinants through the food chain.

Campylobacteriosis is the most reported foodborne disease in the European Union, with more than 100,000 confirmed cases annually. Human infection can be caused by a low infectious dose, and in fragile populations, the food disease can manifest itself in acute and severe forms. This study aims to analyze two cases of campylobacteriosis in fragile people caused by Campylobacter jejuni in 2023 in Tuscany and the actions of the Local Health Competent Authority. From the results of the related investigations, it was possible to attribute both cases of foodborne diseases to unsafe food management during preparation/administration. Given the peculiar characteristics of the etiological agent, it is necessary to focus the attention of the population, especially those who deal with fragile subjects, on the good hygiene practices to be followed both at home and in collective catering., Competing Interests: Conflict of interest: the authors declare that they have no competing interests, and all authors confirm accuracy., (Copyright © 2024, the Author(s).)

In recent years, essential oils (EOs) have received interest due to their antibacterial properties. Accordingly, the present study aimed to investigate the effectiveness of the EOs obtained from seven species of Salvia on three strains of Listeria monocytogenes (two serotyped wild strains and one ATCC strain), a bacterium able to contaminate food products and cause foodborne disease in humans. The Salvia species analysed in the present study were cultivated at the Botanic Garden and Museum of the University of Pisa, and their air-dried aerial parts were subjected to hydrodistillation using a Clevenger apparatus. The obtained EOs were analysed via gas chromatography coupled with mass spectrometry for the evaluation of their chemical composition, and they were tested for their inhibitory and bactericidal activities by means of MIC and MBC. The tested Eos showed promising results, and the best outcomes were reached by S. chamaedryoides EO, showing an MIC of 1:256 and an MBC of 1:64. The predominant compounds of this EO were the sesquiterpenes caryophyllene oxide and β-caryophyllene, together with the monoterpenes bornyl acetate and borneol. These results suggest that these EOs may possibly be used in the food industry as preservatives of natural origins.

Tenebrio molitor (mealworm) is one of the most promising insect species to produce sustainable feed and food with high nutritional value. Insects may harbour microorganisms both in the gut and on the exoskeleton originating from the rearing environment. Staphylococcus aureus is a pathogenic microorganism frequently involved in food poisoning due to its enterotoxin production. This study aimed to evaluate the S. aureus growth and enterotoxins production following an experimental inoculation into the T. molitor rearing substrate (about 7 log CFU/g). Analyses on the substrate and larvae were performed over a testing period of seven days. The microbial population dynamics were also evaluated through total viable count and lactic acid bacteria count. The effects of fasting, washing, and cooking on the microbial loads of mealworms were evaluated. The results highlighted that mealworms and substrates can maintain their microbial loads of S. aureus over the tested period. Moreover, fasting and washing were generally not able to significantly reduce ( p -value > 0.05) S. aureus count in mealworms. On the other hand, cooking significantly reduced ( p -value < 0.001) the microbial load in almost all cases. No production of enterotoxins was revealed during the trial. Therefore, microbiological risks can be reduced by a wise choice of substrate, appropriate control measures, and thermal treatment of larvae.

The Commission Regulation (EU) No. 2021/382 (European Commission, 2021), amending the Regulation (EC) No. 852/2004 (European Commission, 2004), introduced the obligation for companies to establish and maintain a food safety culture (FSC). The methodology to evaluate, implement, and enhance the level of FSC is up to the individual companies. This study aimed to investigate the perception of FSC among the employees of 3 Tuscan medium-sized enterprises in the food sector, producing cured meat (A), dairy products (B), and frozen fish products (C). The survey was conducted through the development and administration of a questionnaire based on a 5 points Likert scale, referring to different aspects of FSC, organized in 6 sections with 5-6 statements each and subjected to a percentage of employees between 76 and 85%, classified also by the length of service (≤3 and >3 years). For all the companies, the minimum median and mode value for scores obtained by the different sections was 4, and the minimum median and mode value for the single statement was 3 (A, B; except for a bimodal value 2-4) and 4 (C). The section awareness and perception of risk showed the highest mean scores in all companies. As for the length of service, senior employees gave lower scores than junior ones in all sections in B and 3 sections in C. Overall, the results of the questionnaires showed a good perception of FSC, even though it was possible to identify some partial weaknesses., Competing Interests: Conflict of interest: The authors declare no potential conflict of interest., (©Copyright: the Author(s).)

A total of 193 wild boars hunted in Tuscany, an Italian region with a high presence of wild ungulates, were examined to assess the occurrence of Campylobacter species in faeces, bile, liver and carcasses, with the aim of clarifying their contribution to human infection through the food chain. Campylobacter spp. were found in 44.56% of the animals, 42.62% of the faecal samples, 18.18% of the carcass samples, 4.81% of the liver tissues and 1.97% of the bile samples. The Campylobacter species genotypically identified were C. coli , C. lanienae , C. jejuni and C. hyointestinalis . The prevalent species transpired to be C. coli and C. lanienae , which were isolated from all the matrices; C. jejuni was present in faeces and liver, while C. hyointestinalis only in faeces. Identification was carried out by matrix-assisted laser desorption/ionisation-time-of-flight mass spectrometry (MALDI-TOF MS) on 66 out of 100 isolates identified genotypically, and the technique yielded unsatisfactory results in the case of C. lanienae , which is responsible for sporadic human disease cases. The level of Campylobacter spp. contamination of meat and liver underlines the need to provide appropriate food safety information to hunters and consumers.

The activity of manuka ( Leptospermum scoparium ) essential oil (EO) on biofilms of foodborne Listeria monocytogenes and Staphylococcus aureus has been studied. Seven strains of L. monocytogenes and 7 of S. aureus (5 methicillin-resistant) were tested. EO minimal inhibitory concentration (MIC), EO minimal bactericidal concentration (MBC) and biofilm production quantification were determined for each strain by microtiter methods. Moreover, EO Minimum Biofilm Inhibitory Concentration (MBIC) and Minimum Biofilm Eradicating Concentration (MBEC) were determined on 2 L. monocytogenes and 3 S. aureus that showed the best biofilm production. Finally, on 4 strains out of 5 (2 L. monocytogenes and 2 S. aureus ) EO Biofilm Reduction Percentage (BRP) vs . untreated controls was assessed after a treatment with EO subinhibitory concentrations. The chemical composition of manuka essential oil was determined by Gas Chromatography- Electron Impact Mass Spectrometry (GCEIMS). The manuka EO demonstrated good antimicrobial activity: L. monocytogenes MIC and MBC were 0.466 mg/ml and 0.933 mg/ml, respectively, whereas S. aureus MIC and MBC were 0.233 mg/ml and 0.466 mg/ml, respectively. Furthermore, L. monocytogenes showed a MBIC of 0.933 mg/ml and a MBEC in the range of 0.933-1.865 mg/ml, whereas S. aureus had a MBIC in the range of 7.461-14.922 mg/ml and a MBEC of 14.922 mg/ml. L. monocytogenes revealed no significant BRP after the treatment with manuka EO, whereas S. aureus showed a BRP higher than 50% with MIC/2 and MIC/4 EO concentrations. These results provide information for feasible manuka EO applications in food production systems., (©Copyright: the Author(s).)

This study aims to evaluate the microbiological status, pH, and water activity of European beaver meat to establish its shelf-life and microbiological safety. In this study, the microbiological profiles of meat and minced meat obtained from the carcasses of beavers were investigated. Microbial evaluation of the chilled meat was performed within 24 h after hunting, on the 7th day and 14th day, and the evaluation of the frozen meat was made during the 11th week of storage. Meat samples were analysed for total viable count (TVC), psychrotrophic bacteria count (PBC), Enterobacteriaceae count (EBC), Escherichia coli count (EC), total staphylococcal count (TSC), lactic acid bacteria count (LABC) and total yeast and mould counts (TYMC). Tests for the presence of pathogenic bacteria from the genus Salmonella and Listeria were also performed. Additionally, the pH and water activity were determined. The initial amount of TVC was 4.94 log CFU/g in meat samples and 4.80 log CFU/g in minced meat. After 14 days of storage, the TVC increased to 8.33 in meat samples and 8.08 log CFU/g in minced meat. Pathogenic bacteria such as Listeria and Salmonella were not found in the beaver meat tested. The microbiological state of meat stored frozen for 11 weeks was comparable to the state found in meat stored refrigerated for seven days regarding the number of microorganisms.

Teat-dipping is one of the most effective methods to prevent mammary infections in ruminants, including sub-clinical mastitis caused by coagulase-negative staphylococci (CoNS). Improper disinfectant application could expose microorganisms to sub-inhibitory concentrations leading to phenotypic variations. In this study, 12 chlorhexidine-digluconate (CHDG)-tolerant (of which 4 qac positive) and 12 benzalkonium chloride (BC)-tolerant (of which 7 qac-positive) CoNS isolates from ovine milk were exposed to sub-inhibitory concentrations of CHDG and BC, respectively. Changes in disinfectant susceptibility against BC and CHDG, antibiotic resistance against 12 antibiotics and biofilm production were then assessed for both groups. After CHDG stress, 67 % and 83 % of the CHDG-stressed isolates doubled their MICs for BC and CHDG, respectively and 2 qac-negative isolates showed a four-fold increase of their MBCs for CHDG. After BC stress, MICs for BC and CHDG doubled in 58 % and 83 % of the BC-stressed isolates, respectively, while one qac-positive isolate increased four-fold the MIC for BC. Cross-resistance to antibiotics was assessed by disc diffusion method. Some qac-positive isolates varied their resistance profile, while a blaZ-positive isolate showed a resistant phenotype against ampicillin only after the exposure to the disinfectant. As for qac-positive isolates, one CHDG-stressed and 2 BC-stressed increased their resistance to kanamycin and cefoxitin, respectively. The Congo Red Agar test was carried out to assess the in vitro slime production: all isolates were negative after stress. In conclusion, sub-inhibitory exposure to disinfectants may affect disinfectant and antibiotic susceptibility, the latter in particular for qac-positive isolates and those hosting unexpressed antibiotic resistance genes., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Neonatal diarrhoea (ND), post-weaning diarrhoea (PWD) and oedema disease (OD) are among the most important diseases affecting pig farming due to economic losses. Among the main aetiological agents, strains of Escherichia coli are identified as the major responsible pathogens involved. Several strategies have been put in place to prevent these infections and, today, research is increasingly studying alternative methods to antibiotics to reduce the antibiotic resistance phenomenon. Essential oils (EOs) are among the alternative tools that are being investigated. In this study, the in vitro effectiveness of winter savory and manuka essential oils and their mixtures in different proportions against strains of E. coli isolated from episodes of disease in pigs was evaluated. The EOs alone demonstrated slight antibacterial effectiveness, whereas the blends, by virtue of their synergistic action, showed remarkable activity, especially the 70%-30% winter savory-manuka blend, showing itself as a potential tool for prevention and therapy.

Rabbit burgers were evaluated for fatty acids profile, oxidative status, antioxidant potential and sensory during storage time of seven days at 4 °C. The aim of the research study was to evaluate if a common culinary spice as garlic could overcome the controversy effects of salt (mostly related to a pro-oxidant effect). Therefore, four formulations were made: only meat (control, C) meat added with garlic powder at 0.25% (G), meat added with salt at 1.00% (S) and meat added with garlic powder (0.25%) and salt (1.00%) (GS). As results, it was highlighted that garlic powder was not capable to reduce significantly the negative effect related to the salt addition. Salt induced different modifications to burgers leading to a more liked product due to its properties to enhance sensory characteristics, on the other hand, induced different oxidation processes that could decrease the nutritional value. Further studies are needed to better elucidate if different garlic products/concentrations could apport higher beneficial effects., Competing Interests: Declaration of Competing Interest Authors confirm that there are no known conflicts of interest associated with this publication and there has been no financial support for this work that could have influenced its outcome., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

The present study investigated the genomic constitution and antimicrobial resistance (AMR) of 238 Campylobacter from pigs and wild boars in Italy between 2012 and 2019. Campylobacter strains were genotyped using multilocus sequence typing (MLST) and whole genome MLST (wgMLST), screened for antimicrobial resistance genes, and tested for phenotypic susceptibility to six different antibiotics. C. coli was detected in 98.31% and 91.66% of pigs and wild boars, while C. jejuni was isolated in the remaining cases. MLST assigned 73 STs and 13 STs in pigs and wild boars, respectively, including 44 novel STs. The predominant ST in pigs was ST-854 (12.36%), followed by ST-9264 (6.18%). ST-1055 and ST-1417 were predominant in wild boars (30% and 13.33%, respectively). The minimum spanning tree using 1,121 global MLST profiles showed specific Italian clusters and a clear separation between pig and wild boar profiles. The wgMLST confirmed the MLST clustering and revealed a high genetic diversity within C. coli population in Italy. Minimum inhibitory concentrations (MIC) of six antibiotics revealed higher resistance in pigs to ciprofloxacin, nalidixic acid, streptomycin and tetracycline, compared to wild boar. In contrast, most strains were susceptible to gentamicin. Worrying levels of multidrug resistance (MDR) were observed mostly in pig isolates. Molecular screening of AMR mechanisms revealed the predominance of gyrA T86I substitution among fluoroquinolone- and quinolone-resistant isolates, and the 23S rRNA A2075G mutation among macrolide-resistant isolates. Other resistance determinants were observed: (i) tet(O) gene was present among tetracycline-resistant isolates; (ii) rpsL and aph (3')-III genes conferring resistance to aminoglycosides, were identified only in streptomycin or gentamicin-resistant pig isolates; (iii) cmeA, cmeB, cmeC, cmeR genes responsible of pump efflux mechanisms, were observed in almost all the strains; (iv) OXA -61, encoding β-lactamase, was found in the half of the strains. Genotypic and phenotypic AMR profiling was fairly correlated for quinolones/fluoroquinolones. Campylobacter infection is common also in wild boar populations in Italy, suggesting that wild boars could be a reservoir of resistant and multi-resistant Campylobacter species, which may be of public health concern. The present study adds to our knowledge on the epidemiological and ecological traits of this pathogen in domesticated and wild swine., (Copyright © 2020 Marotta, Di Marcantonio, Janowicz, Pedonese, Di Donato, Ardelean, Nuvoloni, Di Giannatale and Garofolo.)

The aim of the research study was to evaluate the effects of a common culinary spice such as garlic powder and salt addition on the quality and microbial shelf life of rabbit meat burgers. Rabbit burgers were evaluated for pH, the colour parameters, the water holding capacity and microbial loads during storage time of seven days at 4 °C. Four different formulations of burgers ( n = 180 in total) were tested as control samples (only meat, C), burgers with garlic powder (at 0.25%, G), burgers with salt (at 1.00%, S) and burgers with both garlic powder and salt (0.25% and 1.00%, respectively, GS). As results, it was highlighted that garlic powder and salt addition significant affected pH, water holding capacity and some colour parameters of burgers. In particular, salt affected the pH of the raw burgers, leading to lower values that partially influenced all the colour parameters with higher a* values of S burgers. The mix of garlic powder and salt (GS burgers) showed mixed effects even if more closed to the G burgers than S ones. Salt expressed its properties of binding water molecules reducing drip and cooking losses in S and GS burgers. No variations in microbial loads were highlighted in relation to the formulations. Storage time affected all the parameters, highlighting a deterioration of the burgers' quality and an increase of the microbial loads.

Pseudorabies, or Aujeszky's disease, is a notifiable worldwide infection of domestic and feral swine that causes economic losses for the swine industry. In domestic pigs, the virus is responsible for nervous and/or respiratory symptoms; in pregnant sows, it is one of the major causes of stillbirth, mummification, embryonic death, and infertility (SMEDI). It is known that PRV infection in wild boar is associated with low pathogenicity and attenuated or absent symptomatology, but limited information is available about the ability of the virus to infect the foetuses of infected wild boar pregnant sows. Due to scarce information about the reproductive consequences, we investigate the possible intrauterine vertical transmission of the virus in wild boar pregnant sow living in a highly infected area. A number of 54 hunted wild boar were sampled during 2018-2019, and blood, genital and nasal swabs, placenta, and fetuses were collected for serological and molecular investigations. A seroprevalence of 74% (40/54) was detected, while 1/24 pregnant sow and 1/24 pooled foetuses tested positive by PCR (gene gB). This is the first evidence of viral detection in foetuses from seropositive pregnant wild boar. This finding suggests the possible pathogenetic role of PRV on pregnancy in wild boar and the existence of an additional transmission route.

Propolis antimicrobial activity has been limitedly studied in food, particularly in dairy products. We studied the antimicrobial activity of an alcoholic extract of an Italian propolis in sterile skim milk, pasteurized cow's milk, and cow's and goat's whey cheese (ricotta). Following the determination of the minimal inhibitory concentration on Gram+ and Gram- bacteria, the extract was employed at 2 and 5% (P2, P5), using controls with the same ethanol concentrations (E2, E5) and without any addition. In milk trials, Listeria monocytogenes , Staphylococcus aureus , Bacillus cereus , and Pseudomonas fluorescens were tested. P2 and P5 samples registered significant decreases of Gram+ bacteria in skim milk. The same was true for P5 in cows' milk, but only with S. aureus for P2. Ricotta was inoculated with L. monocytogenes , S. aureus and B. cereus and stored at 8.5°C. In cow's milk ricotta, L. monocytogenes counts in P5 were always lower than control during the storage time, significantly so from the 14 th day. In goat's ricotta, L. monocytogenes counts in P5 were at least one logarithm lower than E5, whereas the extract didn't show a significant effect on S. aureus and B. cereus . The antimicrobial activity of propolis, particularly on L. monocytogenes , could be employed in ready-to-eat refrigerated dairy products., Competing Interests: Conflict of interest: The authors declare no potential conflict of interest., (©Copyright: the Author(s), 2019.)

Today, edible insects represent a hot topic as an emerging and eco-friendly source of protein. The mealworm (Tenebrio molitor L.) is among the most employed insects for human consumption and feed purposes. So far Listeria monocytogenes, have never been detected either in products sold on the market or during the rearing process. In this study, the substrate employed for mealworm rearing was deliberately contaminated with L. monocytogenes and the bacterium was enumerated during the rearing period and after technological treatments of the larvae. L. monocytogenes persisted during the rearing period. Washing the larvae did not produce any significant effect, while fasting the larvae for 24 or 48 h reduced the L. monocytogenes load (P < 0.001). Oven cooking eliminated L. monocytogenes cells from the product, reducing the risk associated to this foodborne pathogen to zero., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Essential oils (EOs) are mixtures of secondary metabolites of plant origin with many useful properties, among which the antimicrobial activity is also of interest for the food industry. EOs can exert their antimicrobial potential both directly, in food products and active packaging, and indirectly, as sanitizing and anti-biofilm agents of food facility surfaces. Aim of this research was to evaluate the antimicrobial activity of four EOs (bergamot, cinnamon, manuka and thyme) against Pseudomonas fluorescens and Staphylococcus aureus isolated from milk and dairy products. The chemical composition of EOs was evaluated by Gas Chromatography-Mass Spectrometry analysis. Minimum Inhibitory Concentration values were determined by a microplate method against 9 Ps. fluorescens from marketed mozzarella with blue discoloration defect, and 3 biofilm-producing S. aureus from milk. Reference ATCC strains were included. Pigment production activity by Ps. fluorescens was assessed both in culture and in cheese. EOs of manuka (leptospermone 23%) and thyme (carvacrol 30%, pcymene 20%, thymol 15%) showed the highest antimicrobial activity against S. aureus , MIC values were 0.012%-0.024% and 0.024% v/v, respectively; meanwhile EOs from thyme and cinnamon (cinnamaldehyde 55%) exhibited the best activity against Ps. fluorescens with MIC values of 0.098%-0.195% and 0.195%-0.391% v/v, respectively. The antimicrobial activity of these EOs is promising and they could be exploited in the dairy production chain., Competing Interests: Conflict of interest: the authors declare no potential conflict of interest.

In the European Union, slaughter without stunning is allowed for religious slaughter to obtain halal and kosher meat. Especially in the case of Jewish slaughtering, cuts which are not deemed as kosher are sold to regular market without any specific labelling. This survey, conducted in Tuscany in 2016, aimed to quantify the carcasses rejected in relation to the type of religious slaughter. 656 bovines were slaughtered without stunning: 538 (82%) for halal and 118 (18%) for kosher. All carcasses slaughtered by the Islamic procedure (dhabiha) were considered halal, while 77.1% of carcasses slaughtered by the Jewish procedure (shechita) did not pass the approval. Carcasses were rejected after chest cavity inspection (50%) and after the lungs control (50%). This study provides an important insight in this field and postulates how to amalgamate the concepts of freedom of religion, as enshrined by the Charter of Fundamental Rights of the EU, with consumer rights and animal welfare., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Mycobacterium avium paratuberculosis (Map) is a pathogen which causes a chronic progressive granulomatous enteritis known as paratuberculosis or Johne's disease and it primarily affects wild and domestic ruminants. The aim of this research was to examine a flock which consisted of 294 goats and was located in Garfagnana district (Tuscany, Italy) performing ELISA tests, culture and IS900 PCR assay; direct diagnostic methods were carried out not only on bulk tank milk and cheese samples but also on individual milk and tissue specimens collected from nine subjects positive to ELISA tests. Out of 294 animals, 20 goats (6.8%) were positive to ELISA surveys. Bulk tank milk samples were negative to culture and to PCR assay carried out on the DNA extracted directly from them, while, with respect to cheese, Map was detected by culture in 2/12 (16.66%) cheeses ripened for 3-7 days, and by PCR in 2/12 (16.66%) cheeses ripened for 3-7 days and in 3/12 (25%) cheeses ripened for 45 days. Regarding individual milk samples, Map was detected by culture in 2/9 (22.22%) specimens and by PCR in 5/9 (55.55%) samples. Furthermore, Map was isolated from the intestine in 9/9 (100%) animals, from the mesenteric lymph nodes in 8/9 (88.88%) subjects, from the liver in 4/9 (44.44%) goats, from the spleen in 5/9 (55.55%) animals, while Map DNA was found in all the tissue samples analyzed.The results demonstrated the presence of paratuberculosis in a goat flock located in Garfagnana district (Tuscany, Italy).

The capability to produce biofilm is an important persistence and dissemination mechanism of some foodborne bacteria. This paper investigates the relationship between some molecular characteristics (SCCmec, ST, spa-type, agr-type, cna, sarA, icaA, icaD, clfA, fnbA, fnbB, hla, hlb) of 22 food-related methicillin-resistant Staphylococcus aureus (MRSA) strains and their ability to form biofilm on stainless steel and polystyrene. Five (22.7%, 5/22) strains were able to synthesize biofilm on polystyrene, and one of these (4.5%, 1/22) strains was also able to synthesize biofilm on stainless steel. The largest amount of biofilm was formed on polystyrene by 2 MRSA strains isolated from cows' milk, thus raising concern about the dairy industry. The majority of MRSA biofilm producers carried SCCmec type IVa, suggesting that the presence of SCCmecIVa and/or agr type III could be related to the ability to form biofilm. In conclusion, in order to achieve an acceptable level of food safety, Good Hygiene Practices should be strictly implemented along the food chain to reduce the risk of colonization and dissemination of MRSA biofilm-producing strains in the food industry., Practical Application: In this study, some assayed isolates of food-related MRSA demonstrated the capacity to form biofilm. Biofilm formation differed according to surface characteristics and MRSA strains. A relationship was observed between some molecular characteristics and the ability to form biofilms. Few studies have investigated the ability of MRSA to form biofilms, and the majority of these studies have investigated clinical aspects. This work was performed to investigate whether or not there is a difference between MRSA food isolates and MRSA clinical isolates in their ability to form biofilm. These initial findings could provide information that will contribute to a better understanding of these aspects., (© 2017 Institute of Food Technologists®.)

Donkey milk is recently gaining attention due to its nutraceutical properties. Its low casein content does not allow caseification, so the production of a fermented milk would represent an alternative way to increase donkey milk shelf life. The aim of this study was to investigate the possibility of employing selected Streptococcus thermophilus and Lactobacillus plantarum isolates for the production of a novel donkey milk fermented beverage. Lysozyme resistance and the ability to acidify donkey milk were chosen as main selection parameters. Different fermented beverages (C1-C9) were produced, each with a specific combination of isolates, and stored at refrigerated conditions for 35days. The pH values and viability of the isolates were weekly assessed. In addition, sensory analysis was performed. Both S. thermophilus and L.plantarum showed a high degree of resistance to lysozyme with a Minimum Bactericidal Concentration>6.4mg/mL for 100% of S. thermophilus and 96% of L. plantarum. S. thermophilus and L. plantarum showed the ability to acidify donkey milk in 24h at 37°C, with an average ΔpH value of 2.91±0.16 and 1.78±0.66, respectively. Four L. plantarum and two S. thermophilus were chosen for the production of fermented milks. Those containing the association S. thermophilus/L. plantarum (C1-C4) reached a pH lower than 4.5 after 18h of fermentation and showed microbial loads higher than 7.00logcfu/mL until the end of the storage period. Moreover, comparing the microbial loads of samples containing both species and those containing S. thermophilus alone (C5), we highlighted the ability of L. plantarum to stimulate S. thermophilus replication. This boosted replication of S. thermophilus allowed to reach an appropriate pH in a time frame fitting the production schedule. This was not observed for samples containing a single species (C5-C9). Thus, L. plantarum strains seem to be good candidates in the production of a novel type of fermented milk, not only for their probiotic potential, but also for the enhancing effect on S. thermophilus growth., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Retail poultry meat is a crucial vehicle for consumers' exposure to Campylobacters, but no official controls are currently applied in Italy. The aim of this study was the evaluation of Campylobacter contamination of a wide range of poultry meats marketed in Italy. N. 472 chicken and turkey meat samples (sectioned meats, offal, meat preparations and products) were taken from slaughterhouses, deboning plants and different retailers and submitted to detection/enumeration of Campylobacter spp. The isolates were identified by phenotypic and biomolecular techniques. Campylobacter spp. was detected in 34.1% of the samples, with general low counts. Higher values were observed in offal (especially liver) and sectioned meats, with significantly higher rates in skin-on samples (86.8% vs 32.7%). Minced meat preparations showed lower prevalence (22.4% vs 58.3%) and counts than whole pieces. Decreasing rates were observed among slaughterhouses (80%), deboning plants (49%), butcher's shops (37%) and large scale retailers (25%). Sectioned chicken meats were significantly more contaminated than turkey meats. Almost all the isolates were identified as C. jejuni or C. coli, with similar prevalences (18.4% and 20.5%, respectively); C. jejuni was predominant only in samples from slaughterhouses/deboning plants. For setting future control programs, meat typology should be considered the main critical factor., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

In this study, the prevalence of Campylobacter spp. in poultry fresh meat and ready‑to‑cook products was evaluated. Seventy‑three samples were collected at retail level from supermarkets and discount stores, obtaining 61.6% positivity. Of 133 Campylobacter isolates, 86 strains (Campylobacter coli, 58.1% and Campylobacter jejuni, 41.9%) were selected for characterisation on the basis of their SmaI and kpnI pulsed field gel‑electrophoresis (PFGE) profiles, to exclude clonal replicates. Campylobacters resulted highly resistant to tetracycline, ciprofloxacin, and nalidixic acid (79.1%, 72.1% and 65.1%, respectively); 50% of C. coli and 13.9% of C. jejuni were resistant to ciprofloxacin and erythromycin, the most important antimicrobials for human campylobacteriosis therapy. Five C. coli were resistant to 5/7 of the tested antimicrobials. HS4c was the prevailing C. jejuni serotype group (22.3%), whereas 8 other serotypes were identified in low percentages. SmaI and kpnI profiles showed a wide variability. The survey showed a high Campylobacter contamination of poultry meat and poultry products at retail level in Tuscany, Italy. A wide strains' heterogeneity and a remarkable level of strains' antimicrobial resistance have been reported, confirming the need for an improvement of specific preventive measures along the production chain.

Extensive use of antimicrobial agents in finfish farming and the consequent selective pressure lead to the acquisition of antibiotic resistance in aquaculture environment bacteria. Vibrio genus represents one of the main pathogens affecting gilthead sea bream. The development of antibiotic resistance by Vibrio represents a potential threat to human health by exchange of resistant genes to human pathogens through food chain. The objective of the present study was to conduct a multisite survey on the antibiotic resistance of Vibrio spp. isolated from gilthead sea bream reared in Italian mariculture. Vibrio spp. strains were isolated from skin, gills, muscles and intestinal content of 240 gilthead sea bream. A random selection of 150 strains was sequenced for species identification. Resistance against 15 antimicrobial agents was tested by the broth microdilution method. Vibrio harveyi and Vibrio alginolyticus accounted for 36.7% and 33.3% of the isolates respectively. 96% of the strains showed multiple resistance to the tested drugs, with two strains, Vibrio aestuarianus and Vibrio harveyi resistant to 10 and 9 antibiotics, respectively. Ampicillin, amoxicillin, erythromycin and sulfadiazine showed low efficacy against Vibrio spp. Rational use of antimicrobial agents and surveillance on antibiotic administration may reduce the acquisition of resistance by microorganisms of aquatic ecosystems.

The aim of this study was to investigate some probiotic properties of 42 wild Lactobacillus plantarum strains isolated from different Italian foods of animal origin. The strains were first screened for their antibiotic resistance profile (chloramphenicol, erythromycin, gentamicin, and tetracycline), subsequently they were tested for their in vitro resistance to lysozyme (100 mg L⁻¹), low pH (3.0, 2.5 and 2.0) and bile salts (0.3, 0.5 and 1.0 %). Moreover, agglutination property was studied (adhesion to Saccharomyces cerevisiae cells), as well as the presence of bsh and msa genes. The strains with the best characteristics were subjected to a further trial in order to evaluate their ability to survive to multiple stresses over time (lysozyme, low pH and bile salts) and the effect of these treatments on adhesion to yeast cells. All the strains were susceptible to chloramphenicol, erythromycin and gentamicin, while 6 strains were excluded from further evaluation because of their resistant phenotype against tetracycline. All the strains were able to grow in presence of lysozyme, as well as in MRS broth at pH 3.0. Only 4 strains showed a growth rate lower than 80 % when grown in MRS broth at pH 2.5, while a relevant growth rate decrease was observed after exposure to pH 2.0. Bile salts didn't affect the viability of the L. plantarum cells. Twenty-one strains out of 33 tested strains were able to adhere to S. cerevisiae cells. Presence of both bsh and msa genes was detected in 6 strains. The strains resistant to all the stresses, positive to agglutination with S. cerevisiae and showing bsh and msa genes were selected for further evaluation and subjected to different stress treatments over time. The assessment of growth rates showed that exposure to lysozyme significantly increased low pH resistance in L. plantarum. This increase ranged from 2.35 to 15.57 %. The consequential lysozyme and low pH exposures didn't affect the growth rate values after bile salts treatment, as well as the ability of the strains to adhere to yeast cells wasn't modified by previous treatments (lysozyme, low pH and bile salts). The present work allows to increase knowledge about non starter lactic acid bacteria from Italian food products. The studied L. plantarum strains showed a good potential for their use as probiotic cultures. However, more in vivo tests are necessary to confirm this potentiality.

Toxoplasma gondii is the causative agent of a major zoonosis with cosmopolitan distribution and is known to be transmitted mainly by the ingestion of undercooked or raw animal products. Drinking unpasteurized goat's milk is a risk factor associated with human toxoplasmosis. However, very little is known about the excretion of DNA in goat milk. Aim of the present study was to determine the seroprevalence of T. gondii infection using a modified agglutination test (MAT), to detect T. gondii DNA by nested-PCR (n-PCR) in samples of blood and milk from seropositive goats, and to genotype DNA isolates using 11 molecular markers in 127 adult lactating goats from 6 farms in Italy. Positive MAT results were found in 60.6% of goats while 13% of blood and milk samples from seropositive goats were positive to n-PCR. A kappa coefficient of 1 indicated a perfect agreement between blood and milk n-PCR. Genetic characterization of isolates revealed the occurrence of genotype III (n = 7), genotype I (n = 1), and atypical genotypes with hints for genotype I (n = 2). Our results suggest that the risk of excretion of Toxoplasma tachyzoites might frequently occur in milk of seropositive goats testing positive to n-PCR on blood.

Microsporum canis is the dermatophyte most commonly responsible for ringworm in cats. The purpose of this paper was to evaluate the in vivo efficacy of oral terbinafine (Lamisil; Sandoz) in the treatment of feline ringworm caused by M canis, and to consider this drug as an alternative to griseofulvin or imidazoles. Fifteen cats infected with M canis were treated orally once daily with 30 mg/kg of terbinafine over a 2-week period. All treated animals were checked for dermatophytes on the last day of treatment, a month later and 3 months after the last administration of the drug. Only 12 cats could be used in the whole trial and 11 of these (92%) showed a complete cure. Terbinafine could be an effective alternative to griseofulvin when fungal resistance or idiosyncrasic intolerance are shown and, compared with griseofulvin, could give a faster rate of cure and less relapses., (Copyright 1999 European Society of Feline Medicine.)

Objective: To determine efficacy of orally administered itraconazole in cats with dermatophytosis caused by Microsporum canis., Design: Uncontrolled clinical trial., Animals: 15 cats with dermatophytosis caused by M canis., Procedure: All cats were treated with itraconazole (1.5 to 3.0 mg/kg [0.7 to 1.4 mg/lb] of body weight, PO, q 24 h, for 15 days). Six cats had been treated with griseofulvin (10 mg/kg [4.5 mg/lb], PO, q 24 h) during a 60-day period, but their clinical condition had not improved. Five cats treated at the highest dosage of itraconazole vomited or became anorectic. Consequently, dosages were progressively decreased for each cat until adverse effects were not evident. After treatment, samples of hair were submitted for fungal cultures, and if appropriate, treatment was repeated when culture results were positive., Results: 8 cats treated with itraconazole recovered completely, as indicated by resolution of lesions and negative results of fungal cultures. Six of these 8 cats received a single 15-day course of treatment, whereas the remaining 2 cats needed prolonged treatment (two 15-day courses of treatment and three 15-day courses of treatment). In 4 other cats that became clinically normal, M canis was isolated from hair samples obtained at the completion of treatment, even though only 1 colony or a small number of colonies was isolated. In the other 3 cats, itraconazole did not cause clinical improvement, and culture results remained positive., Clinical Implications: Oral administration of itraconazole at dosages of 1.5 to 3.0 mg/kg may be useful for the treatment of cats with dermatophytosis attributable to M canis infections.