Antibiotic Resistance in Enterococci and Enterobacteriaceae from Laboratory-Reared Fresh Mealworm Larvae (Tenebrio molitor L.) and Their Frass.

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 ß-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.

Analysis of two cross-contamination cases of Campylobacter jejuni foodborne disease in fragile subjects in the territory of a Local Health Authority in Tuscany, Italy.

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.

Behaviour of Staphylococcus aureus in the Rearing Substrate of Tenebrio molitor Larvae.

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.

Food safety culture in food companies: evaluation of the perception of food safety culture in three Tuscan food companies.

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.

Occurrence of Campylobacter in Faeces, Livers and Carcasses of Wild Boars Hunted in Tuscany (Italy) and Evaluation of MALDI-TOF MS for the Identification of Campylobacter Species.

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.

Chemical Composition and Antimicrobial Activity against the Listeria monocytogenes of Essential Oils from Seven Salvia Species.

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 ß-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.

Antimicrobial and anti-biofilm activity of manuka essential oil against Listeria monocytogenes and Staphylococcus aureus of food origin.

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.

Microbiological Changes in Meat and Minced Meat from Beavers (Castor fiber L.) during Refrigerated and Frozen Storage.

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), Escherichiacoli 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.

Coagulase negative staphylococci from ovine bulk-tank milk: Effects of the exposure to sub-inhibitory concentrations of disinfectants for teat-dipping.

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.

In Vitro Antibacterial Activity of Manuka (Leptospermum scoparium J.R. et G. Forst) and winter Savory (Satureja montana L.) Essential Oils and Their Blends against Pathogenic E. coli Isolates from Pigs.

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.

Genotyping and Antibiotic Resistance Traits in Campylobacter jejuni and coli From Pigs and Wild Boars in Italy.

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 ß-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.

Effects of Garlic Powder and Salt on Meat Quality and Microbial Loads of Rabbit Burgers.

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.

Effects of garlic powder and salt additions on fatty acids profile, oxidative status, antioxidant potential and sensory properties of raw and cooked rabbit meat burgers.

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.

Detection of Pseudorabies Virus in Wild Boar Foetus.

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.

Listeria monocytogenes contamination of Tenebrio molitor larvae rearing substrate: Preliminary evaluations.

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.

Effect of an Italian propolis on the growth of Listeria monocytogenes, Staphylococcus aureus and Bacillus cereus in milk and whey cheese.

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 14th 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.

Biofilm Formation and Its Relationship with the Molecular Characteristics of Food-Related Methicillin-Resistant Staphylococcus aureus (MRSA).

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.

Meat from cattle slaughtered without stunning sold in the conventional market without appropriate labelling: A case study in Italy.

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.

Lactobacillus plantarum and Streptococcus thermophilus as starter cultures for a donkey milk fermented beverage.

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.

Prevalence, phenotypic and genetic diversity of Campylobacter in poultry fresh meat and poultry products on retail sale in Tuscany (Italy).

In this study, the prevalence of Campylobacter spp. in poultry fresh meat and ready­to­cook products was evaluated. Seventy­three 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 gel­electrophoresis (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.