Occurrence of Helicobacter pullorum in Retail Chicken Meat: A One-Health Approach to Consumer Health Protection.

Helicobacter pullorum is an emerging foodborne pathogen that commonly colonizes the gastrointestinal tract of poultry, causing gastroenteritis. It has been related to several clinically important infections, including colitis and hepatitis, inflammatory bowel disease, recurrent diarrhea, and bacteremia in the human population. The bacterium may be transmitted to humans through undercooked poultry meat. In order to investigate the occurrence of H. pullorum in raw retail chicken meat (thighs and breasts), we analyzed 240 samples: 120 chicken thigh and 120 chicken breast samples. The samples were analyzed by means of an isolation protocol using Steele and McDermott's modified filtration technique on Brucella agar supplemented with 5% of defibrinated sheep's blood. The presumptive colonies were biochemically identified and analyzed using a previously described conventional PCR test based on the 16S rRNA gene. In total, 35% of analyzed samples were positive using the microbiological protocol and 45% were positive by PCR. These results suggest that H. pullorum can be transmitted to humans through the handling and consumption of raw poultry meat, representing a risk for food business operators and consumers. Efforts to control H. pullorum in broiler meat should prioritize the implementation of stringent hygienic practices across all stages of the food chain, from the farm to the consumer.

Effect of rearing system (free-range vs cage) on gut and muscle histomorphology and microbial loads of Italian White breed rabbits.

OBJECTIVE: The growing consumers' interest on animal welfare has raised the request of products obtained by alternative rearing systems. The present study was conducted to assess the influence of housing system on gut and muscle morphology and on microbial load in rabbits reared under free-range (FR) and cage system (CS). METHODS: A total of forty weaned (35 days of age) male Italian White breed rabbits were allotted according to the rearing system, and at 91 days of age were randomly selected and slaughtered for the morphological evaluation of tissue from duodenum and longissimus lumborum. Morphometric analysis of the villus height, villus width, crypt depth, villus height/crypt depth ratio, and villus surface was performed. The microbial loads on hind muscle was determined by total mesophilic aerobic count (TMAC), Escherichia coli and Enterobacteriaceae; whereas, total anaerobic bacteria count (TABC) and TMAC, E. coli and Enterobacteriaceae was determined on caecal content. RESULTS: Rearing system did not interfere with the duodenum and muscle histomorphology in both rabbit groups. Similarly, microbial load of caecal content showed no significant differences on the TABC and TMAC. Conversely, significant difference was found for E. coli strains in caecal content, with the lower counts in FR compared to CS rabbits (p<0.01). Microbiological assay of muscle revealed significant lower TMAC in FR vs CS rabbits (p< 0.05). All rabbit meat samples were negative for E. Coli and Enterobacteriaceae. CONCLUSION: Free-range could be considered a possible alternative and sustainable rearing system in rabbits to preserve gut environment and muscle quality.

Preliminary survey on the occurrence of microplastics in bivalve mollusks marketed in Apulian fish markets.

Microplastics (MPs) are a relevant threat to food safety because they are ingested by humans through various foods. Bivalves are at high risk of microplastic contamination due to their filter-feeding mechanism and pose a risk to consumers as they are ingested whole. In this work, microplastics were detected, quantified, identified, and classified in samples of mussels (Mytilus galloprovincialis) and oysters (Crassostrea gigas) marketed in the Apulia region. The total number of plastic debris was 789 particles in the mussel samples and 270 particles in the oyster samples, with size ranging from 10 to 7350 µm. Fragments with size within the category of 5-500 µm were the predominant findings in both species, with blue as the predominant color in mussels and transparent in oysters; most of the debris was polyamide and nylon polymers in the mussels and chlorinated polypropylene in the oysters. These results show that mussel and oyster samples purchased at fish markets are contaminated with microplastics. The sources may be diverse and further studies are needed to assess the impact of the marketing stage on microplastic contamination in bivalves to better define the human risk assessment associated with microplastic exposure from bivalves consumption.

Occurrence and Characterization of Microplastics in Commercial Mussels (Mytilus galloprovincialis) from Apulia Region (Italy).

Microplastics are a ubiquitous pollutant whose spreading is a growing concern worldwide. They can pose a threat to food safety and consumer health as they are ingested through various foods. Bivalves are considered the most contaminated, as they filter large amounts of seawater and enter consumers' diet ingested whole. The aim of this study was to detect, quantify, identify and classify microplastics in mussels (Mytilus galloprovincialis) marketed in fishery stores in Bari and its surroundings (Apulia, Italy). A total of 5077 particles were isolated from our samples, with an average value of 1.59 ± 0.95 MPs/g and 6.51 ± 4.32 MPs/individual. Blue fragments, sized 10-500 µm, were the prevalent findings; most of them belonged to Polyamide (PA) polymers. The results of this study help to show that mussels represent a source of microplastics for consumers and a direct risk to their health, even considering that they may contain many chemical compounds and microorganisms that may or may not be pathogenic to humans. Further research is needed to assess the role of commercialization in bivalve molluscs contamination.

DNA metabarcoding for identification of species used in fish burgers.

The absence of morphological identification characters, together with the complexity of the fish supply chain make processed seafood vulnerable to cases of species substitution. Therefore, the authentication and the traceability of such products play a strategic role in ensuring quality and safety. The aim of the present study was to detect species used in the production of multi-species fish burgers and to evaluate mislabelling rates, using a DNA metabarcoding approach by sequencing a fragment of the 16S rRNA mitochondrial gene. The study highlighted the presence of 16 marine and 2 mammalian taxa with an overall mislabelling rate of 80%, including cases of species substitution, the undeclared presence of molluscs and of taxa whose use is not permitted by current Italian legislation. The presence of swine DNA as well as the inclusion of undeclared taxa potentially causing allergies raise concerns regarding consumer safety and protection regarding ethical or religious issues. Overall, the study shows that the application of DNA metabarcoding is a promising approach for successfully enforcing traceability systems targeting multi-species processed food and for supporting control activities, as a guarantee of an innovative food safety management system.

Levels of Mercury, Methylmercury and Selenium in Fish: Insights into Children Food Safety.

Total mercury (THg), methylmercury (MeHg), and selenium (Se) concentrations were measured in various commercially important fish species. The benefit-risk binomial associated with these chemicals was assessed in children through the probability of exceeding the provisional tolerable weekly intakes (PTWIs) of the contaminants and the Se recommended dietary allowance (RDA). The Se:Hg molar ratios, selenium health benefit values (HBVSe), and monthly consumption rate limits (CRmm) for each species were also calculated. THg and Se were analyzed by atomic absorption spectrophotometer (Shimadzu, Milan, Italy), while MeHg was determined by Trace Ultra gas chromatograph connected with a PolarisQ MS (Thermo Fisher Scientific, Waltham, MA, USA). None of the analyzed fish had Hg levels above the European Community regulatory limits, while most large predators had MeHg levels over the threshold concentration set by US EPA. The estimated weekly intakes of THg and MeHg exceeded in many cases the PTWIs and the Se estimated daily intakes were provided from 0.71% to 2.75% of the RDA. Se:Hg molar ratios above 1 and positive HBVSe index suggested that Se in fish could be enough to alleviate the potential toxic effect of Hg. However, high-risk groups as children should consume fish in moderation because a large consumption pattern, especially of swordfish and tunas, might be of concern for health.

Helicobacter pylori: Survival in cultivable and non-cultivable form in artificially contaminated Mytilus galloprovincialis.

Several studies report the presence of Helicobacter pylori (H. pylori) in seawater either free or attached to planktonic organism. After considering the role played by plankton in the food chain of most aquatic ecosystems and the possible role that seafood products can assume in the transmission of H. pylori to humans, the aim of this study was to assess the survival of H. pylori in artificially contaminated Mytilus galloprovincialis (M. galloprovincialis). A traditional culture method and a reverse transcriptase-PCR (RT-PCR) assay were employed to detect the mRNA of known virulence factor (VacA) which can be considered use a marker of bacterial viability. The obtained results clearly show that H. pylori is able to survive in artificially contaminated mussels for 6 days (2 days in a cultivable form and 4 days in a non-cultivable form).

Food safety in collective catering: knowledge, attitudes and correct application of GHP/GMP knowledge among foodservice workers.

Food safety critically reliant on the behaviors adopted by foodservice personnel, involved in various stages of meal preparation, from the supply of raw food materials until its distribution to the consumer. The purpose of this work was to collect information on the level of knowledge on food safety, the correct application of the Good Hygiene Practices (GHP) and Good Manufacturing Practices (GMP) and the attitudes of the operators involved in collective catering. The survey was conducted among 15 social catering companies in Apulia (Italy) using a modified Sharif and Al-Malki (2010) anonymous questionnaire intended to assess the knowledge, attitudes and practices of the food handlers. The results highlight the need to improve both knowledge and practices regarding food hygiene and safety in order to minimize the risk of transmitting foodborne diseases. More careful planning, implementation, monitoring and evaluation of training programs for foodservice workers is required, taking into consideration the provisions of European food laws.

Dioxin and PCB residues in meats from Italy: Consumer dietary exposure.

In order to investigate PCB and PCDD/F concentrations and potential human health risk a study has been conducted in meat samples (beef, pork, chicken and turkey) purchased from Italian supermarkets. PCBs were dominant (41.8-77.7 ng g-1 l.w.) with respect to PCDD/Fs (20.1-91.1 pg g-1 l.w.). The levels were variable and largely dependent upon the type of meat. Accumulation pattern showed a distribution typically reported for meat. PCBs tended to decrease from lowest to highest congeners, while for PCDD/Fs the concentration profile was dominated by highly chlorinated dioxins and furans. Concentrations of PCDD/Fs plus dl-PCBs and indicator PCBs exceeding the EU maximum permissible levels were found in 23.3% and 53.3% of the samples, respectively. Exposure estimates to PCDD/Fs plus dl-PCBs (0.08-4.16 pg WHO-TEQs kg-1 b.w. w.-1) were within the new limit proposed by the EFSA's expert panel, except for pork sausage, showing a twofold higher value. Concerning the exposure derived from indicator PCBs, the maximum level set by different European countries (10 ng kg-1 b.w. d.-1) was surpassed solely via consumption of pork sausage (17.22 ng kg-1 b.w. d.-1), thought also the estimated intake from hamburger consumption (6.88 ng kg-1 b.w. d.-1) merits attention being close to guideline limit.

Helicobacter pylori: A foodborne pathogen?

Helicobacter pylori (H. pylori) is an organism that is widespread in the human population and is sometimes responsible for some of the most common chronic clinical disorders of the upper gastrointestinal tract in humans, such as chronic-active gastritis, duodenal and gastric ulcer disease, low-grade B-cell mucosa associated lymphoid tissue lymphoma of the stomach, and gastric adenocarcinoma, which is the third leading cause of cancer death worldwide. The routes of infection have not yet been firmly established, and different routes of transmission have been suggested, although the most commonly accepted hypothesis is that infection takes place through the faecal-oral route and that contaminated water and foods might play an important role in transmission of the microorganism to humans. Furthermore, several authors have considered H. pylori to be a foodborne pathogen because of some of its microbiological and epidemiological characteristics. H. pylori has been detected in drinking water, seawater, vegetables and foods of animal origin. H. pylori survives in complex foodstuffs such as milk, vegetables and ready-to-eat foods. This review article presents an overview of the present knowledge on the microbiological aspects in terms of phenotypic characteristics and growth requirements of H. pylori, focusing on the potential role that foodstuffs and water may play in the transmission of the pathogen to humans and the methods successfully used for the detection of this microorganism in foodstuffs and water.

Estimated Dietary Intake of Trace Metals from Swordfish Consumption: A Human Health Problem.

Trace element (Hg, Pb, Cd, Zn, Cu, Ni, and Cr) occurrence was determined in the muscle tissue of swordfish collected in the Mediterranean Sea to assess whether the intakes complied with the recommended levels for essential metals and permissible levels for toxic elements. Metals were analyzed by an atomic absorption spectrophotometer (Shimadzu AA 7000). The methodology of Target Hazard Quotient (THQ) was also evaluated. The ranking order of toxic metal concentration was Hg > Cd > Pb, while for essential elements the distribution pattern followed the sequence Zn > Cu > Ni > Cr. The Estimated Weekly Intakes (EWI) as well as THQ for Cd and Pb indicated that swordfish consumption did not pose a risk to human health, whereas the major concern was for Hg. Fish size-related changes in Hg concentrations resulted in high EWI and THQ values relative to larger fish consumption, implying a potential risk to human health. For consumer protection, catches of swordfish approximately above 44 kg should be avoided as these fish have a higher risk of containing toxic levels of Hg.

Methicillin-resistant Staphylococcus aureus (MRSA) in slaughtered pigs and abattoir workers in Italy.

Methicillin-resistant Staphylococcus aureus (MRSA) is a pathogen present in the hospital environment (HA-MRSA), in the community (CA-MRSA) and in livestock, including pigs (LA-MRSA). MRSA may enter the human food chain during slaughtering and may infect humans coming into direct contact with pigs or pork products. This study aimed to determine the prevalence and characteristics of MRSA isolated from pigs and workers at industrial abattoirs in southern Italy. A total of 215 pig nasal swabs were screened for the presence of MRSA using PCR. An MRSA isolate was detected from each mecA/nuc PCR-positive sample and characterized by spa-typing, Multi-Locus Sequence Typing, SCC-mec and Panton-Valentine Leukocidin (PVL), and also tested for the production of staphylococcal enterotoxins (SEs). Eighty-one MRSA isolates (37.6%) were obtained from the 215 pig nasal swabs; 37 of these isolates were further characterized, and showed 18 different spa-types and 8 different STs. The most frequently recovered STs were ST398 (CC398-t034, t011, t899, t1939 - 43.2%) followed by ST8 (CC8-t008, t064, t2953, t5270 - 24.3%) and ST1 (CC1-t127, t174, t2207 - 10.8%). Nine MRSA isolates were obtained from the 113 human swabs; the isolates showed 5 different spa-types and 5 different STs, including the novel ST2794 (t159). The most representative STs recovered were ST1 (CC1-t127) and ST398 (CC398-t034) (33.3%). None of the MRSA isolates showed the ability to produce SEs and PVL and all resulted resistant to two or more classes of antimicrobials. This study shows the great genetic diversity of MRSA strains in slaughtered pigs and in abattoir employees in Italy, and clearly demonstrates the need for improved hygiene standards to reduce the risk of occupational and food-borne infection linked to the handling/consumption of raw pork containing MRSA.

Microbiological quality of Burrata cheese produced in Puglia region: southern Italy.

Burrata cheese is a popular typical Italian food product, produced in Puglia (an administrative region of southern Italy), and this study investigated the microbiological quality of 404 samples of this cheese. The samples were analyzed in order to quantify Escherichia coli and to detect the presence of Staphylococcus aureus, Salmonella spp., and Listeria monocytogenes. No sample exceeded the values of E. coli set by EC Regulation 1441/07 for some dairy products, while 15 (3.7%) samples tested coagulase-positive staphylococci positive, with values greater than 10(3) CFU/g. One strain of S. aureus was identified and characterized from each of these positive samples, and of these strains, 7 (46.6%) produced staphylococcal enterotoxin A, 5 (33.3%) produced staphylococcal enterotoxin C, 2 (13.3%) produced staphylococcal enterotoxin D, and 1 (6.6%) produced both staphylococcal enterotoxins A and D. All strains were mecA negative. The 15 S. aureus isolates were tested for their antimicrobial resistance patterns, and all analyzed strains showed antimicrobial resistance properties for at least one of the tested antibiotics. Testing for the other pathogens mentioned above gave negative results. The results of our study mean that the microbiological quality of Burrata cheese can be assumed to be good, although care must be taken with raw materials and good hygiene during processing in order to guarantee greater food safety.

Toxic metals (Hg, Cd, and Pb) in fishery products imported into Italy: suitability for human consumption.

Mercury, cadmium, and lead concentrations were determined in various fishery products (fishes, cephalopod molluscs, and crustaceans) imported into Italy from many European and non-European coastal countries. Considerable differences were found in the concentrations of these metals among the products tested. The highest mean Hg concentration was found in fishes (0.21 µg g(-1) wet weight), whereas cephalopods had the highest mean Cd concentration (0.35 µg g(-1) wet weight). Swordfish (0.80 µg g(-1) wet weight), longtail tuna (0.53 µg g(-1) wet weight), and thornback ray (0.52 µg g(-1) wet weight) had the highest concentrations of Hg, whereas maximum Cd concentrations were found in samples of common cuttlefish (0.85 µg g(-1) wet weight) and common octopus (0.64 µg g(-1) wet weight). The majority of the samples analyzed were in compliance with European Union legislation, except for a few cases. The calculated mean weekly intakes of Hg, Cd, and Pb through consumption of the fishery products tested were all below the legislated respective provisional tolerable weekly intakes. In general, the samples analyzed were considered safe to eat with regard to the metal concentrations found and the allowable intakes based on legislation. Nevertheless, the consumption of some species may be of significant importance for consumer health.

Verocytotoxin-producing Escherichia coli O26 in raw water buffalo (Bubalus bubalis) milk products in Italy.

Escherichia coli 026 is known as a verocytotoxin-producing E. coli (VTEC) organism that causes severe foodborne diseases such as hemorrhagic colitis and hemolytic uremic syndrome. Although cattle are the most important reservoir of VTEC, only a few reports on the role of water buffalo (Bubalus bubalis) as a reservoir of VTEC and on the presence of these organisms in their milk are available. However, in Southern Italy, where water buffalo are intensively reared, an outbreak of hemolytic uremic syndrome due to E. coli 026 has recently been reported, in which the consumption of typical dairy products was considered to be a common risk factor. The aims of this work were to assess the prevalence of E. coli O26 in raw water buffalo milk, to characterize the virulence gene profiles of the isolates, and to evaluate their phenotypic antimicrobial resistance pattern. Of 160 analyzed samples, 1 (0.6%) tested positive for E. coli O26, and the isolate showed the stx1+/stx2+/eae-/hlyA+ genotypic profile. The strain showed resistance against glycopeptides, macrolides, and penicillins. The presence of VTEC organisms in raw water buffalo milk could be considered to be a potential threat to consumers; however, the strict adherence to the processes used in the preparation of the most common buffalo dairy products could strongly mitigate the foodborne risk. To our knowledge, this article reports the first isolation and characterization of E. coli O26 VTEC in raw water buffalo milk.