Italian Journal of Food Safety

Shotgun metagenomic investigation of foodborne pathogens and antimicrobial resistance genes in artisanal fermented meat products from the Mediterranean area

2024-05-10T09:32:36+00:00

In this pilot study, we compared the metagenomic profiles of different types of artisanal fermented meat products collected in Italy, Greece, Portugal, and Morocco to investigate their taxonomic profile, also in relation to the presence of foodborne pathogens and antimicrobial resistance genes. In addition, technical replicates of the same biological sample were tested to estimate the reproducibility of shotgun metagenomics. The taxonomic analysis showed a high level of variability between different fermented meat products at both the phylum and genus levels. Staphylococcus aureus was identified with the highest abundance in Italian fermented meat; Escherichia coli in fermented meat from Morocco; Salmonella enterica in fermented meat from Greece; Klebsiella pneumoniae and Yersinia enterocolitica in fermented meat from Portugal. The fungi Aspergillus, Neosartoria, Emericella, Penicillum and Debaryomyces showed a negative correlation with Lactococcus, Enterococcus, Streptococcus, Leuconostoc and Lactobacillus. The resistome analysis indicated that genes conferring resistance to aminoglycoside, macrolide, and tetracycline were widely spread in all samples. Our results showed that the reproducibility between technical replicates tested by shotgun metagenomic was very high under the same conditions of analysis (either DNA extraction, library preparation, sequencing analysis, and bioinformatic analysis), considering both the degree of overlapping and the pairwise correlation.

Microsporidian heavy infection in a batch of salted and dried cod

2024-05-03T06:55:54+00:00

The aim of this work is the description and characterization of a severe microsporidian infection in a batch of salted and dried cod. Particularly, the case involves a batch of approximately 800 kg obtained from Gadus macrocephalus (Food and Agriculture Organization Zone 61 - Northwest Pacific Ocean), which, after rehydration and sectioning operations, underwent routine company checks before packaging. In about 20% of the samples, the presence of whitish nodules with a diameter ranging from 1 to 2 mm was observed on the surface of the fillets and in cross-section. The lesions ranged from a few units to 10 per cm². Some samples were subjected to fresh microscopic observation with the stereomicroscope, confirming the nodular nature of the lesions, which were often confluent, alternating with empty spaces, giving the tissue a honeycombing aspect. The histological examination at low magnification allowed us to observe the heavy vacuolization of nodular lesions irregularly surrounded by a spongy-like wall. The observation at higher magnification of other sections allowed us to identify intra-myofibrillar cists containing presumptive microsporidian elements. The tissue damage derived from the technological processes and gravity of lesions did not allow a morphological characterization of presumptive protozoans. The molecular examination of the nodular lesions and the analysis of the sequence of an 897 bp fragment of the small subunit 16S rRNA revealed 100% identity with Microsporidium theragrae (GenBank accession number MT928885-89) first isolated from the skeletal muscles of Gadus chalcogrammus specimens from the Sea of Okhotsk. This finding confirms the importance of selecting suppliers and raw materials in the seafood industry, as well as the usefulness of an effective traceability system.

Official controls on the distance sale of dairy products in the territory of the Modena Local Competent Authority: an analysis of websites

2024-04-18T07:10:24+00:00

The internet and digital technologies pose new, specific challenges to competent authorities, whose activity fields are limited to their own jurisdictions. Consequently, these authorities must operate a system of controls adapted to online sites and e-sales, and official control must be strengthened. To address these challenges and protect consumers from misleading practices that can lead to the consumption of unsafe food, it is necessary to adapt official controls to new distance-selling techniques. The food inspection scheme as conceived in the European legal system cannot be easily applied to online food sales. To this purpose, the competent authorities need to equip themselves to guarantee effective control and compliance with the European Union laws regarding food sold online. It emerged that there are different levels of non-compliance with online food: the non-registration of the food business operator (registrant) and the non-compliance with the obligations as the law prescribes (obligations on food hygiene, electronic commerce, and information to consumers about the food). The focus of this survey on the distance sale of dairy products in the Modena Local Competent Authority territory is on the pattern suggested by the Food Standards Agency in 2016. This analysis is based on two different phases. The first one is the study of the websites, and the second one is the physical visit to the address of the online food business. This survey shows how to collect the first phase’s data and organize the official controls.

One-year monitoring of potentially pathogenic microorganisms in the waters and sediments of the Lesina and Varano lagoons (South-East Italy)

2024-05-07T14:04:28+00:00

In this study, two Mediterranean coastal lagoons (Lesina and Varano) of southern Italy, located in the north of the Apulia region, were investigated for the presence of Shiga toxin Escherichia coli (STEC) and potentially enteropathogenic Vibrio species in parallel with norovirus (NoV), hepatitis A virus (HAV), hepatitis E virus (HEV), and severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). This study aimed to evaluate the presence of potentially pathogenic bacteria and viruses in the water and sediments of these ecosystems. From March 2022 to February 2023, a total of 98 samples were collected: 49 water samples and 49 sediment samples. STEC strains were isolated in three samples (3.1%), of which one (2%) was water (stx1 and stx2 positive) and two (4.1%) were sediment (both stx2 positive) samples. Vibrio spp. were detected in twenty samples (20.4%), of which nine were water (18.4%) and eleven were sediment (22.4%) samples. The species detected included V. parahaemolyticus, V. cholerae, and V. vulnificus. NoV was detected in 25 (25.5%) samples, while none of the water or sediment samples were positive for HAV, HEV, and SARS-CoV-2. The results of this study provide an overview of the presence of potentially pathogenic microorganisms in areas influenced by anthropogenic pressure. Monitoring the circulation of these pathogens could be useful to evaluate the water flowing into the lagoons, in particular discharge waters (i.e., urban, agricultural, and livestock runoff), considering the presence of fish and shellfish farms in these sites.

Implementation of a pilot checklist for control standardization on fishery goods consignments at the Livorno border control post

2024-04-11T09:53:14+00:00

To standardize control activities, it is necessary to introduce checklists to support the control of consignments entering the European Union through border control posts (BCPs). This study aimed to develop a pilot checklist for the control of fishery consignments, preliminarily identified as the predominant group of goods entering the Livorno (Italy) BCP. The design of the pilot checklist was preceded by i) a revision of the current European and national legislation on the general and specific objectives of border control activities on fishery products and ii) a comparative analysis of two checklists (one of the Ministry of Health and one of the former Livorno border inspection post) developed on the basis of the repealed legislation. This comparison aimed to define the pilot checklist structure, verification objectives, and selection of assessment scores to be included in defining consignment compliance and acceptability. Once developed, the clarity and ease of use of the first draft of the pilot checklist were verified through its use in a field test during the control of 64 fishery product consignments. 22 regulatory sources (18 European and 4 national) were selected as reference legislation. The pilot checklist was structured as a dynamic “read-do” document based on the workflow of control activities described in the current legislation. The field test was useful in improving the clarity of the verification objectives within the documentary, identity, and physical control sections and in facilitating the use of the checklist and the collection of evidence during the control activity. This study, which focused on fishery products, can provide a practical approach for the development of checklists for all the other categories of goods under the responsibility of BCPs.

Evaluation of hepatitis E virus RNA persistence in experimentally contaminated cured pork liver sausages

2024-04-02T08:42:10+00:00

Hepatitis E is a disease sustained by RNA viruses, which have four different genotypes, all of which are responsible for acute forms of hepatitis. Genotypes 1 and 2 infect only humans, causing epidemics mainly transmitted by contaminated water, while genotypes 3 and 4 are zoonotic, and the infection is linked to the consumption of raw or undercooked meat or meat products. Hepatitis E virus (HEV) genotypes 3 and 4 have been detected in domestic Suidae, considered the asymptomatic reservoir of HEV, and in wild animals such as wild boar and deer. Despite scientific studies that have highlighted the presence of HEV in cured meat products, such as pork liver sausages, the viral persistence in the different production steps of curing has not been evaluated. Therefore, this study aimed to evaluate the persistence of HEV genotype 3 during the different curing and storage times of experimentally contaminated pork liver sausages using biomolecular methods. The sausages tested positive at all curing and storage times. This study confirms the potential risk attributed to pork liver sausages in HEV transmission. However, to guarantee an efficient risk assessment, future studies will be performed to correlate the presence of HEV RNA with infectious viral particles.

Occurrence and genomic characterization of antimicrobial-resistant and potential pathogenic Escherichia coli from Italian artisanal food productions of animal origin

2024-03-25T08:56:49+00:00

Escherichia coli can harbor a broad repertoire of virulence and antimicrobial resistance (AMR) genes, which can be exchanged across the human gastrointestinal microflora, thus posing a public health risk. In this study, 6 batches of artisanal soft cheese and a 6-month ripened fermented dried sausage were investigated to assess the occurrence, phylogeny, and genomic traits (AMR, virulence, and mobilome) of E. coli. 30 and 3 strains isolated from salami and cheese food chains, respectively, were confirmed as E. coli by whole genome sequencing. The accumulation of single nucleotide polymorphism differences within small clusters of strains encompassing batches or processing stages, combined with high serotype and phylogroup diversity, suggested the occurrence of different contamination phenomena among the facilities. A total of 8 isolates harbored plasmid-mediated resistance genes, including one cheese strain that carried an IncQ1 plasmid carrying AMR determinants to macrolides [mph(B)], sulfonamides (sul1, sul2), trimethoprim (dfrA1), and aminoglycosides [aph(3”)-Ib and aph(6)-Id]. A pool of virulence-associated genes in the class of adhesion, colonization, iron uptake, and toxins, putative ColV-positive iron uptake systems sit, iro, or iuc (8 salami and 2 cheese), plasmid-encoded hemolysin operon hlyABCD (one salami), and potential atypical enteropathogenic E. coli (3 salami environment) were reported. Overall, our findings underscore the importance of routine surveillance of E. coli in the artisanal food chain to prevent the dissemination of AMR and virulence.

Analysis of the original notifications in the Tuscany region “Rapid Alert System for Food and Feed” in the seven-year period 2015-2021

2024-04-04T08:00:55+00:00

The original notifications (n=1355) managed by the Rapid Alert System for Food and Feed of the Tuscany region in the seven-year period 2015-2021 were analyzed. 68.9% of them were classified as alert notifications, and they mostly originated (56.3%) from official controls on the market, followed by the company’s own checks (29.1%). Italy was the most represented notifying country (73.3%) and the most represented country of origin of the notified products (64.5%). Out of the 28 considered food categories, “fish and products thereof” (F), “cereals and bakery products” (C), “bivalve mollusks and products thereof” (B), and “meat and meat products (other than poultry)” (M) were the most notified. F were especially notified for the presence of heavy metals (45.7%), C for undeclared allergens (32.1%), B for microbial contaminants (Escherichia coli, 49.6%), and M for pathogenic microorganisms (55.9%), mostly Salmonella spp. and Listeria monocytogenes. This study provides an updated framework for the main food safety issues at the regional level. The Rapid Alert System for Food and Feed should be periodically analyzed to evaluate historical trends and emerging or poorly known hazards. This is especially important for the planning of official control activities and the prevention of risks through the food supply chain.

Molecular evidence of Toxoplasma gondii from the tissue and blood of naturally infected sheep

2024-03-28T10:39:35+00:00

Toxoplasmosis is a cosmopolitan zoonotic infection that has significant effects on public health and causes economic losses in the livestock industry. The current study was designed to detect the Toxoplasma parasite in sheep blood samples and tissue samples of slaughtered sheep at the Sulaimani abattoir using molecular technique. A total of 300 peripheral sheep blood samples were randomly collected from 20 small ruminant flocks at 4 locations in the Sulaymaniyah province, northern Iraq. Also, 150 meat samples from thigh muscle, heart, and diaphragm were collected from slaughtered sheep. All collected blood samples were subjected to polymerase chain reaction (PCR) amplification to confirm Toxoplasma infection; in addition, meat samples were also analyzed for Toxoplasma by PCR following the digestion process. Of the 300 amplified blood samples, 94 were considered positive for Toxoplasma gondii, with a prevalence rate of 31.3%. The overall prevalence of Toxoplasma among meat samples was 34%. The diaphragm reported a higher infection rate (46%) than the heart (32%), while the femoral muscle reported an infection rate of 24%. Aged animals (older than 24 months) presented a higher infection rate (32.8%) than younger animals (28.9%). Contact with or consumption of uncooked meat from infected sheep increases the chance of parasite transmission to humans.

A study on textile microfiber contamination in the gastrointestinal tracts of Merluccius merluccius samples from the Tyrrhenian Sea

2024-03-28T08:25:37+00:00

The increased demand and consumption of synthetic textiles have contributed to microplastic pollution in the form of microfibers. These particles are widely spread in the aquatic environment, leading to the exposure of marine biota, including edible species. The current study aimed to assess the extent of microfiber contamination in a commercially relevant fish species, Merluccius merluccius, which is considered a small-scale bioindicator for the monitoring of plastic ingestion in the Mediterranean coastal environment. The frequency of ingestion, abundance, and composition of textile microfibers isolated from the fish gut were characterized. Results showed the occurrence of microfibers in 75% of the samples, with a mean number of 10.6 microfibers/individual, of which 70% were classified as natural microfibers. The spectroscopic analyses confirmed both the visual identification of microfibers and the prevalence of cellulosic fibers. The obtained findings provided evidence of both natural/artificial and synthetic microfiber exposure in an important commercial fish species that, considering the consumption of small individuals without being eviscerated, may be a potential route of microfiber exposure in humans. Monitoring programs for fishery products from markets are needed to assess contamination levels and human health risks. In addition, measures to control microfiber pollution need to occur at multiple levels, from textile industries to international governments.

Qualitative characteristics of sheep’s and goat’s milk in Albania

2024-03-19T10:25:14+00:00

Cost-effective methods are crucial in small-scale operations to assess the physicochemical properties of milk, identify potential clinical or subclinical mastitis, and test β-lactams in small ruminants. The aim was to evaluate the importance of physicochemical characteristics, somatic cells, and antibacterial substances as factors for monitoring the quality and safety of bulk tank milk from sheep and goats. Furthermore, we aimed to elucidate the influence of grazing capacity on the composition of sheep milk. The fat content of sheep milk (n=119) was found to be 7.7%, with a range of 5.3% to 9.9%. The average fat content of dairy goats (n=33) was found to be 4.9%, with a range of 2.8-6.7%. The results indicate that 2.8% of sheep milk samples and 7.8% of goat milk samples exhibited clear positive reactions when assessed using the California mastitis test. β-lactam antibiotics were present in 0.7% of sheep milk samples, but not in any of the goat milk samples using lateral flow strips. This measurement method can be useful to detect adulterated milk and better understand the quality and safety of small ruminant milk before processing.

Monitoring of bivalve mollusk harvesting areas: the relevance of Salmonella spp.

2024-03-15T09:11:37+00:00

The microbiological monitoring of bivalve mollusk harvesting areas in the Marche region is based on the parameters of Escherichia coli and Salmonella spp. However, Regulation EU/2019/627 stated criteria based on E. coli only to determine the health status of these areas. Therefore, the reason for Salmonella spp. monitoring, as provided in the Marche region, could be aimed at reducing the risk of placing on the market contaminated bivalve mollusks. This study, using the results of microbiological monitoring carried out in the Marche region from 2015 to 2022 and the methods based on Bayes’ theorem and Poisson’s distribution, evaluated the effectiveness and efficiency of Salmonella spp. monitoring in reducing the risk to the consumer. The results show that i) the use of a single sample unit significantly reduced the possibility of detecting non-compliance with the microbiological safety criterion; ii) the time taken to report positive results (average of approximately 10 days) did not allow the timely implementation of control measures; iii) the prevalence of positive outcomes was quite sporadic: a random trend of positivity is recognizable on a geographical and monthly basis for mussels and a geographical basis for striped clams; iv) considering the predictive value of E. coli against Salmonella spp., the specificity is very high and the negative predictive value versus Salmonella spp. would be >80%. In conclusion, the study shows that the monitoring of Salmonella spp. has a limited effect on reducing the risk to the consumer; however, in the cost/benefit assessment, other aspects not covered by this study should be considered.

Staphylococcal food poisoning outbreaks occurred in Sicily (Italy) from 2009 to 2016

2024-03-13T10:54:44+00:00

Notification of foodborne outbreaks has been mandatory in Europe since 2005, and surveillance is carried out along the entire food chain. Here we report the results obtained from laboratory investigations about four cases of foodborne outbreaks that occurred in Sicily between 2009 and 2016, deemed to be related to staphylococcal enterotoxins (SEs) and coagulase-positive Staphylococci (CPS) by the Local Public Health Authority. Primosale cheese samples were processed by culture methods for enumeration of CPS and immunoenzymatic assays for detection and differentiation of the SEs possibly contained in food samples. In all cases, the mistrusted foods were found to be contaminated by CPS at bacterial loads between 5 and 8 log CFU/g and contained SE type C (SEC). The reported data confirm the risk of staphylococcal food poisoning associated with the consumption of raw milk cheese. SEC is the most commonly occurring SE in goat milk and dairy products and the most represented enterotoxin in Sicilian dairy products. Our results highlighted the need for improving the current monitoring efficiency and implementing the available laboratory methods to collect more faithful epidemiological data on the current prevalence of staphylococcal toxins in the food chain, including SEs currently not detectable by validated analytical methods.

A systematic review of the role of integrated farming and the participation of universities in ensuring food security: Malaysia’s effort

2024-03-29T08:36:27+00:00

Food security is critical for promoting health and well-being and achieving sustainable development, especially in developing countries. Despite the recent efforts to improve it, food security is still a concern due to the rapid increase in populations, conflicts, and natural disasters worldwide. Universities, particularly agricultural universities, play an essential role in addressing food security issues by researching, developing new technologies, and providing education and training to farmers and other stakeholders. The main objective of this review is to discuss the role of universities and integrated farming in ensuring food security, specifically in Malaysia. It includes a brief overview of the different types of integrated farming methods that can be used to improve food security and finally discusses the Student Farmer Entrepreneur program, which can be crucial for promoting food security by increasing agricultural productivity, promoting local food production, encouraging sustainable agriculture practices, and supporting rural development. This review also considers the significant impact of the Malaysian government and universities on food security. With integrated farming and the Student Farmer Entrepreneur program, food security can be further improved.