Quantitative risk assessment of human salmonellosis and listeriosis related to the consumption of raw milk in Italy.

Two quantitative risk assessment (RA) models were developed to describe the risk of salmonellosis and listeriosis linked to consumption of raw milk sold in vending machines in Italy. Exposure assessment considered the official microbiological records monitoring raw milk samples from vending machines performed by the regional veterinary authorities from 2008 to 2011, microbial growth during storage, destruction experiments, consumption frequency of raw milk, serving size, and consumption preference. Two separate RA models were developed: one for the consumption of boiled milk and the other for the consumption of raw milk. The RA models predicted no human listeriosis cases per year either in the best or worst storage conditions and with or without boiling raw milk, whereas the annual estimated cases of salmonellosis depend on the dose-response relationships used in the model, the milk storage conditions, and consumer behavior in relation to boiling raw milk or not. For example, the estimated salmonellosis cases ranged from no expected cases, assuming that the entire population boiled milk before consumption, to a maximum of 980,128 cases, assuming that the entire population drank raw milk without boiling, in the worst milk storage conditions, and with the lowest dose-response model. The findings of this study clearly show how consumer behavior could affect the probability and number of salmonellosis cases and in general, the risk of illness. Hence, the proposed RA models emphasize yet again that boiling milk before drinking is a simple yet effective tool to protect consumers against the risk of illness inherent in the consumption of raw milk. The models may also offer risk managers a useful tool to identify or implement appropriate measures to control the risk of acquiring foodborne pathogens. Quantification of the risks associated with raw milk consumption is necessary from a public health perspective

Four-Year Monitoring of Foodborne Pathogens in Raw Milk Sold by Vending Machines in Italy

Prevalence data were collected from official microbiological records monitoring four selected foodborne pathogens (Salmonella spp., Listeria monocytogenes, Escherichia coli O157:H7 and Campylobacter jejuni) in raw milk sold by self-service vending machines in seven Italian Regions (n. 60907 samples from 1239 vending machines) during the years 2008 to 2011. Data of samples analyzed both by culture-based and real-time PCR methods were collected in one Region. A total of 100 raw milk consumers in four regions were interviewed while purchasing raw milk from vending machines. One hundred and seventy eight samples out of 60907 were positive for one of the four foodborne pathogens investigated; overall, 18 samples were positive for Salmonella spp., 83 for L. monocytogenes, 24 for E. coli O157:H7 and 53 for C. jejuni in the seven Regions investigated. There were no significant differences in prevalence among Regions, but a significant increase in C. jejuni prevalence was observed over the years. A comparison of the two different analysis methods showed that real-time PCR is from 2.71 to 9.40 times more sensitive than culture-based method. Data on consumer habits showed that some behaviors may enhance the risk of infection due to raw milk consumption: 37% of consumers do not boil milk before consumption, 93% never use an insulated bag to transport raw milk home, and raw milk is consumed by children under five years of age. The study emphasizes that end-product controls alone are not sufficient to guarantee an adequate level of consumer protection. The beta distribution of positive samples in this study and the data on raw milk consumer habits are useful and appropriate for the development of a National Quantitative Risk Assessment of Salmonella spp., L. monocytogenes, E. coli O157 and C. jejuni related to raw milk consumption

Distribution of Transferable Antibiotic Resistance Genes in Laboratory-Reared Edible Mealworms (Tenebrio molitor L.)

In the present study, the distribution of antibiotic resistance genes in laboratory-reared fresh mealworm larvae (Tenebrio molitor L.), their feeding substrates (carrots and wheatmeal), and frass was assessed. Microbial counts on selective media added with antibiotics highlighted the presence of lactic acid bacteria resistant to ampicillin and vancomycin and, more specifically, enterococci resistant to the latter antibiotic. Moreover, staphylococci resistant to gentamicin, erythromycin, tetracycline, and vancomycin were detected. Enterobacteriaceae resistant to ampicillin and gentamicin were also found, together with Pseudomonadaceae resistant to gentamicin. Some of the genes coding for resistance to macrolide-lincosamide-streptogramin B (MLSB) [erm(A), erm(C)], vancomycin [vanA, vanB], tetracycline [tet(O)], and β-lactams [mecA and blaZ] were absent in all of the samples. For the feeding substrates, organic wheatmeal was positive for tet(S) and tet(K), whereas no AR genes were detected in organic carrots. The genes tet(M), tet(K), and tet(S) were detected in both mealworms and frass, whereas gene aac-aph, coding for resistance to amynoglicosides was exclusively detected in frass. No residues for any of the 64 antibiotics belonging to 10 different drug classes were found in either the organic wheatmeal or carrots. Based on the overall results, the contribution of feed to the occurrence of antibiotic resistance (AR) genes and/or antibiotic-resistant microorganisms in mealworm larvae was hypothesized together with vertical transmission via insect egg smearing

A Model for Coxiella burnetii Monitoring on a Sheep Dairy Farm

The aim of our study was to develop a model for farm monitoring, to be used in the case of the suspected presence of C. burnetii. Such a model could also be used to identify possible threats to human health. Thus, a C. Burnetii positive sheep farm was selected for this pilot study. Samples were taken from various areas on the farm and tested for the presence of C. burnetii using molecular methods. A possible relationship between C. burnetii presence and human seroconversion in farm workers was also investigated. Results showed that air and surface samples taken in areas of the farm used to house pregnant animals and for parturition could represent a useful tool for monitoring C. burnetii. Seroprevalence was found in farm workers, but not in veterinarians, who occasionally spent limited time on the farm. This suggests that the threat of infection is restricted to workers who are continuously present on site

Presence of Escherichia coli O157, Salmonella spp., and Listeria monocytogenes in Raw Ovine Milk Destined for Cheese Production and Evaluation of the Equivalence Between the Analytical Methods Applied

Italy is one of the main producers and exporters of cheese made from unpasteurized sheep milk. Since raw milk and its derived products are known sources of human infections, cheese produced from raw sheep milk could pose a microbiological threat to public health. Hence, the objectives of the study were: to characterize the potential risk of the presence of pathogens Escherichia coli O157, Listeria monocytogenes, and Salmonella in raw ovine milk destined for cheese production obtained from all the sheep farms (n = 24) in the Marches region (Central Italy) that directly transform raw milk into cheeses and to evaluate the equivalence between the analytical methods applied. A three-step molecular method (simultaneous culture enrichment, species-specific DNA magnetic isolation, and multiplex real-time polymerase chain reaction) was used for milk (n = 143) and cheese (n = 5) analysis over a 3- year period. L. monocytogenes was not detected on any of the farms, while E. coli O157 was found on three farms, although only using the molecular method. Four farms tested positive for Salmonella spp., and Salmonella enterica subsp. diarizonae serovar 61:k:1,5,7 was isolated in one of those cases. This information highlights the need to develop preventative measures to guarantee a high level of consumer safety for this specific product line, and the molecular method could be a time-saving and sensitive tool to be used in routine diagnosis

Human campylobacteriosis related to the consumption of raw milk sold by vending machines in Italy: quantitative risk assessment based on official controls over four years

open18noA quantitative risk assessment (RA) model was developed to describe the risk of campylobacteriosis linked to consumption of raw milk sold in vending machines in Italy. Exposure assessment was based on the official microbiological records of raw milk samples from vending machines monitored by the regional Veterinary Authorities from 2008 to 2011, microbial growth during storage, destruction experiments, consumption frequency of raw milk, serving size, consumption preference and age of consumers. The differential risk considered milk handled under regulation conditions (4°C throughout all phases) and the worst time-temperature field handling conditions detected. Two separate RA models were developed, one for the consumption of boiled milk and the other for the consumption of raw milk, and two different dose-response (D-R) relationships were considered. The RA model predicted no human campylobacteriosis cases per year either in the best (4°C) storage conditions or in the case of thermal abuse in case of boiling raw milk, whereas in case of raw milk consumption the annual estimated campylobacteriosis cases depend on the dose-response relationships used in the model (D-R I or D-R II), the milk time-temperature storage conditions, consumer behaviour and age of consumers, namely young (with two cut-off values of ≤5 or ≤6 years old for the sensitive population) versus adult consumers. The annual estimated cases for young consumers using D-R II for the sensitive population (≤5 years old) ranged between 1013.7/100,000 population and 8110.3/100,000 population and for adult consumers using D-R I between 79.4/100,000 population and 333.1/100,000 population. Quantification of the risks associated with raw milk consumption is necessary from a public health perspective and the proposed RA model represents a useful and flexible tool to perform future RAs based on local consumer habits to support decision-making on safety policies. Further educational programmes for raw milk consumers or potential raw milk consumers are required to encourage consumers to boil milk to reduce the associated risk of illness.openFederica Giacometti; Paolo Bonilauri; Simonetta Amatiste; Norma Arrigoni; Manila Bianchi; Marina Nadia Losio; Stefano Bilei; Giuseppe Cascone; Damiano Comin; Paolo Daminelli; Lucia Decastelli; Giuseppe Merialdi; Renzo Mioni; Angelo Peli; Annalisa Petruzzelli; Franco Tonucci; Silvia Piva; Andrea Serraino.Federica Giacometti; Paolo Bonilauri; Simonetta Amatiste; Norma Arrigoni; Manila Bianchi; Marina Nadia Losio; Stefano Bilei; Giuseppe Cascone; Damiano Comin; Paolo Daminelli; Lucia Decastelli; Giuseppe Merialdi; Renzo Mioni; Angelo Peli; Annalisa Petruzzelli; Franco Tonucci; Silvia Piva; Andrea Serraino

Trends in the microbial contamination of bovine, ovine and swine carcasses in three small-scale abattoirs in central Italy: A four-year monitoring

The microbial contamination of animal carcasses with respect to the limits established by Regulation (EC) No. 2073/2005 was investigated. Bovine, ovine, and swine carcasses (n=536 samples) fromthree small-scale abattoirs were sampled using abrasive sponges and tested for aerobic colony counts (ACC) and Enterobacteriaceae in the period 2010–2013. Mean ACC values reached 1.96 log cfu/cm2 on bovine carcasses and 2.27 log cfu/cm2 on both swine and ovine carcasses; Enterobacteriaceae counts of 0.01, 0.20 and 0.27 log cfu/cm2 were found for bovine, swine and ovine carcasses, respectively. Abattoir 1 showed the highest values of ACC; no differences among abattoirs were highlighted for Enterobacteriaceae. Compared with swine and ovine carcasses, bovine carcasses showed significantly lower means for both ACC and Enterobacteriaceae. The data collected indicated that the management of the three abattoirs met high quality standards, thereby proving that it is feasible to achieve good microbiological quality in abattoirs when adequate Good Hygiene Practices are applied

Revealing the microbiota of marketed edible insects through PCR-DGGE, metagenomic sequencing and real-time PCR

The present study aimed to identify the microbiota present in six species of processed edible insects produced in Thailand and marketed worldwide via the internet, namely, giant water bugs (Belostoma lutarium), black ants (Polyrhachis), winged termites (alates, Termitoidae), rhino beetles (Hyboschema contractum), mole crickets (Gryllotalpidae), and silkworm pupae (Bombyx mori). For each species, two samples of boiled, dried and salted insects were purchased. The microbial DNA was extracted from the insect samples and subjected to polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE), high-throughput sequencing and qualitative real-time PCR assays. The microbiota of the analyzed samples were widely characterized by the presence of spore-forming bacteria mainly represented by the genera Bacillus and Clostridium. Moreover, the genera Anaerobacillus, Paenibacillus, Geobacillus, Pseudomonas, Stenotrophomonas, Massilia, Delftia, Lactobacillus, Staphylococcus, Streptococcus, Vagococcus, and Vibrio were also detected. Real-time PCR allowed for ascertainment of the absence of Coxiella burnetii, Shiga toxin-producing E. coli (STEC), and Pseudomonas aeruginosa in all samples. The results of this study confirm the importance of combining different molecular techniques to characterize the biodiversity of complex ecosystems such as edible insects. The presence of potential human pathogens suggests the need for a careful application of good manufacturing practices during insect processing. This study provides further data that will be useful in risk analyses of edible insects as a novel food source

Microbiological quality assessment of meals and work surfaces in a school-deferred catering system

Microbiological contamination is one of the main parameters that must be assessed to assure the safety of foods produced by a catering system. The aim of this study conducted between 2011 and 2015 was to assess the microbiological quality of meals (n =620), work surfaces (n =621) and the operators’ hands (n =319) in a school-deferred catering system. A general positive trend regarding the microbial contamination of food samples and surfaces was observed. Salmonella spp. and Listeria monocytogenes were never detected in prepared foods; moreover, the total mesophilic aerobes and the sulfite-reducing clostridia were always below the reference limit. The occurrence of Bacillus cereus in raw and cooked vegetables highlighted the need for corrective action and suggested that continuous monitoring of this toxin-forming microorganism was necessary. A critical evaluation of the data obtained from the microbiological analyses allowed us to obtain an overview of improvements and emerging critical issues