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SARS-CoV-2 can be detected in the feces of infected people, consequently in wastewater, and in bivalve mollusks, that are able to accumulate viruses due to their ability to filter large amounts of water. This study aimed to monitor SARS-CoV-2 RNA presence in 168 raw wastewater samples collected from six wastewater treatment plants (WWTPs) and 57 mollusk samples obtained from eight harvesting sites in Campania, Italy. The monitoring period spanned from October 2021 to April 2022, and the results were compared and correlated with the epidemiological situation. In sewage, the ORF1b region of SARS-CoV-2 was detected using RT-qPCR, while in mollusks, three targets—RdRp, ORF1b, and E—were identified via RT-dPCR. Results showed a 92.3% rate of positive wastewater samples with increased genomic copies (g.c.)/(day*inhabitant) in December–January and March–April 2022. In the entire observation period, 54.4% of mollusks tested positive for at least one SARS-CoV-2 target, and the rate of positive samples showed a trend similar to that of the wastewater samples. The lower SARS-CoV-2 positivity rate in bivalve mollusks compared to sewages is a direct consequence of the seawater dilution effect. Our data confirm that both sample types can be used as sentinels to detect SARS-CoV-2 in the environment and suggest their potential use in obtaining complementary information on SARS-CoV-2. [ABSTRACT FROM AUTHOR]

Salmonella is one of the most common agents of foodborne illness. The genus Salmonella includes two species (Salmonella bongori and S. enterica) and six subspecies (enterica I, salamae II, arizonae IIIa, diarizonae IIIb, houtenae IV, and indica VI), each of which contains multiple serotypes associated with animal and human infections. The aim of the study was to evaluate the presence of Salmonella spp. in carcasses of food-producing animals and foods in southern Italy and the serovar distribution among different sources. From 2011 to 2021, a total of 12,246 foods and 982 samples from animal carcasses were collected and analyzed. The overall percentage of positive samples was 5.84% (N = 773) and a significant increase in prevalence was observed by comparing the years 2011–2015 (257, 3.27%) and 2016–2021 (516, 9.61%; p < 0.05). The highest percentage of positive food samples was observed in “Meat and Meat Products” (N = 327/2,438, 13.41%) followed by “Fish and fishery products” (N = 115/1,915, 6.01%). In carcasses, the highest percentage of positive samples was reported from broilers (N = 42/81, 51.85%) followed by buffalo (N = 50/101, 49.50%) and pork (N = 140/380, 36.84%). After typing, the isolates were assigned to the species S. enterica and to the subspecies: enterica (N = 760, 98.32%), diarizonae (N = 8, 1.03%), salamae (N = 3, 0.39%) and houtenae (N = 2, 0.26%). S. Infantis was the most frequently detected (N = 177, 24.76%), followed by S. Derby (N = 77, 10.77%), monophasic S. Typhimurium (N = 63, 8.81%), S. Typhimurium (N = 54, 7.55%), and S. Rissen (N = 47, 6.57%). By comparing the sampling period 2011–2015 with that of 2016–2021, an increase in the prevalence of S. Infantis and monophasic S. Typhimurium and a decrease of S. Typhimurium were recorded (p < 0.05). Thus, present data suggest that, despite the implementation of national and European control strategies to protect against Salmonella, the prevalence of this pathogen in southern Italy is still increasing and a change of national control programs to protect against Salmonella are necessary. [ABSTRACT FROM AUTHOR]

Bacillus cereus, a bacteria frequently associated with food spoilage, is responsible for 2 different foodborne illness in humans: a diarrheal disease, associated with cytotoxin K, hemolysin BL and a non-hemolytic enterotoxin, and an emetic syndrome, associated with the cereulide toxin. The aim of this study is to investigate the occurrence of B. cereus in 515 hard and soft cheeses collected in southern Italy. The strains were molecularly characterized for the presence of hblA, hblC, hblD, nheA, nheB, nheC, cytK, and entFM genes, related to the production of enterotoxins (trimeric hemolysin BL, trimeric non-hemolytic enterotoxin, cytotoxin K and enterotoxin FM). One hundred thirty eight samples (26.8%) were found contaminated by B. cereus. One hundred of the contaminated samples (72.5%) showed a level of B. cereus contamination ≤103 CFU/g, whereas 9 (6.5%) samples showed a contamination >105 CFU/g. Seven different molecular profiles were found among the 138 strains of B. cereus isolated. The most common profile was nheABC-entFM, found in 42 strains, followed by the hblACD-nheABC-cytK-entFM, found in 33 strains, the nheABC-cytK-entFM, found in 24 strains, the hblACDnheABC-entFM, found in 11 strains, the hblACD-nheAC-entFM, found in 15 strains, and the nheC-entFM, found in 12 strains. Eleven strains did not harbored any of these genes. In 7.2% (37/515) of samples, characterized by a bacterial contamination >103 CFU/g, we found B. cereus strains harboring genes encoding for at least one complete enterotoxin. [ABSTRACT FROM AUTHOR]

In this study, the presence of Coxiella burnetii (Cb) in cheese produced in Southern Italy from milk of cow, buffalo, and small ruminants has been evaluated. Two tests based on real-time PCR assay targeting CB IS1111 element were performed. First an assay based on the use of Taq-Man probe was performed to screen the samples. The positive samples were confirmed using both a SyBr Green test and the evaluation of the melting temperature of the amplicons. In addition, all cheese samples were also tested to determine the milk species utilized (Regulation EC 273/2008). The samples of cheese produced with a milk mix from different species were not included into the study. A total of 169 cheese samples were tested, and the obtained results showed an overall prevalence of Cb of 21.3 % with variation between species. A positivity rate of 39 % was observed in cow's cheese while Cb DNA was detected in 26 % of cheese samples made from small ruminants' milk. However, the bacterium was found only in 6.9 % of buffalo's cheese samples. A direct association between prevalence and milk used for the production was highlighted ( χ = 19.12). The statistical analysis of the prevalence in the samples from cattle and small ruminants compared with those from buffalo shows an OR of 8.4 and 4.9, respectively. [ABSTRACT FROM AUTHOR]

Simple Summary: Toxoplasmosis is a significant public health issue worldwide, caused by the intracellular protozoan Toxoplasma gondii. It has a heteroxenous life cycle in which felines act as definitive reservoirs and a wide range of warm-blooded animals, including humans, act as intermediate hosts. Due to the complex life cycle, monitoring, prevention and control of this parasite are very difficult. A thorough analysis of the epidemiology of T. gondii in humans, animals and food as well as the risk factors associated with the infection are needed to plan adequate control strategies in a given geographical area. Based on this, an integrated approach for monitoring toxoplasmosis was developed and conducted in an endemic area of southern Italy. The main tasks of this approach were based on the following strategies: parasitological and risk factor analysis for T. gondii in livestock farms, serological and molecular monitoring in meat-producing livestock at slaughterhouses, hospital discharge records (HDRs) analysis and outreach activities. The findings of this study confirmed the spread of T. gondii infection in southern Italy with high prevalence values in ruminants and the need of valid control strategies based on comprehensive and transdisciplinary actions according to the One Health approach. Toxoplasmosis is a widespread worldwide zoonotic infection caused by the intracellular protozoan Toxoplasma gondii. This protozoan infection is considered one of the most important food-borne parasitic zoonoses globally. Beyond its impact on public health, toxoplasmosis has also important veterinary implications, because it causes miscarriage or congenital malformations in livestock with negative economic impacts. An integrated monitoring programme aimed to deepen the epidemiological data on toxoplasmosis and to identify the risk factors that may favour T. gondii infections in animals and humans was conducted in an endemic area of southern Italy. The monitoring activities were based on the following tasks: (i) parasitological analysis and risk factors for T. gondii in livestock (sheep, goat, cattle and water buffalo) farms; (ii) serological and molecular monitoring at slaughterhouse in meat-producing livestock; (iii) analysis of hospital discharge records (HDRs); (iv) outreach activities (information, dissemination and health education) to farmers, vet practitioners and school-age children. The present study confirmed a very high seroprevalence of T. gondii infection in livestock farms (e.g., up to 93.1% in sheep farms) in southern Italy and highlighted the potentially significant public health risk in this area. [ABSTRACT FROM AUTHOR]

Yersinia enterocolitica (Ye) is a foodborne pathogen isolated from humans, food, animals, and the environment. Yersiniosis is the third most frequently reported foodborne zoonosis in the European Union. Ye species are divided into six biotypes 1A, 1B, 2, 3, 4, and 5, based on biochemical reactions and about 70 serotypes. Biotype 1A is non-pathogenic, 1B is highly pathogenic, and biotypes 2-5 have moderate or low pathogenicity. The reference analysis method for detecting pathogenic Ye species underestimates the presence of the pathogen due to similarities between Yersinia enterocolitica-like species and other Yersiniaceae and/or Enterobacteriaceae, low concentrations of distribution pathogenic strains and the heterogeneity of Yersinia enterocolitica species. In this study, the real-time PCR method ISO/TS 18867 to identify pathogenic biovars of Ye in bivalve molluscs was validated. The sensitivity, specificity and accuracy of the molecular method were evaluated using molluscs experimentally contaminated. The results fully agree with those obtained with the ISO 10273 method. Finally, we evaluated the presence of Ye in seventy commercial samples of bivalve molluscs collected in the Gulf of Naples using ISO/TS 18867. Only one sample tested resulted positive for the ail gene, which is considered the target gene for detection of pathogenic Ye according to ISO/TS 18867. Additionally, the presence of the ystB gene, used as target for Ye biotype 1A, was assessed in all samples using a real-time PCR SYBR Green platform. The results showed amplification ystB gene aim two samples., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Coronaviruses (CoVs), a subfamily of Orthocoronavirinae, are viruses that sometimes present a zoonotic character. Severe Acute Respiratory Syndrome Coronavirus-2 (SARS-CoV-2) is responsible for the recent outbreak of COVID-19, which, since its outbreak in 2019, has caused about 774,593,066 confirmed cases and 7,028,881 deaths. Aereosols are the main route of transmission among people; however, viral droplets can contaminate surfaces and fomites as well as particulate matter (PM) in suspensions of natural and human origin. Honey bees are well known bioindicators of the presence of pollutants and PMs in the environment as they can collect a great variety of substances during their foraging activities. The aim of this study was to evaluate the possible role of honey bees as bioindicators of the prevalence SARS-CoV-2. In this regard, 91 samples of honey bees and 6 of honey were collected from different apiaries of Campania region (Southern Italy) in four time periods from September 2020 to June 2022 and were analyzed with Droplet Digital RT-PCR for SARS-CoV-2 target genes Orf1b and N. The screening revealed the presence of SARS-CoV-2 in 12/91 in honey bee samples and in 2/6 honey samples. These results suggest that honey bees could also be used as indicators of outbreaks of airborne pathogens such as SARS-CoV-2.

This study was undertaken to detect polymorphisms in the goat and sheep mannose-binding lectin encoding gene (MBL2) and to explore allelic variability of this gene in these two species. The analysis and comparison of the sequences obtained from sheep showed 13 polymorphic sites, six in the promoter and seven in exon 1, four of which were of the missense type. In the goats, 12 polymorphic sites were detected, five intronic, five in the promoter, and one exonic. The exon site was responsible for an amino acid change. Mutations detected at the MBL2 locus in the sheep are of particular interest, being potentially responsible for the alterations of gene expression. A population survey involved 102 ewes of the Sardinian breed and 218 goats of the Nicastrese breed, all reared in southern Italy.