Serotype diversity and antimicrobial susceptibility profiles of Actinobacillus pleuropneumoniae isolated in Italian pig farms from 2015 to 2022.

Actinobacillus pleuropneumoniae (APP) is a bacterium frequently associated with porcine pleuropneumonia. The acute form of the disease is highly contagious and often fatal, resulting in significant economic losses for pig farmers. Serotype diversity and antimicrobial resistance (AMR) of APP strains circulating in north Italian farms from 2015 to 2022 were evaluated retrospectively to investigate APP epidemiology in the area. A total of 572 strains isolated from outbreaks occurring in 337 different swine farms were analysed. The majority of isolates belonged to serotypes 9/11 (39.2%) and 2 (28.1%) and serotype diversity increased during the study period, up to nine different serotypes isolated in 2022. The most common resistances were against tetracycline (53% of isolates) and ampicillin (33%), followed by enrofloxacin, florfenicol and trimethoprim/sulfamethoxazole (23% each). Multidrug resistance (MDR) was common, with a third of isolates showing resistance to more than three antimicrobial classes. Resistance to the different classes and MDR varied significantly depending on the serotype. In particular, the widespread serotype 9/11 was strongly associated with florfenicol and enrofloxacin resistance and showed the highest proportion of MDR isolates. Serotype 5, although less common, showed instead a concerning proportion of trimethoprim/sulfamethoxazole resistance. Our results highlight how the typing of circulating serotypes and the analysis of their antimicrobial susceptibility profile are crucial to effectively manage APP infection and improve antimicrobial stewardship.

The role of co-infections in M. hyopneumoniae outbreaks among heavy fattening pigs: a field study.

Little is known about how co-infections and genotype dynamics affect Mycoplasma hyopneumoniae infection in fattening pigs. This study was aimed at assessing the role of co-infections in M. hyopneumoniae outbreaks, their influence on the presence of M. hyopneumoniae genotypes and their impact on consequent lung lesions. Tracheobronchial swabs (TBS) from 300 finishers were collected from 10 farms at the onset of enzootic pneumonia outbreaks and 1 month later, sampling of 3 groups per farm: Group A showed clinical signs first, Group B was housed near Group A, and Group C was located in a different building. Pigs' lungs were scored at the slaughterhouse. TBS were tested for the main pathogens involved in respiratory diseases, and samples positive for M. hyopneumoniae were genotyped by multiple-locus variable-number tandem repeat analysis (MLVA). Pigs in Group A showed the highest prevalence and load of M. hyopneumoniae. A positive association was detected between M. hyopneumoniae and Mycoplasma hyorhinis, whereas Actinobacillus pleuropneumoniae was more frequent when the M. hyopneumoniae load was higher. Nevertheless, co-infection had no effect on lung lesion scores. The presence of multiple MLVA types (mixed infections) increased in time only in pigs from Group C and was positively associated with porcine reproductive and respiratory syndrome virus infection. Lung lesions were more severe in pigs with at least one TBS positive for M. hyopneumoniae and in pigs with a history of mixed infections. The central role of M. hyopneumoniae and relevance of mixed infections suggest that increased biosecurity might be beneficial for lung lesion sequelae.

The role of transportation in the spread of Brachyspira hyodysenteriae in fattening farms.

BACKGROUND: Direct and indirect contact among animals and holdings are important in the spread of Brachyspira hyodysenteriae. The objective of this study was to investigate the role of slaughterhouse vehicles in spreading B. hyodysenteriae between unconnected farms. RESULTS: Multilocus sequence typing (MLST) and Multiple Locus Variable number tandem repeat Analysis (MLVA) were used to characterize B. hyodysenteriae strains isolated from trucks. Before cleaning, 976 batches of finishing pigs transported by 174 trucks from 540 herds were sampled. After cleaning, 763 of the 976 batches were also sampled. Sixty-one of 976 and 4 of 763 environmental swabs collected from trucks before and after cleaning and disinfection operations, respectively, were positive for B. hyodysenteriae. The 65 isolates in this study originated from 48 farms. Trucks were classified into five categories based on the number of visited farms as follows: category 1: 1-5 farms, category 2: 6-10 farms, category 3: 11-15 farms, category 4: 16-20 farms, category 5: >21 farms. Although the largest number of vehicles examined belonged to category 1, the highest percentage of vehicles positive for B. hyodysenteriae was observed in categories 3, 4 and 5. Specifically, 90.9% of trucks belonging to category 5 were positive for B. hyodysenteriae, followed by categories 4 and 3 with 85.7% and 83.3%, respectively. The results of MLST and MLVA suggest that trucks transporting pigs from a high number of farms also play a critical role in spreading different B. hyodysenteriae genetic profiles. STVT 83-3, which seems to be the current dominant type in Italy, was identified in 56.25% of genotyped isolates. The genetic diversity of isolated strains from trucks was high, particularly, in truck categories 3, 4 and 5. This result confirmed that MLST and MLVA can support the study of epidemiological links between different B. hyodysenteriae farm strains. CONCLUSIONS: This study highlights the potential role of shipments in B. hyodysenteriae spread. Moreover, it emphasizes the importance of strict vehicle hygiene practices for biosecurity programmes.

An Integrated Analysis of Abattoir Lung Lesion Scores and Antimicrobial Use in Italian Heavy Pig Finishing Farms.

Respiratory diseases significantly affect intensive pig finishing farms, causing production losses and increased antimicrobial use (AMU). Lesion scoring at slaughter has been recognized as a beneficial practice to evaluate herd management. The integrated analysis of abattoir lesion scores and AMU data could improve decision-making by providing feedback to veterinarians and farmers on the effectiveness of antimicrobial treatments, thus rationalizing their use. This study compared lung and pleural lesion scores collected at Italian pig slaughterhouses with on-farm AMU, estimated through a treatment index per 100 days (TI100). Overall, 24,752 pig carcasses, belonging to 236 batches from 113 finishing farms, were inspected. Bronchopneumonia and chronic pleuritis were detected in 55% and 48% of the examined pigs, respectively. Antimicrobials were administered in 97% of the farms during the six months prior to slaughter (median TI100 = 5.2), notwithstanding compliance with the mandatory withdrawal period. EMA category B (critical) antimicrobials were administered in 15.2% of cases (median TI100 = 0.06). The lung score was not associated with the total AMU, but significant, positive associations were found with the past use of critical antimicrobials (p = 0.041) and macrolides (p = 0.044). This result highlights the potential of abattoir lung lesion monitoring to rationalize antimicrobial stewardship efforts, contributing to AMU reduction.

Effects of vaccination timing and target pathogens on performances and antimicrobial use in long-transported Charolais beef cattle from France to Italy - A retrospective study.

Antimicrobial use (AMU) in the livestock sector is a major driver of antimicrobial resistance. Italian beef industry strongly relies on the import of young cattle from France, which are commingled in sorting facilities before transportation to Italy. Both commingling and transportation are stressors for animals and lead to higher risk of bovine respiratory disease (BRD), which in turn increases the risk of AMU. This study aimed to investigate how the timing of first BRD vaccination and the different vaccination target pathogens affect AMU and performance of young Charolais beef cattle imported from France to Italy. Information on animal performance, antimicrobial treatments, and vaccinations was available for 60,726 Charolais cattle belonging to 1449 batches in 33 Italian specialised fattening farms between January 2016 and December 2021. Antimicrobial use was estimated using the treatment incidence 100 adapted for Italy (TI100it). A mixed linear model was used to quantify the effects of the vaccination and the time of first administration on slaughter age, carcase weight, and average daily carcase gain. Similarly, a generalised linear mixed model was used to analyse the TI100it. The vaccination programme was usually applied the first day after the animals' arrival to the Italian fattening farms. Most animals were vaccinated with a polyvalent vaccine against infectious bovine rhinotracheitis (IBR), bovine parainfluenza type 3 virus (PI-3), bovine viral diarrhoea virus type 1 and 2 (BVDV), and bovine respiratory syncytial virus (BRSV). The most used class of antimicrobials to treat BRD were the macrolides, followed by aminoglycosides, amphenicols, tetracyclines, aminopenicillins, and fluoroquinolones. Animals that got vaccinated against any of the considered BRD pathogens upon arrival had significantly lower TI100it, greater average daily carcase gain, and reached slaughter age earlier than animals that got vaccinated later. Animals that received the vaccination against BVDV had lower TI100it and greater average daily carcase gain, and animals that received the vaccination against BRSV were younger at slaughter than unvaccinated animals. The vaccination against Mannheimia haemolytica significantly decreased the slaughter age and increased the carcase weight and average daily carcase gain, and the vaccination against PI-3 and Histophilus somni significantly increased the slaughter age. Thus, even if the vaccination programme is essential to tackle BRD, this practice is questionable if applied at arrival to the Italian fattening farms and it is advisable that the vaccination programme is planned before the commingling procedure in France.

The Characterisation of Lactococcus garvieae Isolated in an Outbreak of Septicaemic Disease in Farmed Sea Bass (Dicentrarchus labrax, Linnaues 1758) in Italy.

In aquaculture, Lactococcus garvieae is a common fish pathogen that can cause significant economic losses in several fresh and saltwater species. Despite the extensive range of hosts, L. garvieae infection in sea bass (Dicentrarchus labrax) has rarely been reported. During the summer of 2023, an outbreak occurred in an inland farm in the Gulf of Follonica (Tuscany, Italy). Fish of various sizes were affected, showing apathy, inappetence, erratic swimming and eye lesions, while the mortality was low (2-3% per month). Anatomopathological examinations suggested a septicaemic infection characterised by melanosis, diffuse redness (skin and fins), paleness (gills and internal organs), haemorrhages and splenomegaly. Seventy swabs from the viscera of 14 subjects were collected and colonies similar to Streptococcus spp. grew from all the samples. Lactococcus garvieae was identified via the biochemical tests, API20STREP, MALDI-TOF, 16S rDNA and whole genome sequencing. Genetical characterisation revealed remarkable differences between this isolate and the strains previously isolated in Italian fish farms. Feed treatments with flumequine and erythromycin were ineffective. Considering the limited effects of antimicrobials, preventive measures, such as vaccination and biosecurity, should be implemented.

Regional-Scale Analysis of Antimicrobial Usage in Smallholder Cattle Herds (Aosta Valley, Italy): Why Surveillance Matters.

Optimising antimicrobial usage (AMU) in livestock is pivotal to counteract the emergence of antimicrobial resistance. We analysed AMU in more than 1000 cattle herds over 11 years (2008-2018) in the Aosta Valley (Italy), a region where 80% of farms house less than 50 cattle. Dairy cows accounted for over 95% of AMU. AMU was estimated using the defined daily dose animal for Italy (DDDAit) per biomass for the whole herd and a treatment incidence 100 (TI100) for cows. Average annual herd-level AMU was low, with 3.6 DDDAit/biomass (range: 3.2-4.0) and 1.2 TI100 in cows (range: 1.1-1.3). Third and fourth generation cephalosporins, which are critical for human medicine, represented almost 10% of usage, and intramammary antimicrobials accounted for over 60%. We detected significant downward temporal trends in total AMU, as well as a positive relationship with herd size. The magnitude of such effects was small, leaving scant room for further reduction. However, the frequent use of critical antimicrobials and intramammary products should be addressed, following the principles of prudent AMU. Our findings highlight the importance of monitoring AMU even in low-production, smallholding contexts where a low usage is expected, to identify any deficiencies and implement interventions for further AMU optimisation.

Infectious Agents Associated with Abortion Outbreaks in Italian Pig Farms from 2011 to 2021.

The present study retrospectively analyzed the infectious agents associated with 829 abortion outbreaks occurring from 2011 to 2021 in northern Italy. Foetuses were subjected to necropsies, and organ samples were analyzed by direct PCR to screen for six swine pathogens. In 42.0% of the examined outbreaks, at least one infectious agent was found. Porcine reproductive and respiratory syndrome virus (PRRSV) (24.9%) and porcine circovirus-2 (PCV2) (11.5%) were the most frequently detected among the known abortion-inducing pathogens. Chlamydia spp. (5.6%), porcine parvovirus (PPV) (4.0%), and Leptospira spp. (2.6%) were less common. Although its role in swine reproductive disorders is still unclear, PCV3 was detected in 19.6% of the cases. Coinfections were detected in 25.0% of positive outbreaks, and the most frequent coinfection was represented by PRRSV and PCV2 (32.2%), followed by PRRSV and PCV3 (23%). PCV2 prevalence showed a slight but consistent reduction during the study period, while PCV3 increased in frequency. Our data suggest an overall reduction in abortion outbreaks during the study period. PRRSV was confirmed as the main abortion agent detected in the examined area, while PCV2 prevalence showed a decline. Conversely, PCV3 detection has been increasing, supporting its potential role as an abortion agent. Our results highlight the importance of implementing a consistent and standardized sampling procedure, as well as a thorough diagnostic protocol, to reduce the incidence of inconclusive diagnoses.

Mortality in farmed European eel (Anguilla anguilla) in Italy due to Streptococcus iniae.

BACKGROUND: Streptococcal infections are one of the main causes of fish disease. During the last decade, Streptococcus iniae has become one of the most important aquatic pathogens worldwide, causing high losses in marine and freshwater finfish. Clinical signs in farmed fish include loss of appetite, lethargy and grouping at the bottom of the tank. Gross changes comprise darkening of the skin and haemorrhage at the basis of fins and opercula. To date, S. iniae has been isolated from several wild and farmed fish species but never in the European eel (Anguilla anguilla). In Europe, eel production from aquaculture is around 4500 tonnes and Italy is the third largest producer. This communication represents the first report of an outbreak of S. iniae infection in European eels. CASE PRESENTATION: The outbreak occurred at an eel farm in northern Italy between May 2021 and September 2021. The outbreak caused about 2% mortality per month, resulting in the loss of about 10% of the farmed fish. The diseased eels showed apathy, lethargy, inactivity and inappetence. In July 2021, three eels were necropsied. Necropsy revealed skin and branchial hyperaemia, a few skin ulcers, and diffuse peritoneal congestion with a few haemorrhagic-like spot lesions. Swab samples for bacteriology were taken from the kidneys, liver, spleen, and brain. Additionally, four eels were opened and swap samples as above were taken. All the investigated eels were found dead. Bacteriological examination revealed growth of Streptococcus spp. from all samples. Identification of S. iniae was done by biochemical characterization, the API20STREP microsystem, 16S rDNA sequencing, and MALDI-TOF. Antimicrobial therapy (oxytetracycline and erythromycin) was ineffective. CONCLUSIONS: This is the first report of S. iniae infection in the European eel. Although this may be an isolated outbreak, it is of concern due to the losses associated with this pathogen in fish worldwide and because the European eel is an endangered species. Due to the difficulties of controlling the disease with antimicrobials, it is advisable to plan other effective control measures, such as improving water quality and the environmental conditions, reducing fish density, improving biosecurity, and by using immunostimulants and, when possible, vaccines.

First Detection of mcr-9 in a Multidrug-Resistant Escherichia coli of Animal Origin in Italy Is Not Related to Colistin Usage on a Pig Farm.

The emergence of colistin resistance raises growing concerns because of its use as a last-resort antimicrobial for the treatment of severe gram-negative bacterial infections in humans. Plasmid-borne mobile colistin resistance genes (mcr) are particularly worrisome due to their high propensity to spread. An mcr-9-positive Escherichia coli was isolated from a piglet in Italy, representing the first isolation of this gene from an E. coli of animal origin in the country. Whole genome sequencing (WGS) revealed that mcr-9 was borne by an IncHI2 plasmid carrying several other resistance genes. The strain was indeed phenotypically resistant to six different antimicrobial classes, including 3rd and 4th generation cephalosporins. Despite the presence of mcr-9, the isolate was susceptible to colistin, probably because of a genetic background unfavourable to mcr-9 expression. The lack of colistin resistance, coupled with the fact that the farm of origin had not used colistin in years, suggests that mcr-9 in such a multidrug-resistant strain can be maintained thanks to the co-selection of neighbouring resistance genes, following usage of different antimicrobials. Our findings highlight how a comprehensive approach, integrating phenotypical testing, targeted PCR, WGS-based techniques, and information on antimicrobial usage is crucial to shed light on antimicrobial resistance.

Complex interplay between PRRSV-1 genetic diversity, coinfections and antimicrobial use influences performance parameters in post-weaning pigs.

Porcine reproductive and respiratory syndrome (PRRS) is one of the main diseases of pigs, leading to large economic losses in swine production worldwide. PRRSV high mutation rate and low cross-protection between strains make PRRS control challenging. Through a semi-longitudinal approach, we analysed the relationships among performance parameters, PRRSV-1 genetic diversity, coinfections and antimicrobial use (AMU) in pig nurseries. We collected data over the course of five years in five PRRS-positive nurseries belonging to an Italian multisite operation, for a total of 86 batches and over 200,000 weaners involved. The farm experienced a severe PRRS outbreak in the farrowing unit at the onset of the study, but despite adopting vaccination of all sows, batch-level losses in nurseries in the following years remained constantly high (mean±SE: 11.3 ± 0.5 %). Consistently with previous studies, our phylogenetic analysis of ORF 7 sequences highlighted the peculiarity of strains circulating in Italy. Greater genetic distances between the strain circulating in a weaners' batch and strains from the farrowing unit and the previous batch were associated with increased mortality (p < 0.0001). All the respiratory and enteric coinfections contributed to an increase in losses (all p < 0.026), with secondary infections by Streptococcus suis and enteric bacteria also inducing an increase in AMU (both p < 0.041). Our findings highlight that relying solely on sows' vaccination is insufficient to contain PRRS losses, and the implementation of rigorous biosecurity measures is pivotal to limit PRRSV circulation among pig flows and consequently minimise the risk of exposure to genetically diverse strains that would increase production costs.

PRRS Monitoring by Processing Fluids on Italian Swine Breeding Farms.

The porcine reproductive and respiratory syndrome (PRRS) control strategy within swine breeding farms is based on herd classification relative to PRRSV infection status. This study aims to assess the efficacy of a monitoring plan based on processing fluids (PFs) by comparing it with the classification of herds based on the analysis of blood serum. Twenty-five breeding herds were enrolled in the study, with at least five consecutive batches sampled from each herd. Each batch was tested for PRRSV by RT-PCR performed on (i) pre-weaning blood serum from 30 piglets and (ii) PFs from all the male piglets in the batch. PRRS categories following the Holtkamp classification were assigned based on the results of each testing protocol. The two protocols assigned the same category to 18 out of 25 herds: while they showed perfect agreement in identifying positive unstable and stable herds, we observed some discrepancy in discriminating between low- and high-prevalence classes within unstable herds. PFs are thus a reliable sample to assign PRRS categories in Italian breeding herds characterized by widespread PRRSV circulation. However, in case of an unstable epidemiological scenario, we recommend the adoption of an integrated monitoring strategy that combines blood sampling with PFs.

1H NMR Metabolomics on Pigs' Liver Exposed to Antibiotics Administration: An Explorative Study.

An untargeted Nuclear Magnetic Resonance (NMR) spectroscopy-based metabolomics approach was applied as a first attempt to explore the metabolome of pigs treated with antibiotics. The final goal was to investigate the possibility of discriminating between antibiotic-treated (TX group) and untreated pigs (CTRL group), with the further perspective of identifying the authentication tools for antibiotic-free pork supply chains. In particular, 41 samples of pig liver were subjected to a biphasic extraction to recover both the polar and the non-polar metabolites, and the 1H NMR spectroscopy analysis was performed on the two separate extracts. Unsupervised (principal component analysis) and supervised (orthogonal partial least squares discriminant analysis) multivariate statistical analysis of 1H NMR spectra data in the range 0-9 ppm provided metabolomic fingerprinting useful for the discrimination of pig livers based on the antibiotic treatment to which they were exposed. Moreover, within the signature patterns, significant discriminating metabolites were identified among carbohydrates, choline and derivatives, amino acids and some lipid-class molecules. The encouraging findings of this exploratory study showed the feasibility of the untargeted metabolomic approach as a novel strategy in the authentication framework of pork supply chains and open a new horizon for a more in-depth investigation.

A Preliminary Study on the Relationship between Gastric Lesions and Anti-Inflammatory Drug Usage in Heavy Pigs.

Gastric lesions in pigs cause welfare and economic losses. Their prevalence in heavy pigs reared for premium products (e.g., Parma ham) requires further investigation. Stress, nutrition, and farm management are known risk factors, but the effects of steroidal and non-steroidal anti-inflammatory drugs (NSAIDs) are largely unknown. The aim of this study was to evaluate the prevalence of gastric lesions in Italian heavy pigs and their possible association with the use of anti-inflammatory drugs. A total of 9371 pig stomachs from 76 farms were evaluated. Among these, 20.3% showed no lesions, while 30.7%, 42.1%, and 6.8% were scored 1, 2 and 3, respectively. A tendency for an inverse relationship with farm size emerged. The use of steroids and NSAIDs was estimated by calculating a treatment incidence per 1000 (TI1000) in a subset of 36 farms. At least one prescription for NSAIDs and/or steroids was found in 80.6% of the farms (55.6% used NSAIDs and 63.9% used steroids). Median TI1000 was 0.07 (range: 0-30.1) and 0.18 (range: 0-6.2) for NSAIDs and steroids, respectively. Gastric scores were positively associated with NSAID use, but not with steroid use. Although the role of these drugs in gastric lesions needs to be further clarified, these findings suggest a cautious use of non-selective NSAIDs.

EPI-Net One Health reporting guideline for antimicrobial consumption and resistance surveillance data: a Delphi approach.

Strategic and standardised approaches to analysis and reporting of surveillance data are essential to inform antimicrobial resistance (AMR) mitigation measures, including antibiotic policies. Targeted guidance on linking full-scale AMR and antimicrobial consumption (AMC)/antimicrobial residues (AR) surveillance data from the human, animal, and environmental sectors is currently needed. This paper describes the initiative whereby a multidisciplinary panel of experts (56 from 20 countries-52 high income, 4 upper middle or lower income), representing all three sectors, elaborated proposals for structuring and reporting full-scale AMR and AMC/AR surveillance data across the three sectors. An evidence-supported, modified Delphi approach was adopted to reach consensus among the experts for dissemination frequency, language, and overall structure of reporting; core elements and metrics for AMC/AR data; core elements and metrics for AMR data. The recommendations can support multisectoral national and regional plans on antimicrobials policy to reduce resistance rates applying a One Health approach.

Promoting Judicious Antimicrobial Use in Beef Production: The Role of Quarantine.

Judicious antimicrobial stewardship in livestock industry is needed to reduce the use of antimicrobials (AMU) and the associated risk of antimicrobial resistance. Biosecurity measures are acknowledged for their role against the spread of diseases and the importance in reducing AMU in different species. However, their effectiveness in beef production has been scarcely considered. The aim of this study was to investigate the effect of the quarantine strategy on AMU in beef cattle. A total of 1206 Charolaise animals in five farms were included in the trial. Roughly half of the animals followed the standard procedure of the fattening cycle (no-quarantine; NO-QUA group) and half followed a 30-day period of quarantine (QUA group) since their arrival. Performance and antimicrobial data were recorded and a treatment incidence 100 (TI100it) per animal was calculated. Penicillins was the most used class of antimicrobials. Differences between groups were significant for males only, with NO-QUA group having greater TI100it (3.76 vs. 3.24; p < 0.05) and lower body weight at slaughter (713.4 vs. 723.7 kg; p < 0.05) than QUA group. Results suggest that quarantine strategy can reduce AMU in males without compromising their performance, whereas further investigation is needed for females.

Wastewater-based surveillance in Italy leading to the first detection of mcr-10-positive Klebsiella quasipneumoniae.

Wastewater-based surveillance enabled the first detection of the mobile colistin resistance gene mcr-10 in Italy. This plasmid-borne resistance gene was found in strains of Klebsiella quasipneumoniae isolated from samples of human raw sewage collected over several months. Although the isolates were phenotypically susceptible to colistin, the emergence of mcr-10 is concerning due to the highly variable expression of the gene and the potential for horizontal transfer to other species. In addition, the strains also carried an extended-spectrum ß-lactamase gene and were phenotypically resistant to several beta-lactams. This study highlights the value of wastewater-based surveillance as an effective tool to monitor the emergence of antimicrobial resistance in strains circulating in the community and the environment.

Rapid displacement of SARS-CoV-2 variant Delta by Omicron revealed by allele-specific PCR in wastewater.

On November 26, 2021, the B.1.1.529 COVID-19 variant was classified as the Omicron variant of concern (VOC). Reports of higher transmissibility and potential immune evasion triggered flight bans and heightened health control measures across the world to stem its distribution. Wastewater-based surveillance has demonstrated to be a useful complement for clinical community-based tracking of SARS-CoV-2 variants. Using design principles of our previous assays that detect SARS-CoV-2 variants (Alpha and Delta), we developed an allele-specific RT-qPCR assay which simultaneously targets the stretch of mutations from Q493R to Q498R for quantitative detection of the Omicron variant in wastewater. We report their validation against 10-month longitudinal samples from the influent of a wastewater treatment plant in Italy. SARS-CoV-2 RNA concentrations and variant frequencies in wastewater determined using these variant assays agree with clinical cases, revealing rapid displacement of the Delta variant by the Omicron variant within three weeks. These variant trends, when mapped against vaccination rates, support clinical studies that found the rapid emergence of SARS-CoV-2 Omicron variant being associated with an infection advantage over Delta in vaccinated persons. These data reinforce the versatility, utility and accuracy of these open-sourced methods using allele-specific RT-qPCR for tracking the dynamics of variant displacement in communities through wastewater for informed public health responses.

Occurrence of Salmonella Typhimurium and its monophasic variant (4, [5],12:i:-) in healthy and clinically ill pigs in northern Italy.

BACKGROUND: The serovar Typhimurium (4, [5],12:i:1,2), is the most frequently isolated serovar in case of salmonellosis in pigs in Europe and its monophasic variant (4, [5],12:i:-) has been increasingly responsible for Salmonella outbreaks in humans. A total of 25,215 samples were collected, during the years 2002-2017, from 1359 pig farms located in Northern Italy. Samples were collected from different material sources including fecal samples, rectal swabs, gut content and different organs. RESULTS: Salmonella was isolated in 15.80% of samples and, among the isolates, 733 were typed as Salmonella Typhimurium (ST) or its monophasic variant (MST). Over time, there was an increase of isolation of MST which outnumbered ST. Most of the strains were isolated in animals during the weaning stage and the growing - fattening period whereas the clinical cases were mainly present in young pigs after weaning. CONCLUSIONS: This study confirms the presence of ST and MST in pig farms although, considering the total of isolated serotypes, with lower percentages than previously reported. In the last few years, ST has increasingly been replaced by MST suggesting that MST has a competitive advantage over ST, probably due to its different antigenicity and pathogenicity which renders the infection stealthier to recognize and control.

ESBL/AmpC-Producing Escherichia coli in Wild Boar: Epidemiology and Risk Factors.

The complex health problem of antimicrobial resistance (AMR) involves many host species, numerous bacteria and several routes of transmission. Extended-spectrum ß-lactamase and AmpC (ESBL/AmpC)-producing Escherichia coli are among the most important strains. Moreover, wildlife hosts are of interest as they are likely antibiotics free and are assumed as environmental indicators of AMR contamination. Particularly, wild boar (Sus scrofa) deserves attention because of its increased population densities, with consequent health risks at the wildlife-domestic-human interface, and the limited data available on AMR. Here, 1504 wild boar fecal samples were microbiologically and molecularly analyzed to investigate ESBL/AmpC-producing E. coli and, through generalized linear models, the effects of host-related factors and of human population density on their spread. A prevalence of 15.96% of ESBL/AmpC-producing E. coli, supported by blaCTX-M (12.3%), blaTEM (6.98%), blaCMY (0.86%) and blaSHV (0.47%) gene detection, emerged. Young animals were more colonized by ESBL/AmpC strains than older subjects, as observed in domestic animals. Increased human population density leads to increased blaTEM prevalence in wild boar, suggesting that spatial overlap may favor this transmission. Our results show a high level of AMR contamination in the study area that should be further investigated. However, a role of wild boar as a maintenance host of AMR strains emerged.