Third-generation cephalosporin (3GC) resistance and its association with Extra-intestinal pathogenic Escherichia coli (ExPEC). Focus on broiler carcasses.

The worldwide spread of Extra-intestinal Pathogenic Escherichia coli (ExPEC), together with the antimicrobial resistance linked with extended-spectrum ß-lactamases (ESBLs) and plasmid-mediated AmpC ß-lactamases (pAmpCs) are pressing threats for public health. This study aimed to investigate the presence of ExPEC genes in third-generation cephalosporin (3 GC)-resistant E. coli and to study their distribution in broiler carcasses at the slaughterhouse after the chilling process. To this purpose, isolates from a collection of 3 GC-resistant E. coli from carcasses of broilers originating from twelve broiler farms and three production chains were investigated. Several multivariate statistical approaches were adopted to elucidate the relationships among features. Phylogroup F was predominant in all broiler batches and was mainly associated with blaTEM and ESBL genes but less correlated to ExPEC genes. Another remarkable finding was the predominance of ExPEC strains assigned to uncommon phylogroups, such as B2, D, E and Clade I, commonly found into the environment. This study represents a first step for a comprehensive characterization of ExPEC genes harboured by 3 GC-resistant E. coli. These findings may be valuable for the identification of potential risks associated to broiler carcasses as source of uncommon E. coli phylogroups.

A review on the current situation and challenges of colistin resistance in poultry production.

Colistin has been re-assessed as a critically important antimicrobial in humans due to its efficacy against multi-drug resistant Gram-negative bacteria, in particular P. aeruginosa, A. baumannii, and K. pneumoniae. The recent discovery of mobile colistin resistance (mcr) determinants in humans and animals has brought concerns regarding the future of this antimicrobial. In this paper, we aim to highlight the current challenges with colistin resistant bacteria and to summarize reliable global data on colistin resistance in poultry production. In addition, we present and compare data from a screening for colistin resistance carried out on a collection of clinical Escherichia coli isolated from poultry in Italy. In Europe, resistance rates for Salmonella and E. coli are in general low with sporadic incidence of high colistin resistance levels. Absence of resistance or very low rates have been recorded in countries where colistin is either not employed (e.g. Norway) or used in minimal amounts (e.g. Denmark) in food-producing animals. In large poultry meat producing countries, such as China and Brazil, the widespread use of colistin has resulted in the dissemination of resistance determinants in diverse bacterial species. Worryingly, these bacteria are often co-resistant to other critically important antimicrobials, such as extended-spectrum cephalosporins. The data gap for many countries and for zoonotic bacteria, the role of the "phantom resistome" and the circulation of mcr-carriers expressing resistance phenotypes close or below the current ECOFF values, should be considered in future investigations. The importance of poultry as a cheap protein source and the global effort to mitigate colistin resistance and preserve this essential antimicrobial require a thorough re-assessment of colistin use in poultry.

Multilocus sequence typing of Campylobacter jejuni and Campylobacter coli to identify potential sources of colonization in commercial turkey farms.

Poultry are the main reservoir for thermophilic Campylobacter spp., which is the most common causative agent of human bacterial gastroenteritis. The epidemiology of Campylobacter in poultry, particularly in turkeys, is not completely understood. This study aimed at identifying potential sources and transmission routes of thermophilic Campylobacter spp. in commercial turkey farms. C. jejuni and C. coli isolates from breeders (n = 29, 20 C. jejuni and 9 C. coli) and their progeny (n = 51, 18 C. jejuni and 33 C. coli) reared in two different farms for three sequential production cycles were analysed by multilocus sequence typing (MLST). Strains (n = 88, 42 C. jejuni and 46 C. coli) isolated from environmental (i.e. anteroom and in-house overshoes), water (i.e. drinkers and water line), and pest (i.e. flies, Alphitobius diaperinus, and mice) sources were also examined. MLST of C. jejuni and C. coli isolates resulted in 13 and 12 different sequence types (STs) belonging to six and one previously-described clonal complexes (CCs), respectively. Three novel STs were identified. Genetic similarities were detected between isolates from fattening turkeys and the considered environmental, water, and pest sources, and with the breeders to a lesser extent. Source attribution analysis estimated that environmental and water sources accounted for most (∼75%) of fattening turkey isolates and were therefore identified as the most likely sources of flock colonization, followed by pests (∼20%) and breeders (∼5%). These sources may thus be targeted by control measures to mitigate the risk of Campylobacter colonization in commercial turkeys. RESEARCH HIGHLIGHTS High occurrence of C. jejuni and C. coli in commercial turkey flocks. High genetic diversity of C. jejuni and C. coli in commercial turkey flocks. Horizontal transmission responsible for Campylobacter colonization of commercial turkey flocks. Environmental and water sources involved in Campylobacter colonization of commercial turkey flocks. Strategies for prevention and control of Campylobacter colonization of commercial turkey flocks are needed.

Versatile nano-platform for tailored immuno-magnetic carriers.

Custom immuno-magnetic devices are desirable tools for biomedical and biotechnological applications. Herein, surface active maghemite nanoparticles (SAMNs) are proposed as a versatile platform for developing tailored immuno-magnetic nano-carriers by simple wet reactions. Two examples for conjugating native and biotinylated antibodies were presented along with their successful applications in the recognition of specific foodborne pathogens. Nanoparticles were functionalized with rhodamine B isothiocyanate (RITC), leading to a fluorescent nano-conjugate, and used for binding anti-Campylobacter fetus antibodies (SAMN@RITC@Anti-Cf). The microorganism was selectively captured in the presence of two other Campylobacter species (C. jejuni and C. coli), as verified by PCR. Alternatively, SAMNs were modified with avidin, forming a biotin-specific magnetic nano-carrier and used for the immobilization of biotinylated anti-Listeria monocytogenes antibodies (SAMN@avidin@Anti-Lm). This immuno-magnetic carrier was integrated in piezoelectric quartz crystal microbalance (QCM) sensor for the detection of L. monocytogenes in milk, showing a detection limit of 3 bacterial cells. The present work presents a new category of customized immuno-magnetic nano-carriers as a competitive option for suiting specific applications. Graphical abstract ᅟ.

A first molecular characterization of Listeria monocytogenes isolates circulating in humans from 2009 to 2014 in the Italian Veneto region.

Listeriosis is a disease usually associated with the consumption of low-processed ready-to-eat food products contaminated by Listeria monocytogenes. In Italy, listeriosis has an incidence of 0.19-0.27 cases per 100000 persons. Since detailed information concerning the molecular characterization of listeriosis in the Italian Veneto region is currently lacking, we analyzed 36 L. monocytogenes clinical isolates collected between 2009 and 2014. Results show that the serotype 1/2a was the most represented among the tested samples. No antimicrobial resistance was detected in selected isolates representing the main pulsotypes.

Campylobacter geochelonis sp. nov. isolated from the western Hermann's tortoise (Testudo hermanni hermanni).

During a screening study to determine the presence of species of the genus Campylobacter in reptiles, three putative strains (RC7, RC11 and RC20T) were isolated from different individuals of the western Hermann's tortoise (Testudo hermanni hermanni). Initially, these isolates were characterized as representing Campylobacterfetus subsp. fetus by multiplex PCR and partial 16S rRNA gene sequence analysis. Further whole- genome characterization revealed considerable differences compared to other Campylobacter species. A polyphasic study was then undertaken to determine the exact taxonomic position of the isolates. The three strains were characterized by conventional phenotypic tests and whole genome sequencing. We generated robust phylogenies that showed a distinct clade containing only these strains using the 16S rRNA and atpA genes and a set of 40 universal proteins. Our phylogenetic analysis demonstrates their designation as representing a novel species and this was further confirmed using whole- genome average nucleotide identity within the genus Campylobacter (~80 %). Compared to most Campylobacter species, these strains hydrolysed hippurate, and grew well at 25 °C but not at 42 °C. Phenotypic and genetic analyses demonstrate that the three Campylobacter strains isolated from the western Hermann's tortoise represent a novel species within the genus Campylobacter, for which the name Campylobactergeochelonis sp. nov. is proposed, with RC20T (=DSM 102159T=LMG 29375T) as the type strain.

A novel real-time PCR assay for quantitative detection of Campylobacter fetus based on ribosomal sequences.

BACKGROUND: Campylobacter fetus is a pathogen of major concern for animal and human health. The species shows a great intraspecific variation, with three subspecies: C. fetus subsp. fetus, C. fetus subsp. venerealis, and C. fetus subsp. testudinum. Campylobacter fetus fetus affects a broad range of hosts and induces abortion in sheep and cows. Campylobacter fetus venerealis is restricted to cattle and causes the endemic disease bovine genital campylobacteriosis, which triggers reproductive problems and is responsible for major economic losses. Campylobacter fetus testudinum has been proposed recently based on genetically divergent strains isolated from reptiles and humans. Both C. fetus fetus and C. fetus testudinum are opportunistic pathogens for immune-compromised humans. Biochemical tests remain as the gold standard for identifying C. fetus but the fastidious growing requirements and the lack of reliability and reproducibility of some biochemical tests motivated the development of molecular diagnostic tools. These methods have been successfully tested on bovine isolates but fail to detect some genetically divergent strains isolated from other hosts. The aim of the present study was to develop a highly specific molecular assay to identify and quantify C. fetus strains. RESULTS: We developed a highly sensitive real-time PCR assay that targets a unique region of the 16S rRNA gene. This assay successfully detected all C. fetus strains, including those that were negative for the cstA gene-based assay used as a standard for molecular C. fetus identification. The assay showed high specificity and absence of cross-reactivity with other bacterial species. The analytical testing of the assay was determined using a standard curve. The assay demonstrated a wide dynamic range between 102 and 107 genome copies per reaction, and a good reproducibility with small intra- and inter-assay variability. CONCLUSIONS: The possibility to characterize samples in a rapid, sensitive and reproducible way makes this assay a good option to establish a new standard in molecular identification and quantification of C. fetus species.

Class 1 and class 2 integrons in avian pathogenic Escherichia coli from poultry in Italy.

The aim of this study was to investigate the occurrence of class 1 and 2 integrons in avian pathogenic Escherichia coli (APEC) from poultry in northern Italy. Strains were tested for phenotypic resistance to aminoglycosides and sulphonamides, and the association between the presence of integrons and the resistance to these antimicrobials was evaluated. A total of 299 isolates (158 from turkeys, 110 from broilers, and 31 from layer hens) were collected from 200 industrial farms. Antimicrobial susceptibility test by the disk diffusion method was performed in accordance with the Clinical and Laboratory Standards Institute (CLSI) guidelines. All strains were screened for the presence of class 1 and 2 integrons by PCR and sequencing. About 55% of APEC contained integrons (class 1, 49.8%; class 2, 10.4%). Different variants of the aadA (5 variants) and the dfrA (4 variants) genes, encoding for streptomycin and trimethoprim resistance respectively, were detected in integron-positive isolates. Less common gene cassettes, such as sat, estX, and orfF, were also identified. Fifteen and 4 gene cassette arrays were found among class 1 and 2 integrons, respectively. High levels of resistance were observed for triple sulphonamides (79.3%), streptomycin (67.2%), and sulfamethoxazole combined with trimethoprim (62.2%), whereas resistance against gentamycin (16.7%), kanamycin (14.7%), and apramycin 3.0%) was low. Integron positivity was significantly higher in isolates phenotypically resistant to aminoglycosides (63.6% vs. 37.8%, P<0.001) and sulfonamides (64.1% vs. 21.1%, P<0.001) than in susceptible ones. Integron-borne aminoglycoside and sulfonamide resistance in APEC represents a concern for the poultry industry in Italy, since they are among the most commonly used antimicrobials in poultry therapy.

Multilocus sequence typing of Campylobacter jejuni and Campylobacter coli from humans and chickens in North-Eastern Italy.

This paper reports the multilocus sequence typing (MLST) of 57 C. jejuni and C. coli isolates from humans and chickens in Italy and the identification of 17 new sequence types (STs). A high genetic diversity was detected among C. jejuni/C. coli and human/chicken isolates, with a predominance of clonal complexes CC21 and CC828. Although human STs were not the same as those found in chickens, 3 CCs overlapped between human and chicken isolates. Genotyping of Campylobacter strains by MLST should be encouraged in order to implement surveillance and con- trol of infection in humans and in animal reservoirs in Italy.

A systematic review and meta-analysis on the efficacy of vaccination against colibacillosis in broiler production.

Colibacillosis, a disease caused by Escherichia coli in broiler chickens has serious implications on food safety, security, and economic sustainability. Antibiotics are required for treating the disease, while vaccination and biosecurity are used for its prevention. This systematic review and meta-analysis, conducted under the COST Action CA18217-European Network for Optimization of Veterinary Antimicrobial Treatment (ENOVAT), aimed to assess the efficacy of E. coli vaccination in broiler production and provide evidence-based recommendations. A comprehensive search of bibliographic databases, including, PubMed, CAB Abstracts, Web of Science and Agricola, yielded 2,722 articles. Following a defined protocol, 39 studies were selected for data extraction. Most of the studies were experimental infection trials, with only three field studies identified, underscoring the need for more field-based research. The selected studies reported various types of vaccines, including killed (n = 5), subunit (n = 8), outer membrane vesicles/protein-based (n = 4), live/live-attenuated (n = 16), and CpG oligodeoxynucleotides (ODN) (n = 6) vaccines. The risk of bias assessment revealed that a significant proportion of studies reporting mortality (92.3%) or feed conversion ratio (94.8%) as outcomes, had "unclear" regarding bias. The meta-analysis, focused on live-attenuated and CpG ODN vaccines, demonstrated a significant trend favoring both vaccination types in reducing mortality. However, the review also highlighted the challenges in reproducing colibacillosis in experimental setups, due to considerable variation in challenge models involving different routes of infection, predisposing factors, and challenge doses. This highlights the need for standardizing the challenge model to facilitate comparisons between studies and ensure consistent evaluation of vaccine candidates. While progress has been made in the development of E. coli vaccines for broilers, further research is needed to address concerns such as limited heterologous protection, practicability for application, evaluation of efficacy in field conditions and adoption of novel approaches.

Supporting Measures to Improve Biosecurity within Italian Poultry Production.

This paper describes the selection and validation of supporting measures (SMs) aimed at enhancing biosecurity compliance within Italian poultry farms. A tailored methodology, based on a stakeholders' survey involving farmers and advisors, included a virtual farm tour, group discussion, and farmer coaching. Virtual farm tours and group discussions were delivered during two meetings targeting meat and egg production stakeholders, separately. Coaching was validated in 26 pilot farms (PFs) by assessing farmers' attitudes towards change (i.e., ADKAR®) and farms' biosecurity score (i.e., Biocheck.UgentTM) before and after a minimum six-month period. A total of 20 out of 26 farmers agreed to implement at least one action plan (AP). Full implementation of the agreed APs was observed in ten farms, while others only partially implemented (n = 7) or did not implement (n = 3) the improvement. Most APs focused on enhancing house hygiene locks (n = 7), followed by bacterial auto-control after cleaning and disinfection (n = 4). Scoring tools indicated minimal or no variations in farmers' attitudes towards change and farm biosecurity. Virtual farm tours and group discussions were found to be effective in fostering interaction and facilitating the exchange of experiences and knowledge among farmers and stakeholders of poultry production. Coaching indicated that farmers might prefer implementing minor changes possibly influenced by time and cost constraints associated with structural interventions. These limitations could have also impacted the scores of the farmer/farm. The findings of this study provide a foundation for further application of SMs to improve biosecurity in Italian poultry farms.

Factors Affecting Poultry Producers' Attitudes towards Biosecurity.

Poultry producers' attitudes towards biosecurity practices were assessed by using the ADKAR® (Awareness, Desire, Knowledge, Ability, and Reinforcement) behavioral change model. Conventional poultry producers (n = 155) from different production types including broilers (n = 35), layers (n = 22), breeders (n = 24), turkeys (n = 19), ducks (n = 23), free-range broilers (n = 11), free-range layers (n = 11), and hatcheries (n = 10) from seven European countries were scored for each ADKAR element (1 = total absence to 5 = perfect fulfilment). Each country performed selected interventions (e.g., coaching, participatory meetings, etc.) to improve biosecurity compliance. After the interventions, significant change was observed in three of the four attitude elements. The overall mean scores (x¯ ± SD) obtained during the initial assessment (n = 130) were 4.2 ± 0.6 for Awareness, 4.1 ± 0.7 for Desire, 3.8 ± 0.8 for Knowledge, and 4.0 ± 0.7 for Ability, whereas after intervention, the scores were A = 4.3 ± 0.6, D = 4.2 ± 0.7, K = 4.1 ± 0.7, and Ab = 4.1 ± 0.7. The Reinforcement component was only evaluated after the change and obtained a score of 3.7 ± 0.7 on average. Identifying the elements influencing poultry producers and their behavior related to farm management decisions was useful in guiding our educational interventions to effectively change their behavior.

Biosecurity implementation on large-scale poultry farms in Europe: A qualitative interview study with farmers.

Biosecurity is an essential tool for rearing healthy animals. Biosecurity measures (BMs) are well known in poultry production, but it is difficult to assess actual implementation on farms. The aims of this qualitative study were (1) to provide an overview of biosecurity implementation according to poultry farmers in Europe; and (2) to better understand the reported reasons and potential obstacles for not implementing the measures. In seven European Union Member States, 192 farmers (118 under contract with a company and 68 independents) working in seven different categories of poultry production were interviewed on 62 BMs to determine the frequency of implementation and the reasons for non-implementation. Most of the replies (n = 7791) concerning BM implementation were reported by the farmers as "always" implemented (81%), statistically higher for breeders (87%) and layers (82%) and lower for independent farms versus farms under contract with a company (79.5% and 82.5%, respectively). Regardless the poultry production category, the most frequently implemented BMs declared by the farmers were daily surveillance of birds, rodent control and feed storage protection. Standard hygiene practices were also mentioned as high-implementation measures for most production categories, with some deficiencies, such as rendering tank disinfection after each collection and, for meat poultry, disinfection of the feed silo and bacterial control of house cleaning and disinfection between each cycle. The entry of vehicles and individuals onto poultry farms, especially during critical points of eggs collection for breeders and layers, as well as the presence of other animals, such as the "all in/all out" practice, particularly in layers and ducks, were also reported as the least commonly practiced measures. The main reasons for not implementing the measures (n = 1683 replies) were low awareness and poor knowledge of the expected benefits of biosecurity ("no known advantages" 14%, and "not useful" 12%), the lack of training ("not enough training" 5% and "advice" 7%), lack of time (19%), and financial aspects (17%). Despite the good overall biosecurity mentioned by the farmers, these findings highlight certain deficiencies, suggesting room for improvement and the need for targeted and tailored support of poultry farmers in Europe.

Development and use of Biocheck.UGent™ scoring system to quantify biosecurity in conventional indoor (turkey, duck, breeder) and free-range (layer and broiler) poultry farms.

To assess and enhance the application of biosecurity measures in poultry farming, an objective measurement tool (Biocheck.UGent™) was already available for broiler and layer. This study describes the development, validation and application of a risk-based weighted scoring tool for breeder, turkey, duck, free-range layer and free-range broiler production. In collaboration with an expert panel (n= 38), five different questionnaires were developed, following the format of the existing Biocheck.UGent scoring tools. Weights were attributed to external (7-9 subcategories) and internal (3-4 subcategories) biosecurity categories, as well as to the corresponding individual questions within each subcategory. The biosecurity measures were prioritized and weighed based on their relative importance in preventing disease transmission. Upon completion of the questionnaire, and upload of all answers to the Biocheck.UGent website, the algorithm generates a biosecurity score varying between ''0'' which equals the total absence of any biosecurity measure up to ''100'' which refers to full application of all biosecurity measures. The final scoring systems are available online (https://biocheckgent.com/en) for free and have been used to assess biosecurity in 70 breeders, 100 turkeys, 23 ducks, 16 free-range broilers, and 15 free-range layer farms originating from 12 countries so far. On average, the overall biosecurity score (mean ± std. dev) was 78 ± 7 % for breeders, 73 ± 11 % for turkeys, 71 ± 8 % for ducks, 73 ± 8 % for free-range layers and 70 ± 13 % for free-range broilers. There were significant differences (p < 0.05) in biosecurity (both at the overall and subcategory levels) across different poultry types. The overall farm biosecurity score for breeders was significantly higher than that for turkey (p <.001) and duck production (p = 0.001). External biosecurity levels were highest in breeders in comparison to turkeys (p < 0.001), ducks (p = 0.008) and broiler free-range (p = 0.005). There was a notable difference in internal biosecurity levels between duck and turkey (p = 0.041) production as well. The study contributed to the poultry biosecurity database which allows benchmarking of the biosecurity levels of the users' farm results to national or international averages, indicating room for improvement and aiding to motivate stakeholders to enhance their biosecurity levels.

Biofilm Formation Ability of ESBL/pAmpC-Producing Escherichia coli Isolated from the Broiler Production Pyramid.

Escherichia coli able to produce extended spectrum ß-lactamases (ESBLs) and plasmid-mediated AmpC ß-lactamases (pAmpCs) represents a serious threat to public health, since these genes confer resistance to critically important antimicrobials (i.e., third generation cephalosporins) and can be transferred to non-resistant bacteria via plasmids. E. coli are known to be able to form a biofilm, which represents a favorable environment for the exchange of resistance determinants. Here, we assessed the ability of 102 ESBL/pAmpC-producing E. coli isolated from the broiler production pyramid to form a biofilm and to identify genetic factors involved in biofilm formation. All but one of the ESBL/pAmpC-producing E. coli were able to form a biofilm, and this represents a great concern to public health. E. coli belonging to phylogroups D, E, and F, as well as strains harboring the blaCTX-M-type gene, seem to be associated with an increased biofilm capability (p < 0.05). Furthermore, virulence genes involved in adherence and invasion (i.e., csgBAC, csgDEFG, matABCDEF, and sfaX) seem to enhance biofilm formation in E. coli. Efforts should be made to reduce the presence of ESBL/pAmpC- and biofilm-producing E. coli in the broiler production pyramid and, therefore, the risk of dissemination of resistant bacteria and genes.

Stakeholders' Perceptions of Biosecurity Implementation in Italian Poultry Farms.

The level of implementation of biosecurity measures (BMs), the reasons for not implementing BMs and the effectiveness of BMs were assessed according to the perceptions of stakeholders (i.e., farmers and advisors) in Italian poultry farms. For this purpose, data were collected using a questionnaire administered to advisors (n = 37) and farmers (n = 30) of conventional broiler (n = 13) and layer (n = 13), free-range broiler (n = 8) and layer (n = 10), turkey (n = 13), duck (n = 3) and breeder (n = 7) farms between April and September 2021. The frequency of the implementation of BMs was 66.97% and 81.14% according to the answers provided by the advisors and farmers, respectively, with the breeder sector showing the highest level of implementation (85.71%). "Not knowing advantages" (21.49% for advisors) and "other/specific reasons" (21.49% for advisors and 38.32% for farmers) were the most common answers regarding the lack of implementation of BMs for all poultry sectors. Only 31.09% of farmers acknowledged the effectiveness of not-implemented BMs in contrast to 61.02% of advisors, with the layers' stakeholders being the most aware. The findings of this study may be useful for identifying failures in biosecurity and failures to develop intervention strategies to fulfil the biosecurity gaps still present in Italian poultry farms.

Association between ability to form biofilm and virulence factors of poultry extra-intestinal Campylobacter jejuni and Campylobacter coli.

Campylobacter species are known to be able to produce biofilm, which represents an ideal protective environment for the maintenance of such fragile bacteria. Since the genetic mechanisms promoting biofilm formation are still poorly understood, in this study we assessed the ability of C. jejuni (n = 7) and C. coli (n = 3) strains isolated from diseased poultry, and previously characterized by whole genome sequencing, to form biofilm. The in vitro analyses were carried out by using a microtiter based protocol including biofilm culturing and fixation, staining with crystal violet, and measurement of the optical density (OD570). The ability to form biofilm was categorized into four classes (no, weak, moderate, and strong producers). Potential correlations between OD570 and the presence/absence of virulence determinants were examined. The C. jejuni were classified as no (n = 3), weak (n = 2), and moderate (n = 2) biofilm producers; however, all possessed genes involved in chemotaxis, adhesion, and invasion to the host cells. No genes present exclusively in biofilm producers or in non-biofilm producers were identified. All C. coli were classified as weak producers and showed a similar set of virulence genes between each other. A trend of increased mean OD570 was observed in the presence of flaA and maf7 genes. No association between biofilm production classes and the explanatory variables considered was observed. The results of this study suggest that further investigations are needed to better identify and characterize the genetic determinants involved in extra-intestinal Campylobacter biofilm formation.

SARS-CoV-2 and Companion Animals: Sources of Information and Communication Campaign during the COVID-19 Pandemic in Italy.

This study analyzed data on the sources and the level of Italians' awareness on the risk of infection by SARS-CoV-2 at the human-animal interface. Data were collected through a survey-type investigation on a representative sample of the Italian population. Forty-five percent of the interviewees were aware that companion animals could be infected by SARS-CoV-2. However, 29.8% were familiar with preventive measures to adopt to avoid viral transmission between infected humans and companion animals, and only 20.7% knew which companion animals could be at risk of infection. Higher awareness regarding the risk of SARS-CoV-2 transmission between animals and humans (51.7%) and the measures to prevent it (33.3%) was detected among companion animals' owners. Notably, 40.4% of interviewees were not informed at all. Television broadcasts (26.4%) represented the main source of information, while only 3.5% of the interviewees relied on veterinarians, of which 31.9% considered this source of information as the most trustworthy. Overall, 72.4% of Italians recognized that the communication campaign on COVID-19 and companion animals was inadequate. This survey highlights the need for increasing the public awareness of the risk of companion animals being infected with SARS-CoV-2 and the involvement of professionals in the public communication on zoonoses.

Monitoring biosecurity in poultry production: an overview of databases reporting biosecurity compliance from seven European countries.

Compliance with required on-farm biosecurity practices reduces the risk of contamination and spread of zoonotic and economically important diseases. With repeating avian influenza epidemics in the poultry industry, the need to monitor and improve the overall level of biosecurity is increasing. In practice, biosecurity compliance is assessed by various actors (e.g., academic, private and public institutions), and the results of such assessments may be recorded and gathered in databases which are seldom shared or thoroughly analyzed. This study aimed to provide an inventory of databases related to the assessment of biosecurity in poultry farms in seven major poultry-producing European countries to highlight challenges and opportunities associated with biosecurity data collection, sharing, and use. The institutions in charge of these databases were contacted and interviewed using a structured questionnaire to gather information on the main characteristics of the databases and the context of their implementation. A total of 20 databases were identified, covering the gamut of poultry species and production types. Most databases were linked to veterinary health authorities or academia, and to a lesser extent interbranch organizations. Depending on the institutions in charge, the databases serve various purposes, from providing advice to enforcing regulations. The quality of the biosecurity data collected is believed to be quite reliable, as biosecurity is mostly assessed by trained farm advisors or official veterinarians and during a farm visit. Some of the databases are difficult to analyze and/or do not offer information concerning which biosecurity measures are most or least respected. Moreover, some key biosecurity practices are sometimes absent from certain databases. Although the databases serve a variety of purposes and cover different production types, each with specific biosecurity features, their analysis should help to improve the surveillance of biosecurity in the poultry sector and provide evidence on the benefits of biosecurity.

A longitudinal study on thermophilic Campylobacter spp. in commercial turkey flocks in northern Italy: occurrence and genetic diversity.

Poultry are recognized as a main reservoir of thermophilic campylobacters, but few studies have been carried out on commercial meat turkeys. This study was aimed at assessing the occurrence of thermophilic Campylobacter spp., their genetic diversity, and the trend of the infection during the whole production cycle of three turkey flocks from different farms in Northern Italy. Flocks were monitored from the time of housing 1-day-old poults to slaughter time by collecting samples (meconium and cloacal swabs) at weekly intervals up to the recovery of Campylobacter spp. and then twice a month. A conventional culture method and a multiplex PCR assay were used for Campylobacter detection and identification. A subset of isolates was genetically characterized by random amplified polymorphic DNA-PCR (RAPD-PCR) and flagellin gene A short variable region (flaA-SVR) sequencing. Although at different times, all flocks became colonized by Campylobacter jejuni or Campylobacter coli (or both) that persisted throughout the entire production cycle. Overall, nine RAPD types and 14 flaA-SVR types were detected with differences in their distribution among flocks and sampling times. Moreover, changes in the Campylobacter genotypes colonizing turkeys were observed over time within each flock. These findings suggest that Italian commercial turkeys might be widely colonized by different genotypes of C. jejuni and C. coli and also suggest that differences in the distribution and epidemiologic dynamics of these microorganisms might occur among flocks.