First Report of Caseous Lymphadenitis by Corynebacterium pseudotubercolosis and Pulmonary Verminosis in a Roe Deer (Capreolus capreolus Linnaeus, 1758) in Italy.

Caseous lymphadenitis is a chronic debilitating disease typical of small ruminants, but it is also noted in several other domestic and wild species. In this report, we present the first documented case in Italy of pseudotuberculosis in a roe deer (Capreolus capreolus, Linnaeus 1758) found dead in the mountains of Forlì-Cesena province, Emilia Romagna region. The carcass underwent necropsy according to standard protocols, revealing generalized lymphadenopathy and severe apostematous pneumonia with multifocal and encapsulated abscesses. Corynebacterium pseudotuberculosis was isolated from the lung parenchyma, lymph nodes and abscesses. Additionally, severe parasitic bronchopneumonia of the caudal lobes and gastrointestinal strongyle infestation were detected. To our knowledge, this is the first documented case of CLA referable to C. pseudotubercolosis in a roe deer in Italy.

Assessment of aflatoxin M1 enrichment factor in cheese produced with naturally contaminated milk.

Aflatoxin M1 (AFM1) is a well-known carcinogenic compound that may contaminate milk and dairy products. Thus, with the regulation 1881/2006, the European Union established a concentration limit for AFM1 in milk and insisted on the importance of defining enrichment factors (EFs) for cheese. In 2019, the Italian Ministry of Health proposed four different EFs based on cheese's moisture content on a fat-free basis (MMFB) for bovine dairy products. This study aimed to define the EFs of cheese with different MFFB. The milk used for cheesemaking was naturally contaminated with different AFM1 concentrations. Results showed that all the EF average values from this study were lower than those of the Italian Ministry of Health. Hence, the current EFs might need to be reconsidered for a better categorization of AFM1 risk in cheese.

Screening of antibiotic residues in raw bovine milk in Lombardy, Italy: Microbial growth inhibition assay and LC-HRMS technique integration for an accurate monitoring.

Antibiotic residues in food of animal origin is a great concern for public health worldwide in terms of antibiotic resistance development, potential allergic reactions and disruption of intestinal flora equilibrium. In this study the presence of antibiotic residues in raw bovine milk samples collected from farms located in Lombardy region in Italy from 2018 to 2022 was assessed in the context of the national milk quality payment system. Samples were screened with microbiological growth inhibition test Delvotest ® SP NT and a very low positivity rate ranging from 0.1% to 0.07% over the four years was determined. A total of 79 positive samples were further analysed by LC-HRMS screening technique to confirm positivity and detect the specific antibiotic compound contaminating the sample. The ß-lactam antibiotics resulted to be the most frequently detected, with the penicillin G being the most abundant compound. The data suggested that low levels of antibiotic contamination are consistently maintained over the last four years and the integration of the techniques used in this study is a valuable tool for a deep and precise monitoring of antibiotic residues in milk.

Prevalence of Different Salmonella enterica Subspecies and Serotypes in Wild Carnivores in Emilia-Romagna Region, Italy.

Salmonella is a pathogen of considerable health concern, given its zoonotic potential, and, in Italy, is the most frequently reported causative agent for foodborne outbreaks. Wild animals and in particular wild carnivores may be carriers of different Salmonella enterica subspecies and serotypes. Given their potential role as reservoirs, surveillance activities are necessary. This study aims to investigate the presence of different Salmonella subspecies and serotypes in wild carnivores in the Emilia-Romagna Region. A total of 718 fox (Vulpes vulpes), 182 badger (Meles meles) and 27 wolf (Canis lupus) carcasses, submitted between 2016-2022, were included for the present work. Gender and age data were collected along with geographical coordinates of carcass' discovery site. Contents of the large intestine were sampled and cultured according to ISO 6579-1 and both serogroup and serotype identification were performed according to ISO/TR 6579-3:2014. Salmonella was retrieved from 42 foxes (6%), 21 badgers (12%) and 3 wolves (12%), respectively. Isolated Salmonella enterica strains belonged to 4 different subspecies and 25 different serotypes. S. veneziana and S. typhimurium were the most frequent serotypes found (11/67 and 10/67, respectively). In conclusion, zoonotic serotypes were found in all these species of wildlife, thus confirming their potential role in the ecology of Salmonella spp.

Efficacy of live attenuated, vector and immune complex infectious bursal disease virus (IBDV) vaccines in preventing field strain bursa colonization: A European multicentric study.

Infectious bursal disease virus (IBDV) is among the most relevant and widespread immunosuppressive agents, which can severely damage poultry farming by causing direct losses, predisposing the host to secondary diseases and reducing the efficacy of vaccination protocols against other infections. IBDV has thus been the object of intense control activities, largely based on routine vaccination. However, the need for protecting animals from the infection in the first period of the production cycle, when the bursa susceptibility is higher, clashes with the blanketing effect of maternally derived antibodies. To overcome this issue, other strategies have been developed besides live attenuated vaccines, including vector vaccines and immune complex (icx) ones. The present study aims to investigate, in field conditions, the efficacy of these approaches in preventing IBDV infection in laying chickens vaccinated with either live attenuated, vector or immune complex (icx) vaccines. For this purpose, a multicentric study involving 481 farms located in 11 European countries was organized and IBDV infection diagnosis and strain characterization was performed at 6 weeks of age using a molecular approach. Vaccine strains were commonly detected in flocks vaccinated with live or icx vaccines. However, a significantly higher number of field strains (characterized as very virulent IBDVs) was detected in flocks vaccinated with vector vaccines, suggesting their lower capability of preventing bursal colonization. Different from vector vaccines, live and icx ones have a marked bursal tropism. It can thus be speculated that vaccine virus replication in these sites could limit vvIBDV replication by direct competition or because of a more effective activation of innate immunity. Although such different behavior doesn't necessarily affect clinical protection, further studies should be performed to evaluate if vvIBDV replication could still be associated with subclinical losses and/or for viral circulation in a "vaccinated environment" could drive viral evolution and favor the emergence of vaccine-escape variants.

Virulence Factors and Antimicrobial Resistance Profile of Escherichia Coli Isolated from Laying Hens in Italy.

Colibacillosis is the most common bacterial disease in the poultry industry. The isolation of Escherichia coli (E. coli) strains with multiple resistance to various classes of antimicrobials has been increasing in recent years. In this study, antimicrobial resistance features, serotyping and the presence of avian pathogenic Escherichia coli (APEC) virulence genes were investigated on a total of 71 E. coli strains isolated during outbreaks of colibacillosis in laying hens. The correlation between these features was evaluated. The most frequently isolated serogroups were O2 and O88. Resistance was often detected with nalidixic acid (49%) and ampicillin (38%), while all strains were sensitive to ceftiofur and florfenicol. Overall, 25% of the isolates showed resistance to at least three or more antimicrobial classes (multidrug-resistant strains), and 56% of the isolates were defined as APEC strains (due to the presence of at least five virulence genes). Correlation between the different parameters (virulence genes, serogroup and antimicrobial resistance) did not reveal relevant associations. The comparison of the obtained results with those of similar studies highlighted the importance of continuous monitoring in order to have a better understanding of colibacillosis. An evaluation of the national epidemiological situation would allow, especially with regard to antimicrobial resistance, to focus on the right measures in order to prioritize the available resources for effective disease control.

Potential Applications of Essential Oils for Environmental Sanitization and Antimicrobial Treatment of Intensive Livestock Infections.

The extensive use of antibiotics has contributed to the current antibiotic resistance crisis. Livestock infections of Salmonella spp, Clostridium spp. and E. coli antimicrobial-resistant bacteria represent a public threat to human and animal health. To reduce the incidence of these zoonoses, essential oils (EOs) could be effective antibiotic alternatives. This study aims at identifying EOs safe for use, effective both in complementary therapy and in the environmental sanitization of intensive farming. Natural products were chemo-characterized by gas chromatography. Three S. Typhimurium, three C. perfringens and four E. coli strains isolated from poultry and swine farms were used to assess the antimicrobial properties of nine EOs and a modified GR-OLI (mGR-OLI). The toxicity of the most effective ones (Cinnamomum zeylanicum, Cz; Origanum vulgare, Ov) was also evaluated on porcine spermatozoa and Galleria mellonella larvae. Cz, Ov and mGR-OLI showed the strongest antimicrobial activity; their volatile components were also able to significantly inhibit the growth of tested strains. In vitro, Ov toxicity was slightly lower than Cz, while it showed no toxicity on G. mellonella larvae. In conclusion, the study confirms the importance of evaluating natural products to consolidate the idea of safe EO applications in reducing and preventing intensive livestock infections.