Background and Aim: Pseudomonas aeruginosa is a relevant opportunistic and difficult to treat pathogen due to its widespread environmental diffusion, intrinsic resistance to many classes of antimicrobials, high ability to acquire additional resistance mechanisms, and wide range of pathogenic factors. The present study aimed to investigate the prevalence of P. aeruginosa in canine clinical samples, the antimicrobial susceptibility against antipseudomonal antibiotics, and the presence of extracellular pathogenic factors of the isolates, as well as their ability to produce biofilm. Materials and Methods: Overall, 300 clinical specimens from dogs with pyoderma or abscesses (n=58), otitis (n=59), and suspected bladder infection (n=183) were analyzed by standard bacteriological methods. P. aeruginosa isolates were tested for their antimicrobial susceptibility by disk and gradient diffusion methods to determine the minimum inhibitory concentrations. The ability of the isolates to produce biofilm was investigated by a microtiter plate assay, while virulence genes coding for elastase (lasB), exotoxin A (toxA), alkaline protease (aprA), hemolytic phospholipase C (plcH), and exoenzyme S (ExoS) were detected by polymerase chain reaction method. Results: A total of 24 isolates of P. aeruginosa were found in clinical specimens (urine n=3, skin/soft tissue n=6, and ear canal n=15). No resistance was found to ceftazidime, gentamicin, aztreonam, and imipenem (IMI), while low levels of resistance were found to enrofloxacin (ENR) (4.2%) and piperacillin-tazobactam (8.3%). However, 41.7% and 29.2% of the isolates showed intermediate susceptibility to ENR and IMI, respectively. Disk and gradient diffusion methods showed high concordance. The majority of the isolates revealed a weak (33.3%) or intermediate (45.8%) ability to form biofilm, while the strong biofilm producers (20.8%) derived exclusively from the ear canal samples. All isolates (100%) were positive for lasB, aprA, and plcH genes, while exoS and toxA were amplified in 21 (87.5%) and 22 (91.7%) isolates, respectively. Conclusion: In the present study, P. aeruginosa isolates from canine clinical samples were characterized by low levels of antimicrobial resistance against antipseudomonal drugs. However, the high presence of isolates with intermediate susceptibility for some categories of antibiotics, including carbapenems which are not authorized for veterinary use, could represent an early warning signal. Moreover, the presence of isolates with strong ability to produce biofilm represents a challenge for the interpretation of the antimicrobial susceptibility profile. In addition, the high prevalence of the extracellular pathogenic factors was indicative of the potential virulence of the isolates.

The slaughterhouse can act as a valid checkpoint to estimate the prevalence and the economic impact of diseases in farm animals. At present, scoring lesions is a challenging and time-consuming activity, which is carried out by veterinarians serving the slaughter chain. Over recent years, artificial intelligence(AI) has gained traction in many fields of research, including livestock production. In particular, AI-based methods appear able to solve highly repetitive tasks and to consistently analyze large amounts of data, such as those collected by veterinarians during postmortem inspection in high-throughput slaughterhouses. The present study aims to develop an AI-based method capable of recognizing and quantifying enzootic pneumonia-like lesions on digital images captured from slaughtered pigs under routine abattoir conditions. Overall, the data indicate that the AI-based method proposed herein could properly identify and score enzootic pneumonia-like lesions without interfering with the slaughter chain routine. According to European legislation, the application of such a method avoids the handling of carcasses and organs, decreasing the risk of microbial contamination, and could provide further alternatives in the field of food hygiene.

Diseases of the respiratory system are known to negatively impact the profitability of the pig industry, worldwide. Considering the relatively short lifespan of pigs, lesions can be still evident at slaughter, where they can be usefully recorded and scored. Therefore, the slaughterhouse represents a key check-point to assess the health status of pigs, providing unique and valuable feedback to the farm, as well as an important source of data for epidemiological studies. Although relevant, scoring lesions in slaughtered pigs represents a very time-consuming and costly activity, thus making difficult their systematic recording. The present study has been carried out to train a convolutional neural network-based system to automatically score pleurisy in slaughtered pigs. The automation of such a process would be extremely helpful to enable a systematic examination of all slaughtered livestock. Overall, our data indicate that the proposed system is well able to differentiate half carcasses affected with pleurisy from healthy ones, with an overall accuracy of 85.5%. The system was better able to recognize severely affected half carcasses as compared with those showing less severe lesions. The training of convolutional neural networks to identify and score pneumonia, on the one hand, and the achievement of trials in large capacity slaughterhouses, on the other, represent the natural pursuance of the present study. As a result, convolutional neural network-based technologies could provide a fast and cheap tool to systematically record lesions in slaughtered pigs, thus supplying an enormous amount of useful data to all stakeholders in the pig industry.

Contagious bovine pleuropneumonia (CBPP) is a severe disease caused by Mycoplasma mycoides subsp. mycoides (Mmm). Knowledge on CBPP pathogenesis is fragmented and hampered by the limited availability of laboratory animal and in vitro models of investigation. The purpose of the present study is to assess respiratory explants as useful tools to study the early stages of CBPP. Explants were obtained from trachea, bronchi and lungs of slaughtered cattle, tested negative for Mycoplasma spp. and for the major bacterial and viral respiratory pathogens. The interaction of Mmm with explant cells was studied by immunohistochemistry (IHC), double-labelling indirect immunofluorescence (DLIIF) and laser scanning confocal microscopy (LSCM). Mmm capability to survive and proliferate within the explants was evaluated by standard microbiological procedures. Finally, the putative cellular internalization of Mmm was further investigated by the gentamicin invasion assay. IHC and DLIIF indicated that Mmm can colonize explants, showing a marked tropism for lower airways. Specifically, Mmm was detected on/inside the bronchiolar and alveolar epithelial cells, the alveolar macrophages and the endothelial cells. The interaction between Mmm and explant cells was abolished by the pre-incubation of the pathogen with bovine anti-Mmm immune sera. Mmm was able to survive and proliferate in all tracheal, bronchial and lung explants, during the entire time course of the experiments. LSCM and gentamicin invasion assay both confirmed that Mmm can enter non-phagocytic host cells. Taken together, our data supports bovine respiratory explants as a promising tool to investigate CBPP, alternative to cattle experimental infection.