Three different genotypes of porcine circovirus 2 (PCV-2) identified in pigs and warthogs in Namibia.

Porcine circovirus 2 (PCV-2) is the causative agent of porcine circovirus diseases (PCVD). A study was undertaken to determine whether PCV-2 was present in samples collected from commercial pigs (n = 46) and warthogs (n = 42) in Namibia between 2019 and 2020. Twenty-three of the collected samples were positive by PCR (13 from pigs and 10 from warthogs), and a phylogenetic analysis of ORF2 identified three genotypes (PCV-2b and PCV-2d in pigs and PCV-2c in warthogs). This is the first time that PCV-2 has been identified in warthogs and in Namibia. It is also the first report of PCV-2c in Africa.

Scoring pleurisy in slaughtered pigs using convolutional neural networks.

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: A Comprehensive Overview.

Contagious bovine pleuropneumonia (CBPP) is a respiratory disease of cattle that is listed as notifiable by the World Organization for Animal Health. It is endemic in sub-Saharan Africa and causes important productivity losses due to the high mortality and morbidity rates. CBPP is caused by Mycoplasma mycoides subsp. mycoides (Mmm) and is characterized by severe fibrinous bronchopneumonia and pleural effusion during the acute to subacute stages and by pulmonary sequestra in chronic cases. Additional lesions can be detected in the kidneys and in the carpal and tarsal joints of calves. Mmm infection occurs through the inhalation of infected aerosol droplets. After the colonization of bronchioles and alveoli, Mmm invades blood and lymphatic vessels and causes vasculitis. Moreover, Mmm can be occasionally demonstrated in blood and in a variety of other tissues. In the lung, Mmm antigen is commonly detected on bronchiolar and alveolar epithelial cells, in lung phagocytic cells, within the wall of blood and lymphatic vessels, inside necrotic areas, and within tertiary lymphoid follicles. Mmm antigen can also be present in the cytoplasm of macrophages within lymph node sinuses, in the germinal center of lymphoid follicles, in glomerular endothelial cells, and in renal tubules. A complete pathological examination is of great value for a rapid presumptive diagnosis, but laboratory investigations are mandatory for definitive diagnosis. The purpose of this review is to describe the main features of CBPP including the causative agent, history, geographic distribution, epidemiology, clinical course, diagnosis, and control. A special focus is placed on gross and microscopic lesions in order to familiarize veterinarians with the pathology and pathogenesis of CBPP.

Follicular dendritic cell disruption as a novel mechanism of virus-induced immunosuppression.

Arboviruses cause acute diseases that increasingly affect global health. We used bluetongue virus (BTV) and its natural sheep host to reveal a previously uncharacterized mechanism used by an arbovirus to manipulate host immunity. Our study shows that BTV, similarly to other antigens delivered through the skin, is transported rapidly via the lymph to the peripheral lymph nodes. Here, BTV infects and disrupts follicular dendritic cells, hindering B-cell division in germinal centers, which results in a delayed production of high affinity and virus neutralizing antibodies. Moreover, the humoral immune response to a second antigen is also hampered in BTV-infected animals. Thus, an arbovirus can evade the host antiviral response by inducing an acute immunosuppression. Although transient, this immunosuppression occurs at the critical early stages of infection when a delayed host humoral immune response likely affects virus systemic dissemination and the clinical outcome of disease.

Congenital lung displacement in an English bulldog puppy.

The English bulldog is notoriously predisposed to a number of disease conditions, including congenital defects, likely due to extreme genetic selection which has reduced genetic variability within this breed. The present report describes, for the first time, the main clinical and pathological features of congenital lung displacement in an English bulldog puppy. The defect involved the entire left lung, which was normally developed but rotated and shifted to the right hemithorax. The pathogenesis of congenital anomalies is largely unknown. characterisation of the genetic bases of these malformations is crucial for a reform of the breeding selection which is also aimed at correcting phenotypic abnormalities with a strong impact on individual health.

Respiratory explants as a model to investigate early events of contagious bovine pleuropneumonia infection.

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.

Lung lesion score system in cattle: proposal for contagious bovine pleuropneumonia.

Contagious bovine pleuropneumonia (CBPP) is a severe infectious disease caused by Mycoplasma mycoides subsp. mycoides. The peculiar pathological features of CBPP make desirable the assessment of ad hoc score methods to grade the disease in the affected animals. Thus, the present work aims to assess a new lung score system for CBPP. Our results indicate that the present score system strongly correlates with that previously published by Turner and could be effectively used in CBPP-affected animals.

Depletion of follicular dendritic cells in tonsils collected from PMWS-affected pigs.

Post-weaning multisystemic wasting syndrome (PMWS) is a relevant, worldwide disease caused by porcine circovirus type 2 (PCV2). Microscopically, PMWS is mainly characterized by lymphocytic depletion, macrophage infiltration and syncytia in lymphoid tissues. Some data suggest that follicular dendritic cells (FDCs) could be infected by PCV2, thus likely playing a role in the pathogenesis of PMWS. The present paper aims at assessing, qualitatively and quantitatively, the FDCs' network in the soft palate tonsils of clinically healthy and PMWS-affected pigs. Consecutive tissue sections were tested by immunohistochemistry to detect PCV2, FDCs and macrophages. FDCs and PCV2 antigens were quantitatively assessed by means of the Image J software and results submitted to statistical analysis. Our data demonstrated that FDCs are significantly reduced in PMWS-affected pigs compared with healthy pigs and that FDCs' depletion should be considered among microscopic features of PMWS. It is reasonable to hypothesize that depletion of FDCs further compromises the immune response and enhances the occurrence and the severity of secondary infections, which are relevant for the clinical manifestation of PMWS.

Detection and genetic characterization of hepatitis E virus (HEV) genotype 3 subtype c in wild boars in Italy.

Hepatitis E virus (HEV) was detected in stools collected from wild boars in Italy, with an overall prevalence of 1.5 % (3/196). The sequence of a ~3.0-kb portion at the 3' end of the genome of one such strain, HEV/WB/P6-15/ITA, was determined. In the full-length ORF2, which encodes the capsid protein, the virus was genetically closest to wild boar and human HEV strains currently classified as genotype 3 subtype c. Interestingly, the 3' end of ORF2 of the WB/P6-15/ITA matched the 340-nucleotide (nt) sequence (94.0 % nt identity) of the human strain PeGe, identified in 2015 from a patient with acute hepatitis E in Genoa, Italy, suggesting that similar HEV strains are circulating in the same geographical setting in humans and animals.

Role of palatine tonsils as a prion entry site in classical and atypical experimental sheep scrapie.

Atypical and classical scrapie-infected sheep brain tissue was monolaterally injected into the tonsils of lambs to investigate their role as a prion entry point. We first detected classical PrP(Sc) within the inoculated tonsil and in the ipsilateral retropharyngeal lymph node at 3 months postinoculation (p.i.). At 7 months p.i., PrP(Sc) colonized other lymphoid tissues bilaterally, including ileal Peyer's patches. The earliest PrP(Sc) deposition within the brain was ipsilaterally observed at 9 months p.i. in the substantia reticularis of the medulla oblongata. At 12 months p.i., PrP(Sc) deposition was present bilaterally in the nucleus parasympathicus nervi vagi, as well as in the intermediolateral cell column of the thoracolumbar spinal cord. No PrP(Sc) was detected in the lambs inoculated with atypical scrapie. These findings suggest that neuroinvasion may naturally occur from the tonsil after a widespread prion replication within the lymphoid tissues during classical scrapie only, thus mimicking the pathogenesis after oral ingestion.

Intestinal Emphysema and Gut Bacterial Microbiota Composition.

Pneumatosis cystoides intestinalis, or intestinal emphysema, is a condition characterized by the presence of multiple cystic structures within the gut wall and on the serosal surface of the intestine. Intestinal emphysema represents an accidental finding in swine, although it can be clinically relevant in humans. Its etiology is unknown, and many theories have been proposed. Among them, a bacterial etiology is considered the most likely. Therefore, in this study, the V3-V4 region of the 16S rRNA gene was sequenced from 19 swine ileal tracts, 12 with intestinal emphysema and 7 without lesions, to detect a possible bacterial agent. In parallel, prevalence was estimated. Escherichia-Shigella (13.15%), Clostridium_sensu_stricto_1; s__uncultured_bacterium (7.09%), and Fusobacterium; s_uncultured bacterium (6.60%) were the most abundant species identified. No statistically relevant differences were observed between the pathological and physiological groups. Prevalence ranged from 1.25 to 5.12% depending on the batch. Our results suggest that the gut wall bacterial microbiota greatly match the normal gut microbiota, and that the etiological agent of intestinal emphysema may be (1) undetectable due to the chronicity of the lesions, (2) not considered statistically relevant in comparing the two groups (p < 0.05) and likewise in causing lesions, and (3) undetectable due to contamination. Regarding prevalence, the condition is moderately frequent.

Stiffness estimated by strain elastography reflects canine testicular spermatogenesis and histology.

Ultrasound elastography was proposed for the evaluation of testicular focal lesions, but no studies verified the agreement between the whole histological architecture of the testis and the stiffness measured by elastography. The present study explored the use of strain elastography in the evaluation of testis with normal or abnormal spermatogenesis, classified based on epididymal sperm attributes, and the consistency between elastographic parameters and the testicular histological feature. Strain elastography was performed during the routine andrological examination in 22 dogs presented for elective orchiectomy. Epididymal sperm attributes and testicular histology were analyzed after orchiectomy. Based on the epididymal sperm characteristics, testes were classified according to normal or abnormal spermatogenesis, and strain elastographic attributes were compared between groups. Possible correlations between strain elastography and histological features were also explored. Consistent with the literature in humans, testes with abnormal spermatogenesis were stiffer (mean strain elastographic index 3.6 ± 0.6) compared with normal testes (mean strain elastographic index 1.9 ± 0.2; P < 0.01). The strain elastographic index was negatively correlated with the area occupied by seminiferous tubules (Pearson's rho = -0.716; P = 0.0003), the mean diameter (Pearson's rho = -0.742; P = 0.0002), and thickness of the seminiferous tubule (Pearson's rho = -0.728; P = 0.0002). Surprisingly, no correlations were found between the area occupied by connective tissue in histological sections and elastographic attributes, suggesting that the increased stiffness was not related to the increased amount of connective tissue. This study demonstrated that strain elastography could be used to support the andrological examination, but measurements should be acquired in specific regions to be reliable.

Canaries' Microbiota: The Gut Bacterial Communities along One Female Reproductive Cycle.

Investigations of bacterial communities are on the rise both in human and veterinary medicine. Their role in health maintenance and pathogenic mechanisms is in the limelight of infectious, metabolic, and cancer research. Among the most considered, gut bacterial communities take the cake. Their part in animals was assessed mainly to improve animal production, public health, and pet management. In this regard, canaries deserve attention, being a popular pet and source of economic income for bird-keepers, for whom breeding represents a pivotal point. Thus, the present work aimed to follow gut bacterial communities' evolution along on whole reproductive cycle of 12 healthy female canaries. Feces were collected during parental care, molting, and resting phase, and submitted for 16S rRNA sequencing. Data were analyzed and a substantial presence of Lactobacillus aviarius along all the phases, and a relevant shift of microbiota during molting and rest due to an abrupt decrease of the Vermiphilaceae family were detected. Although the meaning of such change is not clear, future research may highlight unforeseen scenarios. Moreover, Lactobacillus aviarius may be deemed for normal bacteria flora restoration in debilitated birds, perhaps improving their health and productivity.

Condemnation of Porcine Carcasses: A Two-Year Long Survey in an Italian High-Throughput Slaughterhouse.

Ante-mortem and post-mortem inspection at slaughter are performed by Official Veterinarians and it is essential to identify alterations/lesions, which can make organs/carcasses unsuitable for human consumption. Obviously, carcass condemnation must be regarded as a highly undesirable event for the entire swine industry chain, as it represents the total failure of a long period of challenging work. Therefore, it seems valuable to estimate the prevalence and causes of carcasses condemnation, in order to preserve consumers' health and pig farming profitability. Bearing that in mind, the present study aimed at providing a reliable picture of the condemnation of porcine carcasses in Italy, with a special emphasis being placed upon pathological findings. Data were collected in a high-throughput abattoir located in northern Italy, where a total of 2,062,278 pigs were slaughtered during the period of study (2021-2022). Overall, 1362 pigs were considered unfit for slaughtering after ante-mortem inspection, mostly because of death during transportation or in resting pens, after their arrival at the abattoir. Moreover, 2007 carcasses were condemned after post-mortem inspection. The most common causes of condemnation were severe and diffuse peritonitis, disseminated abscesses, jaundice, and erysipelas. In our opinion, the present survey may provide useful and updated information about the condemnation of pig carcasses in Italy. At the same time, it highlights the need to collect data in a more systematic and standardized way, thus making possible their comparison over time and among different geographic areas.

Behavioral Disorders in Mares with Ovarian Disorders, Outcome after Laparoscopic Ovariectomy: A Case Series.

Owner complaints of estrus-related behavior in mares are a common cause of referral for laparoscopic ovariectomy. Granulosa cell tumors are a common neoplastic condition affecting the equine ovary, causing behavioral changes at rest and reduced performance. The reported success rate of ovariectomy in treating behavioral disorders is 64-86%. The aim of this study was to retrospectively evaluate the long-term follow-up of laparoscopic ovariectomy in mares in our case series, focusing on the owner's perspective of the behavior of the mares after surgery. In addition, the histopathological features of the removed ovaries were investigated. The clinical records of mares that underwent laparoscopic ovariectomy between 2015 and 2022 were retrospectively reviewed. Owners complaining of poor behavior were interviewed about the main behavioral problem leading to referral and its eventual resolution after surgery. Eleven mares were included. The most common complaints were increased sensitivity on both flanks (10/11, 91%) and general riding problems (9/11, 82%). In 5/11 cases (45%), both ovaries were removed by laparoscopic ovariectomy. Histopathologic findings consistent with GCT/GTCT were found in five out of six examined ovaries (five granulosa theca cell tumors, GTCT). According to owner interviews, the scores assigned to each behavior improved significantly after surgery, regardless of histological findings. Although many factors can influence the behavior of horses, granulosa cell tumors (GCTs) proved to be a common cause and, as reported by the owners, ovariectomy resulted in improvement or complete resolution of the abnormal behavior.

Scoring Enzootic Pneumonia-like Lesions in Slaughtered Pigs: Traditional vs. Artificial-Intelligence-Based Methods.

Artificial-intelligence-based methods are regularly used in the biomedical sciences, mainly in the field of diagnostic imaging. Recently, convolutional neural networks have been trained to score pleurisy and pneumonia in slaughtered pigs. The aim of this study is to further evaluate the performance of a convolutional neural network when compared with the gold standard (i.e., scores provided by a skilled operator along the slaughter chain through visual inspection and palpation). In total, 441 lungs (180 healthy and 261 diseased) are included in this study. Each lung was scored according to traditional methods, which represent the gold standard (Madec's and Christensen's grids). Moreover, the same lungs were photographed and thereafter scored by a trained convolutional neural network. Overall, the results reveal that the convolutional neural network is very specific (95.55%) and quite sensitive (85.05%), showing a rather high correlation when compared with the scores provided by a skilled veterinarian (Spearman's coefficient = 0.831, p < 0.01). In summary, this study suggests that convolutional neural networks could be effectively used at slaughterhouses and stimulates further investigation in this field of research.

Bovine Papillomavirus Type 1 Infection in an Equine Congenital Papilloma.

Papillomas are benign epithelial lesions protruding on the epithelial surfaces as finger-like or warty projections. These lesions are often caused by papillomavirus (PV) infection. Congenital papillomas have been reported in foals. However, to date, no evidence of PV infection has been provided. In the present paper, we describe the main clinical-pathological features of a congenital papilloma observed in a foal. In addition, biomolecular tests demonstrated BPV1 infection in the case under study. Such data stimulate further investigations, even on archived samples, aiming to clarifying the etiology of equine congenital papilloma and the clinical relevance, if any, of BPV1 vertical transmission in horses.

Epithelial-mesenchymal transition in an EcPV2-positive vulvar squamous cell carcinoma of a mare.

BACKGROUND: Vulvar squamous cell carcinoma (VSCC) has been recently associated with Equus caballus papillomavirus type 2 (EcPV2) infection. Still, few reports concerning this disease are present in the literature. OBJECTIVE: To describe a case of naturally occurring EcPV2-induced VSCC, by investigating tumour ability in undergoing the epithelial-to-mesenchymal transition (EMT). STUDY DESIGN: Case report. METHODS: A 13-year-old Haflinger mare was referred for a rapidly growing vulvar mass. After surgical excision, the mass was submitted to histopathology and molecular analysis. Histopathological diagnosis was consistent with a VSCC. Real-time qPCR, real-time reverse transcriptase (RT)-qPCR and RNAscope were carried out to detect EcPV2 infection and to evaluate E6/E7 oncogenes expression. To highlight the EMT, immunohistochemistry (IHC) was performed. Expression of EMT-related and innate immunity-related genes was investigated through RT-qPCR. RESULTS: Real-time qPCR, RT-qPCR and RNAscope confirmed EcPV2 DNA presence and expression of EcPV2 oncoproteins (E6 and E7) within the neoplastic vulvar lesion. IHC highlighted a cadherin switch together with the expression of the EMT-related transcription factor HIF1α. With RT-qPCR, significantly increased gene expression of EBI3 (45.0 ± 1.62, p < 0.01), CDH2 (2445.3 ± 0.39, p < 0.001), CXCL8 (288.7 ± 0.40, p < 0.001) and decreased gene expression of CDH1 (0.3 ± 0.57, p < 0.05), IL12A (0.04 ± 1.06, p < 0.01) and IL17 (0.2 ± 0.64, p < 0.05) were detected. MAIN LIMITATIONS: Lack of ability to generalise and danger of over-interpretation. CONCLUSION: The results obtained were suggestive of an EMT event occurring within the neoplastic lesion.

Mandibular Osteoma in a Haflinger Mare: Clinical and Pathological Features.

A 11-year-old, Haflinger breed mare developed a slowly growing mass, protruding on the ventral aspect of the mandible. On palpation, the lesion was pedunculated, very firm, cold and not painful. Radiographic examinations showed a radiopaque, multi-lobular, "mushroom-like" lesion, which arose from the mandibular cortex without involving the bone marrow cavity. Thereafter, the mass was excised for cosmetic purposes. Microscopically, the lesion mainly consisted of bone trabeculae, covered by a thick layer of connective tissue resembling the periosteum. History and clinical findings allowed the temptative diagnosis of mandibular osteoma, which was further supported by diagnostic imaging and pathological features. In particular, the presence of pedunculated lesions arising from the bone surface appeared as highly suggestive of osteoma. Equine osteomas usually affect paranasal sinuses and the rostral mandible and share many similarities with other neoplastic and neoplastic-like conditions, such as ossifying fibroma and fibrous dysplasia. The prognosis is good whenever the neoplasm is fully removed, that depending on the localization and the size of the mass.

Aspiration Pneumonia with Prominent Alveolar Mineralization in a Dairy Cow.

A 2-years-old Jersey breed cow showed severe respiratory distress and prolonged lateral recumbency about 1 week after parturition. The cow was regularly vaccinated against the main respiratory pathogens and was given a calcium salt solution per os shortly after calving in order to prevent milk fever. Despite therapy with broad-spectrum antibiotics and anti-inflammatory drugs, the cow spontaneously died one week later and was necropsied. On gross examination, a severe, acute, diffuse fibrinonecrotic tracheitis was observed. In addition, the cranioventral portions of both lungs appeared firm and severely congested, while the pleural surface was covered by a discrete amount of fibrinous exudate. Microscopically, the following lesions were observed: tracheal hemorrhages, acute, fibrinonecrotic and suppurative tracheitis, pulmonary hemorrhages, fibrinous bronchopneumonia and fibrinous pleuritis. Noteworthy, multiple foci of mineralization were observed, scattered throughout the lung parenchyma and occasionally within the tracheal mucosa. The presence of calcium deposits was confirmed by means of Von Kossa staining method. Based on clinical history, clinical signs and pathological findings, aspiration pneumonia caused by the accidental inhalation of liquid calcium salt supplement was diagnosed. The present case report highlights the relevance of the staff training to optimize animal production and welfare.