Survivin, ß-catenin, and ki-67 immunohistochemical expression in canine perivascular wall tumors: Preliminary assessment of prognostic significance.

High survivin expression has been correlated with poor outcomes in several canine tumors but not in soft tissue tumors (STTs). Survivin is a target gene of the Wnt/ß-catenin pathway, which is involved in human STT oncogenesis. Immunohistochemistry for survivin, ß-catenin, and Ki-67 was performed on 41 canine perivascular wall tumors (cPWTs), and statistical associations of protein expression and histopathologic and clinical variables with clinical outcomes were investigated. Immunohistochemically, there was nuclear positivity (0.9%-12.2% of tumor cells) for survivin in 41/41 (100%), cytoplasmic positivity (0 to > 75% of tumor cells) for survivin in 31/41 (76%), nuclear positivity (2.9%-67.2% of tumor cells) for ß-catenin in 24/41 (59%), and cytoplasmic positivity (0% to > 75% of tumor cells) for ß-catenin in 23/41 (56%) of cPWTs. All tumors expressed nuclear Ki-67 (2.2%-23.5%). In univariate analysis and multivariate analysis (UA and MA, respectively), every 1% increase of nuclear survivin was associated with an increase of the instantaneous death risk by a factor of 1.15 [hazard ratio (HR) = 1.15; P = .007]. Higher nuclear survivin was associated with grade II/III neoplasms (P = .043). Expression of cytoplasmic survivin, nuclear and cytoplasmic ß-catenin, and nuclear Ki-67 were not significantly associated with prognosis in UA nor MA. Tumor size was a significant prognostic factor for local recurrence in UA [subdistribution HR (SDHR) = 1.19; P = .02] and for reduced overall survival time in MA. According to UA and MA, a unitary increase of mitotic count was associated with an increase of the instantaneous death risk by a factor of 1.05 (HR = 1.05; P = .014). Nuclear survivin, mitotic count, and tumor size seem to be potential prognostic factors for cPWTs. In addition, survivin and ß-catenin may represent promising therapeutic targets for cPWTs.

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.

MEK1/2 regulate normal BCR and ABL1 tumor-suppressor functions to dictate ATO response in TKI-resistant Ph+ leukemia.

Resistance to tyrosine kinase inhibitors (TKIs) remains a clinical challenge in Ph-positive variants of chronic myeloid leukemia. We provide mechanistic insights into a previously undisclosed MEK1/2/BCR::ABL1/BCR/ABL1-driven signaling loop that may determine the efficacy of arsenic trioxide (ATO) in TKI-resistant leukemic patients. We find that activated MEK1/2 assemble into a pentameric complex with BCR::ABL1, BCR and ABL1 to induce phosphorylation of BCR and BCR::ABL1 at Tyr360 and Tyr177, and ABL1, at Thr735 and Tyr412 residues thus provoking loss of BCR's tumor-suppression functions, enhanced oncogenic activity of BCR::ABL1, cytoplasmic retention of ABL1 and consequently drug resistance. Coherently, pharmacological blockade of MEK1/2 induces dissociation of the pentameric MEK1/2/BCR::ABL1/BCR/ABL1 complex and causes a concurrent BCRY360/Y177, BCR::ABL1Y360/Y177 and cytoplasmic ABL1Y412/T735 dephosphorylation thereby provoking the rescue of the BCR's anti-oncogenic activities, nuclear accumulation of ABL1 with tumor-suppressive functions and consequently, growth inhibition of the leukemic cells and an ATO sensitization via BCR-MYC and ABL1-p73 signaling axes activation. Additionally, the allosteric activation of nuclear ABL1 was consistently found to enhance the anti-leukemic effects of the MEK1/2 inhibitor Mirdametinib, which when combined with ATO, significantly prolonged the survival of mice bearing BCR::ABL1-T315I-induced leukemia. These findings highlight the therapeutic potential of MEK1/2-inhibitors/ATO combination for the treatment of TKI-resistant leukemia.

Use of the international classification of diseases (ICD)-11 method applied to veterinary forensic pathology for coding the cause and manner of death in wildlife.

The growth of human population has led, in recent years, to increasingly frequent contacts with the wild animals with which we share the territory, sometimes leading to negative interactions with them. The purpose of the study is to apply the codes contained in the 11th Revision of the International Classification of Diseases (ICD-11) method to investigate the cause and the manner of death, also to entrust the veterinarian with the task of recognizing and describing a suspected animal abuse as a sentinel indicator of violence toward humans and non-humans, thus expanding the concept of "One Health" from a forensic investigation perspective. The subjects recruited are wild mammals submitted for autopsy to the Pathology Unit of the Department of Veterinary Science, University of Parma, Italy, from 2015 to 2018. The manner and the cause of death of 167 wild animals of 16 different species have been investigated. When possible, an on-site inspection where the corpse was found was performed. Injuries were classified according to the on-line 11th Revision of the International Classification of Diseases method. Section 22 (Injury, poisoning or certain other consequences of external causes) was used to record the "immediate cause of death" (cause of death) and Section 23 (External causes of morbidity or mortality) was used to record the "underlying cause of death" (manner of death) for each animal. In most cases, death occurred as a result of road trauma but in some cases, abuse and voluntary killing were investigated. The recognition of non-accidental injuries is particularly important for both the defense in court of animals and for the connection between crimes committed against animals and against humans, known as "The Link". The use of the ICD-11 method, as a sort of summary of the autopsy report, was confirmed to be of great value for the clarity and simplicity of processing the data collected also by veterinary pathologists. The veterinary pathologists can use this evidence-based method with the aim of creating a national register and therefore, to understand the real extent of the human impact on wildlife and document it in a scientific and statistically usable way.

An Overview of Epithelial-to-Mesenchymal Transition and Mesenchymal-to-Epithelial Transition in Canine Tumors: How Far Have We Come?

Historically, pre-clinical and clinical studies in human medicine have provided new insights, pushing forward the contemporary knowledge. The new results represented a motivation for investigators in specific fields of veterinary medicine, who addressed the same research topics from different perspectives in studies based on experimental and spontaneous animal disease models. The study of different pheno-genotypic contexts contributes to the confirmation of translational models of pathologic mechanisms. This review provides an overview of EMT and MET processes in both human and canine species. While human medicine rapidly advances, having a large amount of information available, veterinary medicine is not at the same level. This situation should provide motivation for the veterinary medicine research field, to apply the knowledge on humans to research in pets. By merging the knowledge of these two disciplines, better and faster results can be achieved, thus improving human and canine health.

Nipah Virus Disease: Epidemiological, Clinical, Diagnostic and Legislative Aspects of This Unpredictable Emerging Zoonosis.

Nipah virus (NiV) infection is a viral disease caused by a Henipavirus, belonging to the Paramyxoviridae family, responsible for a zoonosis. The course of the disease can be very serious and lead to death. NiV natural hosts are fruit bats (also known as megabats) belonging to the Pteropodidae family, especially those of the Pteropus genus. Natural infection in domestic animals has been described in farming pigs, horses, domestic and feral dogs and cats. Natural NiV transmission is possible intra-species (pig-to-pig, human-to-human) and inter-species (flying bat-to-human, pig-to-human, horse-to-human). The infection can be spread by humans or animals in different ways. It is peculiar how the viral transmission modes among different hosts also change depending on the geographical area for different reasons, including different breeding methods, eating habits and the recently identified genetic traits/molecular features of main virus proteins related to virulence. Outbreaks have been described in Malaysia, Singapore, Bangladesh, India and the Philippines with, in some cases, severe respiratory and neurological disease and high mortality in both humans and pigs. Diagnosis can be made using different methods including serological, molecular, virological and immunohistochemical methods. The cornerstones for control of the disease are biosecurity (via the correct management of reservoir and intermediate/amplifying hosts) and potential vaccines which are still under development. However, the evaluation of the potential influence of climate and anthropogenic changes on the NiV reservoir bats and their habitat as well as on disease spread and inter-specific infections is of great importance. Bats, as natural reservoirs of the virus, are responsible for the viral spread and, therefore, for the outbreaks of the disease in humans and animals. Due to the worldwide distribution of bats, potential new reports and spillovers are not to be dismissed in the future.

Peripheral Nerve Sheath Tumors Resembling Human Atypical Neurofibroma in Goldfish (Carassius auratus, Linnaeus, 1758).

Skin spindle cell tumors (SSTs) frequently occur in fishes, with peripheral nerve sheath tumors (PNSTs) being the most commonly reported neoplasms in goldfish. However, distinguishing PNSTs from other SCTs is not always possible when relying exclusively on routine cytological and histopathological findings. Therefore, the aim of this study is to characterize six skin nodules, resembling atypical neurofibromas in humans, found in six cohabiting goldfish (Carassius auratus), and to determine a minimal subset of special stains required to correctly identify PNSTs in this species. Routine cytology and histopathology were indicative of an SCT with nuclear atypia in all cases, with randomly distributed areas of hypercellularity and loss of neurofibroma architecture. Muscular and fibroblastic tumors were excluded using Azan trichrome staining. Alcian blue and Gomori's reticulin stains revealed the presence of intratumoral areas of glycosaminoglycans or mucins and basement membrane fragments, respectively. PAS and PAS-diastase stains confirmed the latter finding and revealed intra- and extracellular glycogen granules. Immunohistochemistry displayed multifocal, randomly distributed aggregates of neoplastic cells positive for S100 protein and CNPase, intermingled with phosphorylated and non-phosphorylated neurofilament-positive axons. Collectively, these findings are consistent with a PNST resembling atypical neurofibroma in humans, an entity not previously reported in goldfish, and suggest that Azan trichrome staining, reticulin staining, and immunohistochemistry for S100 protein and CNPase represent a useful set of special stains to identify and characterize PNSTs in this species.

Epithelial to Mesenchymal Transition (EMT) in a Laryngeal Squamous Cell Carcinoma of a Horse: Future Perspectives.

Squamous cell carcinoma (SCC) is one of the most frequent tumors of skin and muco-cutaneous junctions in the horse. Equine papillomavirus type 2 (EcPV2) has been detected in equine SCC of the oral tract and genitals, and recently also in the larynx. As human squamous cell carcinoma of the larynx (SCCL), it is strongly etiologically associated with high-risk papillomavirus (h-HPV) infection. This study focuses on tumor cells behavior in a naturally occurring tumor that can undergo the so-called epithelial to mesenchymal transition (EMT). A SCCL in a horse was investigated by immunohistochemistry using antibodies against E-cadherin, pan-cytokeratin AE3/AE1, ß-catenin, N-cadherin, vimentin, ZEB-1, TWIST, and HIF-1α. EcPV2 DNA detection and expression of oncogenes in SCC were investigated. A cadherin switch and an intermediate filaments rearrangement within primary site tumor cells together with the expression of the EMT-related transcription factors TWIST-1, ZEB-1, and HIF-1α were observed. DNA obtained from the tumor showed EcPV2 positivity, with E2 gene disruption and E6 gene dysregulation. The results suggest that equine SCCL might be a valuable model for studying EMT and the potential interactions between EcPV2 oncoproteins and the EMT process in SCCL.

Endocanalicular transendothelial crossing (ETC): A novel intravasation mode used by HEK-EBNA293-VEGF-D cells during the metastatic process in a xenograft model.

In cancer metastasis, intravasation of the invasive tumor cell (TCi) represents one of the most relevant events. During the last years, models regarding cancer cell intravasation have been proposed, such as the "endocanalicular transendothelial crossing" (ETC) theory. This theory describes the interplay between two adjacent endothelial cells and the TCi or a leukocyte during intravasation. Two endothelial cells create a channel with their cell membranes, in which the cell fits in without involving endothelial cell intercellular junctions, reaching the lumen through a transendothelial passage. In the present study, ten SCID mice were subcutaneously xenotransplanted with the HEK-EBNA293-VEGF-D cell line and euthanized after 35 days. Post-mortem examinations were performed and proper specimens from tumors were collected. Routine histology and immunohistochemistry for Ki-67, pAKT, pERK, ZEB-1, TWIST-1, F-actin, E-cadherin and LYVE-1 were performed followed by ultrastructural serial sections analysis. A novel experimental approach involving Computed Tomography (CT) combined with 3D digital model reconstruction was employed. The analysis of activated transcription factors supports that tumor cells at the periphery potentially underwent an epithelial-to-mesenchymal transition (EMT)-like process. Topographical analysis of LYVE-1 immunolabeled lymphatics revealed a peritumoral localisation. TEM investigations of the lymphatic vessels combined with 3D digital modelling enhanced the understanding of the endotheliocytes behavior during TCi intravasation, clarifying the ETC theory. Serial ultrastructural analysis performed within tumor periphery revealed numerous cells during the ETC process. Furthermore, this study demonstrates that ETC is an intravasation mode more frequently used by the TCi than by leukocytes during intravasation in the HEK-EBNA293-VEGF-D xenograft model and lays down the potential basis for promising future studies regarding intravasation blocking therapy.

Association Between Infectious Agents and Lesions in Post-Weaned Piglets and Fattening Heavy Pigs With Porcine Respiratory Disease Complex (PRDC).

Porcine Respiratory Disease Complex (PRDC) is a multifactorial syndrome that causes health problems in growing pigs and economic losses to farmers. The etiological factors involved can be bacteria, viruses, or mycoplasmas. However, environmental stressors associated with farm management can influence the status of the animal's health. The role and impact of different microorganisms in the development of the disease can be complex, and these are not fully understood. The severity of lesions are a consequence of synergism and combination of different factors. The aim of this study was to systematically analyse samples, conferred to the Veterinary Diagnostic Laboratory (IZSLER, Brescia), with a standardized diagnostic protocol in case of suspected PRDC. During necropsy, the lungs and carcasses were analyzed to determine the severity and extension of lesions. Gross lung lesions were classified according to a pre-established scheme adapted from literature. Furthermore, pulmonary, pleural, and nasal lesions were scored to determine their severity and extension. Finally, the presence of infectious agents was investigated to identify the microorganisms involved in the cases studied. During the years 2014-2016, 1,658 samples of lungs and carcasses with PRDC from 863 farms were analyzed; among them 931 and 727 samples were from weaned piglets and fattening pigs, respectively. The most frequently observed lesions were characteristic of catarrhal bronchopneumonia, broncho-interstitial pneumonia, pleuropneumonia, and pleuritis. Some pathogens identified were correlated to specific lesions, whereas other pathogens to various lesions. These underline the need for the establishment of control and treatment programmes for individual farms.

Profile of gamma-delta (γδ) T lymphocytes in the peripheral blood of crossbreed dogs during stages of life and implication in aging.

BACKGROUND: Data on gamma-delta (γδ) T lymphocytes in the peripheral blood of dogs are scant, related only to healthy pure breed dogs and limited to a restricted age range. The aim of the study was to investigate the modulation of the γδ T lymphocyte (TCRγδ+) subpopulation in peripheral blood of crossbreed healthy dogs according to five identified stages of life: Puppy, Junior, Adult, Mature, Senior and to determine its implication in aging. A rigorous method of recruitment was used to minimize the influence of internal or external pressure on the immune response. Twenty-three intact female and twenty-four intact male dogs were enrolled. Blood samples were collected and immunophenotyping of peripheral blood T lymphocytes and γδ T cell subpopulations was performed. RESULTS: The percentage of γδ T cells in peripheral blood lymphocytes was comparable with the value of 2.5% published by Faldyna and co-workers (2001), despite the percentage reported was investigated in less arranged age range groups and coming from four different dog pure breeds, whereas our data were recorded on wider age range groups and coming from crossbreed dogs. Therefore, the γδ T cell percentage (2.5%) is consistent and points out that such value is breed-independent. Statistical analysis highlighted differences in both percentage and absolute γδ T cells according to the stage of life. γδ T cells decreased significantly in the peripheral blood of elder dogs (Senior group) in comparison with previous stages of life (Puppy, Junior, and Adult groups). Differences in γδ T cells are significant and they are reported, for the first time, related to dog aging. CONCLUSIONS: The study confirms dogs to be among the animals with a low TCRγδ+ cell profile. A decrease of the TCRγδ+ subpopulation percentage was observed in elder dogs. TCRγδ+ cells of group S were different from those of groups P, J, and A. The differences are reported for the first time in dog aging. Identifying the stage of life when the decrease of γδ T lymphocytes starts can be useful for providing a rationale for drafting a wellness plan trial to support thymus immune functions and mitigate its functional exhaustion.

Mesenchymal to epithelial transition driven by canine distemper virus infection of canine histiocytic sarcoma cells contributes to a reduced cell motility in vitro.

Sarcomas especially of histiocytic origin often possess a poor prognosis and response to conventional therapies. Interestingly, tumours undergoing mesenchymal to epithelial transition (MET) are often associated with a favourable clinical outcome. This process is characterized by an increased expression of epithelial markers leading to a decreased invasion and metastatic rate. Based on the failure of conventional therapies, viral oncolysis might represent a promising alternative with canine distemper virus (CDV) as a possible candidate. This study hypothesizes that a CDV infection of canine histiocytic sarcoma cells (DH82 cells) triggers the MET process leading to a decreased cellular motility. Immunofluorescence and immunoblotting were used to investigate the expression of epithelial and mesenchymal markers followed by scratch assay and an invasion assay as functional confirmation. Furthermore, microarray data were analysed for genes associated with the MET process, invasion and angiogenesis. CDV-infected cells exhibited an increased expression of epithelial markers such as E-cadherin and cytokeratin 8 compared to controls, indicating a MET process. This was accompanied by a reduced cell motility and invasiveness. Summarized, these results suggest that CDV infection of DH82 cells triggers the MET process by an increased expression of epithelial markers resulting in a decreased cell motility in vitro.

Oxidative Stress in Canine Histiocytic Sarcoma Cells Induced by an Infection with Canine Distemper Virus Led to a Dysregulation of HIF-1α Downstream Pathway Resulting in a Reduced Expression of VEGF-B in vitro.

Histiocytic sarcomas represent malignant tumors which require new treatment strategies. Canine distemper virus (CDV) is a promising candidate due to its oncolytic features reported in a canine histiocytic sarcoma cell line (DH82 cells). Interestingly, the underlying mechanism might include a dysregulation of angiogenesis. Based on these findings, the aim of the present study was to investigate the impact of a persistent CDV-infection on oxidative stress mediated changes in the expression of hypoxia-inducible factor (HIF)-1α and its angiogenic downstream pathway in DH82 cells in vitro. Microarray data analysis, immunofluorescence for 8-hydroxyguanosine, superoxide dismutase 2 and catalase, and flow cytometry for oxidative burst displayed an increased oxidative stress in persistently CDV-infected DH82 cells (DH82Ond pi) compared to controls. The HIF-1α expression in DH82Ond pi increased, as demonstrated by Western blot, and showed an unexpected, often sub-membranous distribution, as shown by immunofluorescence and immunoelectron microscopy. Furthermore, microarray data analysis and immunofluorescence confirmed a reduced expression of VEGF-B in DH82Ond pi compared to controls. In summary, these results suggest a reduced activation of the HIF-1α angiogenic downstream pathway in DH82Ond pi cells in vitro, most likely due to an excessive, unusually localized, and non-functional expression of HIF-1α triggered by a CDV-induced increased oxidative stress.

Feline coronavirus-associated myocarditis in a domestic longhair cat.

CASE SUMMARY: A 9-month-old entire male domestic longhair indoor cat presented with a 3-week history of fluctuating fever, weight loss and small intestine diarrhoea, which was unresponsive to antibiotics and supportive treatment. Abdominal ultrasound revealed severe jejunal and ileocolic junction intestinal wall thickening with loss of layering. An enterectomy was performed and histopathology revealed severe pyogranulomatous enteritis with vasculitits, compatible with the diagnosis of feline infectious peritonitis (FIP). Four days after surgery, the cat re-presented with anorexia and acute onset of expiratory dyspnoea. Echocardiography showed left ventricular hypertrophy and bilateral atrial enlargement. Congestive heart failure caused by hypertrophic cardiomyopathy was suspected and treatment with furosemide was started, which led to amelioration of the clinical signs. The following day, four-limb ataxia, hypermetria and bilateral uveitis were evident. Given the persistent anorexia and worsening of the clinical signs, the cat was humanely euthanized and a post-mortem examination was performed. Necropsy revealed multifocal pyogranulomatous lesions involving multiple organs (adrenal glands, kidneys, lungs, brain, myocardium, lymph nodes, liver), compatible with the diagnosis of FIP. Immunohistochemistry performed on the myocardium revealed feline coronavirus-positive macrophages associated with pyogranulomatous lesions, justifying a diagnosis of feline coronavirus-associated myocarditis. RELEVANCE AND NOVEL INFORMATION: To the authors' knowledge, the case described here represents the first published report of feline coronavirus-associated myocarditis. This should be considered as a possible differential diagnosis in cats presenting with cardiac-related signs and other clinical signs compatible with FIP.

Functional interplay between NF-κB-inducing kinase and c-Abl kinases limits response to Aurora inhibitors in multiple myeloma.

Considering that Aurora kinase inhibitors are currently under clinical investigation in hematologic cancers, the identification of molecular events that limit the response to such agents is essential for enhancing clinical outcomes. Here, we discover a NF-κB-inducing kinase (NIK)-c-Abl-STAT3 signaling-centered feedback loop that restrains the efficacy of Aurora inhibitors in multiple myeloma. Mechanistically, we demonstrate that Aurora inhibition promotes NIK protein stabilization via downregulation of its negative regulator TRAF2. Accumulated NIK converts c-Abl tyrosine kinase from a nuclear proapoptotic into a cytoplasmic antiapoptotic effector by inducing its phosphorylation at Thr735, Tyr245 and Tyr412 residues, and, by entering into a trimeric complex formation with c-Abl and STAT3, increases both the transcriptional activity of STAT3 and expression of the antiapoptotic STAT3 target genes PIM1 and PIM2. This consequently promotes cell survival and limits the response to Aurora inhibition. The functional disruption of any of the components of the trimer NIK-c-Abl-STAT3 or the PIM survival kinases consistently enhances the responsiveness of myeloma cells to Aurora inhibitors. Importantly, concurrent inhibition of NIK or c-Abl disrupts Aurora inhibitor-induced feedback activation of STAT3 and sensitizes myeloma cells to Aurora inhibitors, implicating a combined inhibition of Aurora and NIK or c-Abl kinases as potential therapies for multiple myeloma. Accordingly, pharmacological inhibition of c-Abl together with Aurora resulted in substantial cell death and tumor regression in vivo The findings reveal an important functional interaction between NIK, Abl and Aurora kinases, and identify the NIK, c-Abl and PIM survival kinases as potential pharmacological targets for improving the efficacy of Aurora inhibitors in myeloma.

Correction to: The synergistic effect of organic acids, phytochemicals and a permeabilizing complex reduces Salmonella Typhimurium 1,4,[5],12:i-shedding in pigs.

This article was originally published with all author names incorrectly listed. All author names have now been transposed and appear correctly above.

The synergistic effect of organic acids, phytochemicals and a permeabilizing complex reduces Salmonella Typhimurium 1,4,[5],12:i-shedding in pigs.

Salmonella Typhimurium (including S.Typhimurium 1,4,[5],12:i-) and other enteric pathogens cause acute infection in pigs during the weaning stage, often evolving into chronic infections responsible for the introduction of zoonotic bacteria into the slaughterhouse and thus determining carcass contamination. In addition to being zoonotic hazards, these pathogens are responsible for economic losses in affected farms. Traditionally, antibiotic treatments have been largely administered in order to reduce the infection burden but it favored, as a direct consequence, an increase in the number of multi-drug resistance strains. In order to overcome antibiotic-resistance concerns, new alternative control strategies should be developed. In this context, a blend of organic acids, phytochemicals and a permeabilizing complex, administered in feed (Group A - 459 piglets) or water (Group B - 458 piglets), was tested in field conditions for its capability of reducing Salmonella-infection in weaned piglets of an endemic farm. Data recorded were compared to results of a control group (Group C - 456 piglets). Zootechnical parameters were recorded in all animals, while microbiological, serological and PCR analyses were conducted in 15 piglets for each group. Results demonstrated that additive administered in feed improved animal weight gain (better average daily gain [A.D.G.] and increment), and rapidly reduced Salmonella-shedding in feces. Administration of additive in feed gave better results than in water.

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.

Phenotypic characterization of a highly pathogenic Italian porcine reproductive and respiratory syndrome virus (PRRSV) type 1 subtype 1 isolate in experimentally infected pigs.

Highly pathogenic (HP) isolates of the PRRS virus started emerging in North America and Asia in the late 1990s. More recently, they have emerged in Europe. These isolates are characterized by high viral loads, severe general clinical signs and high mortality, in sows, weaners and growers. Their genome shows a discontinuous aminoacids deletion in the non-structural protein 2 (NSP2). The present study was aimed at characterizing the clinical, pathological and immunological features of a highly pathogenetic, Italian PRRSV-1 subtype 1 isolate (PRRSV1_PR40/2014), following experimental infection in conventional 4-weeks-old pigs. The PRRSV1_PR40/2014 infected group showed severe clinical signs (high fever and dispnoea). Pathological lesions, including severe lymphocytopenia in bronchial lymph-nodes and thymus were also recorded. Higher serum PRRSV genome copies and lower virus neutralizing antibody titer were observed in the PR40 group, when compared to the group infected with a conventional PRRSV strain. The genetic analysis of the strain, and the phenotypic features observed in the field and reproduced in the experimental study, confirmed the high pathogenicity of the Italian PRRSV-1 subtype 1 PR40 isolate.

Prime-boost vaccination with attenuated Salmonella Typhimurium ΔznuABC and inactivated Salmonella Choleraesuis is protective against Salmonella Choleraesuis challenge infection in piglets.

BACKGROUND: Salmonella enterica serovar Choleraesuis (S. Choleraesuis) infection causes a systemic disease in pigs. Vaccination could represent a solution to reduce prevalence in farms. In this study, we aimed to assess the efficacy of an attenuated strain of Salmonella enterica serovar Typhimurium (S. Typhimurium ΔznuABC) against S. Choleraesuis infection. The vaccination protocol combined priming with attenuated S. Typhimurium ΔznuABC vaccine and boost with an inactivated S. Choleraesuis vaccine and we compared the protection conferred to that induced by an inactivated S. Choleraesuis vaccine. METHODS: The first group of piglets was orally vaccinated with S. Typhimurium ΔznuABC and boosted with inactivated S. Choleraesuis, the second one was intramuscularly vaccinated with S. Choleraesuis inactivated vaccine and the third group of piglets was unvaccinated. All groups of animals were challenged with a virulent S. Choleraesuis strain at day 35 post vaccination. RESULTS: The results showed that the vaccination protocol, priming with S. Typhimurium ΔznuABC and boosted with inactivated S. Choleraesuis, applied to group A was able to limit weight loss, fever and organs colonization, arising from infection with virulent S. Choleraesuis, more effectively, than the prime-boost vaccination with homologous S. Choleraesuis inactivated vaccine (group B). CONCLUSION: In conclusion, these research findings extend the validity of attenuated S. Typhimurium ΔznuABC strain as a useful mucosal vaccine against S. Typhimurium and S. Choleraesuis pig infection. The development of combined vaccination protocols can have a diffuse administration in field conditions because animals are generally infected with different concomitant serovars.