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Cattle are stoic by nature and have long been subject to evolutionary pressure to mask pain to avoid becoming prey. Quantifying pain via biomarkers allows researchers to capture changes that are not easily detected by the human eye. Negative public perception of routine husbandry practices such as dehorning and castration is growing, increasing the need for the development of practices to relieve pain and suffering in cattle. The objective was to quantify pain associated with induced bacterial pneumonia, scoop and hot-iron dehorning, surgical castration, and hot-iron branding, and to investigate analgesic effectiveness. Calves were experimentally inoculated with Mannheimia haemolytica, scoop or cautery dehorned, surgically castrated, or hot-iron branded. Outcome variables were collected at baseline and following husbandry procedures and included serum cortisol; infrared thermography (IRT); mechanical nociceptive threshold (MNT); substance P; kinematic gait analysis; a visual analog scale; chute defense and behavior scoring; clinical illness score; computerized lung score; average activity and rumination; prostaglandin E2 metabolite; plasma serum amyloid A and rectal temperature. Responses were analyzed using repeated measures with calf nested in treatment designated as a random effect, and treatment, time, and their interaction designated as fixed effects. A combination of reduced activity levels, decreased force on calves’ right front limb, and increased visual analog scale pain scores all support that bacterial pneumonia in cattle is painful (P ≤ 0.01). Differences in right front force were observed in calves challenged with M. haemolytica and treated with flunixin transdermal (96.5 kg) and those given a placebo (85.5 kg), indicating that flunixin transdermal may attenuate specific pain biomarkers in cattle with respiratory disease (P < 0.01). Administration of a bupivacaine liposome suspension block at the time of dehorning and castration was as effective at controlling pain as a multi-modal approach of lidocaine and meloxicam (as evidenced by treatment differences in MNT, cortisol and gait analysis (P ≤ 0.04)). Oral meloxicam administration at branding reduced IRT differences from the branding and control site and reduced lying bouts (P < 0.01). Breed and sex effects were observed across a wide range of biomarkers and should be investigated in future pain studies. The need for long-acting analgesic options for cattle that demonstrate pain alleviation across multiple biomarkers is apparent and would be beneficial to alleviating pain from routine husbandry procedures like dehorning, castration, and branding, as well as painful disease conditions such as bovine respiratory disease.

Pleuritic chest pain from bacterial pneumonia is often reported in human medicine. However, studies investigating pain associated with bovine respiratory disease (BRD) are lacking. The objectives of this study were to assess if bacterial pneumonia elicits a pain response in calves with experimentally induced BRD and to determine the analgesic effects of transdermally administered flunixin. A total of 26 calves, 6–7 mo of age, with no history of BRD were enrolled into one of three treatment groups: 1) experimentally induced BRD + transdermal flunixin at 3.3 mg/kg twice, 24 h apart (BRD + FTD); 2) experimentally induced BRD + placebo (BRD + PLBO); and 3) sham induction + placebo (CNTL + PLBO). Calves induced with BRD were inoculated with Mannheimia haemolytica via bronchoalveolar lavage. Outcomes were collected from −48 to 192 h post-treatment and included serum cortisol, infrared thermography, mechanical nociceptive threshold, substance P, kinematic gait analysis, visual analog scale (VAS), clinical illness score, computerized lung score, average activity and rumination level, prostaglandin E2 metabolite, plasma serum amyloid A, and rectal temperature. Outcomes were evaluated using either a generalized logistic mixed model for categorical variables or a generalized linear mixed model for continuous variables. Right front force differed by treatment (P = 0.01). The BRD + PLBO had lower mean force applied to the right front limb (85.5 kg) compared with BRD + FTD (96.5 kg; P < 0.01). Average VAS differed by a treatment by time interaction (P = 0.01). The VAS scores differed for BRD + PLBO at −48 (3.49 mm) compared with 168 and 192 h (13.49 and 13.64 mm, respectively) (P < 0.01). Activity for BRD + PLBO was higher at −48 h (27 min/h) compared with 48, 72, 120, and 168 h (≤ 22.24 min/h; P < 0.01). Activity differed by a treatment by time interaction (P = 0.01). Activity for BRD + FTD was higher at −48 and 0 h (28.2 and 28.2 min/h, respectively) compared to 48, 72, 96, and 168 h (≤23.7 min/h; P < 0.01). Results show a combination of reduced activity levels, decreased force on the right front limb, and increased VAS pain scores all support that bacterial pneumonia in cattle is painful. Differences in right front force indicate that flunixin transdermal may attenuate certain pain biomarkers in cattle with BRD. These findings suggest that BRD is painful and analgesic drugs may improve the humane aspects of care for cattle with BRD.

Bovine respiratory disease (BRD) is the most economically significant disease for cattle producers in the U.S. Cattle with advanced lung lesions at harvest have reduced average daily gain, yield grades, and carcass quality outcomes. The identification of biomarkers and clinical signs that accurately predict lung lesions could benefit livestock producers in determining a BRD prognosis. Receiver operating characteristic (ROC) curves are graphical plots that illustrate the diagnostic ability of a biomarker or clinical sign. Previously we used the area under the ROC curve (AUC) to identify cortisol, hair cortisol, and infrared thermography imaging as having acceptable (AUC 0.7) diagnostic accuracy for detecting pain in cattle. Herein, we used ROC curves to assess the sensitivity and specificity of biomarkers and clinical signs associated with lung lesions after experimentally induced BRD. We hypothesized pain biomarkers and clinical signs assessed at specific time points after induction of BRD could be used to predict lung consolidation at necropsy. Lung consolidation of 10% was retrospectively assigned at necropsy as a true positive indicator of BRD. Calves with a score of 10% were considered negative for BRD. The biomarkers and clinical signs analyzed were serum cortisol; infrared thermography (IRT); mechanical nociceptive threshold (MNT); substance P; kinematic gait analysis; a visual analog scale (VAS); clinical illness score (CIS); computerized lung score (CLS); average activity levels; prostaglandin E2 metabolite (PGEM); serum amyloid A; and rectal temperature. A total of 5,122 biomarkers and clinical signs were collected from 26 calves, of which 18 were inoculated with M. haemolytica. All statistics were performed using JMP Pro 14.0. Results comparing calves with significant lung lesions to those without yielded the best diagnostic accuracy (AUC 0.75) for right front stride length at 0 h; gait velocity at 32 h; VAS, CIS, average activity and rumination levels, step count, and rectal temperature, all at 48 h; PGEM at 72 h; gait distance at 120 h; cortisol at 168 h; and IRT, right front force and serum amyloid A, all at 192 h. These results show ROC analysis can be a useful indicator of the predictive value of pain biomarkers and clinical signs in cattle with induced bacterial pneumonia. AUC values for VAS score, average activity levels, step count, and rectal temperature seemed to yield good diagnostic accuracy (AUC 0.75) at multiple time points, while MNT values, substance P concentrations, and CLS did not (all AUC values 0.75).Bovine respiratory disease (BRD) is the most economically significant disease for cattle producers in the United States, affecting 16.2% of cattle on feed. Cattle with advanced lung lesions at harvest have reduced average daily gain, yield grades, and carcass quality outcomes. The identification of biomarkers and clinical signs that accurately predict lung lesions could benefit livestock producers in determining a BRD prognosis. Herein, we used receiver operating characteristic curves to assess the predictive value of biomarkers and clinical signs associated with lung lesions after experimentally induced BRD. In the first 72 h after onset of BRD, right front stride length, gait velocity, visual analog scale score, clinical illness score, average activity level, step count, and rectal temperature yielded the best diagnostic accuracy (AUC 0.75) for predicting calves with significant lung lesions (10% consolidation) at necropsy. Biomarkers and clinical signs with the best diagnostic accuracy early in the disease process would likely be the most valuable in field conditions. These results can be used to guide refinement of the optimal time points and biomarkers for the diagnosis of significant lung lesions after BRD.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) descriptions of infection and transmission have been increasing in companion animals in the past year. Although canine susceptibility is generally considered low, their role in the COVID-19 disease cycle remains unknown. In this study, we detected and sequenced a delta variant (AY.3) from a 12-year-old Collie living with owners that previously tested positive for SARS-CoV-2. It is unclear if the dogs’ symptoms were related to SARS-CoV-2 infection or underlying conditions. The whole genome sequence obtained from the dog sample had several unique consensus level changes not previously identified in a SARS-CoV-2 genome that may play a role in the rapid adaptation from humans to dogs. Within the spike coding region, 5/7 of the subconsensus variants identified in the dog sequence were also identified in the closest in-house human reference case. Taken together, the whole genome sequence, and phylogenetic and subconsensus variant analyses indicate the virus infecting the animal originated from a local outbreak cluster. The results of these analyses emphasize the importance of rapid detection and characterization of SARS-CoV-2 variants of concern in companion animals.

Because black-tailed prairie dogs (Cynomys ludovicianus) are used as a model for research on gallstones and bacterial infections, performing urinary evaluations can provide invaluable data. This case report involves 5 prairie dogs that developed moist necrotic skin lesions after urine collection by cystocentesis. The information presented here serves as a resource regarding a potential adverse event that may develop after cystocentesis in black-tailed prairie dogs.

A 13-yr-old male cheetah (Acinonyx jubatus) presented for an acute history of lateral recumbency and anorexia. Upon physical examination under general anesthesia, severe icterus was noted. A serum biochemical profile confirmed markedly elevated total bilirubin and alanine transaminase. Based on ultrasound-guided liver aspirates and cytology, a presumptive diagnosis of large granular lymphocyte hepatic lymphoma was reached. Abdominal and thoracic radiographs did not assist in reaching an antemortem diagnosis. Postmortem examination and histopathology provided a definitive diagnosis of hepatic lymphoma with acute massive hepatocelluar necrosis and hemorrhage, as well as concurrent lesions of gastric ulcers, ulcerative and sclerosing enteritis, myocardial hypertrophy, and splenic myelolipomas. Immunohistochemistry of the liver yielded CD-3 positive and CD-20 negative results, confirming lymphocytes of a T-cell lineage. Due to concern for possible retrovirus-associated disease, enzyme-linked immunosorbent assays for feline leukemia virus and feline immunodeficiency virus were performed retrospectively on a banked serum sample and yielded negative results, thus diminishing concern for the male conspecific housed in the same exhibit.

A 10-yr-old, captive, intact male Asian small-clawed otter (Amblonyx cinereus) with a history of bilateral nephrolithiasis was presented for acute-onset lethargy and inappetance of 5 days duration. On physical examination, the otter was about 8% dehydrated and a palpable fluid wave was present in the abdomen. An abdominal ultrasound revealed hydronephrosis of the left kidney and a hyperechoic structure present within the lumen of the left ureter, causing an obstruction. A urinalysis revealed struvite crystalluria, bacteriuria, and an elevated pH. Following 4 days of antibiotic therapy, a left ureteronephrectomy was performed. Upon opening the kidney to retrieve calculi, a large amount of purulent material was noted within the renal pelvis. To the authors' knowledge, this is the first documented case of a nephrectomy in an Asian small-clawed otter. Nephrectomy should be considered as a viable option for treatment of ureteral obstruction, hydronephrosis, or severe pyelonephritis.

Feline cytauxzoonosis is a highly fatal tick-borne disease caused by a hemoparasitic protozoan, Cytauxzoon felis. This disease is a leading cause of mortality for cats in the Midwestern United States, and no vaccine or effective treatment options exist. Prevention based on knowledge of risk factors is therefore vital. Associations of different environmental factors, including recent climate were evaluated as potential risk factors for cytauxzoonosis using Geographic Information Systems (GIS).There were 69 cases determined to be positive for cytauxzoonosis based upon positive identification of C. felis within blood film examinations, tissue impression smears, or histopathologic examination of tissues. Negative controls totaling 123 were selected from feline cases that had a history of fever, malaise, icterus, and anorexia but lack of C. felis within blood films, impression smears, or histopathologic examination of tissues. Additional criteria to rule out C. felis among controls were the presence of regenerative anemia, cytologic examination of blood marrow or lymph node aspirate, other causative agent diagnosed, or survival of 25 days or greater after testing. Potential environmental determinants were derived from publicly available sources, viz., US Department of Agriculture (soil attributes), US Geological Survey (land-cover/landscape, landscape metrics), and NASA (climate). Candidate variables were screened using univariate logistic models with a liberal p value (0.2), and associations with cytauxzoonosis were modeled using a global multivariate logistic model (p0.05). Spatial heterogeneity among significant variables in the study region was modeled using a geographically weighted regression (GWR) approach.Total Edge Contrast Index (TECI), grassland-coverage, humidity conditions recorded during the 9(th) week prior to case arrival, and an interaction variable, "diurnal temperature range × percent mixed forest area" were significant risk factors for cytauxzoonosis in the study region. TECI and grassland areas exhibited significant regional differences in their effects on cytauxzoonosis outcome, whereas others were uniform.Land-cover areas favorable for tick habitats and climatic conditions that favor the tick life cycle are strong risk factors for feline cytauxzoonosis. Spatial heterogeneity and interaction effects between land-cover and climatic variables may reveal new information when evaluating risk factors for vector-borne diseases.

Equine multinodular pulmonary fibrosis, equine herpesvirus 5 (EHV-5), and multicentric lymphoma were discovered in one patient. Review of gamma herpesvirus activity in humans revealed a propensity for lymphoproliferative disorders associated with infection. The objective was to determine the frequency of EHV-5 in lymphoma tissues and compare with the frequency found in the lymph nodes of clinically normal horses. Case control investigation of lymphoma-positive tissues and analysis via polymerase chain reaction (PCR) for EHV-5 was performed on 12 horses. Prospective collection and PCR analysis of lymph nodes (mesenteric or submandibular) for EHV-5 was performed on 21 control horses. Thirteen samples of lymphoma-positive tissues and fluid were submitted for PCR analysis for EHV-5. Of these, 67% was positive. In the control horse population, 14% was positive for EHV-5 ( P = .004). Neoplastic samples positive for EHV-5 were classified as T-cell rich B-cell lymphoma (three), T-cell lymphoma (one), one was nondifferentiated, and two were not stained. Gamma herpesviruses in humans have been associated with lymphoproliferative diseases such as Kaposi sarcoma and Burkitt lymphoma. This study reveals an increased frequency of EHV-5 (gamma herpesvirus) in horses diagnosed with lymphoma compared with healthy control horses. Although the exact role this virus plays in the initiation or perpetuation of lymphoproliferative neoplasia is unknown, EHV-5 may be an etiologic agent associated with the development of some types of equine lymphoma.

A 9-year-old mare presented with a 2-week history of partial anorexia, multiple swellings in the area of the throatlatch, and purulent nasal discharge. On initial presentation, the horse had submandibular and retropharyngeal lymphadenopathy, mild ventral edema, and weight loss. Thoracic radiographs revealed a pulmonary interstitial pattern. Necropsy revealed enlarged lymph nodes throughout the body. The lung parenchyma contained multiple random, well-circumscribed nodules, which, on cut section, were pale, tan colored, and very firm with a distinct line of demarcation from the surrounding normal parenchyma. The subendocardium of the left ventricle, left atrium, and, multifocally, the right ventricle contained white, gritty areas of mineralization. There was marked subintimal mineralization of the aorta and pulmonary artery. Histopathology of the lymph nodes revealed effacement of the parenchyma by a neoplasm composed of large numbers of small mature lymphocytes, fewer large lymphocytes, and scattered moderate numbers of histiocytes. Immunohistochemistry tests for CD3, CD79a, and CD20, confirmed the lymphoma was T-cell-rich B-cell lymphoma. The lungs contained marked interstitial fibrosis with alveolar histiocytosis. Polymerase chain reaction test results of lymph node and lungs were positive for equine herpesvirus-5 (EHV-5), a gammaherpesvirus. Gammaherpesvirus infection has been associated with lymphoma and pulmonary fibrosis in other species. This report describes the association between EHV-5 and both pulmonary fibrosis and lymphoma.

Summary Cerebellar abiotrophy (CA) is an uncommon neurological disease that most commonly affects Arabian horses. Affected horses are typically identified within the first 6 months of life. Intention tremor, wide based stance and ataxia are common clinical signs observed in affected individuals. No treatment is available for resolution of clinical signs. Definitive diagnosis is based on histopathological examination of cerebellar tissue, which is characterised by loss of Purkinje cell layer. This report describes a case of cerebellar abiotrophy that had a delayed diagnosis until 6 years of age.

Five Bulldog pups, 4 weeks of age or younger, were presented over a 2-day period for postmortem examination and diagnostic evaluation. The pups originated from 2 different litters but had been cared for at a common facility since their birth. All 5 pups died after exhibiting symptoms consisting of lethargy, dyspnea, nasal discharge, anorexia, vomiting, diarrhea, and abdominal pain. Necropsy examination revealed locally extensive to diffusely red, firm, consolidated lungs in all pups. Histopathologically, the lungs were variably effaced by multifocal areas of necrosis. The alveolar lumens contained fibrin, edema fluid, macrophages, and neutrophils. Many of the bronchioles contained cellular debris and neutrophils admixed with sloughed bronchiolar epithelium, which often contained large intranuclear amphophilic inclusion bodies that peripherally displaced chromatin. Fluorescent antibody testing was positive for Canine adenovirus. An adenovirus isolated via cell culture was positive on direct fluorescent antibody test and was identified as Canine adenovirus serotype 2 via polymerase chain reaction. Electron microscopy revealed typical viral inclusions within bronchiolar epithelial cells. Hemolytic Escherichia coli was also isolated from the lungs in 3 of the 5 pups. The current case demonstrates a natural and rare fatal infection with a viral agent that is typically associated with immunosuppression in both animals and humans.

Sézary syndrome is an uncommon leukemic variant of cutaneous lymphoma in cats. This cat had recurrent dermatitis with erythematous, pruritic plaques. Multiple skin imprints and biopsy samples were obtained over a 6-month period, and histopathological findings were consistent initially with eosinophilic miliary dermatitis and later with erythema multiforme. One week before death, Sézary cells were identified in the peripheral blood that expressed cluster of differentiation (CD)3 and CD8 antigens. Massive infiltration of CD3+ lymphocytes was noted in the skin and multiple internal tissues by histopathological examination. This case demonstrates the difficulty in diagnosing cutaneous lymphoma early in the disease course.

A 15‐month‐old spayed female ferret (Mustela putorius furo) presented for lethargy and weight loss of 2 weeks duration. Upon physical examination, a 2‐mm‐diameter focal area of opacity was noted in the left cornea. In addition, the ferret was quiet, in poor body condition, and dehydrated. A complete blood count and plasma biochemistry revealed a severe nonregenerative anemia, azotemia, hyperproteinemia, hypoalbuminemia, and mild hyperphosphatemia and hyperchloremia. Urinalysis revealed hyposthenuria. Whole body radiographs showed multifocal thoracic nodular disease, splenomegaly, and renomegaly. Abdominal ultrasonography confirmed bilaterally enlarged kidneys, hypoechoic liver and spleen, and a caudal abdominal hypoechoic mobile nodule. The ferret was humanely euthanized, and a postmortem examination with subsequent histopathology showed multifocal necrotizing pyogranulomas in the lung, spleen, kidneys, mesenteric lymph nodes, and serosa of the duodenum. Pyogranulomatous panophthalmitis was diagnosed in the left eye. The multisystemic granulomatous lesions were suggestive of ferret systemic coronavirus (FRSCV). The presence of coronavirus in the left eye was confirmed by positive immunohistochemistry. Reverse transcriptase polymerase chain reaction (RT‐PCR) on formalin fixed paraffin embedded tissue from the lung, spleen, and kidney was negative for FRSCV and positive for ferret enteric coronavirus (FRECV). Systemic coronavirus disease in ferrets closely resembles feline infectious peritonitis (FIP) in domestic cats, which can manifest with anterior uveitis, chorioretinitis, optic neuritis, and retinal detachment. To the authors’ knowledge, this is the first report of ocular lesions in a ferret with systemic coronavirus disease, suggesting that ferrets presented with similar ocular lesions should also be evaluated for evidence of coronavirus infection.

An 11-year-old spayed-female German Shepherd dog was presented to the Veterinary Medical Teaching Hospital at Kansas State University with a history of weight loss, anorexia, depression, and lethargy for 2-3 weeks. Radiographic examination revealed a mass in the spleen and several round radiodense foci in the liver. CBC results included normocytic normochromic anemia, marked thrombocytopenia, and low numbers of neoplastic cells that frequently had cytoplasmic projections or blebs. A bone marrow aspirate contained about 80% neoplastic megakaryoblasts with the same microscopic features as those observed in peripheral blood. Using flow cytometry, cells of large size were identified in peripheral blood that expressed CD41/61, CD45, CD61, and CD62P (P-selectin) and were negative for markers of T cells, B cells, monocyte/macrophages, and dendritic cells. Because of the poor prognosis, euthanasia and subsequently necropsy were performed. On histopathologic examination, neoplastic megakaryoblasts were identified in spleen, liver, mesenteric lymph node, and the pulmonary vasculature. Using immunohistochemistry, the neoplastic megakaryoblasts weakly expressed von Willebrand factor. Based on microscopic and immunophenotypic findings, a diagnosis of acute megakaryoblastic leukemia (AMegL) was made. To our knowledge, this is the first report of AMegL in a domestic animal in which immunophenotyping by flow cytometry and a panel of antibodies against CD41/61, CD61, and CD62P were used to support the diagnosis.

A 13-year-old spayed female Labrador Retriever with a 1-year history of progressive exercise intolerance was diagnosed with an interventricular mass in the heart via echocardiogram. The animal's general condition progressively declined over the next 8 months, and it was euthanatized. The intracardiac mass, which protruded into the lumen of the right ventricle, was removed at necropsy and fixed in 10% buffered formalin. Histopathologic diagnosis was an ectopic thyroid carcinosarcoma based on the presence of 3 distinct neoplastic tissue types. Intermixed within the tumor were neoplastic thyroid follicles containing colloid and solid nests of thyroid follicular epithelial cells, vascular channels and clefts filled with blood and lined by neoplastic endothelium, and osteoid surrounded by spindle cells and often rimmed by large multinucleated cells. Immunohistochemical reaction for thyroglobulin was positive in the tumor cells forming the colloid-filled follicles and solid nests of epithelial cells. Neoplastic endothelium was positive for factor VIII-related antigen. The thyroid gland was located in its normal anatomic position and was histologically normal, ruling out the possibility that the intracardiac tumor was a metastatic lesion. To the authors' knowledge this is the first reported case of an intracardiac ectopic thyroid carcinosarcoma, and possibly the first ectopic thyroid carcinosarcoma in any location in any species.