Infectious keratoconjunctivitis in free-ranging mule deer in Wyoming: a retrospective study and identification of a novel alphaherpesvirus.

We describe the clinicopathologic findings, relative prevalence, and pathogens associated with infectious keratoconjunctivitis in mule deer ( Odocoileus hemionus) in Wyoming. Seventeen cases with ocular lesions were identified among 1,036 mule deer postmortem submissions (1.6%) in an ~16 y period. Sixteen cases were observed in winter and most were in male (15 cases) and juvenile (13 cases) deer. Blindness was the most commonly reported clinical sign (10 cases). A herpesvirus was detected only in the 4 cases of bilateral necrotizing bulbar conjunctivitis. Phylogenetic analysis of glycoprotein amino acid sequences consistently identified this virus as a novel alphaherpesvirus. In 2 of these herpesvirus-positive cases, Actinomyces sp. and Moraxella ovis were also identified. Trueperella pyogenes was identified in 4 cases of unilateral ulcerative keratitis, keratoconjunctivitis, and panophthalmitis. M. ovis was cultured from 3 cases of bilateral conjunctivitis and keratoconjunctivitis. In the remaining cases, isolates included Moraxella bovis (1 case), Staphylococcus sp. and Streptococcus sp. (2), Flavobacterium sp. and Pseudomonas sp. (2), Escherichia coli and Enterobacter sp. (1), and bovine viral diarrhea virus 1 (1). No pathogens were identified in 2 cases. The relative prevalence of keratoconjunctivitis in mule deer in Wyoming appears to be low, and this disease is most commonly associated with infection by a novel alphaherpesvirus, T. pyogenes, and M. ovis.

Canine distemper outbreak in pet store puppies linked to a high-volume dog breeder.

Canine distemper is uncommon in the pet trade in the United States, in large part due to effective vaccines against Canine distemper virus (CDV). This is a report of CDV affecting 24 young dogs of multiple breeds shortly after sale by 2 pet stores in Wyoming during August-October 2010. Cases were diagnosed over 37 days. Diagnosis was established by a combination of fluorescent antibody staining, reverse transcription polymerase chain reaction, negative stain electron microscopy, and necropsy with histopathology. Viral hemagglutinin gene sequences were analyzed from 2 affected dogs and were identical (GenBank accession no. JF283477). Sequences were distinct from those in a contemporaneous unrelated case of CDV in a Wyoming dog (JF283476) that had no contact with the pet store dogs. The breeding property from which the puppies originated was quarantined by the Kansas Animal Health Department. Puppies intended for sale were tested for CDV. Canine distemper was diagnosed on site in November 2010. At that point 1,466 dogs were euthanized to eliminate dispersal of the disease through commercial channels. The investigation underscores the risks inherent in large-scale dog breeding when vaccination and biosecurity practices are suboptimal.

Vascular mineralization in the brain of horses.

Vascular mineralization (siderocalcinosis) in the brain of horses has been usually assumed to be an incidental age-related finding with no clinic significance. In the present study, eight 15-32-year-old horses of different breeds with cerebral siderocalcinosis were studied. Four of these horses had acute and severe central nervous system clinical signs of unknown etiology, 2 horses had neurological signs of known cause, and 2 horses did not have neurological signs. Gross examination of the brains in 4 animals revealed symmetrical foci of malacia in the cerebellar white matter. Histologically, moderate to severe mineralization of blood vessels and parenchyma were observed in all 8 horses, occasionally associated with necrosis of the adjacent tissue. Some horses were tested by virus isolation, polymerase chain reaction, immunohistochemistry, and serology to investigate Rabies virus; West Nile virus; Equid herpesvirus 1 and 4; Eastern, Western, Venezuelan, and Saint Louis encephalitis virus; and Sarcocystis neurona infection. These tests were negative in all samples analyzed. Brain cholinesterase activity and heavy metal screening were also unremarkable. The significance of the vascular and parenchymal mineralization in the brains of some of these horses remains undetermined. However, the severity of the lesions observed in the brains of some of the animals in the present study, coupled with the negative results for other common causes of neurological disease in horses, suggests a possible relationship between siderocalcinosis and the clinical signs observed.

Restricted enzooticity of hepatitis E virus genotypes 1 to 4 in the United States.

Hepatitis E is recognized as a zoonosis, and swine are known reservoirs, but how broadly enzootic its causative agent, hepatitis E virus (HEV), is remains controversial. To determine the prevalence of HEV infection in animals, a serological assay with capability to detect anti-HEV-antibody across a wide variety of animal species was devised. Recombinant antigens comprising truncated capsid proteins generated from HEV-subgenomic constructs that represent all four viral genotypes were used to capture anti-HEV in the test sample and as an analyte reporter. To facilitate development and validation of the assay, serum samples were assembled from blood donors (n = 372), acute hepatitis E patients (n = 94), five laboratory animals (rhesus monkey, pig, New Zealand rabbit, Wistar rat, and BALB/c mouse) immunized with HEV antigens, and four pigs experimentally infected with HEV. The assay was then applied to 4,936 sera collected from 35 genera of animals that were wild, feral, domesticated, or otherwise held captive in the United States. Test positivity was determined in 457 samples (9.3%). These originated from: bison (3/65, 4.6%), cattle (174/1,156, 15%), dogs (2/212, 0.9%), Norway rats (2/318, 0.6%), farmed swine (267/648, 41.2%), and feral swine (9/306, 2.9%). Only the porcine samples yielded the highest reactivities. HEV RNA was amplified from one farmed pig and two feral pigs and characterized by nucleotide sequencing to belong to genotype 3. HEV infected farmed swine primarily, and the role of other animals as reservoirs of its zoonotic spread appears to be limited.

Experimental reproduction of tumbleweed shield lichen (Xanthoparmelia chlorochroa) poisoning in a domestic sheep model.

Xanthoparmelia chlorochroa, commonly called tumbleweed lichen, is found throughout the Rocky Mountain region. This particular species of lichen was incriminated in the poisoning of cattle and sheep in Wyoming during the 1930s. More than 70 years elapsed before another case was reported. There is virtually no information in the veterinary literature regarding toxicity of this lichen. This report describes X. chlorochroa poisoning in domestic sheep fed lichen collected from different locales and at different times of the year. Affected animals voided red urine and displayed incoordination. A transient spike in serum creatine kinase activity occurred in all ewes during the course of the feeding trial. Histologically, necrosis of a few individual appendicular skeletal myocytes was observed in 1 ewe, but grossly discernible myonecrosis was absent. The severity of clinical signs varied depending on the location and/or time of year the lichen was collected, indicating that toxicity of the lichen may be influenced by environmental conditions. Results demonstrate that domestic sheep are a useful model for further investigation of X. chlorochroa intoxication. The current study should act as a starting point for elucidating the pathogenesis of X. chlorochroa poisoning and aid in the development of a diagnostic assay to confirm lichen poisoning.

Tularemia in range sheep: an overlooked syndrome?

Abortion and death caused by Francisella tularensis were well recognized in range flocks of domestic sheep in Idaho, Montana, and Wyoming in the first 6 decades of the 20th century. The current report describes 4 episodes of tularemia in 3 range flocks in Wyoming and South Dakota in 1997 and 2007 (1 flock was affected twice). Flock owners reported that ticks were unusually numerous and commonly present on sheep during outbreaks. Tularemia presented as late-term abortions (3 episodes) or listlessness and death in lambs and, to a lesser extent, ewes (1 episode). Lesions were multifocal pinpoint necrotic foci in tissues, particularly spleen, liver, and lung. An immunohistochemical procedure demonstrated F. tularensis, particularly in necrotic foci. The diagnosis was corroborated by bacterial isolation and, in individual cases, by serology, fluorescent antibody assay, and/or polymerase chain reaction detection of F. tularensis. Diagnosticians in endemic areas should include tularemia as a differential diagnosis when investigating late-term abortions or outbreaks of fatal illness in young lambs, particularly in years of high tick activity and when characteristic necrotic foci occur in spleen, liver, and lung.

Acute non-cytopathic bovine viral diarrhea virus infection induces pronounced type I interferon response in pregnant cows and fetuses.

Bovine viral diarrhea virus (BVDV) infection occurs in the cattle population worldwide. Non-cytopathic (ncp) BVDV strains cause transient infection (TI) or persistent infection (PI) depending on the host's immune status. Immunocompetent adult animals and fetuses in late gestation resolve the infection. Fetal infection in early gestation results in PI with chronic viremia and life-long viral shedding, ensuring virus perpetuation in the population. Eighteen pregnant heifers, divided into three groups, were intranasally inoculated with ncp BVDV2 virus early (day 75) and late (day 175) in gestation, or kept BVDV-naïve. Fetuses were retrieved on day 190. Antiviral activity in blood of dams and fetuses, maternal expression of interferon (IFN) stimulated gene 15kDa (ISG15), virological and serological status of heifers and fetuses, and fetal growth were studied. A pronounced antiviral activity in blood of heifers and TI fetuses during acute BVDV infection was accompanied by drastic up-regulation of ISG15 mRNA in maternal blood. Only one PI fetus expressed low IFN response 115 days post inoculation despite high BVDV antigen and RNA levels. PI fetuses presented with growth retardation. Infection of pregnant heifers with ncp BVDV2 early in gestation adversely affects fetal development and antiviral responses, despite protective immune responses in the dam.

Serologic and reproductive findings after a herpesvirus-1 abortion storm in goats.

CASE DESCRIPTION: An abortion storm occurred in a goat herd, resulting in 75 aborted kids and 1 neonatal death from December 2004 to February 2005. CLINICAL FINDINGS: Aborted fetuses ranged from being premature to past term. Laboratory findings in 4 of 5 aborted fetuses were consistent with herpesvirus abortion. A virus that yielded positive results with a fluorescent antibody test for bovine herpesvirus-1 was isolated and identified as caprine herpesvirus-1 (CpHV-1) via DNA sequence analysis. TREATMENT AND OUTCOME: Many does that aborted were rebred for kidding in late summer. Most of the young wethers born in 2005 were sold; however, all of the young does were kept for breeding in fall. In November 2005, all 241 goats in the herd were tested for antibodies against CpHV-1 to identify goats that had seroconverted during the outbreak. No complications attributable to CpHV-1 were identified during kidding in 2006. CLINICAL RELEVANCE: On the basis of serologic findings, infection with CpHV-1 was not associated with reduced reproductive success in the subsequent breeding.

Application of fluorescent in situ hybridisation for demonstration of Coxiella burnetii in placentas from ruminant abortions.

A fluorescent in situ hybridisation (FISH) assay targeting 16S ribosomal RNA was developed for detection of the zoonotic bacterium Coxiella burnetii in formalin-fixed, paraffin-embedded tissue, and applied on placentas from ruminant abortions. The applicability of the FISH assay was compared to immunohistochemistry (IHC) using human positive control serum in 12 cases of C. burnetii-associated placentitis as well as 7 negative control tissue samples. In all 12 cases the bacterium was detected within trophoblasts as well as free in the placental debris by both FISH and IHC. Extensive and significant infection by C. burnetii was revealed in 10 of the cases, whereas a slighter and focal distribution of the bacterium was observed in two cases. 90 aborted placentas from Danish ruminants were investigated by FISH. C. burnetii was detected in one bovine case only, representing the first confirmation of C. burnetii in Danish animals. The study shows that FISH targeting 16S ribosomal RNA is a feasible diagnostic tool for detection of C. burnetii in tissue samples and fully comparable to IHC.

Attenuation of acute rejection in a rat liver transplantation model by a liver-targeted dextran prodrug of methylprednisolone.

BACKGROUND: The use of methylprednisolone (MP) and other corticosteroids for the treatment of acute liver allograft rejection is associated with severe toxicities in nontarget tissues. Therefore, selective delivery of MP to the liver may improve its efficacy and alleviate its side effects. We investigated the effects of a novel liver-targeted dextran prodrug of MP (DMP) in an orthotopic rat liver transplantation (OLT) model. METHODS: The model consisted of a high responder rejection strain combination (Dark Agouti donors and Lewis recipients). Liver recipients were intravenously administered saline or a single subtherapeutic dose of MP (5 mg/kg) as the parent drug (MP) or its prodrug (DMP). Different groups were then monitored for graft survival or euthanized 5 or 9 days posttransplantation. Plasma chemistry, including alkaline phosphatase and bilirubin, allograft histology, and survival duration were determined. RESULTS: Untreated recipients exhibited elevated plasma levels of liver injury markers, progressive portal and venous inflammation and cellular infiltration in liver allografts, and a mean graft survival time (MST) of 10.5 days. MP treatment did not alter any of these parameters. In contrast, a single dose of DMP resulted in a decrease in plasma levels of liver injury markers, a decrease in histological grade of rejection on day 5, and a substantial increase in MST (27.5 days). CONCLUSIONS: These results demonstrate attenuation of acute rejection following local (allograft) immunosuppression with a single subtherapeutic dose of MP delivered as a liver-targeted prodrug. Dextran prodrugs may be useful for selective delivery of immunosuppressants to the liver following liver transplantation.

Prevalence, outcome, and health consequences associated with persistent infection with bovine viral diarrhea virus in feedlot cattle.

OBJECTIVE: To estimate prevalence of cattle persistently infected (PI) with bovine viral diarrhea virus (BVDV) at arrival at a feedlot, prevalence of chronically ill and dead PI cattle, and the magnitude of excess disease attributable to a PI animal. DESIGN: Cross-sectional and cohort studies. ANIMALS: 2,000 cattle at the time they arrived at a feedlot, 1,383 chronically ill cattle from 7 feedlots, and 1,585 dead cattle from a single feedlot. PROCEDURE: Skin biopsy specimens were collected and evaluated via immunohistochemistry. Cattle were characterized as either PI or not PI with BVDV on the basis of characteristic immunostaining. Follow-up was obtained for the 2,000 cattle from which samples were collected at arrival, and health outcomes were determined for cattle exposed and not exposed to a PI animal. RESULTS: Prevalence of PI cattle was 0.3% at arrival, 2.6% in chronically ill cattle, and 2.5% in dead cattle. Risk of initial treatment for respiratory tract disease was 43% greater in cattle exposed to a PI animal, compared with those not exposed to a PI animal. Overall, 15.9% of initial respiratory tract disease events were attributable to exposure to a PI animal. CONCLUSIONS AND CLINICAL RELEVANCE: Relatively few PI cattle arrive at feedlots. However, those cattle are more likely to require treatment for respiratory tract disease and either become chronically ill or die than cattle that are not PI. In addition, they are associated with an increase in the incidence of respiratory tract disease of in-contact cattle.