A retrospective histologic study of 140 cases of clinically significant equine ocular disorders.

Ocular diseases are an important category in equine medicine; however, most articles regarding histologic ocular lesions in horses are exclusive to a specific condition and do not provide a complete review of clinically significant ocular disease frequency in a diagnostic laboratory. We reviewed sections of equine eyes from 140 cases (98 enucleations [biopsies] and 42 autopsies) with clinically relevant ocular alterations at 2 diagnostic centers in the United States. The most common primary conditions were non-traumatic keratitis (36), equine recurrent uveitis (ERU; 31), traumatic injuries (22), ocular and periocular neoplasms (19), and uveitis and/or endophthalmitis resulting from sepsis (18). Congenital anomalies (3) and retinal atrophy and detachment alone (3) were infrequent. Non-traumatic keratitis was frequently accompanied by anterior uveitis (22), corneal rupture (16), pre-iridal fibrovascular membrane formation (13), and secondary mycotic infection (11). ERU was the second and third most prevalent disease in autopsies and enucleations, respectively. This condition was commonly associated with glaucoma (15). Glaucoma (25) and cataract (20) were the most prevalent secondary alterations in the evaluated cases. Keratitis (20) and corneal rupture (16) were among the most prevalent consequences of trauma. Information presented herein may guide clinicians and pathologists, contributing to the early diagnosis of potentially vision-impairing conditions and raising the chances of successful treatment and cure.

Toxicity and Efficacy Evaluation of an Adeno-Associated Virus Vector Expressing Codon-Optimized RPGR Delivered by Subretinal Injection in a Canine Model of X-linked Retinitis Pigmentosa.

Applied Genetic Technologies Corporation (AGTC) is developing a recombinant adeno-associated virus (rAAV) vector AGTC-501, also designated rAAV2tYF-GRK1-hRPGRco, to treat X-linked retinitis pigmentosa (XLRP) in patients with mutations in the retinitis pigmentosa GTPase regulator (RPGR) gene. The vector contains a codon-optimized human RPGR cDNA (hRPGRco) driven by a photoreceptor-specific promoter (G protein-coupled receptor kinase 1 [GRK1]), and is packaged in an AAV2 capsid variant with three surface tyrosine residues changed to phenylalanine (AAV2tYF). We conducted a toxicity and efficacy study of this vector administered by subretinal injection in the naturally occurring RPGR mutant (X-linked progressive retinal atrophy 2 [XLPRA2]) dog model. Sixteen RPGR mutant dogs divided into four groups of three to five animals each received either a subretinal injection of 0.07 mL of AGTC-501 at low (1.2 × 1011 vector genome [vg]/mL), mid (6 × 1011 vg/mL), or high dose (3 × 1012 vg/mL), or of vehicle control in the right eye at early-stage disease. The left eye remained untreated. Subretinal injections were well tolerated and were not associated with systemic toxicity. Electroretinography, in vivo retinal imaging, and histological analysis showed rescue of photoreceptor function and structure in the absence of ocular toxicity in the low- and mid-dose treatment groups when compared with the vehicle-treated group. The high-dose group showed evidence of both photoreceptor rescue and posterior segment toxicity. These results support the use of AGTC-501 in clinical studies with patients affected with XLRP caused by RPGR mutations and define the no-observed-adverse-effect level at 6 × 1011 vg/mL.

Theileriosis in Multiple Neonatal White-tailed Deer ( Odocoileus virginianus) in Delaware, USA.

Postmortem examination of 21 neonatal white-tailed deer ( Odocoileus virginianus) from Delaware, US identified six fawns with Theileria spp. organisms or suspected infection.

Evaluation of the genes encoding CD39/NTPDase-1 and CD39L1/NTPDase-2 in horses with and without abnormal hemorrhage and in horses with pathologic evidence of exercise-induced pulmonary hemorrhage.

BACKGROUND: Exercise-induced pulmonary hemorrhage (EIPH) is a common disorder of equine athletes. The role of polymorphisms in genes encoding hemostasis-regulatory proteins in horses with abnormal hemorrhage is unknown. OBJECTIVES: The goal of this study was to evaluate the genes encoding 2 ectonucleotidases, CD39/NTPDase-1 and CD39L1/NTPDase-2, and one ecto-5' nucleotidase, CD73, in horses with abnormal hemorrhage or pathologic changes consistent with EIPH. METHODS: Twenty-three horses with histories of abnormal hemorrhage, 8 horses with gastrointestinal signs, and 45 healthy horses were evaluated using polymerase chain reaction-based techniques. Formalin-fixed tissues from 21 horses with pathologic changes consistent with EIPH were also evaluated. RESULTS: Three single nucleotide polymorphisms (SNPs) were identified in the gene encoding CD39 and one SNP was identified in the gene encoding CD39L1. No SNPs were identified in the gene encoding CD73. CD39 SNPs were identified in 19 of 20 (95%) horses with unexplained hemorrhage and 20 of 21 (95%) horses with pathologic features consistent with EIPH. CD39L1 SNPs were identified in 6 of 20 (30%) horses with unexplained hemorrhage and 8 of 21 (38%) horses with pathologic features consistent with EIPH. CD39 and CD39L1 SNPs were identified in 5 of 8 (62.5%) and one of 8 (12.5%) horses, respectively, presenting with colic or weight loss. CD39 and CD39L1 SNPs were identified in 28 of 45 (62%) and 13 of 45 (28.8%) healthy horses, respectively. CONCLUSIONS: CD39 and CD39L1 are critically important in maintaining normal hemostasis and limiting inflammation. Further studies are needed to evaluate their role in the pathogenesis of equine EIPH.

Hepatocellular carcinomas in Vietnamese pot-bellied pigs (Sus scrofa).

Various neoplasms have been reported in Vietnamese pot-bellied pigs (Sus scrofa) with few reports of hepatocellular tumors. Twenty-two pot-bellied pigs diagnosed with hepatocellular carcinoma at necropsy over a 3-year period at one institution are described, representing 29% of the total pot-bellied pigs necropsied. The average age of affected pigs was 16.6 years with 15 males and 7 females. The most common clinical signs were decreased appetite (16/22) and weight loss (7/22). Grossly, the majority were massive tumors (13/22) with fewer nodular tumors (8/22) and 1 diffuse tumor. Massive tumors were typically multilobulated, very large, and encompassing 1 or more adjacent liver lobes, and were soft to firm and tan-yellow to orange-brown. Nodular tumors had multiple, 1-15 cm in diameter, discrete nodules in multiple liver lobes. Gross evidence of abscesses, necrosis, hemorrhage, or cysts associated with the tumor was occasionally described. Half of the cases had possible intrahepatic metastasis, and extrahepatic metastasis was identified in 3 cases, including to the hepatic lymph node (1/3), lung (2/3), spleen (1/3), and kidney (1/3). Histologically, all tumors had a trabecular or solid pattern, or a combination. An adenoid pattern was only identified in small regions of a few tumors. The neoplastic cells were relatively well-differentiated with moderate pleomorphism and a low mitotic index. Other histologic features within the tumors included intracellular glycogen or lipid accumulation, extramedullary hematopoiesis, foci of coagulative necrosis, and bile stasis. Aged pot-bellied pigs can be predisposed to hepatocellular carcinomas, which are locally aggressive and can metastasize within the liver and to other organs.

Intraocular nematodiasis in a llama (Lama glama).

This report describes a unique case of presumed migration of Parelaphastrongylus tenuis through the spinal cord into the eye of a llama where it survived and matured within the ocular environment. Blindness of the eye was most likely attributable to migration of the parasite through the central nervous tissue.

Generalized cerebral atrophy seen on MRI in a naturally exposed animal model for Creutzfeldt-Jakob disease.

BACKGROUND: Magnetic resonance imaging has been used in the diagnosis of human prion diseases such as sCJD and vCJD, but patients are scanned only when clinical signs appear, often at the late stage of disease. This study attempts to answer the questions "Could MRI detect prion diseases before clinical symptoms appear?, and if so, with what confidence?" METHODS: Scrapie, the prion disease of sheep, was chosen for the study because sheep can fit into a human sized MRI scanner (and there were no large animal MRI scanners at the time of this study), and because the USDA had, at the time of the study, a sizeable sample of scrapie exposed sheep, which we were able to use for this purpose. 111 genetically susceptible sheep that were naturally exposed to scrapie were used in this study. RESULTS: Our MRI findings revealed no clear, consistent hyperintense or hypointense signal changes in the brain on either clinically affected or asymptomatic positive animals on any sequence. However, in all 37 PrPSc positive sheep (28 asymptomatic and 9 symptomatic), there was a greater ventricle to cerebrum area ratio on MRI compared to 74 PrPSc negative sheep from the scrapie exposed flock and 6 control sheep from certified scrapie free flocks as defined by immunohistochemistry (IHC). CONCLUSIONS: Our findings indicate that MRI imaging can detect diffuse cerebral atrophy in asymptomatic and symptomatic sheep infected with scrapie. Nine of these 37 positive sheep, including 2 one-year old animals, were PrPSc positive only in lymph tissues but PrPSc negative in the brain. This suggests either 1) that the cerebral atrophy/neuronal loss is not directly related to the accumulation of PrPSc within the brain or 2) that the amount of PrPSc in the brain is below the detectable limits of the utilized immunohistochemistry assay. The significance of these findings remains to be confirmed in human subjects with CJD.

Effect of subcutaneous administration of a killed Mycobacterium avium subsp paratuberculosis vaccine on colonization of tissues following oral exposure to the organism in calves.

OBJECTIVE-To evaluate the effect of vaccination of calves with a killed Mycobacterium avium subsp paratuberculosis (MAP) vaccine on colonization of tissues following oral MAP exposure. ANIMALS-12 healthy Holstein calves. PROCEDURES-At 14 days after birth, calves received the MAP vaccine (1.0 mL, SC) or saline (0.9% NaCl) solution (1.0 mL, SC [control treatment]). Each calf received 1.2 x 10(9) CFUs of live MAP orally 21 and 22 days after vaccination. Prior to vaccination and at subsequent intervals, a blood sample was collected for ELISA detection of antibodies against MAP and for whole blood, antigen-specific, interferon (IFN)-gamma-release assay. Nine weeks after MAP challenge, calves were euthanized and various tissue samples were collected for mycobacterial culture. Interferon-gamma production in prescapular lymph node cells was measured following in vitro stimulation with MAP antigens. RESULTS-Calves were seronegative for anti-MAP antibodies at all times. Compared with the findings in control calves, antigen-specific IFN-gamma production in circulating lymphocytes and prescapular lymph node cells from vaccinated calves was significantly higher. Culture of tissues from vaccinated calves yielded significantly fewer CFUs of MAP (2,417 CFUs/g), compared with tissues from control calves (15,709 CFUs/g). Furthermore, significantly fewer tissue samples from vaccinated calves yielded MAP in culture (21.8 tissues/calf), compared with findings in control calves (27.6 tissues/calf). CONCLUSIONS AND CLINICAL RELEVANCE-Inoculation of calves with a killed MAP vaccine was associated with reduced colonization of intestinal tissues following experimental exposure to MAP. Use of the vaccine could potentially reduce transmission of MAP to calves in infected herds.

Agreement in histologic assessments of the pituitary pars intermedia in aged horses.

OBJECTIVE: To evaluate concordance among veterinary pathologists in the assessment of histologic findings in the pars intermedia of pituitary gland sections from aged horses with mild signs suggestive of pituitary pars intermedia dysfunction (PPID). Sample Population-10 pituitary glands from aged horses. PROCEDURE: 7 pathologists were provided with signalment, clinical signs, and a single H&E-stained pituitary gland section from 10 aged horses with mild signs suggestive of PPID. Pathologists described histologic findings for each section and stated whether findings were consistent with PPID. Agreement among pathologists and with antemortem diagnostic test results was calculated. RESULTS: Overall, only fair agreement was found among the pathologists as to which horses had histologic findings consistent with disease (mean +/- SE kappa value, 0.34 +/- 0.069). Interpretation of individual sections varied, with minimal agreement (4 or 5/7 pathologists) for 5 of 10 sections evaluated. Postmortem assessment was in agreement with an antemortem endocrine diagnostic test result 79% of the time. CONCLUSIONS AND CLINICAL RELEVANCE: Validation of antemortem diagnostic testing for PPID in horses often relies on the results of postmortem histologic evaluation. The lack of consensus in histologic interpretation of pituitary glands from aged horses with mild clinical signs in our study indicates that postmortem histologic evaluation of pituitary glands is an inappropriate standard in validation of antemortem diagnostic tests for detection of early PPID. Caution should be used when interpreting diagnostic test results in horses in which early PPID is suspected.

Zinc phosphide intoxication of wild turkeys (Meleagris gallopavo).

Zinc phosphide (Zn3P2) is a rodenticide used to control a variety of small mammal species. It is available over-the-counter or as a restricted-use pesticide depending on how it is to be applied. The toxicity of Zn3P2 is dependent on the species exposed, whether the animal is able to vomit or not, and whether it is ingested on a full or empty stomach. Nontarget species can be exposed through inadvertent or intentional product misapplication. In this article we describe four mortality events in which wild turkeys (Meleagris gallopavo) were believed to have been intoxicated following the ingestion of baits containing Zn3P2.

Myonecrosis and cutaneous infarction associated with Salmonella serovar Infantum infection in a horse.

A 5-year-old Quarter Horse mare was referred for evaluation of oral ulcers, limb edema, weight loss, and weakness. There was marked diffuse swelling extending from the stifle region to the tarsal region of the left hind limb, and the horse had a left hind limb lameness. Firm swellings ranging from 2 to 15 cm in diameter and consisting of nodules, plaques, and discrete masses were palpated on both sides of the neck, over the right shoulder region, over the left elbow region, and over the left caudoventral aspect of the abdomen. Laboratory abnormalities included hypoproteinemia, neutrophilia, and hyperfibrinogenemia. Results of ultrasonographic examination of the left hind limb and masses were suggestive of muscle edema, necrosis, and hemorrhage. Histologic examination of a biopsy specimen from a subcutaneous mass revealed necrotizing, suppurative myositis. The horse's condition gradually deteriorated, and the horse was euthanatized. Necropsy revealed myonecrosis, cutaneous infarcts, hepatic abscesses, and cholangitis. Salmonella serovar Infantum was cultured from liver and muscle lesions, and a diagnosis of Salmonella myonecrosis was made.

Efficacy of commercial and field-strain Mycobacterium paratuberculosis vaccinations with recombinant IL-12 in a bovine experimental infection model.

The efficacy of commercial (Strain 18) and field-isolate paratuberculosis vaccine preparations was investigated. The effect of prior exposure to Mycobacterium paratuberculosis and the adjuvant effect of rIL-12 on vaccine efficacy were also tested. Both Strain 18 and field-isolate vaccines induced strong local, systemic and enteric IFN-gamma responses. A significant reduction in mycobacterial colonization was observed when calves were vaccinated with the field-isolate prior to challenge, but not following vaccination with Strain 18 vaccine. Vaccination with rIL-12 prevented infection in some calves but its overall effect on IFN-gamma response and total mycobacterial load was not statistically significant. Efficacy of paratuberculosis vaccines may be enhanced if calves are vaccinated prior to M. paratuberculosis exposure with field-isolate vaccine instead of Strain 18 vaccine currently in use.