Effect of synthetic agonists of toll-like receptor 9 on canine lymphocyte proliferation and cytokine production in vitro

Synthetic agonists of TLR9 containing novel DNA structures and R'pG (wherein R=1-(2'-deoxy-beta-d-ribofuranosyl)-2-oxo-7-deaza-8-methyl-purine) motifs, referred to as immune modulatory oligonucleotides (IMOs), have been shown to stimulate T(H)-1-type-immune responses and potently reverse allergen-induced T(H)-2 responses to T(H)-1 responses in vitro and in vivo in mice. In order to investigate the immunomodulatory potential of IMOs in dogs, canine peripheral blood mononuclear cells (PBMC) from healthy dogs were stimulated with three different IMOs and a control IMO, alone or in combination with concanavalin A (ConA). Lipopolysaccharide (LPS) was used as a positive control for B lymphocyte activation. Carboxyfluorescein diacetate succinimidyl ester and phenotype staining was used to tag proliferating T and B lymphocytes (CD5(+) and CD21(+)) by flow cytometry. Real-time PCR and ELISA were processed to assay cytokine production of IFN-gamma, IL-10, TGF-beta, IL-6 and IL-10. Like LPS, IMOs alone induced neither proliferation of CD5(+) T cells nor CD21(+) B cells, but both LPS and IMO had the capacity to co-stimulate ConA and induced proliferation of B cells. In combination with ConA, one of the IMOs (IMO1) also induced proliferation of T cells. IMO1 also significantly enhanced the expression of IFN-gamma on the mRNA and protein level in canine PBMC, whereas expression of IL-10, TGF-beta and IL-4 mRNAs was not induced by any of the IMOs. These results indicate that in canine PBMC from healthy dogs, IMO1 was able to induce a T(H)-1 immune response including T- and B-cell proliferation.     

Adverse Effects of Zilpaterol Administration in Horses: Three Cases

Three healthy horses were fed the beta-adrenergic agonist feed additive zilpaterol at a dosage of 0.17 mg/kg body weight to study zilpaterol elimination kinetics. Soon after ingestion of zilpaterol, the horses developed skeletal muscle tremors and tachycardia. A 75 to 87.5% reduced dose of zilpaterol was fed to the horses 24 hours after the initial dose; administration was discontinued thereafter. The horses exhibited restlessness, muscle tremors, and profuse sweating 20 to 25 minutes after ingestion of zilpaterol. Tachycardia developed within 40 minutes and took up to 2 weeks to resolve. Muscle tremors lasted up to 1 week. The most pronounced derangements in serum biochemistry were increased activities of lactic dehydrogenase, creatine kinase, and aspartate transferase, indicating muscle damage. The most severely affected horse also had transient azotemia, hematuria, and proteinuria, suggesting renal damage. All three horses recovered without treatment and were clinically normal 2 to 3 weeks after the initial dose of zilpaterol. Because of their anabolic properties, beta-adrenergic feed additives are considered a risk for abuse in performance horses, despite the absence of Food and Drug Administration approval for such use. Oral administration of zilpaterol to horses at the dosage indicated for use in cattle may result in prolonged adverse effects, including tachycardia, muscle tremors, and renal damage.     

Interlaboratory testing of porcine sera for antibodies to porcine circovirus type 2.

A panel of 20 porcine sera was distributed to 5 laboratories across Europe and Canada. Each center was requested to test the sera for the presence of porcine circovirus type 2 antibodies using the routine assays, indirect immunofluorescence assay (IFA) and indirect immunoperoxidase monolayer assay (IPMA), and to determine the titer of each serum. Results from all centers were then compiled and correlated. They demonstrate a wide variation in the titers obtained between laboratories. These differences were dependent on the assay used and the choice of fixative. In general, IPMA gave higher titers than did IFA, and paraformaldehyde gave higher titers than did acetone or ethyl alcohol. This report highlights the need for standardized procedures and biologicals for this virus.     

Identification of genomic alterations with clinical impact in canine splenic hemangiosarcoma

Canine hemangiosarcoma (HSA) is an aggressive cancer of endothelial cells with short survival times. Understanding the genomic landscape of HSA may aid in developing therapeutic strategies for dogs and may also inform therapies for the rare and aggressive human cancer angiosarcoma. The objectives of this study were to build a framework for leveraging real-world genomic and clinical data that could provide the foundation for precision medicine in veterinary oncology, and to determine the relationships between genomic and clinical features in canine splenic HSA. One hundred and nine dogs with primary splenic HSA treated by splenectomy that had tumour sequencing via the FidoCure® Precision Medicine Platform targeted sequencing panel were enrolled. Patient signalment, weight, metastasis at diagnosis and overall survival time were retrospectively evaluated. The incidence of genomic alterations in individual genes and their relationship to patient variables including outcome were assessed. Somatic mutations in TP53 (n = 44), NRAS (n = 20) and PIK3CA (n = 19) were most common. Survival was associated with presence of metastases at diagnosis and germline variants in SETD2 and NOTCH1. Age at diagnosis was associated with somatic NRAS mutations and breed. TP53 and PIK3CA somatic mutations were found in larger dogs, while germline SETD2 variants were found in smaller dogs. We identified both somatic mutations and germline variants associated with clinical variables including age, breed and overall survival. These genetic changes may be useful prognostic factors and provide insight into the genomic landscape of hemangiosarcoma.     

A prospective genetic evaluation of familial dilated cardiomyopathy in the Doberman pinscher

BACKGROUND: The Doberman Pinscher is one of the most common breeds of dogs to develop dilated cardiomyopathy (DCM), a primary heart muscle disorder characterized by myocardial dysfunction, cardiac arrhythmias, and congestive heart failure. In the Doberman Pinscher, the disease is typically adult onset, and a familial etiology has been suggested. HYPOTHESIS: DCM in the Doberman Pinscher, is a familial disease linked to a specific genetic marker. ANIMALS: The study comprised an extended family of Doberman Pinschers with a history of DCM. METHODS: Participating dogs were prospectively evaluated over an 8-year period. Phenotype of participating dogs was determined by annual echocardiography and ambulatory electrocardiography, and the pedigree was evaluated to determine a specific mode of inheritance. Three hundred seventy-two microsatellite markers were selected and genotyped to cover the 38 autosomal chromosomes. Phenotyping, genotyping, and pedigree information was entered into a database, and parametric, 2-point analysis was performed. Markers were considered to be linked to the development of DCM if the logarithm of odds LOD score was >/= 3.0. RESULTS: An autosomal dominant mode of inheritance was defined by the appearance of the disease in multiple generations, equal gender representation (P = .973) and male-to-male transmission. A maximum LOD score of 1.31 was obtained for I marker on chromosome 20, a score not high enough to be associated with DCM. CONCLUSION: DCM in the Doberman Pinscher is a familial disease inherited as an autosomal dominant trait. The causative gene(s) responsible for this condition remain unresolved. Association studies by means of array technology may provide new insights into gene identification.     

Antibiotic-resistant Corynebacterium jeikeium urinary tract infection in a cat

A 10-year-old, castrated male, domestic longhaired cat with a history of urinary tract disease and perineal urethrostomy was presented for evaluation of persistent urinary tract inflammation. Prior to referral, diphtheroid organisms had been cultured from a urine sample obtained by cystocentesis, and they were interpreted as sample contamination. Subsequent urine culture and gene sequencing identified Corynebacterium jeikeium, which was resistant to antibiotics and appeared to be the cause of the urinary tract infection.     

Histologic effect of semiconductor diode laser transscleral cyclophotocoagulation on the normal equine eye

Abstract Objective To determine the acute histologic effects of semiconductor diode laser transscleral cyclophotocoagulation (TSCP) on the normal equine eye. Animal studied Part 1: eight eyes of four horses. Part 2: 10 eyes of five horses. Materials and methods Part 1: TSCP was performed on four eyes at 4 mm and four eyes at 6 mm posterior to the limbus with 15 sites treated in four quadrants at 1800 mW for 1500 ms. The globes were sectioned transversely or sagitally to examine all quadrants and histologic sections were taken every 1 mm for the entire globe. Part 2: Based on the results from Part 1, TSCP was performed at 20 sites 4 mm posterior to the dorsotemporal limbus with a constant energy varying from 0.75 to 4 J/site. Histologic sections were taken every 1 mm for a total of 10 sections per eye and 20 sections per energy level group. Results Part 1: At 4 mm posterior to the limbus, coagulation of the nonpigmented epithelium (NPE) of the pars plicata was observed in the temporal (14%) and dorsal quadrants (12%). Retinal detachment was observed in the nasal quadrant (12%). Hemorrhage was common in the nasal (19%) and temporal (12%) quadrants. At 6 mm posterior to the limbus, coagulation of the NPE of the pars plicata was observed in the dorsal (14%), ventral (16%), nasal (2%), and temporal (2%) quadrants. Retinal detachment was observed in the dorsal (8%), ventral (18%), nasal (20%) and temporal (2%) quadrants. Part 2: Settings of 0.75 J/site were ineffective; 1.5, 2.25 and 3 J/site damaged the pars plicata without disruption of anatomy; and 4 J/site caused disruption of normal architecture. Conclusions The most appropriate site for equine TSCP appears to be 4 mm posterior to the dorso‐ and ventrotemporal limbus avoiding the 3 and 9 o’clock positions and using an initial energy setting of 2.25 J/site. This results in effective damage to the pars plicata while minimizing surgical complications such as retinal detachment and hemorrhage.     

Clinical characteristics of horses and foals diagnosed with cleft palate in a referral population: 28 cases (1988–2011)

The objective of this case series was to characterize the population, case presentations, and outcomes of 28 equids diagnosed with cleft palate over a 25-year period. The incidence of cleft palate was 0.04%. The median age at presentation was 2 mo (range: 1 d to 3 y). Fifty percent of the animals were < 2 mo old, 21% were ≥ 2 mo but < 1 y old, and 29% were 1 y of age or older. Males and females were nearly equally represented. Short-term outcomes included euthanasia in 50%, surgical repair in 11%, supportive care in 4%, and no treatment in 32% of cases; 46% of the animals survived to discharge. Defects involving both the hard and soft palate and/or aspiration pneumonia generally had less favorable outcomes. Though cleft palate is rare in horses, it should be considered as a differential diagnosis in horses of all ages with nasal discharge, a cough, a history of recurrent respiratory infections, poor growth, or chronic submandibular lymphadenopathy. Endoscopic evaluation of the pharynx may aid in earlier diagnosis and prognostication for owners.