Monoclonal antibodies to equine interferon-alpha (IFN-alpha): new tools to neutralize IFN-activity and to detect secreted IFN-alpha

Interferon-alpha (IFN-alpha) is a type I interferon that is secreted during the early stages of the innate immune response and is often induced upon infection with viral pathogens. IFN-alpha production affects multiple downstream events influencing both innate and adaptive immune responses. Here, we describe the expression of an equine rIFN-alpha/IgG4 fusion protein in mammalian cells. The anti-viral activity of rIFN-alpha/IgG4 was found to be 70-fold higher than that of a previously described IFN-gamma/IgG1 as tested by bioassay. The purified rIFN-alpha was subsequently used for the generation of six monoclonal antibodies (mAbs) to equine IFN-alpha. Four of these mAbs inhibited the protective anti-viral effect of equine leukocyte IFN in bioassays. One mAb (clone 240-2) showed a high-neutralizing capacity. An ELISA was established using two anti-equine IFN-alpha mAbs (clones 29B and 240-2) and its analytical sensitivity for was found to be around 800 pg/ml and 3 U/ml for rIFN-alpha and equine leukocyte IFN, respectively. When analyzing samples with a likely dominance of IFN-alpha among type I IFNs, such as supernatants from equine peripheral blood mononuclear cells stimulated with CpG-oligodeoxyribonucleotides, the results obtained by ELISA and IFN bioassay showed a high agreement (r(2)(sp)=0.98). When analyzing samples likely containing a mixture of type I IFNs, such as serum and nasal secretions from virally infected horses, the ELISA only detected some of the IFN-activity recorded in the bioassay. Overall, the data showed that the new anti-equine IFN-alpha mAbs are valuable tools to detect native IFN-alpha for further characterization of the early innate immune response and anti-viral immunity in horses.     

Toxicity of the lichen secondary metabolite (+)-usnic acid in domestic sheep

Toxicity following ingestion of the vagrant, foliose lichen Xanthoparmelia chlorochroa was identified as the putative etiology in the death of an estimated 400-500 elk on the Red Rim-Daley Wildlife Habitat Management Area in Wyoming during the winter of 2004. A single, unsubstantiated report in 1939 attributed toxicity of X. chlorochroa in cattle and sheep to usnic acid, a common lichen secondary metabolite. To test the hypothesis that usnic acid is the proximate cause of death in animals poisoned by lichen, domestic sheep were dosed PO with (+)-usnic acid. Clinical signs in symptomatic ewes included lethargy, anorexia, and signs indicative of abdominal discomfort. Serum creatine kinase, aspartate aminotransferase, and lactate dehydrogenase activities were considerably elevated in symptomatic sheep. Similarly, only symptomatic ewes exhibited appreciable postmortem lesions consisting of severe degenerative appendicular skeletal myopathy. The median toxic dose (ED(50)) of (+)-usnic acid in domestic sheep was estimated to be between 485 and 647 mg/kg/day for 7 days.     

Comparison of the somatotropic axis in free-ranging and rehabilitated harbor seal pups (Phoca vitulina)

The somatotropic axis, including growth hormone (GH), insulin-like growth factor (IGF)-I, and IGF binding proteins (IGFBP), is a bridge between growth physiology, developmental age, and nutritional status in domestic animals. However, the importance of the somatotropic axis in nutrition, growth, and development of harbor seals has not been previously explored. Given the difficulty of conducting longitudinal studies in free-ranging harbor seals, this study focused on the potential use of harbor seals in rehabilitation facilities as a model for free-ranging seals. The purpose of this research was to compare concentrations of components of the somatotropic axis in free-ranging versus rehabilitated harbor seal pups. The hypothesis was that measurements of the somatotropic axis will be similar between individuals of comparable age and nutritional status (fasting versus feeding). To investigate this hypothesis, harbor seal pups (n=8) brought to The Marine Mammal Center (Sausalito, California, USA) or Mystic Aquarium (Mystic, Connecticut, U.S.A.) were initially assessed and determined to be healthy but abandoned. All pups were less than 2 wk of age upon arrival at rehabilitation facilities. Standard length was assessed at the time of arrival and again at release. Body mass was measured every week and blood samples were collected from each pup at 0, 4, and 8 wk of rehabilitation. Blood was collected and morphometrics assessed in free-ranging harbor seal pups (n=8) from the Gulf of Maine. Sera were analyzed for GH, IGF-I, and IGFBP concentrations. Concentrations of GH, IGF-I, and IGFBP-2 and -3 in rehabilitated pups were within a similar range compared with free-ranging pups when considered in the context of presumed nutrient intake. These data suggest that rehabilitated harbor seals may provide a useful model to investigate the effects of nutrient intake on growth and development of harbor seals, and will provide insight into phocid endocrinology and metabolism.     

Paresis in an Asian small clawed otter (Aonyx cinereus) associated with vertebral and ischial osteolysis caused by a malignant lymphangiosarcoma

A 10-yr-old male intact Asian small clawed otter (Aonyx cinerus) was presumptively diagnosed by histopathology and immunohistochemistry with lymphangiosarcoma after bony destruction of the ischium and spinal column from local tumor invasion had caused progressive signs of hind limb lameness and paresis/paralysis, which led to humane euthanasia. At necropsy, the primary tumor was identified as a flocculent mass present under the caudal lumbar vertebrae. Multiple nerves were seen to run from the spinal cord into the wall of the mass. This mass had locally invaded the surrounding muscle, vertebral column, and spinal cord, which led to the clinical signs noted at presentation. Bony destruction was severe with almost complete obliteration of the right ischium and osteolysis of L6, exposing the spinal cord beneath. The tumor had metastasized to at least two different sites within the spleen. The abdominal tumor was confirmed to be of endothelial origin by the use of immunohistochemical staining for factor VIII-related antigen and was confirmed as lymphatic origin versus vascular origin because of the lack of red blood cells within the vessels. The length of time from initial presentation with hind limb lameness to euthanasia because of hind limb paralysis was 4 mo. This is the first report of lymphangiosarcoma, an uncommon malignant neoplasm of lymphatic origin, in a mustelid and the first report of neoplastic disease in an Asian small clawed otter. In addition, the presentation of hind limb paresis associated with bony lysis because of local tumor invasion has not been previously reported with lymphangiosarcoma in humans, domestic animals, or nondomestic animals.     

Dough Strain Hardening Properties as Indicators of Baking Performance

Bread loaf volume is an important economic criterion. Breeders, millers, and bakers need measurements allowing them to evaluate dough performance during processing. Strain hardening is an important dough property. It describes the stability of the gas cell walls and the ability of cells to expand further; therefore, the higher the strain hardening index (n), the better the baking performance. Dough exhibits strain hardening during uniaxial extensibility tests. However, obtaining n from an extensibility test is time consuming. The objectives of this study were to identify the extensibility parameters that contribute to n. Three parameters were retained in the model (R² = 0.90): dough strength (R_max), extensional delay (E_Diff), and initial slope of the curve (E_i). R_max was the largest contributor and was proportional to the dough strain hardening properties. E_Diff and E_i had a detrimental effect on n. The appropriateness of the model was validated with two sample sets (19 genotypes, 12 environments). Significant correlations were found between loaf volume and n (r = 0.52–0.64) for 7 of the 12 environments. The strain hardening index was found to be a good predictor for baking performance as judged by loaf volume.     

Prevalence and risk factors for Salmonella spp. and Campylobacter spp. caecal colonization in broiler chicken and turkey flocks slaughtered in Quebec, Canada

We conducted an observational study to estimate prevalence and risk factors for Salmonella spp. and Campylobacter spp. caecal colonization in poultry. Eighty-one broiler chicken and 59 turkey flocks selected among flocks slaughtered in the province of Quebec, Canada, were included in the study. Flock status was evaluated by culturing pooled caecal contents from about 30 birds per flock. Exposure to potential risk factors was evaluated with a questionnaire. Odds ratios were computed using multivariable logistic regression.

The prevalence of Salmonella-positive flocks was 50% (95% CI: 37, 64) for chickens and 54% (95% CI: 39, 70) for turkeys, respectively. Odds of Salmonella colonization were 2.6 times greater for chicken flocks which failed to lock the chicken house permanently. In turkeys, odds of Salmonella colonization were 4.8–7.7 times greater for flocks which failed to be raised by ≤2 producers with no other visitors allowed onto the premises, or origin from a hatchery.

The prevalence of Campylobacter-positive flocks was 35% (95% CI: 22, 49) for chickens and 46% (95% CI: 30, 62) for turkeys. Odds of colonization were 4.1 times higher for chicken flocks raised on farms with professional rodent control and 5.2 times higher for flocks with manure heap >200 m from the poultry house, and also increased with the number of birds raised per year on the farm and with the age at slaughter. For turkeys, odds of Campylobacter flock colonization were 3.2 times greater in flocks having a manure heap at ≤200 m from poultry house and 4.2 times greater in flocks drinking unchlorinated water.     

Suspected air embolism associated with post-anesthetic pulmonary edema and neurologic sequelae in a horse

A 523 kg Quarter Horse was anesthetized for unilateral eye enucleation. The anesthetic period was unremarkable. During anesthetic recovery the cap on the jugular venous catheter became dislodged. Clinical signs of pulmonary edema associated with moderate arterial hypoxemia subsequently developed. Although pulmonary edema resolved with medical therapy, the day following anesthetic recovery, clinical signs of vestibular disease and blindness developed. Treatment included nasal oxygen insufflation, flunixin meglumine, furosemide, dexamethasone, thiamine, dimethylsulfoxide, antimicrobials, and phenylbutazone. The horse recovered and was discharged from the hospital after 7 days of treatment and was neurologically normal at 6 weeks. While venous air embolism was not confirmed in this case, the catheter cap complication followed by signs of pulmonary edema and neurologic sequelae support the presumptive pathogenesis of this horse's complications. Diagnostic confirmation of air embolism in horses with compatible acute clinical signs should be documented with echocardiography.