Effects of prescription diet on dealing with stressful situations and performance of anxiety-related behaviors in privately owned anxious dogs

The objectives of this study were to evaluate the effects of a diet (CALM CANINE) supplemented with alpha-casozepine and L-tryptophan on the stress response of anxious dogs with a focus on anxiety-related behavioral parameters and physiological parameters. Forty-four privately owned dogs were first fed with the control diet, followed by the study diet, each for an 8-week period, with a transitional period of 1 week between the diets. After 7 weeks on a particular diet, owners reported their dogs' behavior by filling out a questionnaire. Using the Canine Behavioral Assessment and Research Questionnaire, the effect of the study diet was found to be significant for 4 anxiety-related behavioral parameters, possibly mirroring a placebo effect. After 7 weeks on each diet, the urine cortisol-to-creatinine ratio (UCCR) was measured to assess the stress response to a visit to a veterinary practice for toenail clipping. UCCR was measured to complement the interpretation of behavioral data with regard to the stress susceptibility of the dogs. A positive correlation was found between baseline UCCR, as measured in the home situations, and poststressor UCCR. The stressor-induced increase was significantly lower in the dogs when they were fed the study diet than earlier when they were fed the control diet. The supplementation of casozepine or the proportion of protein in a given diet could have been a determining factor. The study diet seems to improve the ability of an individual to cope with stress and may reduce anxiety-related behavior in anxious dogs.     

Caprine enteritis associated with Yersinia pseudotuberculosis infection

Yersiniosis was prevalent among a caprine herd during the late autumn of 2003 in Iwate Prefecture, Japan. The disease affected 29 of about 100 lactating goats, but not dried or nonparous goats, mature male goats or kids. Four animals died within an epidemic period of 20 days. Affected animals developed decreased milk production with subsequent watery diarrhea, neutrophilia with increased band forms and multiple microabscesses characteristic of yersiniosis in the intestinal mucosa from the jejunum to caecum as well as in the mesenteric lymph nodes. Y. pseudotuberculosis serotype III was isolated from intestinal contents and mesenteric lymph nodes. The organism was also cultured from clinically normal dried animals. The outbreak might have been precipitated by multiple stress factors, such as lactation, cold weather, Corynebacterium pseudotuberculosis infection resulting in abscess formation and tapeworm and coccidium parasitisms.     

FC-20 Efficacy of griseofulvin for juvenile cellulitis in dogs

Medical records of 97 dogs with pemphigus foliaceus were evaluated. The average age of onset was 6.3 years (range 0.5–16 years). Crusts were the most common lesions in 79 dogs; pustules were observed in 36 dogs. No gender predisposition was identified. The trunk was the most commonly involved area (51 dogs), followed by the inner pinnae (46), dorsal muzzle (37), footpads (32), periocular area (26), outer pinnae (23) and planum nasale (23). Facial involvement only was noted in 15 dogs. Of the 48 dogs in which cytology was recorded, concurrent infections were identified in 32 dogs, acantholytic cells were seen in 37 dogs, numerous neutrophils in 35 dogs, and numerous eosinophils in eight cases. Final control of the disease was achieved with: glucocorticoids (24 dogs); azathioprine (9); chlorambucil (1); aurothioglucose (1); a combination of glucocorticoids and azathioprine (31); glucocorticoids and aurothioglucose (2); tetracycline/doxycycline and niacinamide (8); prednisolone, tetracycline and niacinamide (1); fatty acid supplementation (2); and tacrolimus (1). One dog was completely tapered off drugs and stayed in remission. Average time to improvement was 6 weeks, and average time to remission was 9.3 months. Forty-three dogs were followed for <12 months, and 12 of these were euthanized: eight for other diseases and four due to a lack of response or adverse effects of treatment. In 54 dogs, the follow up was >12 months; four of these dogs were euthanized (one due to an unrelated cause, one due to neoplastic disease and two related to pemphigus foliaceus).
Funding: Self-funded.     

P‐5 Importance of psychogenic factors in canine recurrent pyoderma

Recurrent pyoderma is a relapsing bacterial infection of the skin. Evaluation of the primary predisposing factors is essential, but they are not always easy to determine. We reported that psychogenic factors should be considered as one cause of canine dermatoses featuring pruritic behavior such as grooming and scratching, and predicted that it could be added as a cause of recurrent pyoderma. The purpose of this study was to evaluate the roles of psychogenic factors in canine recurrent pyoderma. This study examined 21 dogs with recurrent pyoderma, which showed relapsing but antibiotic‐responsive skin lesions. Predisposing factors, especially allergy, infection, endocrine disorders, congenital factors and improper treatments, were carefully ruled out with standard diagnostic procedures. Diagnosis of psychogenic factors was based on fulfillment of the following features: (1) distinctive area of broken hairs or asymmetric erythema, (2) incidental pruritic behavior related to emotionally unstable situation or physiological intervention, (3) existence of environmental antecedent triggers or concurrent psychiatric symptoms, and (4) requirement for psychogenic treatment, consisting of behavior modification and psychopharmacological therapy in some. The incidence of psychogenic factors and the efficacy of the treatment in recurrent pyoderma were evaluated. In these 21 dogs, 10 cases (47.6%) were compatible with the presence of psychogenic factors, and seven cases (33.3%) were improved with behavioral treatment. Psychogenic factors should be considered in the aetiology of so‐called idiopathic canine recurrent pyoderma. Further investigation is needed to understand the relationship between skin barrier dysfunction and psychogenic factors. Funding: Self‐funded.     

Nutritive value of selected grasses in North Sumatra, Indonesia

The nutritive values of seven native grass species collected in North Sumatra, Indonesia, during dry and rainy seasons were evaluated. The chemical composition, in vitro dry matter digestibility (IVDMD), in vitro organic matter digestibility (IVOMD), in vitro crude protein digestibility (IVCPD), macro mineral concentrations of calcium, phosphorus and magnesium, in vitro gas production profile and metabolizable energy (ME) content of the grasses varied greatly among species and seasons. The crude protein content ranged from 6.6 (Andropogon gayanus) to 16.2% dry matter (Cynodon plectostachyus) in the rainy season, with a significant (P < 0.05) reduction in the dry season. Data on the fiber fraction showed that the grasses contained more neutral detergent fiber (NDF) and acid detergent fiber (ADF) in the dry season and it significantly (P < 0.05) decreased in the rainy season, except for Panicum maximum and Pennisetum purpureum for NDF content and C. plectostachyus, P. maximum and Brachiaria decumbens for ADF content. Data on the mineral concentration showed that C. plectostachyus in the dry and rainy seasons had a higher calcium content than those of other species. The overall means of the seven grasses for IVDMD, IVOMD and IVCPD were significantly higher (P < 0.05) in the rainy season than in the dry season. The in vitro gas production and ME concentration were numerically higher in the rainy season than in the dry season. In conclusion, the nutritive value of the observed grasses in North Sumatra was relatively higher during the rainy season compared with the dry season. Among the species of grass, P. purpureum and C. plectostachyus had a higher nutritive value in both the dry and rainy seasons.     

High level expression and purification of bioactive bovine interleukin-18 using a baculovirus system

Bioactive recombinant bovine interleukin-18 (rboIL-18) was expressed using a baculovirus system. Normally, IL-18 is translated as a precursor form of a 24kDa polypeptide and processed by IL-1beta converting enzyme (ICE) to a mature bioactive form of 18kDa protein. Hence, to express active form IL-18, we constructed two recombinant baculoviruses containing boIL-18 and human ICE (hICE) genes, respectively, and superinfected these viruses into insect cells. Superinfection of both recombinant viruses into the cells resulted in the expression of a 24kDa precursor form and an 18kDa mature form detectable in the supernatant by immunoblotting using anti-porcine IL-18 antibody. Culture supernatant from the superinfected cells showed a synergistic effect with recombinant boIL-12 for production of interferon-gamma (IFN-gamma) in bovine peripheral mononuclear cells. By addition of histidine hexamer at the C-terminal of boIL-18, the mature IL-18 was purified. Bioactivity remained after purification.     

Production and in vivo testing of a recombinant bovine IL-12 as an adjuvant for Salmonella typhimurium vaccination in calves

A recombinant bovine interleukin-12 (boIL-12) that contains a histidine hexamer, rboIL-12His, was produced, purified and administered to calves. We first tried the purification of heterodimer IL-12 from a mixture of p40 homodimer, p40 monomer, and p40-p35 heterodimer with a p35 subunit tagged with a histidine hexamar at its C-terminal (p35His). A recombinant baculovirus expressing p35His was generated and used for superinfection with a recombinant baculovirus expressing p40 subunit. The expressed subunits, p40 and p35His, were assembled into a 70kDa heterodimer in insect cells, released into culture medium, and then purified using a nickel chelate column. The purified rboIL-12His was bioactive for induction of IFN-gamma in bovine peripheral blood mononuclear cells (PBMCs) in vitro.The purified rboIL-12His was then administered to calves with inactivated Salmonella Typhimurium (ST). When sera were assayed by ELISA, specific anti-ST IgG1 antibodies were detected in all ST immunized calves, but, specific anti-ST IgG2 antibodies were detected only in calves administered ST along with rboIL-12His, indicating a possible switch to a Th1 response. Administration of commercially available Salmonella vaccine did not elicit IgG2 antibodies in calves. These results suggest that co-administration of IL-12 with inactivated ST cells could induce a Th1-type response in calves.     

Availability of NMR microscopic observation of mouse embryo disorder: examination in malformations induced by maternal administration of retinoic acid

Nuclear magnetic resonance (NMR) microscopy is a magnetic resonance imaging method with enhanced spatial resolution due to the use of a high static magnetic field and high magnetic field gradients. It is considered to be a useful tool for non-invasive and continuous investigation of tissue and organs at the histological level. In this study, we applied NMR microscopy to assessment of morphology in mouse embryos using a developmental disorder model induced by retinoic acid administration. Pregnant mice were given 50 mg/kg all-trans retinoic acid at 8.5 dpc. Embryos were collected at several time points after treatment and examined by NMR microscopy after fixation. Two-dimensional and three-dimensional spin echo sequences were used. Tissue contrast on two-dimensional images changed according to length of repetition time and echo time, and also to developmental stage of embryos. Two-dimensional and three-dimensional images nondestructively demonstrated defects in development of the skeleton and soft tissue, e.g. hypoplasia of vertebrae in the lumbar and tail regions and dysplasia of the spinal cord, in embryos exposed to retinoic acid. These morphological abnormalities were confirmed by conventional assessment after imaging. Although further improvements are required, NMR microscopy will provide a new approach for multi-parameter assessment of embryonic development under physiological and pathological conditions.     

Effects of Temperature and Host on the Generation of Tomato Spotted Wilt Virus Defective Interfering RNAs

ABSTRACT The generation of defective interfering (DI) RNA molecules of tomato spotted wilt tospovirus (TSWV) was studied by serially passaging in-ocula from plant to plant under different controlled conditions. DI RNAs were generated at higher rates in plants at 16 degrees C than in plants incubated at higher temperatures. Another factor promoting the TSWV DI RNA generation was the use of high virus concentrations in the inocula. The solanaceous species Capsicum annuum, Datura stramonium, Lycopersicon esculentum, Nicotiana benthamiana, and N. rustica supported the generation of DI RNAs, whereas the virus recovered from the inoculated composite species, Emilia sonchifolia, remained free of any DI RNA under all conditions tested. This study resulted in a strategy to maintain DI RNA-free TSWV isolates, as well as in an efficient way to produce a large population of different DI RNA species. A single DI RNA species usually became dominant in an isolate after a few rounds of serial inoculations. The possible mechanisms involved in TSWV DI RNA generation under different inoculation circumstances are discussed.