Comparative serological evaluation of avian influenza vaccine in turkeys

Four- and six-week-old turkeys were vaccinated subcutaneously using avian influenza virus (AIV) A/Duck/613/MN/79 (H4N2) killed oil-emulsion vaccine. Sequential serological tests using agar gel precipitin (AGP), hemagglutination inhibition (HI), and enzyme-linked immunosorbent assay (ELISA) for measuring antibodies to AIV were performed up to 4 weeks postvaccination, when birds were challenged intranasally using A/Turkey/MN/80 (H4N2) live AIV. The ELISA was 25 to 1600 times more sensitive than the HI test and was able to detect antibody production earlier than the HI test. All turkeys with an ELISA titer of greater than or equal to 800 were protected against homologous challenge, as measured by virus recovery 3 days postchallenge. Four turkeys out of 20 serologically negative by AGP and HI tests but ELISA-positive were protected.     

A dot-immunobinding assay for infectious bronchitis virus

Common Whatman filter paper grade 1 and nitrocellulose membrane were compared for their sensitivity in a dot-immunobinding assay for detection of serum antibody titers to Arkansas avian infectious bronchitis virus (AIBV). For a blue to purple color detection, serum antibodies were bound to AIBV antigen adsorbed on the filter-paper discs or nitrocellulose membrane. Rabbit anti-chicken IgG horseradish-peroxidase (HRP) conjugate and hydrogen peroxide with 4-chloro-1-naphthol (HRP-color development reagent) were applied. The study indicates that very small amounts of antigen/antisera are needed for the dot-immunobinding assay. The test is sensitive, economical, and easy to run and can be completed within 6-8 hours.     

Leukoencephalomalacia in a horse induced by fumonisin B1 isolated from Fusarium moniliforme

Each of two horses was dosed by stomach tube with culture material on maize of Fusarium moniliforme MRC 826. One horse developed severe hepatosis and mild oedema of the brain after 6 doses of 2.5 g of culture material/kg body mass/day in 7 days. The second horse, in a similar experiment but at a dosage rate of 1.25 g/kg/day, developed mild hepatosis and moderate oedema of the brain. In both animals the brain oedema was particularly noticeable in the medulla oblongata. The mycotoxin fumonisin B1 was extracted and purified from the culture material of F. moniliforme MRC, 826 which contained approximately 1 g/kg of this compound. A horse was injected intravenously 7 times from Day 0-Day 9 with 0.125 mg of fumonisin B1/kg body mass/day. Clinical signs of neurotoxicosis, which appeared on Day 8, included nervousness followed by apathy, a wide-based stance, trembling, ataxia, reluctance to move, paresis of the lower lip and tongue, and an inability to eat or drink. Euthanasia was performed on the horse on Day 10 while the animal was in a tetanic convulsion. The principal lesions were severe oedema of the brain and early, bilaterally symmetrical, focal necrosis in the medulla oblongata. This report provides experimental evidence that fumonisin B1, produced by F. moniliforme, causes equine leukoencephalomalacia.     

Two-dimensional echocardiography for diagnosis of dirofilariasis in nine dogs

Two-dimensional echocardiography (real-time ultrasonography) of the heart was used to detect adult Dirofilaria immitis in the right ventricle and atrium of a 2-year-old Labrador Retriever with occult heartworm disease and in 8 microfilaremic dogs. Adult worms were easily identified as parallel echogenic lines separated by a hypoechoic region. Images of worms fixed in formalin after recovery from dogs with experimentally induced heartworm infection had an identical appearance. The sensitivity of this imaging procedure was tested on the 8 dogs with naturally acquired infection, and worms were observed in the right ventricle of 7 of the dogs. Two-dimensional echocardiography appears to be a sensitive noninvasive procedure, which can be used in combination with thoracic radiography and serotests to improve accuracy of diagnosis of canine occult heartworm disease.     

Measurement of protein turnover in skeletal muscle strips

Isolated porcine and bovine muscle strips were incubated in Krebs Ringer bicarbonate buffer to determine in vitro protein synthesis (PS) and protein degradation (PD) rates to validate the in vitro system for use with livestock species. The addition of 5X plasma concentrations of amino acids to the medium stimulated PS 30%. Addition of 3.5 mM leucine to a leucine-deficient buffer supplemented with amino acids decreased PD 37% and stimulated PS 24%. The addition of .1 U/ml insulin reduced PD 28% and increased PS 30%. Protein degradation was elevated in longitudinally split rat soleus and extensor digitorum longus muscles compared to their contralateral intact muscles. Muscle strips must be removed within 15 min of exsanguination because PD rates become greatly elevated thereafter. ATP concentrations declined during incubation, but the addition of ATP or creatine had no effect on either PD or PS. Neither PD nor PS was affected by the addition of transferrin, fetuin, ascorbate, dexamethasone or indomethacin to the incubation medium. However, muscle strips were sensitive to the addition of triiodothyronine (T3), PD was increased up to 75% as T3 concentration was increased, and PS rates doubled compared to controls. Serum from mature barrows or gilts had no effect on protein turnover, but the addition of 10% and 15% serum from boars increased both PD and PS. With fasted pigs a continual decline in PS occurred over 5 d, whereas PD was elevated at 3 d and then declined to rates comparable to the fed state after 5 d. These data suggest that the in vitro system has application for assessing relative changes that occur in vivo following nutritional, physiological and endocrinological manipulation.     

Immunological cross-reactivity between pseudocapsular antigens of strains of Staphylococcus aureus isolated from cases of bovine mastitis

An assay was developed to measure the degree of immunological cross-reactivity between the pseudocapsules of a vaccine (reference) strain of Staphylococcus aureus and pseudocapsules of strains of S. aureus isolated from cases of bovine mastitis. The field strains were obtained from Australia, New Zealand, Norway, the U.K. and the U.S.A. There was large variation among strains in cross-reactivity of their pseudocapsules with those of the reference strain. For 104 Australian strains, the range of cross-reactivity indices (CRI) was 6.1-63.2% (on a scale of 0-100%, with 0% being complete identity and 100% being nil identity); for 61 overseas strains the range of CRI values was 25.7-72.1%. The data indicated that pseudocapsule antigens of Australian strains were antigenically more closely related to those of the reference strain than were pseudocapsular antigens of strains from the 4 other countries.