Immunogenicity of bovine parainfluenza type 3 virus proteins encapsulated in nanoparticle vaccines,following intranasal administration to mice

Stimulation of long lasting, protective immunity to respiratory viruses is often difficult to achieve with conventional respiratory vaccines. Polymeric nanoparticles, incorporating viral proteins have been shown to offer sustained release of antigen, with consequent prolongued stimulation of the respiratory immune system. In this paper the efficacy of two nanoparticle vaccines (poly-lactide-co-glycolide, PLGA; polymethylmethacrylate, PMMA), incorporating proteins of bovine parainfluenza type 3 virus (BPI-3) was investigated. As a preliminary to experiments in calves, it was considered essential to demonstrate immunogenicity of the experimental vaccine in mice. Mice immunised with PLGA nanoparticles, containing BPI-3 proteins, developed higher levels of virus-specific antibody than mice immunised with the PMMA vaccine or with soluble viral proteins alone. Immunoblotting using serum from the vaccinated mice, demonstrated strong reactions against the major BPI-3 proteins.     

A comparison of catecholamine and cortisol responses of young lambs and calves to painful husbandry procedures

AIM: To compare the changes in plasma concentrations of noradrenaline, adrenaline and cortisol in lambs after ring castration plus tailing and in calves after dehorning with or without prior local anaesthesia. METHODS: Male lambs were castrated and tailed with rings and calves were dehorned by amputation using a scoop with or without prior local anaesthesia. Blood samples were taken before and after treatment and plasma concentrations of noradrenaline, adrenaline and cortisol were determined. RESULTS: Castration plus tailing of lambs resulted in a rapid increase in noradrenaline concentrations, a lack of an adrenaline response and a marked increase in cortisol concentration. There were similar changes in catecholamine concentrations in calves that were dehorned both with and without local anaesthetic, with adrenaline being elevated within 5 min of treatment and noradrenaline exhibiting a more protracted response. Dehorning caused a marked cortisol increase which was reduced to control concentrations by local anaesthesia for as long as the associated nerve blockade lasted. CONCLUSIONS: The very short-lived adrenaline responses in calves were attributed to dehorning-induced nociceptor input leading to sympathetic stimulation of the adrenal medulla. The longer lasting noradrenaline responses in lambs and calves were thought be due to 'wash-out' of noradrenaline from damaged tissue associated with rings and amputation wounds, respectively.     

Effects of food withdrawal and transport on 5- to 10-day-old calves

The effects of food withdrawal for 30 hours and transport for up to 12 hours on 5- to 10-day-old calves were determined by monitoring plasma concentrations of glucose, beta-hydroxybutyrate, urea, lactate and creatine phosphokinase. In addition gamma-glutamyl transferase activity, packed cell volume, total plasma protein concentration, bodyweight and rectal temperature were recorded. Food withdrawal for 30 hours caused hypoglycemia but calves maintained normothermia and although their beta-hydroxybutyrate level increased indicating lipid mobilisation, the changes in blood urea level were minimal indicating low amino acid catabolism. When calves were transported for 12 hours in addition to having food withdrawn for 30 hours, they remained normoglycemic for six hours longer than non-transported calves, probably due to an effect of muscular activity during bracing against truck movements. Stocking calves at a lower density during transport apparently had a beneficial effect, as calves were able to lie down throughout the journey and therefore produced a similar metabolic profile to resting, non-transported animals. Transport and food withdrawal had no obvious effects on calf hydration. The results of this study suggest that food withdrawal for up to 30 hours and transport for up to 12 hours had no detrimental effects on the metabolism of healthy and clinically normal calves. With correct feeding regimes and transport protocols, welfare compromise in young, healthy calves being transported for up to 12 hours can be minimised when they are slaughtered within 30 hours of the start of transport.     

Surgical procedure and postoperative management of a perineal urethrostomy in a chimpanzee (Pan troglodytes).

A chimpanzee (Pan troglodytes) with traumatic loss of the distal penis developed a gradually enlarging ventral urethral swelling and progressive dysuria. Endoscopy identified a urethral diverticulum, and endoscopic resection of the diverticulum wall was performed. Postoperative infection caused extensive necrosis of the penis, which necessitated a perineal urethrostomy. Complications of the urethrostomy included urethral obstruction from recurrent urethral stricture. The stricture was managed by regular dilatation using urethral bougies. Because of considerable postoperative swelling, catheterization was required to allow micturition following both the diverticular resection and polyp debulking. A shortened catheter sutured to the skin was tolerated for up to 10 days. Four yr after the urethrostomy, the animal is healthy and asymptomatic with endoscopic examinations performed at 12 mo intervals. This case demonstrates that with appropriate aftercare, perineal urethrostomy is an effective technique in the treatment of chronic distal urethral obstruction in the chimpanzee and probably other primate species.     

Xylitol intoxication associated with fulminant hepatic failure in a dog

Objective: To describe a case of xylitol intoxication causing fulminant hepatic failure in a dog. Case summary: A 2.5‐year‐old castrated male English Springer Spaniel weighing 26 kg, was presented after ingestion of half of a loaf of bread containing the sweetener xylitol. Toxic effects of the xylitol in this dog included vomiting, mild hypoglycemia and fulminant hepatic failure. Clinical management of acute hepatic failure and subsequent coagulopathy with supportive care and fresh frozen plasma is described. The dog was discharged 3 days after admission after a full clinical recovery. New or unique information provided: This paper describes the clinical consequence and successful treatment of fulminant hepatic failure in a dog following ingestion of xylitol.     

Properties of dry film lubricants prepared by spray application of aqueous starch–oil composites

Aqueous dispersions of starch–soybean oil (SBO) and starch–jojoba oil (JO) composites, prepared by excess steam jet cooking, form effective dry film lubricants when applied as thick coatings to metal surfaces by a doctor blade. This application method necessitates long drying times, is wasteful, requires the addition of sucrose to promote composite adhesion to the metal surface, and restricts the substrate geometry to planar surfaces. These issues represent important barriers to the commercialization of this aqueous biobased dry film lubricant technology. We now report an air-assisted spray method that uses readily available spray equipment to apply aqueous starch–oil composite dispersions as thin coatings (0.15–2.0 mg/cm²) to metal surfaces quickly and efficiently. Aqueous dispersions of waxy maize starch–oil composites containing either SBO, JO or hexadecane (HD), having 0.020–31.7 wt% oil relative to starch, were applied by air-assisted spraying and could be dried to the touch in approximately 30 s. Additionally, sucrose was found unnecessary for adhesion of the sprayed coatings. Tribological ball-on-flat testing of metal specimens spray coated with starch–SBO, –JO, and –HD composites showed the thin films of starch–SBO and –JO performed better at reducing the coefficient of friction (COF) than the starch–hexadecane composites. A low COF ranging between 0.027 and 0.044 was obtained for the starch–SBO and –JO composites containing 4–5 wt% oil relative to starch. Above 4–5 wt% oil loadings, no further COF reductions were realized. Further results revealed that micrometer-sized oil droplets embedded within the dried starch matrix of the composite film are delivered “on demand”. It appears that when pressure is applied to the dry film lubricant, the starch matrix ruptures and releases the entrained oil to the friction surface.     

Chicken anemia agent: an electron microscopic study

Particles of chicken anemia agent (CAA) negatively stained with uranyl acetate were found to be 26.5 nm in diameter. The surface detail evident on the particles indicated that the virus capsid was composed of 32 structural subunits arranged as in a class P = 3 icosahedron with a triangulation number of 3. Using mouse monoclonal antibodies to CAA and a gold-labeled goat anti-mouse IgG, CAA-specific structures were observed by thin-section electron microscopy in infected MDCC-MSB1 cells and in thymic lymphocytes from experimentally infected chicks. These consisted of electron-dense, granular, non-membrane-bound nuclear inclusions, which were often ring-shaped, and cytoplasmic accumulations of microtubules. Aggregates of virus-like particles were sometimes observed in the nuclei of infected MDCC-MSB1 cells. The nucleolar involvement that is characteristic of the morphogenesis of parvoviruses was not observed with CAA.     

Development of an enzyme-linked immunosorbent assay to detect serum antibody to chicken anemia agent

An enzyme-linked immunosorbent assay (ELISA) for the detection of antibodies to chicken anemia agent (CAA) has been developed. This test utilizes a CAA-specific mouse monoclonal antibody to selectively capture virus antigen. Chicken antibodies to CAA bind to the captured antigen and are detected with horseradish peroxidase-labeled anti-chicken immunoglobulin using a conventional indirect ELISA protocol. When 388 chicken sera from specific-pathogen-free and commercial flocks from the United Kingdom, West Germany, the United States and Australia were examined, 98.5% agreement was obtained between the results of the ELISA and the indirect immunofluorescence assay. This ELISA should have worldwide application in testing SPF and commercial chicken flocks for CAA antibodies.     

High-yield preparation of purified Anaplasma marginale from infected bovine red blood cells

Purification of Anaplasma marginale from infected bovine RBC was achieved through enzyme treatment and density-gradient centrifugation. A relative yield of 41.6% was obtained by dividing the number of organisms in the final purified preparation by the number of A marginale-infected RBC. Purified parasites were verified as A marginale by light microscopy, electron microscopy, and immunologic tests. The purified parasites reacted positively with calf and rabbit anti-A marginale sera in interfacial and slide agglutination tests. Anti-bovine RBC serum did not agglutinate purified A marginale, indicating absence of any contaminating RBC stroma. Anaplasma marginale was antigenic, but did not cause infection when the preparation was inoculated into a susceptible calf. The density of A marginale was determined to be 1.19 g/ml and cell diameters ranged from 0.25 to 0.63 micron. This method provided procedures for obtaining A marginale free of bovine RBC antigens for accurate biochemical assays and vaccine production.