Comparison of dot blot hybridization, polymerase chain reaction, and virus isolation for detection of bovine herpesvirus-1 (BHV-1) in artificially infected bovine semen.

Bovine semen samples spiked with bovine herpesvirus 1 (BHV-1) were used to compare dot blot hybridization, polymerase chain reaction (PCR), and virus isolation for detection of BHV-1 in bovine semen. The PCR amplification used primers targeting the BHV-1 thymidine kinase gene and a nucleic acid releasing cocktail (GeneReleaser); the PCR product was used as the DNA probe in dot blot hybridization; virus isolation was done in primary bovine fetal testis (BFT) cell cultures. Semen diluted 1:20 in tissue culture medium had the least cytotoxicity and inhibition of viral cytopathic effects in BFT cells, allowing detection of 1 TCID50/100 microL of BHV-1 suspension by virus isolation. The presence of foreign DNA such as bovine sperm DNA or salmon sperm DNA increased the sensitivity of dot blot hybridization in detecting BHV-1, allowing detection of 20,000 TCID50/100 microL of neat semen. The inhibition of PCR amplification of BHV-1 DNA in bovine semen was eliminated by diluting the samples 1:20 in tissue culture medium. The best PCR amplification was obtained when semen was diluted 1:20 and when a reaction buffer of pH 9.0, with 1.0 mM MgCl2 was used. Under these conditions, the PCR followed by ethidium bromide staining of agarose gels could detect 1 TCID20/100 microL of sample, whereas PCR followed by Southern blot hybridization could detect 0.01 TCID50/100 microL of sample.(ABSTRACT TRUNCATED AT 250 WORDS)     

Laboratory diagnosis and disease occurrence in the current African swine fever eradication program in Spain, 1989–1991

The purpose of this study was to determine the rate of decline and geographical distribution by municipality of clinical and subclinical African swine fever (ASF) in the affected areas of Spain. A second aim was to evaluate the performance of diagnostic tests in the Spanish ASF eradication program. Clinical outbreaks were confirmed using both the direct and indirect immunofluorescence test (and if both were negative, by the hemabsorption test). The serological status of swine was determined by an enzyme-linked immunosorbent assay (ELISA) and suspect serum samples were confirmed by the immunoblot assay.

The number of clinical outbreaks (herds) of ASF for 1989, 1990 and 1991 was 170, 347 and 207, respectively. The numbers of municipalities within each affected province experiencing acute outbreaks for the same time periods were 49, 69 and 48, respectively. Serologically diagnosed animals positive for ASF were 1.1% of animals tested in 1989, 0.5% in 1990 and 0.8% in 1991. The corresponding positive predictive values of the standard ELISA test used were 99.0, 97.9 and 98.8, respectively. Similarly, the number of municipalities within each affected province experiencing serologically positive subclinically infected animals was 269, 178 and 147 for each of the years 1989, 1990 and 1991, respectively.     

Effects of cultivar,pelleting and enzyme addition on nutritive value of barley in poultry diets

1. The effect of pelleting process and Trichoderma viride enzymes (TVE) addition on apparent metabolisable energy, corrected for nitrogen balance (AMEn) and on productive value of practical diets containing 40 and 45% of three different barley cultivars and one wheat were studied in poultry.

2. The effect of the pelleting process on AMEn was inconsistent and was dependent on the cereal included and the addition of enzyme.

3. The growth trial showed a significant effect of enzyme addition to pelleted diets over the whole growth period (0 to 42 d). Addition of TVE improved weight gain and food efficiency by 1.3% and 2.9%, respectively and decreased food intake by 1.6% between 0 and 22 d. In the finisher period (23 to 42 d) TVE improved efficiency by 2.8% and reduced food intake by 2.9%.

4. The incidence of sticky droppings was related to the viscosity of barley used, and enzyme supplementation reduced it. Both pelleting and enzyme addition increased dry matter content of excreta.

5. At the end of the experiment, 14 animals per treatment were slaughtered and carcass yield, viscera weight and abdominal fat were determined.     

Effects of air temperature, air movement and artificial rain on the heat production of brushtail possums (Trichosurus vulpecula): an exploratory study

Two groups of six mature brushtail possums (Trichosurus vulpeculu) were housed in two respiration chambers, and their heat production, whole body conductance and lower critical temperatures were measured under a variety of simulated weather patterns. The possums were subjected to ambient temperatures of 30, 20 and 3 degrees C. At 20 and 3 degrees C, the animals were exposed to near still air and light winds (wind speed 0.8 and 6.7 km/h), both with, and without, simulated rain every 8 hours. The lower critical temperature in near still air lies between 7 and 10 degrees C. This temperature increases by about 2, 6 and 8 degrees C respectively for a wind velocity of 6.7 km/h, simulated rain and a combination of the two factors. Weather in New Zealand, especially in the cooler part of the year, will often produce conditions below the lower critical temperature of the thermoneutral zone of possums. This will necessitate significant increases in metabolic rate and hence food consumption or mobilisation of body fat reserves, which if not sustainable will result in the death of possums. Field studies have shown that this is often the case in the wild. It is proposed that this stress may be sufficient to decrease the resistance (especially cell-mediated immunity) of some possums and allow acceleration of the disease process in those infected with Mycobacterium bovis.     

Intranuclear cisternal and vesicular structures in canine circumanal gland carcinoma cells

Intranuclear cisternal and vesicular structures, bound by a single membrane, were observed for the first time in some glandular "bird's eye" cells of canine circummanal gland carcinomas. The role of these intranuclear structures is not clear. It is suggested that they may be involved in cellular adaptation to an environment of nutrient deficiency, as a result of cell overcrowding.     

Metabolic fate of [14C]endrin in bluegill fish

Bluegill absorbed 85% of 1 ppb of endrin from water within 48 hr under static exposure conditions. The absorbed radiocarbon was eliminated linearly with a half-life of about 4 weeks. Analyses of eliminated radioactivity revealed only conjugated metabolites. 12-anti-Hydroxyendrin and 12-syn-hydroxyendrin were tentatively identified by cochromatography using thin-layer chromatography/autoradiography and gas chromatography. These metabolites were also present as conjugates in the fish organs. Seventy-three percent of the absorbed radioactivity recovered from fish extracts was in the form of unchanged endrin.     

胡桃长足象研究初报

胡桃长足象是核桃果实的大害虫,为害损失很大。1975年以来,我们在四川平武地区进行了生活习性观察和防治试验,初步明确了以下几个问题。1.胡桃长足象一年一代。11月份以成虫越冬,次年4月上旬开始活动,为害果、芽、嫩枝、叶柄,5月上旬交尾产卵于果中,一般一果一粒,8月中旬产卵结束,10月陆续死亡。当年成虫6月中旬羽化,进行为害。2.成虫有假死性和向阳性,飞翔力弱,具有与核桃树芽苞相似的保护色。3.防治胡桃长足象,应抓住越冬成虫大量出现和卵孵化盛期时进行。用每毫升含孢量二至五亿的白僵菌液或50%的三硫磷乳油、50%速灭松乳剂、82%磷胺乳剂加水一千倍喷雾,效果良好。     

A complement fixing antigen in the livers of birds infected with an avian leukovirus (erythroblastosis virus).

Extracts of erythroblastosis affected livers from which much of the structural protein had been removed were prepared by acid denaturation. These extracts contained complement fixing "soluble" antigen in a form which was not associated with viral or subviral particles. Ultracentrifugation can be of assistance in separating complement fixation activity from contaminating material in these extracts.