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71.
72.
Comparison of dot blot hybridization, polymerase chain reaction, and virus isolation for detection of bovine herpesvirus-1 (BHV-1) in artificially infected bovine semen. 总被引:2,自引:0,他引:2 下载免费PDF全文
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) 相似文献
73.
74.
One hundred two fecal specimens from psittacine birds submitted to Veterinary Laboratory Services of the California Department of Food and Agriculture at Petaluma were screened for Chlamydia psittaci by a direct immunofluorescence assay using a fluorescein-labeled monoclonal antibody conjugate specific for Chlamydia sp. Results were compared with those obtained by isolation of chlamydia in cultures of McCoy mouse cells. The relative specificity of the direct fluorescent antibody test on fecal smears was 98.9% and the relative sensitivity was 62.5%. The results of this study suggested that the direct fluorescent antibody test was highly specific, and it proved to be a useful same-day antemortem diagnostic test for birds with symptomatic chlamydial infection. The use of centrifugation in the cell culture assay was found to significantly enhance the level of chlamydial infection in cell culture. 相似文献
75.
Effect of age on absorption and immune responses to weaning or introduction of novel dietary antigens in pigs 总被引:14,自引:0,他引:14
Pigs weaned at three weeks old absorb food protein antigens from the intestine. The amount of antigen absorbed declines over the next three weeks, and this decline is associated with an increasing level of serum antibody to the fed proteins. There was no difference in the rate of immune elimination of intravenously injected antigen in fed and unfed controls. The reduction of serum antigen is thus likely to reflect reduced absorption, possibly mediated by locally produced antibody. Pigs weaned at 10 weeks old also absorbed antigens and produced an antibody response when introduced to soya; and after three weeks of feeding soya the absorption of antigen was substantially reduced. This latter exclusion was specific for soya as a second novel protein (ovalbumin) was absorbed when introduced to the diet at this time. At six months, pigs no longer absorbed soya proteins when they were introduced to the diet. Furthermore, pigs of this age had serum 'antibody' to soya and other proteins such as keyhole limpet haemocyanin to which they had never been exposed. 相似文献
76.
P Dom F Haesebrouck 《Zentralblatt für Veterin?rmedizin. Reihe B. Journal of veterinary medicine. Series B》1992,39(4):303-306
The virulence of a NAD-independent Actinobacillus pleuropneumoniae serotype 2 strain and NAD-dependent serotype 2, 3 and 9 strains was compared under experimental conditions. Hysterectomy-derived piglets were inoculated endobronchially with 50-500 cfu of these strains. All 23 piglets inoculated with the NAD-dependent strains developed acute disease within 12 hours post inoculation. Twenty-two of these piglets died within 24 hours after the first clinical signs. Three of nine piglets inoculated with the NAD-independent strain did not develop clinical disease. In the other six piglets, disease signs were similar as in the piglets inoculated with the NAD-dependent strains. No differences in clinical disease were observed between colostrum deprived piglets and piglets that obtained colostrum from a SPF sow. 相似文献
77.
78.
A three-phase laboratory procedure suitable for predicting protein degradability in the rumen and digestibility of undegraded protein is reported. In the first phase the feed was incubated with starch and buffered rumen fluid. In the incubation mixture the viability of protease-active bacteria was checked by anaerobic culturing, whereas changes in protease activity were monitored by azocasein degradation. In the second and third phase rumen undegradable protein (UDP) was digested with pepsin and pancreatin, respectively. The measurements showed that 63.2, 5.2 and 4.7% of the crude protein of green lucerne was decomposed by rumen fluid, pepsin and pancreatin, respectively. Degradability of the crude protein of extracted sunflower meal was 68.3, 17.7 and 5.5% in the three phases, respectively. Repeated determination yielded crude protein degradabilities of 66.7, 27.1 and 5.1% for the three phases, respectively. 相似文献
79.
M Krajnicáková E Bekeová I Maracek V Hendrichovsky J Elecko 《Veterinární medicína》1992,37(8):449-458
A postparturient period is characterized by low basal secretion of adenohypophysis gonadotropins with the following appropriate changes in ovarian hormones and their response to the morphology of vaginal epithelium. In this study the dynamics of the cytological picture of vaginal swabs and ovarian hormones 17 beta-oestradiol (E2) and progesterone was investigated in the puerpery of ewes. The objective was to obtain and extend the knowledge of cytological changes in vaginal epithelium and levels of ovarian hormones of ewes after parturition and of their relationships from the first several days after lambing until the 51st day of the period of observation. Vaginal swabs for vaginal cytology were taken from nine ewes on days 1, 4, 7, 14, 17, 21, 25, 34, 42 and 51 after parturition. These swabs were fixed in ether-alcohol 1:1, stained according to the Faltínová-Zidovsky method, embedded in Canada balsam and evaluated by differentiation of cells according to Luksh (1953). Blood samples for E2 and P4 determinations were taken from the jugular vein in the same intervals as vaginal swabs. The serum was centrifuged and stored at -18 degrees C until use. E2 and P4 concentrations were determined radioimmunologically, using kits RIA-test ESTRA and RIA-test PROG from URVJT Kosice. A statistically significant decline (P < 0.05) of percentual representation of basal and parabasal cells (Fig. 1, Tab. I) on day 7 after lambing was replaced by their multiplication from day 14 reaching the values of 66.07 +/- 3.95 on day 42. A statistically significant decrease in intermediary flat cells (Fig. 2, Tab. II) was observed on days 14 (P < 0.001), 34 and 42 (P < 0.01; P < 0.001), in comparison with the first day after lambing. An evaluation of intermediary convoluted cells revealed their highest percentage on days 1 and 17 after parturition (34.65 +/- 4.77-20.62 +/- 12.57) and their decline to values in the range of 6.77 +/- 1.46-7.66 +/- 2.25 on the remaining days of the period of observation. Percent occurrence of superficial flat cells (Fig. 3, Tab. I) ranged from 3.9 +/- 1.10 to 10.63 +/- 7.23 from day 1 to day 51 after lambing. The lowest percentual representation (1.32 +/- 0.79-4.10 +/- 1.89) was recorded for superficial convoluted cells. Multiplication of the evaluated cells was observed, reaching the highest but insignificant representation (P > 0.05) on day 25 of postparturient investigation: 4.10 +/- 1.89 (Fig. 3, Tab. I). 17 beta-oestradiol (E2) concentrations were compared to the -1st day before parturition, when its values varied at the level of 2.45 +/- 0.64 nmol/l serum.(ABSTRACT TRUNCATED AT 400 WORDS) 相似文献
80.
Germinating seeds and young plants of winter rape var. Górczañski were vernalized for 56–63 days under conditions of 9-hour day, at the temperature 2 and 5 °C and in continuous darkness at the temperature 2 °C. After vernalization the plants grew under conditions enabling to complete vernalization: in a glass-house at the temperature day/night 15/10 °C and in semi natural conditions of open vegetation hall in the period from June till August. After sub-optimal vernalization further growth of the plants at lowered temperature increased its effectiveness (completion of vernalization). Depending on the degree of the vernalization of the plants the completion of their vernalization was both obligatory, i.e. conditioning the acquisition of the ability of generative development, and facultative i.e. accelerating this development. It has been demonstrated that the population of plants of the examined variety is strongly differentiated not only with respect of vernalization requirements in the particular plants, but also what regards the effectiveness of vernalization completion. New observations have been made indicating that the mechanisms controlling the successive phases of generative development, i.e. phase of forming flower buds and the flowering phase are not identical which may be interpreted as indicating that the "flowering factor" is polymorphous. 相似文献