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

J Q Xia  C V Yason    F S Kibenge 《Canadian journal of veterinary research》1995,59(2):102-109

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   

In vitro evaluation of protein degradability in the rumen and digestibility of undegraded protein     

T Veresegyházy  A Nagy  S Fekete  F Kutas 《Acta veterinaria Hungarica》1989,37(1-2):103-115

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   

Dynamics of changes in the cytological picture of vaginal smears and circulating ovarian hormones during the puerperal period in ewes]     

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   

The effect of repeated administration of carbetocin (Depotocin, Spofa) in the first days postpartum on levels of thyroxine, triiodothyronine, 17 beta-estradiol, progesterone and on fertilization]     

E Bekeová  M Krajnicáková  V Hendrichovsky  I Maracek 《Veterinární medicína》1992,37(7):393-404

Application of new procedures in the sphere of the control of sexual functions requires an extension of present knowledge of postparturient endocrinium or endogenic factors comprised in postparturient physiology of sexual activity. According to recent data, oxytocin, besides its uterotonic and luteolytic activity, acts as an ovarian factor in the local intrafollicular regulation of stereidogenesis and as a modulator of uterine secretion of prostaglandines. Based on present knowledge of oxytocin effects, this study was aimed at investigation of the influence of repeated carbetocin (Depotocin inj. Spofa) administration on the dynamics of changes in thyroxine (T4), triiodothyronine (T3), 17 beta-estradiol (E2), progesterone (P4) concentrations and their mutual correlations from the 36th hour till the 51st day after parturition. Simultaneous study of a possible delayed influence of applied carbetocin on conception of ewes after oestrus evocation on day 51 after lambing was carried out. Nineteen ewes of the Slovak Merino breed, lambed in the first decade of February, were assigned to the experimental (n = 9) and to the control group (n = 10). Experimental ewes were subjected to repeated postparturient carbetocin treatment at the dose 0.07 mg per animal. The first dose was applied i. m. in 24 hours, and the second in 72 hours after parturition. On day 51 oestrus was induced in nine ewes of each group by combined treatment with chlorsuperlutin (Agelin, vaginal pessaries, Spofa) and PMSG (500 I.U./animal). On the day of PMSG application ewes were housed together with rams for the period of the next six days. Samples of blood were taken 24 hours before parturition (-1st day), up to 36 h after parturition and on days 4, 7, 14, 17, 21, 25, 34, 42 and 51 after parturition. Concentrations of T4, T3, E2 and P4 were determined by commercial kits RIA-test-T4; RIA-test-T3; RIA-test-ESTRA and RIA-test-PROG (URVJT Kosice). Animal of the control group showed variations of T4 concentrations (Tab. I, Fig. 1) at the level of original values (59.4 +/- 9.69 nmol.l-1) up to the 21st day with the exception of temporary drop on day 4 and rise on day 7, insignificant compared to the -1st day. T4 concentrations of the control group displayed an intermittent increasing trend with the statistically insignificant peak after 36 h and on day 17, compared to the -1st day. After the 21st day controls revealed a sustained moderate increase while the experimental ewes displayed a decline of its concentrations until the 51st day.(ABSTRACT TRUNCATED AT 400 WORDS)  相似文献   

Ejaculatory failure associated with aortic-iliac thrombosis in two stallions.     

S M McDonnell  C C Love  B B Martin  V B Reef  R M Kenney 《Journal of the American Veterinary Medical Association》1992,200(7):954-957

Two mature breeding stallions were evaluated because of specific ejaculatory dysfunction, and each was found to have aortic-iliac thrombosis occluding 60 to 70% of the aortic lumen. In each case, the stallion had strong libido, normal mounting, and vigorous initial thrusting. With continued exertion, thrusting became weak and dismount was awkward. Treatment aimed at maximizing sexual arousal before mounting and reducing hind limb pain and exertion during breeding allowed these stallions to continue breeding. A program of gradually increasing daily exercise also was associated with improved breeding performance during subsequent breeding seasons. In cases of specific ejaculatory dysfunction, aortic-iliac disease should be included among the differential diagnoses.  相似文献   

Determination of sulfamethazine and trimethoprim in liquid feed premixes by HPLC and diode array detection, with an analysis of the uncertainty of the analytical results.     

B Cancho Grande  M S García Falcón  M Rodríguez Comesa?a  J Simal Gándara 《Journal of agricultural and food chemistry》2001,49(7):3145-3150

Sulfamethazine (SMZ) and trimethoprim (TMP) are antibacterials used in veterinary practice. This paper describes a method for their determination in veterinary liquid feed premixes that is based on liquid chromatography with diode array detection. Gradient elution with methanol and ammonium acetate achieved excellent separation of the two analytes within 15 min without any interference from the matrix. Absorbance of the column effluent was monitored at 264 nm for SMZ and at 230 nm for TMP. Detailed analyses of the uncertainties of determinations afford estimated expanded uncertainties of, respectively, 0.2 and 0.1 w/v % for typical SMZ and TMP concentrations of 10.7 and 2.1 w/v %, respectively. At the lower end of the calibrated range of the method, the dominant source of uncertainty is the preparation of standards and the construction of the calibration line.  相似文献   

Estradiol-17beta in the blood of boars with disorders of sexual potency     

S Navrátil  P Forejtek 《Veterinární medicína》1986,31(11):681-686

The levels of oestradiol-17 beta (E2) in the blood plasma were evaluated in 15 boars exhibiting signs of sexual potency disorders and in 20 boars with no such disorders before and after i. v. administration of chorion gonadotropin (HCG). No significant difference (P greater than 0.05) was found in the E2 levels in the blood plasma of the boars of the two groups before HCG administration. Two hours after the i. v. injection of 500 I. U. of HCG, an insignificant increase in the basal levels of E2 was recorded, reaching on the average 28.9% in the boars with potency disorders and 38.8% in those with no potency changes. Neither were there any significant differences in the E2 levels determined after HCG treatment between the boars with and without sexual potency disorders. It is inferred from the results that deviations in E2 concentration in the blood obviously do not contribute significantly to disorders in the sexual potency of boars.  相似文献   

Seroprevalence of Ornithobacterium rhinotracheale infection in commercial laying hens in the north central region of the United States.     

C J Heeder  V C Lopes  K V Nagaraja  D P Shaw  D A Halvorson 《Avian diseases》2001,45(4):1064-1067

This study was the first to examine the seroprevalence of Ornithobacterium rhinotracheale (ORT) within a commercial egg layer population. Serum samples collected from egg production companies were examined by serum plate agglutination test (SPAT) and outer membrane protein-based enzyme-linked immunosorbent assay (ELISA). Results show that 90% of layer flocks were positive by SPAT and 100% by ELISA. Of the pullet flocks examined, 43% and 52% were positive by SPAT and ELISA, respectively. Our study indicates that the prevalence of ORT antibody is high in the commercial layer population, suggesting that this respiratory pathogen can easily spread through multiple-age layer farms from older flocks to newly housed pullet flocks.  相似文献   

Relationships between catch and effort in Fijian multispecies reef fisheries subject to different levels of exploitation   总被引：2，自引：0，他引：2  

S. JENNINGS  N.V.C. POLUNIN 《Fisheries Management and Ecology》1995,2(2):89-101

Abstract The catch-per-unit-effort (CPUE) and value-of-catch-per-unit-effort (VPUE) of reef-associated fish species from six Fijian native fishing grounds ( qoliqoli ) subject to different fishing intensities were determined using records of fishing activity from a voluntary logbook scheme. Line and spear fishing techniques were used for more than half the total fishing time (h person^-1 d^-1) in all qoliqoli , and yet the favoured technique in a given qoliqoli was frequently less efficient (lower CPUE) than other techniques. The popularity of relatively ineffective fishing methods implies that fishermen did not always attempt to maximize their catch rates. To compare fishing effort in different qoliqoli , all effort was rescaled on the basis of its recorded efficiency (measured as multispecies CPUE) and expressed as hours equivalent to boat-based spear fishing over coral by day to catch fish for sale. Total fishing intensity in the six qoliqoli ranged from 72 to 4310 h km^-2 reef year^-1. The relationship between catch and effort was linear at all fishing intensities, suggesting that the qoliqoli were all fished on a sustainable basis. Furthermore, whilst there was a significant difference in CPUE and VPUE between the one or two qoliqoli with lowest fishing intensity and all the others, there were no significant differences between these other qoliqoli in terms of the VPUE of the saleable multispecies boat catch or the CPUE of species from piscivorous and carnivorous genera ( Epinephelus, Lethrinus and Plectropomus ) which the fishermen prefer to catch.  相似文献   

Teacher training     

Lopresti VC 《Science (New York, N.Y.)》1987,235(4790):730b

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