A specific DNA probe which identifies Babesia bovis in whole blood.

A genomic library of Babesia bovis DNA from the Mexican strain M was constructed in plasmid pUN121 and cloned in Escherichia coli. Several recombinants which hybridized strongly to radioactively labeled B. bovis genomic DNA in an in situ screening were selected and further analyzed for those which specifically hybridized to B. bovis DNA. It was found that pMU-B1 had the highest sensitivity, detecting 25 pg of purified B. bovis DNA, and 300 parasites in 10 microliters of whole infected blood, or 0.00025% parasitemia. pMU-B1 contained a 6.0 kb B. bovis DNA insert which did not cross-hybridize to Babesia bigemina, Trypanosoma evansi, Plasmodium falciparum, Anaplasma marginale, Boophilus microplus and cow DNA. In the Southern blot analysis of genomic DNA, pMU-B1 could differentiate between two B. bovis geographic isolates, Mexican strain M and Thai isolate TS4. Thus, the pMU-B1 probe will be useful in the diagnosis of Babesia infection in cattle and ticks, and in the differentiation of B. bovis strains.     

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)     

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

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)     

The effectiveness of the combined treatment of thrombocytic hemorrhagic diathesis of dogs with prednisolone and blood transfusion in the model of an aspirin-produced thrombocytopathy

The administration of 20 mg/kg of acetylsalicylic acid in 18 clinically healthy dogs resulted in a thrombocytopathy with lengthened capillary bleeding time and irreversible aggregation inhibition. Through the set up of individual dilution series, one could conclude the proportional percentage of aggregation functional transfused thrombocytes. The capillary bleeding time was shortened after the intravenous injection of prednisolone (5 mg/kg) without measurable influence on the thrombocytes. Compared to the singular use of cortisone or blood transfusion alone, the effect on capillary bleeding time became magnified when one combined transfusion and corticosteroids. The fresh blood conserves (12 hours) were, with respect to the haemostyptical properties, superior to blood stored for 5 days. The component of aggregation-functional thrombocytes was, due to the prophylactic cortisone application prior to transfusion, not increased.     

Experience with Cyclosporin-A After Renal Allografting in Two Dogs Research Note

After renal allografting, cyclosporin-A was administered to one partially nonmatched dog that was followed for 79 days. Cyclosporin-A and prednisolone were administered to one nonmatched dog that was followed for 805 days. Side effects encountered with cyclosporin-A included lymphocytic dermatitis, papillomatosis, bacterial and fungal infections, and B lymphocyte hyperplasia.     

Effect of dietary copper and vitamin E supplementation, and extensive feeding with acorn and grass on longissimus muscle composition and susceptibility to oxidation in Iberian pigs

The objective of this study was to assess the effect of dietary copper and/or vitamin E supplementation on composition and oxidation of M. longissimus in Iberian pigs and to compare it with meat samples from pigs produced extensively and fed with acorn and grass. Grass had the highest α-tocopherol content (> 150 mg/kg DM), while acorns had the highest copper concentration (78.1 mg/kg DM). Dietary treatment did not affect copper composition in muscle and no interactions were detected between copper and α-tocopherol. The α-tocopherol content in muscle from pigs fed diets supplemented with α-tocopheryl acetate (100 mg/kg feed) was 1.5 times greater (p < 0.0001) than those from pigs receiving a basel diet. M. longisssimus dorsi from pigs fed extensively had a higher concentration of α-tocopherol than those fed in confinement with the basel level of α-tocopheryl acetate, but lower values than pigs fed supplemented levels. Total n -3 fatty acids (p < 0.02) and hematin (p < 0.0001) concentrations were significantly higher in muscle from pigs fed extensively than when fed in confinement. Muscle homogenates from Iberian pigs fed in extensive conditions showed significantly (p < 0.02) higher susceptibility to oxidation than those from pigs fed mixed diets. Dietary α-tocopheryl acetate supplementation (100 mg/kg feed) significantly (p < 0.05) reduced lipid oxidation of muscle, while dietary copper concentration did not modify susceptibility to lipid oxidation.     

Effect of birthweight, total protein, serum IgG and packed cell volume on risk of neonatal diarrhea in calves on two California dairies.

The objective of the study was to determine if there was a relationship between hematological, immunological and physiological variables of newborn calves and risk of diarrhea during the neonatal period. Four hundred and seventeen heifer calves from two dairies (A and B) in the San Joaquin Valley of California were enrolled at birth and scored daily, to 28 days of age, for evidence and severity of diarrhea (0 to 3). Calves were weighted at birth and blood sampled at two to five days of age to determine packed cell volume (PCV), total protein (TP) and IgG serum concentration. The Cox proportional hazards model was used to determine if age at onset of the first diarrhea episode and length of the first episode were associated with the hypothesized variables (PCV, TP, IgG and birthweight). The IgG concentration was not associated with the age at onset of diarrhea (p = 0.6052, Dairy A; p = 0.4393, Dairy B) but a high IgG concentration was associated with a decreased length of episode (p = 0.0325, Dairy A; p = 0.0912, Dairy B), particularly for calves born in the winter on dairy A (p = 0.0211). For calves born in the winter, those with either a high or a low birthweight had diarrhea at a younger age (p = 0.0102, Dairy A; p = 0.0020, Dairy B). Associations were also found for PCV and TP with both the age at onset and length of the first episode of diarrhea. Results suggest that parameters measurable at, or shortly after birth may have important prognostic value in evaluating risk of calf diarrhea.     

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]

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)     

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

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.     

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

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.