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.     

Coccidioidomycosis in 48 Cats: A Retrospective Study (1984–1993)

Coccidioidomycosis was diagnosed in 48 cats. Forty-one cases were identified within a period of 3 years. Coccidioides immitis was revealed by cytological or histopathological examinations, or culture in 70% of cats. The remaining 30% of cases were diagnosed by appropriate clinical signs, radiographic lesions, and serological test results. The average age of affected cats was 6.2 years with a median age of 5.0 years. Fifty-four percent (n = 26) were female and 46% (n = 22) were male. Domestic shorthaired and longhaired breeds comprised 89% (n = 41) of affected cats. Sixty-seven percent of cases were diagnosed during the 6-month period of December through May. Cats infected with C immitis were presented for evaluation of dermatologic (56%), respiratory (25%), musculoskeletal (19%), and neurological or ophthalmologic signs (19%). Fever, inappetence, and weight loss were present in 44% of the cats. Duration of clinical signs before diagnosis was less than 4 weeks in 85% (n = 42) of cats, with an average of 3.8 weeks and a median of 2 weeks. Agar gel immunodiffusion tests were positive in all 39 cats tested at sometime during the course of their disease. Hyperproteinemia (greater than 7.9 g/dL) was present in 52% (10/23) of cases. The majority of cats (n = 39) were negative for feline leukemia virus. Antibodies to feline immunodeficiency virus were absent in the 19 cats tested. Ketoconazole was the most common antifungal agent used to treat cats with Coccidioidomycosis. Duration of treatment ranged from less than 1 week to 43 months. Thirty-two cats are currently asymptomatic, with or without treatment. Eleven cats died or were euthanized. Five cats were lost to follow-up. Ketoconazole likely is more suppressive than curative because relapses were common after discontinuing therapy.     

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.     

Cross protection studies on Bordetella bronchiseptica in mice using an intracerebral challenge model.

Protective activities of heat-inactivated (60 degrees C for 30 min) merthiolate preserved Bordetella bronchiseptica and B. pertussis bacterins were compared in intraperitoneally immunized mice challenged intracerebrally (i.p./i.c.) or intraperitoneally (i.p./i.p.). In the i.p./i.c. assay (Kendrick test), a B. pertussis bacterin protected mice against challenge with B. pertussis 18-323, as well as against phase I cytotoxic and non-cytotoxic strains of B. bronchiseptica. A B. bronchiseptica bacterin, prepared from a phase I cytotoxic strain, gave protection against two phase I B. bronchiseptica strains, irrespective of their cytotoxin-production. A non-cytotoxic phase I strain of B. bronchiseptica elicited protection against the homologous strain only. Neither cytotoxic nor non-cytotoxic B. bronchiseptica strains protected mice challenged with B. pertussis 18-323. Vaccines prepared from phase III strains of B. bronchiseptica were not protective at all against any of the challenge strains. No such differences in the protective activities of the bacterins could be detected by the i.p./i.p. method. They seem to cross-protect equally well. The results indicate that the Kendrick test may be useful in testing potency of different B. bronchiseptica bacterins.     

In vivo kinetic study on uptake and distribution of intramuscular tritium-labeled polysulfated glycosaminoglycan in equine body fluid compartments and articular cartilage in an osteochondrial defect model

The uptake and distribution of intramuscularly (IM) administered tritium-labeled polysulfated glycosaminoglycan (³H-PSGAG) in serum, synovial fluid, and articular cartilage of eight horses was quantitated, and hyaluronic acid (HA) concentration of the middle carpal joint was evaluated in a pharmacokinetic study. A full-thickness articular cartilage defect, created on the distal articular surface of the left radial carpal bone of each horse served as an osteochondral defect model. ³H-PSGAG (500 mg) was injected IM, between 14 and 35 days after creation of the defects. Scintillation analysis of serum and synovial fluid, collected from both middle carpal joints at specific predetermined times up to 96 hours post-injection, revealed mean ³H-PSGAG concentrations peaked at 2 hours post-injection. ³H-PSGAG was detected in cartilage and subchondral bone 96 hours post-injection in samples from all eight horses. There were no statistically significant differences in ³H-PSGAG concentration of synovial fluid or cartilage between cartilage defect and control (right middle carpal) joints.

HA assay of synovial fluid revealed concentrations significantly increased at 24, 48, and 96 hours post-injection in both joints. The concentration nearly doubled 48 hours post-injection. However, no statistically significant differences were found between synovial concentrations of HA in cartilage defect and control joints.

³H-PSGAG administered IM to horses, was distributed in the blood, synovial fluid, and articular cartilage. HA concentrations in synovial fluid increased after IM administration of polysulfated glycosaminoglycan.     

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.     

Coagulopathic Effects and Therapy of Brodifacoum Toxicosis in Dogs

The clinical signs and laboratory changes of brodifacoum (BDF) intoxicated dogs and their response to vitamin K1 treatment were examined. Brodifacoum, a second-generation anticoagulant rodenticide, was fed to four dogs for 3 consecutive days producing a cumulative dose of 1.1 mg BDF/kg body weight. Clinical observations of the animals were made daily throughout the study. Monitored laboratory parameters included: one-stage prothrombin time (OSPT), activated partial thromboplastin time (APTT), activated coagulation time (ACT), complete blood counts, thrombocyte counts, and serum chemistry values. Response to vitamin K1 therapy was evaluated clinically and by laboratory tests. Serum BDF concentrations were monitored. Inappetence and hemorrhagic tendencies were exhibited by day 5 postrodenticide exposure. One-stage prothrombin time, APTT, and ACT were 25% greater than time zero values at 24, 24, and 72 hours postdosing, respectively. All laboratory parameters returned to normal within 48 hours of initiating vitamin K1 therapy (0.83 mg/kg orally, TID for 5 days). Serum brodifacoum concentrations were highest (1065-1215 ng/mL) during the 3 days after BDF dosing and were detectable (3.0-7.5 ng/mL) until day 24 postexposure. A mean BDF elimination half-life of 6 +/- 4 days was observed.     

Analysis of a Long-term Experiment with Repeated-measurement Models

Long‐term experiments are a classical case of repeated measurements. Traits are measured on the same experimental unit over many years so that correlations arise between the observations made on the same plot in consecutive years. This paper describes the analysis of a three‐crop‐rotation long‐term experiment. We analysed the yields of the crops and the organic carbon content in the topsoil over 30 consecutive years. Several variance–covariance approaches are discussed and the trait‐specific best fit is interpreted. Mixed models are used to describe the structure of the experiment. Both yields and soil organic carbon show a more or less pronounced variance heterogeneity. Especially for yields, the heterogeneity of cycles and years is dominant. The consideration of correlations results in a better model fit in all cases.