Sex determination of bovine embryos using H-Y antibodies

6 days old bovine embryos (n = 126) were obtained from 8 superovulated cows or heifers by flushing the uteri and oviducts either non-surgically or after slaughter. Part of the embryos (n = 72) (morula stages) were placed in Ham's F-10 or PBS supplemented with 10% fetal calf serum (FCS) diluted 1:1 with supernatant from the H-Y antibody producing clone and cultured at 38 degrees C, in 5% CO2/95% air and 100% humidity. Control embryos (n = 54) were cultured in H-Y antibody free medium. After culture the embryos could be separated into a blastocyst--and a morula group. A subsequent colchemid and hypotonic treatment and fixation and Giemsa staining allowed a precise karyotyping, and thus sex determination for 36 H-Y antibody treated embryos and 22 control embryos. The limiting factor for proper karyotyping was lack of metaphases, incomplete methaphases or poor preparation. Among the H-Y antibody treated embryos we found 7 males and 15 females in the blastocyst and 14 males and 0 females in the morula group. A statistical analysis of these proportions led to the conclusion that the H-Y antibody had a significant influence on the sex ratio.     

Characterization of the bound residues of the fungicide cyprodinil formed in plant cell suspension cultures of wheat

The non-extractable residues of the fungicide cyprodinil formed in heterotrophic cell suspension cultures of wheat were studied by application of [2-pyrimidyl-14C] or [2-pyrimidyl-13C]cyprodinil. The main objective was to examine whether solid-state and liquid 13C NMR spectroscopy can be used to examine plant bound residues of pesticides. For 14C experiments, wheat suspensions grown on glucose as carbon source were treated with 10 mg litre(-1) of 14C-cyprodinil. After incubation for 12 days, 20% of applied 14C was detected as non-extractable residues. The cell debris were treated with 0.1 M HCl (reflux), 1.0 M HCl (reflux), buffer, or 2 M NaOH (50 degrees C); Bj?rkman lignin and acidolysis lignin fractions were also prepared from the debris. Radioactivity liberated and solubilized by these procedures was examined by thin-layer chromatography and high-performance liquid chromatography. The results showed that cyprodinil and primary metabolites contributed to the fungicide's bound residues. Most of the residues (12% of applied 14C) remained associated with polar or polymeric/oligomeric endogenous cell materials in a stable manner. For the study with 13C-cyprodinil, wheat suspensions were cultivated on 13C-depleted glucose for four growth cycles, resulting in maximum 13C depletion of the natural cell components to about 0.10%. During the fourth cycle, 13C-labelled cyprodinil was applied, and cells were incubated (12 days). Cell debris was prepared and examined by solid-state 13C NMR spectroscopy. Debris was then treated as described above in the 14C experiment. Solubilized fractions were analyzed by liquid 13C NMR spectroscopy. However, none of the 13C NMR spectra recorded gave utilizable or unambiguous results, and all exhibited large inconsistencies, especially concerning the data from the conventional 14C experiment.     

Effects of racing and nontraining on plasma thyroid hormone concentrations in sled dogs

OBJECTIVE: To determine the effects of racing and nontraining on plasma thyroxine (T4), free thyroxine (fT4), thyroid-stimulating hormone (TSH), and thyroglobulin autoantibody (TgAA) concentrations in sled dogs and compare results with reference ranges established for dogs of other breeds. DESIGN: Cross-sectional study. ANIMALS: 122 sled dogs. PROCEDURE: Plasma thyroid hormone concentrations were measured before dogs began and after they finished or were removed from the Iditarod Trail Sled Dog Race in Alaska and approximately 3 months after the race. RESULTS: Concentrations of T4 and fT4 before the race were less than the reference range for nonsled dogs in 26% and 18% of sled dogs, respectively. Immediately after racing, 92% of sled dogs had plasma T4 concentrations less than the reference range. Three months after the race, 25% of sled dogs had plasma T4 concentrations less than the reference range. For T4, fT4, TSH, and TgAA, significant differences were not detected in samples collected before the race versus 3 months later. CONCLUSIONS AND CLINICAL RELEVANCE: Plasma T4, fT4, and TSH concentrations decreased in dogs that complete a long distance sled dog race. Many clinically normal sled dogs have plasma T4 and fT4 values that are lower than the reference range for nonsled dogs. We suggest that the reference ranges for sled dogs are 5.3 to 40.3 nmol/L and 3.0 to 24.0 pmol/L for plasmaT4 and fT4 concentrations, respectively, and 8.0 to 370 mU/L for TSH.     

A spontaneous and fatal jejunal intussusception in a European brown hare associated with Eimeria leporis

A recently dead adult female European brown hare (Lepus europaeus Pallas) was found in the wild with no evidence of trauma or external injury. Necropsy revealed symptoms consistent with endotoxic shock and a haemorrhagic jejuno-jejunal intussusception with a necrotic intussusceptum. Small, unattached nodules were present extensively in the jejunal lumen and within the intussuscipiens. These nodules proved to be masses of unsporulated Eimeria leporis oocysts. Review of the literature reveals E. leporis to be a common parasite of wild hares, with protracted pathogenic potential under some circumstances. This is the first report of intussusception in this species and the most severe lesion seen associated with E. leporis. This case suggests that E. leporis may be more harmful to the wild European brown hare population than previously supposed.     

Effect of dietary lipoic acid on metabolic hormones and acute-phase proteins during challenge with infectious bovine rhinotracheitis virus in cattle

OBJECTIVE: To determine the effect of dietary supplemental lipoic acid (LA) on serum concentrations of metabolic hormones and acute-phase proteins of steers challenged with infectious bovine rhinotracheitis virus (IBRV). ANIMALS: 32 steers. PROCEDURES: Steers were randomly assigned to 4 treatments: negative control (NC; no LA, no IBRV challenge), control (CON; no LA, IBRV challenge), 16 mg of LA/kg of body weight (BW)/d plus IBRV challenge (LA16), and 32 mg of LA/kg of BW/d plus IBRV challenge (LA32). Following a 21-day adaptation period, CON, LA16, and LA32 steers received IBRV (2 mL/nostril [day 0]); NC steers received saline (0.9% NaCl) solution. Blood samples, nasal swab specimens, BW, and rectal temperatures were obtained 0, 1, 3, 5, 7, 14, and 21 days after challenge. Serum was analyzed for concentrations of haptoglobin, amyloid-A, leptin, and anti-IBRV antibodies. RESULTS: Steers fed LA32 began gaining BW by day 7, whereas BW of CON and LA16 steers declined. Serum haptoglobin concentration of LA32 steers was lower than that of CON and LA16 steers on day 7. Serum neutralization titers for 30 of 32 steers were negative for anti-IBRV antibodies before challenge; however, all steers (including NCs) had antibodies on day 21. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggested that LA supplementation augmented certain aspects of the immune response; LA32 steers had a more rapid recovery from IBRV viral challenge than did others.     

Rapid identification of the animal pathogens Streptococcus uberis and Arcanobacterium pyogenes by fluorescence in situ hybridization (FISH)

Fluorescence in situ hybridization (FISH) has been reported to be an easy and rapid identification method for many human pathogens, but applications for common veterinary pathogens are lacking. Gene probes for FISH of the animal pathogens Streptococcus uberis and Arcanobacterium pyogenes were designed to provide probes for a specific identification of these bacteria from cultures. Specific FISH probes for these species have so far not been published. Both probes recognized all isolates of the target species correctly. With the S. uberis probe SUB 196 no false-positive results were obtained for reference strains as well as for clinical isolates. Probe APYO 183 for A. pyogenes produced false-positive reactions with so far rarely described Arcanobacterium species from animals at standard hybridization conditions. In order to avoid any incorrect classifications of microorganisms as A. pyogenes, two non-labelled competitor probes were designed and successfully evaluated.