An automated micro-method for the enzymatic determination of D-B-hydroxybutyrate in ruminant plasma

1. An enzymatic ‘reaction rate’ micro-method for the rapid routine estimation of D-B-hydroxybutyrate (D-B-HOB) in ruminant plasma, using an I.L. Multistat III centrifugal analyzer, is described.
2. Reaction conditions were optimized to give a linear response for plasma D-B-HOB concentrations between 100 and 2500 μmoles per litre, at 30°C and pH 9.0.
3. For the standardized method the within-run and between-run coefficients of variation for deproteinised ovine plasma were consistently less than 3.5%.
4. There was good agreement between plasma concentrations obtained by the present method and both original U.V. end-point technique (r=0.927b=0.950) and a colorimetric end-point procedure (r=0.937. b=0.879).
5. Utreated ovine and bovine plasma consistently exhibited high ‘blank’ activity and this was directly correlated with plasma lactate dehydrogenase (LDH) activity in both species (r=0.971; p<0.001 and r=0.949; p<0.001 respectively). The distribution of LDH activity in man was similar to sheep but, contrastingly, non-specific interference was extremely low in human plasma and unrelated to LDH. Horse, chicken and rat had negligible ‘blank” activity and comparatively low LDH levels. In both cattle and sheep non-specific interference was abolished by perchloric acid precipitation. In the sheep subtraction of ‘blank’ activity gave D-H-HOB concentrations for untreated plasma comparable to those in deproteinised samples. However, in the bovine, D-B-HOB levels remained significantly (t=6.44; p<0.001) higher even after ‘blank’ correlation. In contrast to man and other non-ruminants, perchloric acid precipitation is essential in ruminants to avoid false overestimation of plasma D-B-HOB levels.
6. Plasma with EDTA as anticoagulant and serum gave concentrations of D-B-HOB approximately 60% lower, than samples containing heparin or oxalate/fluoride. However, heparin was associated with much higher (up to 50%) non-specific NAD rduction than oxalate/fluoride.
7. High levels of acetoacetate (400–1000 μmoles per litre) reduced the recovery of D-B-HOB from ovine plasma by less than 10%. This effect was negated by the inclusion of hydrazine hydrates in the reaction mixture. Perchlorate ion concentrations above 25 μmoles per litre per test dramatically inhibited the assay in ovine plasma, and therefore precipitation conditions must be carefully controlled.
8. Plasma with oxalate/fluoride as anticoagulant showed the greatest stability in storage; 24 hours at room temperature, one week at +4°C and at least one month at ?20°C.

     

An exploration of Cryoprotective compounds for fowl spermatozoa

1. Glycerol is an effective cryoprotective for fowl spermatozoa, but after thawing the frozen semen it must be reduced in concentration from the level adequate to protect spermatozoa during freezing, otherwise it has a contraceptive action.

2. A series of alternative cryoprotective compounds were tested for their effect on fertility when fowl spermatozoa were inseminated fresh in their presence. Under these circumstances dimethylsulphoxide, dimethylaceta‐mide, ethane‐diol, propane‐diol and methylpyrrolidone did not depress fertility when used in concentrations equivalent to that of glycerol or in amounts reported previously to be non‐toxic and adequate to protect cells during freezing.

3. Dimethylacetamide was compared with propane‐diol for use in freezing fowl semen and the former enabled encouraging levels of fertility to be obtained.     

Effect on Perinatal Mortality of a Single Selenium Injection To Sows and Gilts

Selenium treatment of pregnant sows and gilts was conducted as a double-blind placebo-controlled study in 5 commercial herds with a total of 350 sows and gilts (Danish Landrace, Yorkshire and Landrace-Yorkshire crossbreeds). At 3–6 weeks before farrowing the animals were injected once intramuscularly with either Na-selenite (0.05 mg Se per kg) or placebo. Blood samples were taken immediately before treatment and once again at 3–6 weeks after farrowing, to evaluate the Se status. The productivity of the sows and gilts was recorded.Within herds there was no pretreatment difference in Se status between the two treatment groups, while a considerable variation was seen among herds. After farrowing there were only minor differences in Se status between the treatment groups, whereas the variation among herds persisted.Overall, no effect of the Na-selenite injection was seen, in that there was no difference in number of liveborn, stillborn, and weaned piglets between the treatment groups. The number of days from weaning to oestrus and the number of oestrous cycles per pregnancy were not influenced by the Na-selenite treatment.     

Effects of age and prosthesis material on in vitro cartilage retention of laryngoplasty prostheses in horses

Cartilage retention strengths of laryngoplasty prostheses were compared in larynges of 2-, 3-, and 4-year-old horses, using doubled polyester and expanded polytetrafluoroethylene prostheses. Bilateral laryngoplasties were performed on each of 15 (seven 2-year-old, two 3-year-old, and six 4-year-old) larynges, which were collected at an abbatoir. Prostheses were secured to a mechanical testing machine, and tension causing arytenoid cartilage abduction was applied, until total failure of the cartilage or prosthesis resulted. Tension caused cricoid cartilage failure in 1 specimen, and muscular process cartilage failure in the remainder. There was no significant effect of age, prosthetic material, or side of prosthesis placement on cartilage retention of the prostheses. Additionally, frequency of multiple load-displacement peaks, indicating partial muscular process failure, was not affected by age or prosthetic material variables.     

Bovine endometritis: comparative efficacy of alfaprostol and intrauterine therapies, and other factors influencing clinical success

A comparative study of the treatment of 306 severe, moderate, or mild cases of bovine endometritis was carried out over two calving seasons. The cases were treated with alfaprostol, or an intrauterine antibacterial preparation, or with a combination of both therapies. There was no significant difference between the efficacies of these treatments, and a single injection of alfaprostol was effective in 74 per cent of the cases treated. The effectiveness of the treatments was related to the degree of self-cure of the endometritis after parturition, the luteal activity at the time of treatment, and farm management factors affecting the health and condition of the calving cows.     

Surgical Repair of Tibial Tuberosity Avulsion Fractures in Four Horses

Four horses, 2 to 17 years old, were treated for unilateral avulsion fractures of the tibial tuberosity. Two horses were treated successfully with tension band wiring or plating in combination with lag screw fixation. One horse was euthanatized because of implant failure during recovery and one was euthanatized on day 11 because a longitudinal fracture of the tibial tuberosity occurred through the plane of the screws used for stabilization.     

Pharmacokinetics of pipemidic acid in chickens after single intravenous and oral dosings

The pharmacokinetics of pipemidic acid after 2 single doses were studied in broiler chickens. Chickens were given single IV and oral doses of 10 and 30 mg of pipemidic acid/kg of body weight. Blood samples were collected over 8 hours after each dose administration. High-pressure liquid chromatography with UV detection was used to determine concentrations in plasma of pipemidic acid. The plasma concentration-time curves after IV administration followed 2-compartment characteristics, rapid initial distribution phase, and a terminal elimination phase. The pharmacokinetic variables differed significantly between single doses of 10 and 30 mg of pipemidic acid/kg. Mean disposition variables were a half-life at alpha phase of 0.06 hours or 0.33 hours, a half-life at beta phase of 1.18 hours or 1.72 hours, a volume of distribution in the central compartment of 0.12 L/kg or 0.31 L/kg, a volume of distribution during the elimination beta phase of 1.64 L/kg or 1.05 L/kg, and a total plasma clearance of 0.97 L/h.kg or 0.41 L/h.kg, for the 10 or 30 mg/kg dose, respectively. After oral administration, the pipemidic acid plasma profile could be adequately described by a 1-compartment model. After the single oral doses of 10 and 30 mg of pipemidic acid/kg, pipemidic acid was absorbed rapidly (time to maximal concentration of 0.31 hours or 0.71 hours) and eliminated with a mean half-life of 0.86 hours or 0.61 hours, respectively. The bioavailability was 39% at 10 mg of pipemidic acid/kg and 61% at 30 mg of pipemidic acid/kg.     

Experimental transmission of a dermal sarcoma in fingerling walleyes (Stizostedion vitreum vitreum)

Dermal sarcoma is a benign skin tumor of adult walleyes (Stizostedion vitreum vitreum) with a suspected viral etiology. A laboratory study was initiated to determine if the tumor could be experimentally transmitted by inoculating young walleyes with materials prepared from tumors from adult fish. Eighty walleye fingerlings were divided into four groups of 20 fish each. Two groups were inoculated intramuscularly at 4 months of age either with live tumor cells or with cell-free filtrates of sonicated tumor cells. The two other groups were used as controls and were inoculated either with cultured cells from normal walleye fry or with tissue culture media. Neoplasms, similar to the dermal sarcoma affecting adult walleyes, were observed after 4 months only in fingerlings inoculated with cell-free filtrates of sonicated tumor cells. Like the tumor affecting wild adult walleyes, the transmitted tumors were restricted to the dermis and originated from the superficial surface of scales. They never invaded locally and never metastasized. The transmitted tumors differed from tumors of adult walleyes in their severity and the absence of osteoid. The multicentric origin of transmitted walleye dermal sarcoma suggests that the virus spreads systemically and that tumor cells are polyclonal. This successful transmission of the lesion, along with the presence of C-type virus particles budding from tumor cells in two of seven tumor-bearing fingerlings, supports a retroviral etiology.