Flow Cytometry Identification of X- and Y-Chromosome-Bearing Goat Spermatozoa

Flow cytometric sorting technology was used to measure the difference in DNA content between X- and Y-chromosome-bearing spermatozoa in bucks. Spermatozoa were analysed by flow cytometry to characterize X- and Y-chromosome-bearing sperm populations and to quantify the DNA difference between them. Two symmetrical, overlapping and clearly separated peaks, corresponding to X- and Y-bearing spermatozoa, were detected. The difference in fluorescence intensity between the peaks was 4.4 +/- 0.03% without any significant inter- or intra-animal variations. Therefore, the identification and selection of high-purity samples of sperm populations for sex sorting is easier in bucks compared with other domestic species.     

Key Factors Affecting Reproductive Success of Thoroughbred Mares and Stallions on a Commercial Stud Farm

To evaluate factors contributing to fertility of thoroughbred mares, data from 3743 oestrous periods of 2385 mares were collected on a large thoroughbred farm in Ireland. Fourteen stallions (mean age 8.3 years; range 4–15 years) had bred 2385 mares (mean age 9.4 years; range 3–24 years). Maiden mares accounted for 12%, mares with a foal at foot for 64%, and barren, slipped or rested mares for 24% of the total. The mean pregnancy rate per cycle was 67.8% (68.6% in year 1 and 66.9% in year 2). Backward stepwise multivariable logistic regression analysis was utilized to develop two models to evaluate mare factors, including mare age, reproductive status, month of foaling, dystocia, month of cover, foal heat, cycle number, treatments, walk‐in status and stallion factors including stallion identity, stallion age, shuttle status, time elapsed between covers and high stallion usage on the per cycle pregnancy rate and pregnancy loss. Old age (p < 0.001) and cover within 20 days post‐partum (p < 0.003) were associated with lowered pregnancy rates. High mare age (p < 0.05) and barren, slipped or rested reproductive status (p = 0.05) increased the likelihood of pregnancy loss. Uterine inflammation or infection, if appropriately treated, did not affect fertility. Only high usage of stallions (used more than 21 times in previous week) was associated with lowered (p = 0.009) pregnancy rates. However, shuttle stallions were more likely to have increased (p = 0.035) pregnancy survival, perhaps reflecting a bias in stallion selection. In conclusion, mare age exerted the greatest influence on fertility; nonetheless, thoroughbreds can be effectively managed to achieve high reproductive performance in a commercial setting.     

Influence of Bos indicus crossbreeding and cattle age on apparent utilization of a high-grain diet

Ten Bos indicus x MARC III (initial BW = 303 +/- 25 kg) and 10 MARC III (initial BW = 322 +/- 16 kg) steers were used in a 2 x 2 factorial design to determine whether cattle age or Bos indicus crossbreeding influence site of digestion of a high-grain diet. Initially, five Bos indicus x MARC III and five MARC III steers were fitted with duodenal cannulas and adapted to a 95% concentrate diet that was offered for ad libitum consumption for a 237-d feeding period (calves). During the feeding period, duodenal and fecal samples were collected during 4-d periods beginning on d 14, 67, 137, and 228. The remaining 10 steers were fed a forage-based diet for a targeted daily gain of .6 to .7 kg for 210 d (yearlings). Following this period, yearling steers were duodenally cannulated and adapted to the 95% concentrate diet. Yearling steers had ad libitum access to feed for 165 d, and samples were collected during 4-d periods beginning on d 13, 42, 102, and 159. Dry matter intake was 9.8 and 7.6 kg/d and daily gain was 1.35 and 1.16 kg in yearlings and calves, respectively. Apparent OM digestion in the stomach was greater (P < .01) in yearlings than in calves. In contrast, postruminal disappearance as a percentage of OM intake was greater (P = .05) in calves than in yearlings. Duodenal flows of total N, microbial N, nonmicrobial N, and total amino acids and total tract N digestibility were not affected (P > .05) by age or Bos indicus crossbreeding. Fecal N excretion was greater (P < .01) in yearlings than in calves. Results of this experiment suggest little effect of Bos indicus influence on utilization of a high-grain diet. However, more feed is digested in the rumen of yearlings than of calves consuming a high-grain diet.     

Butorphanol and azaperone as a safe alternative for repeated chemical restraint in captive white rhinoceros (Ceratotherium simum).

Anesthesia in the white rhinoceros (Ceratotherium simum) has routinely involved potent narcotic anesthetic agents such as etorphine or carfentanil with their associated adverse side effects. In captive rhinoceroses conditioned to routine handling, a combination of butorphanol and azaperone at mean (+/- SD) doses of 69.3 +/- 18.0 mg and 103.1 +/- 20.9 mg, respectively, was used to produce levels of neuroleptanalgesia ranging from light "standing" sedation to deeper planes of anesthesia producing sternal and lateral recumbency. This combination was used for repeated (minimum repeat frequency of 3 days between events) anesthetic episodes (n = 26) in two animals, with the remaining episode performed in a white rhinoceros with chronic renal disease. The action of butorphanol was satisfactorily reversed with naltrexone (125 mg i.v. and 125 mg i.m.). Results (mean +/- SD) include sternal recumbency achieved in 14.1 +/- 8.1 min after i.m. dosing, standing and ambulation occurred in 1.7 +/- 0.6 min after reversal, heart rate was 62.0 +/- 10.1 beats/min, respiratory rate was 14.7 +/- 5.6 breaths/min, and percentage of oxygen saturation of hemoglobin (Spo2) was 89.2 +/- 3.0%. Without supplementation, the total elapsed time ranged from 44.9 min to 103.0 min, whereas elapsed times up to 214.3 min were achieved with supplementation (mean time to supplementation was 28.0 +/- 13.9 min after initial dosing). Butorphanol and azaperone produced adequate muscle relaxation and apparently adequate analgesia for minor surgical interferences, including abdominal laparoscopy. Respiratory rates and Spo2 measurements were improved compared with reports of using more potent opioids in this species.     

Intralesional cisplatin chemotherapy and topical cryotherapy for the control of choanal squamous cell carcinoma in an African penguin (Spheniscus demersus).

A 3.5-yr-old, 2.5-kg female African penguin (Spheniscus demersus) was diagnosed with a choanal squamous cell carcinoma on the basis of biopsy after a history of choanal ulceration and anorexia with weight loss. Therapeutic modalities included the use of intralesional cisplatin followed by localized, topical cryotherapy with liquid nitrogen. The penguin remained free of clinical disease for a period of 9 mo, at which time a second cryotreatment was performed. The patient has remained free of all signs of choanal carcinoma for a period of 13 mo since the second cryotreatment.     

Effects of delmadinone acetate on pituitary-adrenal function, glucose tolerance and growth hormone in male dogs

Objective To characterise the effects of delmadinone acetate on the pituitary-adrenal axis, glucose tolerance and growth hormone concentration in normal male dogs and dogs with benign prostatic hyperplasia.
Design A prospective study involving nine normal male dogs and seven with prostatic hyperplasia.
Procedure Delmadinone acetate was administered to six normal male dogs and seven dogs with benign prostatic hyperplasia at recommended dose rates (1.5 mg/kg subcuta-neously at 0, 1 and 4 weeks). Three normal controls received saline at the same intervals. Blood concentrations of ACTH, cortisol, glucose, insulin and growth hormone were measured over 50 days. Intravenous glucose tolerance and ACTH response tests were performed before and after treatment in the nine normal animals.
Results A substantial suppression of basal and 2 h post-ACTH plasma cortisol secretion was demonstrated after one dose in all dogs given delmadinone acetate. Individual responses after the second and third administration varied between recovery in adrenal responsiveness to continued suppression. Plasma ACTH concentration was also diminished after one treatment. No effects were evident on glucose tolerance or serum growth hormone concentrations.
Conclusion Delmadinone acetate causes adrenal suppression from inhibition of release of ACTH from the pituitary gland. Treated dogs may be at risk of developing signs of glucocorticoid insufficiency if subjected to stressful events during or after therapy. Neither glucose intolerance nor hyper-somatotropism seems likely in male dogs given delmadinone acetate at the recommended dose rate, but the potential for excessive growth hormone secretion in treated bitches remains undetermined.     

Lipid Droplet Analysis Using In Vitro Bovine Oocytes and Embryos

The aim of this study was to quantify the content of lipid droplets in bovine oocytes and embryos from Bos indicus (Bi), Bos taurus (Bt) and Bos indicus × Bos taurus (Bi × Bt). Oocytes were aspirated post‐mortem and subjected to in vitro maturation, in vitro fertilization and in vitro development; the medium employed at each stage (TCM‐199, TALP, SOF) was supplemented with (i) serum replacement (SR), (ii) foetal calf serum (FCS) or (iii) oestrous cow serum (ECS). The structure and distribution of the lipid droplets were established using electron microscopy, but were quantified using an optical microscope on semi‐fine toluidine blue‐stained sections. The highest percentage of embryos corresponded to those produced with FCS and ECS, which differed from embryos generated with SR (p < 0.05). The highest percentage of morulae and the lowest percentage of blastocysts were obtained with the SR supplement (p < 0.05). The oocytes cultured in FCS demonstrated a higher number of lipid droplets compared to those cultured in SR and ECS (p < 0.05). Less accumulation of lipids was observed in embryos supplemented with SR. The lowest and highest numbers of lipid droplets in oocytes corresponded to the Bi and Bt strain, respectively. The lowest amount of lipid droplets in embryos was observed in Bi (p < 0.05). In conclusion, supplementation of the in vitro development culture medium (synthetic oviduct fluid) with a synthetic substitute serum produced similar results in terms of embryo development compared to those obtained with FCS, but a decreased degree of lipid droplet accumulation was observed in the in vitro‐cultured embryos.     

Genetic divergence of kingfish from Japan, Australia and New Zealand inferred by microsatellite DNA and mitochondrial DNA control region markers

ABSTRACT: Genetic polymorphism in kingfish, collected from coastal waters of Japan, Australia and New Zealand, were examined using microsatellite (MS) DNA and mitochondrial DNA (mtDNA) control region markers. Sixteen to 25.7 alleles per locus were observed in three MS markers, while the average observed (and expected) heterozygosities were 0.782 (0.918), 0.750 (0.809) and 0.650 (0.888) for Australian, Japanese and New Zealand kingfish, respectively. Twelve mtDNA haplotypes were detected by the digestion of control region sequences with five endonucleases: Hae III, Hinf I Mbo I, Rsa I and Taq I. Significant genetic divergence was observed between the kingfish population from Japan and those from Australia–New Zealand. There was no significant differentiation among the Australian and New Zealand population samples.     

Level of nutrition and splanchnic metabolite flux in young lambs

Splanchnic metabolite flux was measured in young lambs given access to a high-concentrate diet either ad libitum (ADLIB) or at a maintenance level (MAINT) for 21 d. Net fluxes of urea N (UN), ammonia N (NH3 N), alpha-amino N (AAN), amino acids, glucose (G), and lactate (L) across the liver and portal-drained viscera (PDV) were measured in 11 crossbred ram lambs (35 kg) surgically fitted with indwelling catheters in the portal, hepatic, and mesenteric veins and mesenteric artery. During the 21-d period, daily N and ME intakes were 24.6 and 10.7 g N/d and 3.02 and 1.28 Mcal/d, respectively, for ADLIB and MAINT lambs. Intakes, thus, were 42% lower for MAINT than for ADLIB lambs. Net portal fluxes of UN, NH3 N, AAN, and L in MAINT lambs were 46%, 84%, 50%, and 74%, respectively, of that in ADLIB lambs. Expressed as a percentage of N intake, the proportion of AAN absorbed by the PDV was higher in MAINT lambs (P less than .05) than in ADLIB lambs. There was no net portal glucose absorption in either group of lambs; however, net hepatic glucose production in MAINT lambs was 48% of that in ADLIB lambs. There was net utilization of glutamine by the PDV; net glutamine flux in MAINT lambs was 49% of that in ADLIB lambs. The liver utilized AAN and NH3 N and produced UN. Splanchnic tissues modulate metabolite flux following changes in feed intake in young ruminants.     

Differences among breed crosses of cattle in the conversion of food energy to calf weight during the preweaning interval

The objective of this study was to determine whether F1 cows that differ in genetic potential for weight at maturity and milk yield vary in the conversion of food energy to calf weight gain. Food intakes and weight change data were recorded by pen for cows and calves from approximately 45 d postpartum. Cows assigned to the study were 7- to 9-yr-old F1s produced by top-crossing Angus, Hereford, Brown Swiss, Chianina, Gelbvieh; Maine Anjou, and Red Poll sires to either Angus or Hereford dams. Calves were sired by Simmentals. Experimental units were pens (10 to 12 cow/calf pairs); pen was replicated within breed of sire in each of 2 yr (n = 24). Calf weight gain and energy consumed by the dams differed among the F1s, as did the ratio of calf weight gain to energy consumed by the calf and cow. Angus or Hereford (35.8), Red Poll (35.7), or Maine Anjou (35.6) F1s produced more calf weight per unit of energy consumed (g/Mcal) by the cow and calf than Chianina (33.1) or Gelbvieh (33.7) F1 females; Brown Swiss cows were intermediate (34.3). Differences in food conversion efficiency exist among breed crosses. These differences seem to be associated with breed cross differences in genetic potential for milk yield and mature weight; an exception to this trend was the Maine Anjou.