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.     

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.     

Modeling the effect of physiological responses to green pruning on net biomass production of Eucalyptus nitens

Green pruning of Eucalyptus nitens (Deane and Maiden) Maiden increases instantaneous rates of light-saturated CO(2) assimilation (A), and changes patterns of total leaf area and foliage distribution. We investigated the importance of such changes on the rate of recovery of growth following pruning. A simple process-based model was developed to estimate daily net biomass production (G(d)) of three-year-old plantation-grown trees over a 20-month period. The trees had been pruned by removal of 0, 50 or 70% of the length of green crown, equivalent to removal of 0, 55 or 88% of leaf area, respectively, when the plantation verged on canopy closure. Total G(d) was reduced by only 20% immediately following the 50%-pruning treatment, as a result of both the high leaf dark respiration and low A in the portion of the crown removed compared to the top of the crown. Pruning at the time of canopy closure preempted a natural and rapid decline in G(d) of the lower crown. Although leaf area index (L) was approximately 6.0 at the time of pruning, high light interception (95%) occurred with an L of 4.0. The 50%-pruning treatment reduced L to 3.5, but the physiological responses to pruning were sufficient to compensate fully for the reduction in intercepted radiation within 110 days of pruning. The 70%-pruning treatment reduced L to 1.9, and reduced G(d) by 77%, reflecting the removal of branches with high A in the mid and upper crown. Physiological responses to the 70%-pruning treatment were insufficient to increase G(d) to the value of unpruned trees during the study. Model sensitivity analysis showed that increases in A following pruning increased G(d) by 20 and 25% in the 50- and 70%-pruned trees, respectively, 20 months after pruning. Changes in leaf area/foliage distribution had a greater effect on G(d) of 50%-pruned trees (47% increase) than did changes in A. However, the reduction in photosynthetic potential associated with the 70%-pruning treatment resulted in only small changes in leaf area/foliage distribution, which consequently had little effect on G(d). The effects of physiological processes occurring within the crown and in response to green pruning on G(d) are discussed with respect to pruning of plantations.     

Growth responses, physiology and decay associated with pruning plantation-grown Eucalyptus globulus Labill. and E. nitens (Deane and Maiden) Maiden

Detailed studies have been undertaken to define pruning regimes for Eucalyptus nitens, but little is known of E. globulus responses to pruning although this is a more commonly planted species. This paper describes experiments that aimed to identify (a) levels of pruning that reduce E. globulus growth, (b) physiological processes contributing to those growth responses, (c) the incidence of decay and factors influencing decay incidence following pruning of E. globulus, and (d) comparative responses of E. nitens and E. globulus to live branch pruning. Results of a field experiment indicated that removal of between 30 and 50% of the crown length was appropriate for E. globulus plantations verging on canopy closure. The significant reduction in height growth associated with removal of 50 or 70% of crown length suggested pruning should remain below 50% of crown length if reduced stem growth of pruned trees was to be avoided. Stem volume was only significantly reduced over the period of the experiment by 70% pruning, but it was estimated that standing volume following removal of 50% of crown length would be reduced by 82 m³ ha⁻¹ over a 20-year rotation if there were no other silvicultural interventions. The growth responses observed were probably related to large reductions in leaf area following 50 or 70% removal of crown length. Trees responded to pruning by changing patterns of biomass partitioning to favour stem growth at the expense of branch growth. A glasshouse study determined that light-saturated net CO₂ uptake (A_max) increased following pruning. E. nitens seedlings had both a higher baseline A_max and higher A_max following pruning than did E. globulus, which could partially explain the greater effect of pruning on E. globulus growth than has been observed for E. nitens in other studies. This result, as well as apparently different patterns of foliage distribution through the crowns of E. globulus and E. nitens, suggested that models of pruning responses need to be parameterised for both species. In addition, a more conservative pruning regime may be appropriate for E. globulus than E. nitens. Pruning increased the frequency of branch traces with decay infection, and there was a trend towards increasing decay outbreaks with increasing pruning severity. Decay outbreaks were more likely to occur following pruning of high angle or larger diameter branches.     

Anatomical and histochemical defence responses induced in juvenile leaves of Eucalyptus globulus and Eucalyptus nitens by Mycosphaerella infection

The responses of juvenile leaves of two Eucalyptus species, with contrasting susceptibility to infection by Mycosphaerella leaf disease, were compared. The anatomical changes, accumulation of phenolics, suberin, lignin and anthocyanin and the retention of chlorophyll were studied in leaf lesions of varying developmental stages caused by species of Mycosphaerella. Enhanced resistance of Eucalyptus nitens in southern Australia was attributed to the formation of an effective lignified and suberized necrophylactic periderm, to restrict pathogen spread. Leaves of E. nitens contained isobilateral palisade which resulted in both abaxial and adaxial cell division and the initiation of a strong reinforced cellular zone from an early lesion stage. Eucalyptus globulus formed a slower, distorted necrophylactic periderm through hypertrophic changes to existing mesophyll and limited cell divisions of the single adaxial palisade layer. Deposits of lignin and suberin did not occur until later in lesion development, which were not effective in preventing further disease development. From this study it is hypothesized that tolerance of eucalypts to Mycosphaerella pathogens may be associated with constitutive mesophyll density.     

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.     

Retrospective study of 60 cases of feline lymphosarcoma

Objective To characterise epidemiological and clinical findings, and diagnostic procedures undertaken, in cats with lymphosarcoma at a veterinary teaching hospital.
Design Retrospective case study.
Procedure Hospital records were reviewed for 7159 cats, sick or healthy, examined during a 10-year period (1984 to 1994). Sixty cats with lymphosarcoma were identified and classified by anatomical location of the tumour. Data on breed, age, sex, clinical signs and diagnostic procedures were collated.
Results The prevalence of feline lymphosarcoma in the hospital population was 0.84%. Siamese cats appeared predisposed to lymphosarcoma but other purebreds were not. Males were somewhat overrepresented amongst affected cats. Similar numbers of cases (12 to 18) were seen in each of the four anatomic categories (multicentric, mediastinal, alimentary and extranodal). Cats with mediastinal lymphosarcoma were mostly young and Siamese. Clinical signs in affected cats were varied, usually multiple and often nonspecific. Two of 22 cases tested positive for feline leukaemia virus antigen in blood and 6 of 13 were positive for feline immunodeficiency virus antibody.
Conclusions Extranodal lymphosarcoma seemed more prevalent in this study than reported elsewhere. Siamese cats in the study population may have had a genetic predisposition to lymphosarcoma. Limited evidence suggested feline leukaemia virus may be less important, and feline immunodeficiency virus more important, in the local population than indicated in overseas reports. Additional studies are needed to investigate breed predisposition and feline leukaemia virus and feline immunodeficiency virus status in Australian cats with lymphosarcoma.