Development of Procedures for Sex-sorting Frozen–Thawed Bovine Spermatozoa

Dairy bull sperm may be sex‐sorted, frozen and used to artificially inseminate heifers with acceptable fertility if the herd is well‐managed. One drawback to the technology is that donor bulls must be located within a short distance of the sorting facility in order to collect semen, which limits the number of bulls from which sorted sperm are available. A successful method used to overcome this limitation in sheep is sex‐sorting from frozen–thawed semen and refreezing for artificial insemination. This technique is attractive to the dairy industry, and therefore a series of three experiments was designed to investigate the optimal methods to prepare, sex‐sort and re‐freeze frozen–thawed bovine sperm. Sperm were prepared for sorting by density gradient separation in either PureSperm^® or BoviPure?, followed by staining in one of three diluents (Androhep^®, Bovine Sheath Fluid + 0.3% BSA or TALP buffer). Sperm were sorted and collected into Test yolk buffer, and frozen in an extender containing 0, 0.25, 0.375 or 0.5% Equex STM Paste. Frozen–thawed sperm were better orientated (p = 0.006) and had fewer damaged membranes (8.7 ± 0.6% vs 19.5 ± 2.4%; p = 0.003) after centrifugation in PureSperm^® rather than BoviPure? gradients. Sperm orientation (p < 0.05) and motility (69.9 ± 3.0 vs 55.6 ± 4.0; p < 0.001) were highest after staining in Androhep^® rather than in TALP buffer. Sperm were more motile (58.2 ± 4.7 vs 38.7 ± 3.5; p < 0.001) and had better acrosome integrity (74.3 ± 2.9 vs 66.8 ± 2.0; p < 0.001) after freezing in an extender containing 0.375% Equex STM Paste than in extender without Equex. Hence, a protocol has been developed to allow frozen–thawed bull sperm to be sex‐sorted with high resolution between the sexes, then re‐frozen and thawed with retention of motility and acrosome integrity.     

A packed lysimeter experiment to investigate the effect of surface sealing on hydrology and pesticide loss from the reconstructed profile of a clay soil

Abstract. A laboratory experiment was designed to assess the impact of surface seal development on herbicide loss from a clay soil. The influence of surface sealing on herbicide loss through vertical macropores and lateral throughflow pathways was of particular interest. Losses of the phenylurea herbicide, isoproturon, increased from 0.025% to 0.5% of the total applied under sealed compared with unsealed conditions, as a result of two different mechanisms. First, an increased flow rate through the A horizon under the wetter, sealed conditions resulted in earlier initiation of, and hence a greater volume of, throughflow containing isoproturon in the lysimeters. Second, increased concentrations of isoproturon in macropore and throughflow under the sealed treatments were attributed to the physical and chemical characteristics of the surface seal. Localized reductions in infiltration capacity, changes in soil composition, and decrease in diffusion depth of herbicide within the surface seal are presented as possible mechanisms by which herbicide losses from the sealed soil lysimeters were enhanced.     

A lysimeter experiment to investigate the effect of surface sealing on hydrology and pesticide loss from the reconstructed profile of a clay soil. 1. Hydrological characteristics

Abstract. A laboratory experiment was designed to assess the impact of surface seal development on the hydrological response of a clay soil. The influence of surface sealing on vertical macropore flow and lateral throughflow was of particular interest. The extent and development of the surface seal in repacked lysimeters was designed to match that recorded over two growing seasons at a clay field site in Essex, and was not extensive enough to reduce significantly the infiltration capacity of the soil. Consequently, the hydrological response of the lysimeters was similar under sealed and unsealed conditions, with a more rapid wetting response under sealed conditions being attributed to the higher soil moisture content required to create the surface seal. Macropore flow was initiated at the A/B soil boundary of the lysimeters, in response to the development of a saturated layer. The rate of macropore and throughflow in the soil was dictated by rainfall intensity at the soil surface as this controlled the depth of water in the perched water table. Simulation of the tensiometer response in the lysimeters demonstrated that it was possible to attribute the rapid movement of water through the A horizon to water displacement processes alone, without recourse to preferential flow processes.     

Prevalence and risk factors for medical events following exercise at Australian Greyhound race meetings

Aim

A prospective, observational study to determine the prevalence of post‐exercise conditions at Australian Greyhound race meetings and to assess association with race performance and other environmental, race‐ and dog‐related factors was undertaken.

Methods

A total of 4020 starters were observed (2813 Greyhounds, 1009 trainers, 536 races, 52 race meets, 48 race dates and 11 race tracks) following a race. The presence of diaphragmatic flutter (DF), ataxia, seizure, collapse or sudden death was recorded. Risk factors were screened by univariable logistic regression prior to multivariable backward stepwise model building.

Results

In this study, 962 starters (n = 768 dogs) had DF (23.9%), 16 starters were ataxic (0.4%) and there were no observed cases of collapse, seizure or sudden death. Race track location, increasing race distance, race grade based on increasing 1st place prize value, lower (earlier) race number at the meeting, age, a previous observation of DF at the last start, females, colour (white) and better finishing position were all associated with an increased risk of a Greyhound being observed with DF. However, when logistic regression assessing the random effect of dog was performed, the presence of previous DF was not significant. In this cohort, DF was common following strenuous exercise in Greyhounds and on its own does not appear to result in reduced performance or distress to the animal.

Conclusion

The incidence of ataxia was low and collapse, seizure and sudden death were not observed. However, even though uncommon, ataxia has welfare concerns for racing Greyhounds that warrants further investigation.     

Partially Molten Middle Crust Beneath Southern Tibet: Synthesis of Project INDEPTH Results

INDEPTH geophysical and geological observations imply that a partially molten midcrustal layer exists beneath southern Tibet. This partially molten layer has been produced by crustal thickening and behaves as a fluid on the time scale of Himalayan deformation. It is confined on the south by the structurally imbricated Indian crust underlying the Tethyan and High Himalaya and is underlain, apparently, by a stiff Indian mantle lid. The results suggest that during Neogene time the underthrusting Indian crust has acted as a plunger, displacing the molten middle crust to the north while at the same time contributing to this layer by melting and ductile flow. Viewed broadly, the Neogene evolution of the Himalaya is essentially a record of the southward extrusion of the partially molten middle crust underlying southern Tibet.     

Equine perioperative fatalities associated with general anaesthesia at a private practice--a retrospective case series

OBJECTIVE: To determine the fatality rate of horses undergoing general anaesthesia at a private equine referral practice using a limited number of anaesthetic protocols. METHODS: A retrospective analysis of records (n = 17 961) from all horses undergoing general anaesthesia for surgical procedures from 1997 to 2001 at Rood and Riddle Equine Hospital, Lexington, Kentucky, USA. Results were reported as percentage of the population, and as crude mortality rates for each procedure (deaths per 1000). RESULTS: The prevalence of equine fatalities directly related to anaesthesia was 0.12% (n = 21) and this rose to 0.24% (n = 42) with the inclusion of horses killed or dying within 7 days post general anaesthesia. Causes of death directly related to anaesthesia were cardiac arrest (n = 10), fracture in recovery stall (n = 8), neuropathy and myopathy necessitating euthanasia (n = 3). Crude mortality rates per procedure were < or =7 deaths per 1000 cases, except arthrodesis/osteotomy cases were 66.7 deaths per 1000 cases. CONCLUSIONS: The anaesthetic fatality rate at this practice is lower than has been reported previously. CLINICAL RELEVANCE: Familiarity with an anaesthetic protocol in combination with reduced anaesthetic time, emergencies of shorter duration between diagnosis and surgery, and adequate preoperative examination appear to minimize the risks associated with general anaesthesia in horses.     

A comparison of two different ketamine and diazepam combinations with an alphaxalone and medetomidine combination for induction of anaesthesia in sheep

Abstract

AIMS: To investigate the perceived adverse effects of a particular batch of ketamine during induction of anaesthesia in sheep and to assess if any adverse effects would make intubation more difficult for the veterinary students.

METHODS: Thirty adult sheep (mean bodyweight 74.5 (SD 9.4) kg) were randomly assigned to one of six groups of five sheep. Sheep in Groups A and B received I/V 0.5 mg/kg diazepam and 10 mg/kg ketamine (Ketamine Injection; Parnell Laboratories NZ Ltd, of the suspect batch); those in Groups C and D received I/V 0.5 mg/kg diazepam and 10 mg/kg ketamine (Ketalar; Hospira NZ Ltd.), and those in Groups E and F received I/V 2 μg/kg medetomidine and 2 mg/kg alphaxalone. In Groups A, C and E, intubation was by an experienced anaesthetist, and in Groups B, D and F intubation was by a veterinary student. Time from injection to successful intubation, the ease of intubation, saturation of haemoglobin with oxygen (SpO₂) and partial pressure of oxygen in arterial blood (PaO₂) were measured before the sheep were connected to an anaesthetic machine and allowed to breath oxygen. Times to extubation, holding its head up and standing, maximum and minimum heart rates, respiratory rates, maximal end tidal CO₂, and the quality of recovery were then recorded.

RESULTS: There were no measurable differences in outcomes between sheep in Groups A and B compared with C and D. Time to intubation was slightly shorter for the experienced anaesthetist than the student, but the difference was not significant. The sheep in Groups E and F took less time to recover than those in Groups A?D (p<0.05), but there were no significant differences between the groups in either the ease of induction or quality of recovery. Most sheep in Groups E and F showed minor excitatory effects, mainly at induction, which did not interfere with induction. Respiratory rates were lower in Groups E and F than Groups A?D (p<0.01), but SpO₂ was higher in Groups E and F than A and B (p<0.05).

CONCLUSIONS: The clinical impression that the batch of Parnell ketamine produced unexpected effects was shown to be incorrect. All the combinations produced anaesthesia that allowed intubation by the veterinary student.

CLINICAL RELEVANCE: All the drug combinations produced satisfactory anaesthesia in sheep, but the alphaxaloneand medetomidine combination resulted in faster recovery.