Pronuclear Embryo Yield in Canindé and Saanen Goats for DNA Microinjection

The objective of this study was to examine the effect of donor breed on pronuclear‐stage embryo yield to be used for DNA microinjection in a transgenesis goat program. Twelve Canindé and twelve Saanen goats were heat synchronized using a progestagen‐cloprostenol treatment. Forty‐eight hours before the sponge removal, superovulation was induced with a total administration of 4.4 mg/kg bodyweight NIH‐FSH‐P1, given twice daily in decreasing doses over 3 days. In addition, goats received 100 μg of GnRH and they were hand‐mated at 36 and 48 h after progestagen removal. Embryo recovery was performed by oviduct flushing at 72 h after sponge removal. Embryos were microinjected with a DNA construct and noticeable swelling of the nuclei was the criterion for successful microinjection. The total diameter, cytoplasm diameter, zona pellucida thickness and pronuclei diameter were measured for each microinjected embryo. A higher (p < 0.05) percentage of fertilized ova was observed in Canindé (89.9%) than Saanen (36.2%) goats. In addition, Canindé donors produced a higher percentage of pronuclear embryos when compared with Saanen: 72.5% vs 20.6% (p < 0.05), respectively. Successful microinjection was verified in 96.7% and 73.3% of times in Canindé and Saanen embryos, respectively (p < 0.05). Significant differences were observed for all morphometric parameters except for cytoplasm diameter. In conclusion, under our study experimental conditions, Canindé were more efficient than Saanen goats concerning the pronuclear embryo yield and manipulation. The use of Canindé goats in transgenesis programs could be increase the interest in their breeding and could be contribute to saving them from extinction.     

Seed dormancy and germination in the summer annual Galeopsis speciosa

This study examined germination and dormancy in Galeopsis speciosa (Lamiaceae), a common summer annual weed in cold‐temperate areas. Seeds collected in southern Sweden were subjected to several experiments. The seeds were dormant at maturity. Seeds sown outdoors after collection produced a small number of seedlings that emerged early in the spring. After long cold stratification or stratification outdoors over two winters, the maximum germination was 40–50%; germination occurring over a wide range of temperatures. Warm stratification preceding cold stratification had no effect on germination, but repeated warm and cold periods seemed to promote germination. Gibberellic acid (GA) stimulated germination, but full germination was only achieved after more than 2 months of incubation at the most suitable temperature regime tested. Excised embryos grew and developed into normal seedlings. With these results, the species does not fit into the currently used system for seed dormancy classifications. The response to GA and the growth of excised embryos indicate non‐deep or intermediate physiological dormancy, but dormancy alleviation by stratification was not in line with the guiding principles for these classifications. Galeopsis speciosa has a strong dormancy that is sufficiently alleviated during the winter to allow germination of only part of a seed batch each year; hence a stepwise germination pattern occurs over a period of several years.     

Connectivity,habitat, and flow regime influence fish assemblage structure: Implications for environmental water management in a perennial river of the wet–dry tropics of northern Australia

1. Environmental water management seeks to balance competing demands between the water needed to sustain human populations and their economic activities and that required to sustain functioning freshwater ecosystems and the species they support. It must be predicated on an understanding of the environmental, hydrological, and biological factors that determine the distribution and abundance of aquatic species.
2. The Daly River of the wet–dry tropics of northern Australia consists of a perennially flowing main stem and large tributaries, as well as many small to large naturally intermittent tributaries, and associated off‐channel wetlands. Increased groundwater abstraction to support irrigated agriculture during the dry season threatens to reduce dry‐season flows that maintain perenniality and persistence of freshwater fishes.
3. Fish assemblages were surveyed at 55 locations during the dry season over a 2‐year period with the goal of establishing the key landscape‐scale and local‐scale (i.e. habitat) drivers of fish species distribution.
4. Longitudinal (upstream/downstream) and lateral (river/floodplain) gradients in assemblage structure were observed with the latter dependent on the position in the river landscape. Underlying these gradients, stream flow intermittency influenced assemblage composition, species richness, and body size distributions. Natural constraints to dispersal were identified and their influence on assemblage structure was also dependent on position within the catchment.
5. Eight distinct assemblage types were identified, defined by differences in the abundance of species within five groups differing in functional traits describing body size, spawning requirements, and dispersal capacity. These functional groups largely comprised species widely distributed in northern Australia.
6. The results of the study are discussed with reference to the environmental flow needs of the Daly River and other rivers of northern Australia. The findings may also be applied to environmental flow management in savannah rivers elsewhere.

     

Crystal and molecular structure of acetylselenocholine iodide

The structure of acetylselenocholine iodide has been determined by x-ray crystallographic analysis. The molecule is in the trans conformation about the C-C bond of the choline residue. This conformation appears to explain the molecule's inability to give a positive cholinergic response when added to an electroplax preparation.     

Evaluation of auditory evoked potentials to predict depth of anaesthesia during fentanyl/fluanisone−midazolam anaesthesia in rats

Objectives To assess a method for monitoring depth of anaesthesia using components of middle latency auditory evoked potential (AEP) waveforms during anaesthesia with fentanyl/fluanisone and midazolam. Study design Prospective observational study. Animals Five female Wistar rats weighing between 210 and 250 g. Methods Implanted electrodes were used to record AEPs in animals receiving five doses of anaesthetic. Recordings were made at 5 minutes post‐injection (deep anaesthesia; no pedal withdrawal response, PWR) and then at 25 minutes (light anaesthesia; strong PWR). Responses showed five characteristic peaks occurring at 11, 14, 23, 42 and 68 ms that were measured for latency of occurrence and peak amplitude. Results Auditory evoked potential peaks P14, N23 and P42 were increased significantly in latency with successive anaesthetic injections [avg. F(1,4) = 12.53, p < 0.001; avg. F(1,4) = 10.6, p < 0.001; avg. F(1,4) = 3.9, p = 0.02, respectively]. Peak N23 showed a significant reduction in latency during the 20 minute recovery period following both the first and second anaesthetic injections (t(3) = 7.52, p = 0.005; t(4) = 5.17, p = 0.007, respectively). Peak P42 occurred significantly earlier 20 minutes following the second anaesthetic injection (t(4) = 4.75, p = 0.009). The mean overall depth of anaesthesia assessed using PWR scores was significantly correlated with the mean latency of peak N23, such that as the strength of PWR increased, N23 occurred significantly earlier (r = ?0.99, p = 0.01). The amplitude difference between peaks N23 and P42 increased after the second and third drug administrations [avg. F(1,4) = 10.65, p = 0.031 and avg. F(1,4) = 11.24, p = 0.028, respectively]. Conclusion The characteristics of these peaks, and in particular latency of peak N23, may provide a useful tool for assessing depth of anaesthesia produced by this, and possibly other anaesthetic agents.     

Effects of oral isosorbide and glycerol on intraocular pressure,serum osmolality,and blood glucose in normal dogs

Objective To evaluate and compare the effects of oral isosorbide and glycerol on intraocular pressure (IOP), serum osmolality (SOSM), and blood glucose (BG) in normal dogs. Methods Ten normal dogs were administered an oral dose of either isosorbide (1.5 g/kg), glycerol (1.5 g/kg) or control (water, 2 mL/kg) in a double blind protocol. Prior to dosing, baseline IOP, SOSM, and BG were measured in all dogs. IOP was subsequently evaluated every 30 min for 6 h post‐dosing. BG and SOSM were reassessed at times 1, 2, 4, and 6 h post‐dosing. After 1‐week washout periods, every dog was subject to each of the three treatments. The dogs were held NPO for 4 h after dosing. Results The maximal decrease in IOP was 17% by 1 h and 13.5% by 30 min after glycerol and isosorbide administration, respectively. However, the overall changes in IOP were not significant when compared to the controls. SOSM increased above baseline after dosing with glycerol but decreased after isosorbide, which difference was significant at 1, 2, and 4 h post‐administration. BG significantly increased after administration of glycerol relative to the control but was not significantly affected by isosorbide. Conclusions Neither glycerol nor isosorbide significantly affected IOP when compared to the control. However, glycerol induced significant elevations in both BG relative to the control and SOSM relative to isosorbide.     

Treatment response and long term follow up in nineteen dogs diagnosed with chronic enteropathy in Australia

Chronic enteropathy (CE) in dogs is common worldwide, but little data is available from Australia. The aim of this study was to describe treatment response and long‐term outcome in a cohort of dogs with CE. Dogs were prospectively enrolled at Murdoch University and the University of Melbourne. After diagnostic investigation to rule out diseases other than CE, dogs underwent sequential therapeutic trials until achieving a clinical response (diet then antibiotics, and finally immunosuppressants). Success was defined as 75% reduction of clinical severity for a minimum of five weeks. A total of 21 dogs were enrolled, and 19 completed the study. One dog was euthanised for lack of response to treatment and one excluded for lack of owner compliance. Most dogs responded to diet (n = 10), followed by antibiotics (n = 7) and immunosuppressants (n = 2). Long‐term remission (median 21.1 months, [3.0‐44.7]) was achieved in eight out of ten dietary responders without additional treatment. In contrast, only two dogs with antibiotic response remained in long‐term remission, of which one needed on‐going antibiotic treatment. Longer term remission was achieved in the two dogs treated with immunosuppressants with on‐going low dose therapy. This study concludes that most dogs referred for CE in Australia respond to dietary treatment (even after previous dietary interventions), and remission is long‐term compared to dogs treated with an antibiotic. Furthermore, the need for long‐term antibiotics in some dogs to maintain response may lead to antibiotic resistance. This study supports adequate dietary trials for CE in dogs, and a need for alternative second‐line treatments.