Recovery of sperms bearing X chromosomes with different concentrations of magnetic nanoparticles in ram

Pre-conceptual sex selection is still a highly debatable process whereby X and Y chromosome bearing spermatozoa are isolated before oocyte fertilization. Recently, magnetic nanoparticles (MNP) have been used to determine X and Y chromosomes bearing spermatozoa as a result of searching for a cheap, highly efficient method using non-toxic materials. This study aimed to recover the sperm bearing X chromosomes in ram with different concentrations of MNP and then evaluate the success of this method using polymerase chain reaction (PCR). Ram sperms were divided into four groups, treated with 0 (control), 50, 100 and 200 μg/ml MNP, respectively. MNP was used to restore sperm cells bearing X chromosomes. Upon recovery, the PCR was performed to identify the X and Y sperms, Methyl ThiazoleTetrazolium (MTT), to assess MNP toxicity and sperm viability and acridine orange (AO) to evaluate sperm DNA integrity. The results of PCR revealed that the treatment of spermatozoa- bearing X chromosomes with 50 μg/ml MNP had the highest effects on the recovery of X sperm rather than the other concentrations of MNP. However, the concentrations of MNP did not have any toxic effects on spermatozoa, sperm viability and, DNA integrity, but the high concentration of MNP (200 μg/ml) significantly reduced DNA integrity. According to MTT and AO results, the concentrations of MNP used in this study had no toxic effects on spermatozoa and did not reduce the sperm viability and DNA integrity, except that 200 μg/ml MNP significantly reduced DNA integrity.     

批式与两相高温厌氧消化厨余和杂草废弃物对比研究

以厨余和杂草废弃物混合物为发酵底物进行了批式和两相厌氧发酵试验。经过25 d的批式厌氧消化后,污染负荷(以挥发固体(VS)质量浓度计)为6.5、12.5、16.0和20.0 g/L的沼气产率分别为1 012、863、879和467mL/g,甲烷产率分别为595、442、440和316 mL/g,其中接种污泥对沼气产率和甲烷产率的贡献分别是304和170mL/g。试验结果表明:污染负荷为6.5 g/L时达到最大产气效率,80%的气体在反应的前8 d产生;污染负荷为12.5和16.0 g/L的产气率在统计学上无显著差别;污染负荷在20.0 g/L时出现产甲烷抑制,甲烷体积分数在反应的前2 d为20%左右。两相发酵试验采用厌氧固体床反应系统,包括4个1 L的固体床反应器和1个2.2 L的厌氧批式反应器。经过12 d的消化,沼气和甲烷产率分别为530和351 mL/g,系统总固体(TS)和VS去除率分别为78%和82%。与批式消化比较,该两相系统污染负荷高,产气稳定,周期短,是处理该类型有机废弃物的有效方法。     

Short‐term physiological role of thyroxine administration on plasma metabolites and growth performance of Sterlet sturgeon Acipenser ruthenus

Thyroid hormones (THs) play important roles in regulating growth, development and physiological functions in vertebrates. To study the role of short‐term effects of thyroxine (T4) on metabolism and growth in female Sterlet sturgeon, thirty fish with a weight of 707.97 ± 37.15 g were divided into six tanks. Fish were injected intraperitoneal with (a) coconut oil alone (control), (b) 1 mg T4 kg per BW in coconut oil (T₁) and (c) 10 mg T4 kg per BW in coconut oil (T₁₀). Blood samples were collected at 0, 4, 7, 14, 28 and 60 days for plasma biochemical analysis, and body length and mass were determined at day 60. Based on results, higher growth was indicated in T₁₀ group. The highest T4 levels were observed on day 4 in the T₁ and T₁₀ treatments and gradually declined during the trial. Plasma T3 levels were highest on days 4 and 7 in the T₁ and T₁₀ groups, respectively. Plasma triglyceride levels were significantly elevated on day 28 in the T₁ group and on day 60 in the T₁₀ group. Plasma glucose concentrations were significantly elevated on days 7 and 14 in T₁₀ group. Overall, the present results suggest that a single injection of T4 can improve growth via changes in the metabolic profile of Sterlet sturgeon.     

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.     

Epidemiological studies of pig diseases: 1. Use of French protocols for risk factor assessment to predict the health status of Australian herds

Objective To determine in Australian pig herds the accuracy of French protocols for risk factor assessment.
Procedure Data on health indicators and risk factors were collected for three syndromes, 'pre-weaning diarrhoea', 'post-weaning diarrhoea' and 'respiratory problems', using the French protocols. The protocols were used on 118 occasions in 32 Western Australian pig herds during 3 years (1988 to 1991).
Results There was a wide variation in pre-weaning performance, for example growth rate was 107 to 273 g/day (< 200 g/day in 33% of herds). Respiratory lesions at weaning were associated with poor pre-weaning performance. Post-weaning (21 days after weaning) growth rate was 114 to 408 g/day (< 250 g/day in 54% of herds). In the grower herds, 91% of herds had pneumonia, and growth rate was 439 to 625 g/day (< 550 g/day in 54% of herds). Pleurisy as well as pneumonia was associated with reduced growth rate. The risk factor most closely associated with respiratory health status was air volume per pig.
Conclusion Risk factors were most accurate at predicting the health status in post-weaning problems. A weaning weight of at least 7.9 kg and weaning age of 30 days optimised weaner performance. Stocking densities and shed designs providing at least 3 m³ air volume and 0.6 m² floor space per pig throughout the growing phase should be considered for an improved respiratory health status. Australian pig sheds often do not provide a satisfactory environment for optimum health. The technique of risk factor assessment as an aid to the maintenance of health in pig herds is applicable in Australia, but further research is necessary to determine the most important Australian risk factors.     

Bioactive constituents of Leptadenia arborea

The aerial part of Leptadenia arborea has been shown to contain pinoresinol (1), syringaresinol (2), leucanthemitol (3) and E-ferulaldehyde (4). These known compounds are being reported for the first time from this plant. Among them, syringaresinol has shown an inhibitory effect against acetylcholinesterase. The IC(50) (the concentration of 50% enzyme inhibition) value of this compound was 200 microg/ml.     

The Post‐Cervical Insemination does not Impair the Reproductive Performance of Primiparous Sows

The study evaluated the reproductive performance of primiparous sows submitted to post‐cervical insemination (PCAI) compared with cervical artificial insemination (CAI). Difficulty with catheter introduction, the occurrence of bleeding or semen backflow during insemination, and volume and sperm cell backflow up to 60 min after insemination were also evaluated. Sows were homogenously distributed, according to body weight loss in lactation, lactation length, weaned piglets, weaning‐to‐oestrus interval and total born in previous farrowing, in two treatments: PCAI (n = 165) with 1.5 × 10⁹ sperm cells in 45 ml (2.4 ± 0.04 doses per sow) and CAI (n = 165) with 3 × 10⁹ sperm cells in 90 ml (2.5 ± 0.04 doses per sow). During PCAI, sows were inseminated in the absence of boars. Transabdominal real‐time ultrasonography was performed at oestrus onset, immediately before the first insemination and at 24 h after last insemination. There was no difference (P > 0.05) between treatments in farrowing rate (91.5% × 89.1%) and litter size (12.5 × 11.9 piglets born, respectively for PCAI and CAI sows). Successful passage of the intrauterine catheter in all the inseminations was possible in 86.8% (165/190) of sows initially allocated to PCAI treatment. Difficulty of introducing the catheter in at least one insemination did not affect the reproductive performance of PCAI sows (P > 0.05). Bleeding during insemination did not affect (P > 0.05) the farrowing rate in both treatments, but litter size was reduced in CAI and PCAI sows (P ≤ 0.06). Percentage of spermatozoa present in backflow within 1 h after insemination was greater in CAI than PCAI sows (P < 0.01). More than 85% of primiparous sows can be successfully post‐cervical inseminated with doses containing 1.5 × 10⁹ sperm cells in the absence of the boar during insemination without impairing the reproductive performance.