Effects of α-tocopherol and Ascorbic Acid on Equine Semen Quality after Cryopreservation

This study investigated the effects of ascorbic acid and α-tocopherol supplementation on semen quality parameters of equine thawed-frozen semen. Semen was divided in seven different treatments in a final concentration of 100 × 10⁶ sperm/mL by using Gent extender containing no supplements (control) and the following supplements with three different concentrations: α-tocopherol (0.5, 1, and 2 mM) and ascorbic acid (0.45, 0.9, and 1.8 g/L). After thawing, all samples were maintained at 37°C, while analyses were performed at 0, 60, and 120 minutes. Evaluation of viability and acrosome status (using Pisum sativum agglutinin conjugated to fluorescein isothiocyanate and propidium iodide), mitochondrial membrane potential (5,5′,6,6′-tetrachloro-1,1′,3,3′tetraethylbenzimidazolyl carbocyanine iodine [JC-1]), membrane lipid peroxidation (LPO; C₁₁-BODIPY^581/591), and stability of the plasmatic membrane (merocyanine 540 and Yo-Pro-1) of each sample was determined by flow cytometry. Relative to the control group, supplementation with α-tocopherol improved (P ≤ .05) postthaw membrane LPO, yet the higher concentrations of ascorbic acid (0.9 and 1.8 g/L, respectively) showed a negative effect on membrane LPO. Neither antioxidant significantly increased (P > .05) the acrosome integrity and mitochondrial membrane potential of frozen-thawed spermatozoa, although supplementation with α-tocopherol and ascorbic acid (0.9 and 1.8 g/L, respectively) had a positive effect on membrane integrity and stability (P ≤ .05). For all semen parameters, the lower concentration of ascorbic acid (0.45 g/L) did not show significant differences (P > .05) compared with the control. In conclusion, α-tocopherol seems to be an efficient antioxidant for reducing the oxidative stress provoked by cryopreservation, decreasing lipid peroxidation on equine spermatozoa.     

氨基酸络合铁对生长猪生长性能及有关指标的影响

为了比较评估氨基酸络合铁对生长猪生长性能、铁表观消化率、肤色和血液指标的影响,试验选择70日龄初始体重(27.34±2.55)kg的杜×长×大生长猪108头,分为6个处理,每个处理6个重复,每个重复3头猪。试验用玉米-豆粕型基础日粮。在基础日粮中添加100 mg/kg无机铁(一水硫酸亚铁)构成对照组。在对照组基础上分别添加40、80、120、160 mg/kg氨基酸络合铁构成4个试验组,在对照组基础上添加160 mg/kg无机铁(一水硫酸亚铁)构成正对照组,试验期5周。结果表明:试验0~2周、3~5周及全期,随着日粮中氨基酸铁添加量由40 mg/kg增加到160 mg/kg,生长猪日增重和饲料转化效率均得到改善,并呈极显著的线性关系(P<0.01)。与无机铁日粮组相比,添加160 mg/kg氨基酸络合铁试验组极显著地改善了平均日增重(P<0.01)和饲料转化效率(P<0.05);试验第14天和第35天,随着日粮中氨基酸络合铁添加水平的增加,生长猪对日粮中铁元素的表观消化率均呈显著的线性增加关系(P<0.05),添加160 mg/kg氨基酸络合铁试验组铁元素的表观消化率极显著高于对照组和正对照组(P<0.01);试验第14天和第35天,随着日粮中氨基酸络合铁添加水平的提高,红细胞压积、血红蛋白浓度、血清铁和血清铁与总铁结合力之比都呈显著的线性增加关系(P<0.05),并高于无机铁日粮组(P<0.01),而血清总铁结合力呈显著的线性降低关系(P<0.05);在试验期末,160 mg/kg氨基酸络合铁日粮组显著改善生长猪的肤色状况(P<0.05)。与添加硫酸亚铁相比,在生长猪日粮中添加氨基酸铁络合物可提高生长猪的生长性能和铁表观消化率,改善皮肤红度及血液生化指标。     

Investigation of the Role of Androstenedione-19-oic Acid in the Presence of 19-Norandrostenedione in Intact Male Horse Plasma Using Liquid Chromatography–Tandem Mass Spectrometry

A liquid chromatography–tandem mass spectrometry method was developed to confirm the presence of androstenedione-19-oic acid in intact male equine plasma and to show the source of 19-norandrostenedione in equine plasma. Androstenedione-19-oic acid was recovered from acidified plasma by liquid–liquid extraction using methyl tert-butyl ether and separated on an Ace 5 C₈ column. A triple quadrupole mass spectrometer was used to detect the analytes in negative electrospray ionization mode. Limits of detection, quantification, and confirmation of the method were 0.1, 0.5, and 1.0 ng/mL, respectively. The linear dynamic range of quantification was 0.5–50 ng/mL. The presence of androstenedione-19-oic acid was confirmed in all plasma samples obtained from intact male horses but not those from gelded and female horses; the average concentration was 3.1 ± 1.6 ng/mL, suggesting androstenedione-19-oic acid is an endogenous compound only in intact male horse plasma samples. The conversion of androstenedione-19-oic acid to 19-norandrostenedione in equine plasma was demonstrated by spiking androstenedione-19-oic acid into blank plasma and monitoring the generation of 19-norandrostenedione and its increase in concentration during storage. Results indicated that androstenedione-19-oic acid was readily converted into 19-norandrostenedione; the higher the storage temperature, the faster the conversion. The conversion was not affected by the types of plasma samples collected from gelded and female horses or by anticoagulants used in blood collection to harvest plasma. Compared with other matrices such as water, methanol, and phosphate-buffered saline, the conversion of androstenedione-19-oic to 19-norandrostenedione in equine plasma was faster, suggesting that there is an unknown factor(s) in equine plasma that enhances the conversion.     

颈静脉灌注不同模式的氨基酸混合物对泌乳奶牛乳蛋白合成的影响

本试验旨在比较颈静脉灌注酪蛋白模式和理想模式的氨基酸混合物对泌乳中期荷斯坦奶牛乳产量、乳成分以及乳腺对氨基酸摄取利用的影响。选择8头泌乳中期[泌乳天数:(82±11)d]荷斯坦奶牛作为试验动物,试验采用随机区组设计,将试验牛随机分为2组,分别颈静脉灌注160g/d酪蛋白模式(Casein组)和理想模式的氨基酸混合物(R组)。2个试验组分别以各自灌注前作为空白对照组(C1组为Casein组的空白对照组,C2组为R组的空白对照组)。预试期14d,灌注期5d。试验采用全混合日粮(TMR)饲喂,以玉米、豆粕、棉籽粕、玉米青贮、苜蓿干草和羊草为主要原料,参照NRC(2001)奶牛饲养标准配制。结果表明:灌注酪蛋白模式的氨基酸混合物后,乳蛋白产量和含量较灌注前呈上升趋势(乳蛋白产量上升7.14%,P=0.078;乳蛋白含量上升3.27%,P=0.072);并且,奶牛动脉血浆中异亮氨酸(Ile)、亮氨酸(Leu)、赖氨酸(Lys)和组氨酸(His)的浓度较灌注前有不同程度的上升(Ile的浓度提高31.5%,P=0.097;Leu的浓度提高65.9%,P=0.041;Lys的浓度提高36.9%,P=0.088;His的浓度提高40.1%,P=0.010),而苏氨酸(Thr)、缬氨酸(Val)、蛋氨酸(Met)、苯丙氨酸(Phe)和精氨酸(Arg)的浓度在数值上虽较灌注前高但无显著差异(P0.05)。灌注酪蛋白模式的氨基酸混合物后,奶牛乳腺对天冬氨酸(Asp)和半胱氨酸(Cys)的摄取率显著升高(Asp的摄取率提高95.2%,P=0.031;Cys的摄取率提高49.6%,P=0.031),而奶牛乳腺对甘氨酸(Gly)的摄取率显著降低(降低158.3%,P=0.041)。灌注理想模式的氨基酸混合物后,乳蛋白含量比灌注前有上升趋势(提高5.78%,P=0.064),而乳脂产量显著低于灌注前(降低8.57%,P=0.015);并且,奶牛动脉血浆中Arg的浓度有上升趋势(提高18.0%,P=0.093),而酪氨酸(Tyr)的浓度呈下降趋势(降低47.8%,P=0.074)。灌注理想模式的氨基酸混合物后,奶牛乳腺对谷氨酸(Glu)、Cys和Ile的摄取率显著上升(Glu的摄取率提高118.7%,P=0.015;Cys的摄取率提高77.4%,P=0.032;Ile的摄取率提高46.0%,P=0.012),而奶牛乳腺对Ser的摄取率呈下降趋势(降低56.2%,P=0.052)。灌注氨基酸混合物后,Casein组乳脂产量增量显著高于R组(P=0.012),且Casein组的乳产量增量(P=0.095)和乳糖产量增量(P=0.091)较R组有升高的趋势,而2组间其他指标增量无显著差异(P0.05)。由此得出,在本试验条件下,颈静脉灌注酪蛋白模式和理想模式的氨基酸混合物均可提高泌乳奶牛的乳蛋白含量,而灌注酪蛋白模式氨基酸混合物同时可以促进乳蛋白产量的升高,因此,颈静脉灌注酪蛋白模式氨基酸混合物的效果优于灌注理想模式氨基酸混合物。     

No easy solution for the fractionation of faecal nitrogen in captive wild herbivores: results of a pilot study

Faecal nitrogen (FN) concentration is used as a marker for habitat quality and digestive efficiency in free‐ranging herbivores. In herbivores, FN can be separated into undigested plant N (analysed as the N concentration of the neutral detergent residue) and metabolic faecal N (MFN). It has been suggested that by differential analysis of the faecal fibre‐bound N, the MFN fraction can be further split into a bacterial N and an endogenous N fraction [Hesta et al., Br. J. Nutr. 90 (2003) 1007]. We applied these methods to 96 faecal samples of 48 mammalian herbivore species from zoos. Species were grouped into coprophageous and non‐coprophageous hindgut fermenters and ruminating and non‐ruminating foregut fermenters. Diet was not controlled. The FN decreased with body mass, possibly reflecting higher proportions of concentrates in diets of smaller animals. The proportion of MFN increased with FN, indicating that higher quality food might enhance the gastrointestinal bacterial flora. The only outlier to this pattern was the lesser panda (Ailurus fulgens), confirming the low relevance of fermentative digestion in this herbivorous ‘carnivore’. No relevant differences between the four digestion types were noted. The proportion of endogenous faecal N (32–80% of FN) was always higher than that of bacterial faecal N (7–30%), which contradicts basal understanding of herbivore digestive physiology. Thus, the method of Hesta et al. (2003) does not appear applicable to herbivores. While the results do not exclude the possibility that detailed differences might occur between digestion types, they indicate a high degree of similarity between herbivores that rely on bacterial fermentation, regardless of their digestion type, with respect to metabolic faecal losses.     

Effect of retinoic acid on gene expression profiles of bovine intramuscular preadipocytes during adipogenesis

To investigate genes involved in intramuscular adipogenesis in ruminants, 16 genes with dramatic variable expression were selected. These were selected from the differentiation‐ and proliferation‐phase libraries of our previous serial analysis of gene expression (SAGE) studies of a clonal bovine intramuscular preadipocyte (BIP) cell line. We harvested the BIP cells over 12 days after adipogenic stimulation with all‐trans retinoic acid (ATRA). Quantitative real‐time PCR confirmed the earlier SAGE study results of the expression patterns of 15 of the genes. On day 6, TG accumulation increased significantly in the BIP cells but was completely inhibited in the 3T3‐L1 cells (the monogastric reference). ATRA enhanced expression levels of six genes whereas it suppressed expression of eight genes on day 3 of adipogenesis in the BIP cells. Forty‐eight hours after transfection, the messenger RNA expression level of the adipose differentiation‐related protein (ADFP), encoded by one of the upregulated genes, in the ADFP small interference RNA (siRNA)‐transfected cells was 3.5% of that in negative control‐transfected cells. Also, 6 days after induction the TG level in the ADFP siRNA‐transfected cells was 21.8% lower than that in negative control‐transfected cells. This analysis of gene expression profiles after ATRA treatment will contribute to our understanding of the molecular mechanisms involved in bovine intramuscular adipogenesis.     

无氮日粮纤维水平对猪内源氨基酸测值的影响

以醋酸纤维素为纤维源配制成粗纤维水平分别为２．０％、５．０％、８．０％的３种无氮日粮,采用３头回—直肠吻合猪,分３期试验,通过３×３拉丁方设计研究在等进食量（７５０ｇ／ｄ）条件下日粮的纤维水平对回—直肠吻合猪消化道内源氨基酸测值的影响。结果表明：除缬氨酸、异亮氨酸和胱氨酸外,无氮日粮的粗纤维水平对回肠末端食糜干物质中其他内源氨基酸含量（％）无显著影响（Ｐ＞０．０５）。而对回肠末端内源氨基酸的总排泄量影响显著（Ｐ＜０．０５）或极显著（Ｐ＜０．０１）。相关分析表明,粗纤维水平与各种内源氨基酸的排泄总量之间存在显著（Ｐ＜０．０５）或极显著（Ｐ＜０．０１）的强相关关系,随无氮日粮中粗纤维水平的升高,各种内源氨基酸排泄量呈直线增加。     

不同氧化方法对羽毛粉中胱氨酸生成磺基丙氨酸效果的影响

采用均匀设计方法研究了过甲酸法、过乙酸法氧化羽毛粉中胱氨酸为磺基丙氨酸的效果，结果表明，两种方法的效果无显著差异（Ｐ＞０．０５）。从实用经济角度来考虑，以使用过乙酸氧化法为宜。使用此法的最适条件为：过乙酸用量（Ｘ１）０．１ｍＬ／ｇ羽毛粉，过乙酸组成（Ｘ２）（过氧化氢∶乙酸）１∶８；过乙酸的浓度（Ｘ３）９０％；氧化温度１１０℃（Ｘ４）；氧化时间（Ｘ５）８ｈ。在此条件下，磺基丙氨酸最佳得率为８．０２％。氧化条件（Ｘ）与生成磺基丙氨酸（Ｙ）之间的回归方程：ｙ＝０．９４２２＋０．０６８６Ｘ１＋０．５７８７Ｘ２＋０．００４９Ｘ３＋０．００５２Ｘ４＋０．２９５９Ｘ５。     

奶牛乳房炎诊断技术的研究进展

奶牛乳房炎是影响奶牛生产的主要疾病之一。病原菌的免疫抑制以及日趋严重的耐药性,致使乳房炎的发病率增高且治愈困难,给世界奶牛养殖业造成了巨大的经济损失。实践表明,疾病的诊断是疾病病因和治疗之间最重要的一步。经济、快速、可靠的诊断工具是乳房健康管理的基础。本文主要论述了奶牛乳房炎不同诊断方法的优缺点以及新兴诊断技术的研究进展。