超低温冷冻对奶牛胚胎超微结构的影响

奶牛冷冻胚胎移植成活率较鲜胚移植明显低.为明确低温冷冻造成胚胎损伤的机理、为冷冻保存方法的进一步改进提供理论依据,利用透射电子显微镜观察了奶牛胚胎常规冷冻和玻璃化冷冻前后的超微结构变化.结果表明,奶牛鲜胚细胞分为两种类型:一种位于内细胞团(ICM)外层,胞质电子致密度低而着色浅,并含有大量的线粒体和内质网;另一种位于ICM中心区域,着色较深,溶酶体的数量多,其余细胞较较少.常规冷冻后,透明带和卵周隙明显变宽,细胞间接触变得松散.囊胚中含有退化的胞核和凋亡小体,内质网变大、变粗,脂滴明显增多.玻璃化冷冻后,ICM细胞的结构发生破坏,部分细胞发生崩解,细胞碎片大量分布于细胞间隙和卵周隙中,并可见大量的脂滴,凋亡细胞和凋亡小体增多.冷冻过程对早期胚胎细胞器的损伤可能是移植后成活率下降的主要原因.     

Effect of Feeding and Body Condition Score on Multiple Ovulation and Embryo Production in Zebu Cows

The aim of this study was to standardize the feeding regimen and the body condition score (BCS) for maximum superovulatory responses in indigenous zebu cows. Ten regularly cycling 5–8‐year‐old dry cows, weighing 176–260 kg with BCS 2.5–4.5 were divided into two equal groups at random. The groups were maintained on either a good‐nutrition or a high‐nutrition diet. The feedstuffs were analysed by proximate feed analysis and the metabolizable energy content was estimated. After 3 months feeding, individual cows were injected (i.m) with 1500 IU pregnant mare's serum gonadotrophin (PMSG) at day 10 or day 11 of the oestrous cycle (day of oestrus = day 0). Alfaprostol (6 mg) was injected (i.m) 48 h after the injection of PMSG to induce oestrus. At day 6 or day 7 (day of insemination = day 1), the contents of individual uterine horns were flushed with 150–200 ml of phosphate‐buffered saline + 0.2% bovine serum albumin using a two‐way Foley catheter. The embryos were identified, evaluated and graded as excellent, good, fair or poor under a stereomicroscope. For the good‐ and high‐nutrition diets, the daily intake of green grass, straw, concentrate, dry matter, crude protein and estimated metabolizable energy by individual cows were 5 and 6 kg, 3 and 3 kg, 1.5 and 3.5 kg, 4.87 and 6.82 kg, 0.39 and 0.74 kg, and 39.60 and 59.12 MJ, respectively. The protein content was 8 and 11% in the good‐ and high‐nutrition diets, respectively. The two groups of cows on different nutritional diets differed significantly with regard to body weight, body condition score and number of palpated corpora lutea (p < 0.01). For cows on the good‐nutrition diet, the median number of recovered embryos and transferable quality embryos were three and two, respectively. The recovery rate of embryos was 79.30% of palpated corpora lutea. Cows on the high‐nutrition diet did not yield any embryos. The indigenous zebu cows fed on the good‐nutrition diet with BCS 2.5–3 were considered suitable for the induction of superovulation, the cows on the high‐nutrition diet with BCS 4–4.5 were unsatisfactory and were more prone to cyst formation in the ovaries.     

半胱氨酸天冬氨酸特异性蛋白酶介导的细胞凋亡对精子发生发育及冷冻保存的影响

半胱氨酸天冬氨酸特异性蛋白酶(cysteinyl aspartate specific proteinase, Caspase)是直接导致凋亡细胞解体的蛋白酶家族,在细胞凋亡机制网络中处于中心地位。在精子发生发育过程中,机体通过生殖细胞凋亡与细胞增殖平衡机制来确保正常数量和质量的精子分化与成熟。精子冷冻保存会对精子造成不可逆的冷冻损伤,冷冻－解冻后,一些标志性凋亡现象会显著增多。作者对Caspase介导的细胞凋亡对精子发生发育及冷冻保存的影响机制进行了综述,表明精子发生发育和冷冻损伤与精子细胞凋亡存在密切的联系。     

季节对杜泊羊体内胚胎生产与移植效果的影响

为了研究季节对杜泊羊体内胚胎生产效率及胚胎移植妊娠率的影响,于2016年选取内蒙古赛诺草原羊业有限公司种羊场经产纯种杜泊母羊作为供体,采用同期发情、超数排卵及腹腔镜人工输精等方法对供体羊进行处理后,利用手术冲胚的方法获得纯种杜泊羊胚胎,同时对受体羊进行同期发情处理和胚胎移植。经统计分析,2016年全年该公司共有放栓供体4 241只,超排处理4 180只,配种供体4 131只,冲胚供体3 987只,供体可用率为94.01%;冲出胚胎总数为23 516枚,可用胚胎18 002枚,胚胎可利用率为76.55%,平均每只供体能获得可用胚胎4.52枚。供体在10月、11月、12月冲胚所获得的平均冲胚数和平均可用胚数明显高于4月、5月、6月(P<0.05),其中10月的平均冲胚数和平均可用胚数最高(P<0.01),分别为7.04枚和5.62枚。将获得的部分胚胎用于胚胎移植,移植单胚怀孕受体数为4 446只,妊娠率为53.67%;移植双胚怀孕受体数805只,妊娠率65.02%。此外,受体在1月、3月、6月、12月受胎率较其他月份高(P<0.05)。该研究可为杜泊种羊超数排卵、胚胎移植和肉羊的产业化生产提供一定的数据参考。     

水解度对AM-DB/BS14共聚物耐温抗盐性能的影响

本文通过自由基共聚合丙烯酰胺(缩写为AM)和双键硼化物(缩写为DB/BS14)获得共聚物P(AMDB/BS14),水解度分别在20%、40%和60%的条件下对共聚物进行后水解,测试在不同水解度条件下得到的共聚物溶液粘度随体系温度和氯化钠盐度的变化情况。实验表明,在20%、40%和60%三种水解度条件下的共聚物耐温抗盐性能有明显的不同,其中共聚物在60%水解度时耐温和抗盐性能最优,在20%水解度时耐温抗盐性能最差。     

HPLC-ELSD法测定黄芪精质量标准的研究

采用HPLC-ELSD法对黄芪精中黄芪甲苷的含量进行分析。色谱柱为AgilentC18(5μm,4.6mm×250mm),柱温为35℃;流动相:乙腈—水(34:66);流速:1.0mL/min;ELSD检测器参数:漂移管温度为65℃;空气压力为30psi。黄芪甲苷在32.12～321.20μg范围内线性关系良好(r=0.9998),平均回收率为99.26%,RSD=2.75%(n=6)。该法简便、准确、重现性好,可作为黄芪精质量控制方法。     

不同冻存液对奶牛乳腺上皮细胞冻存质量的影响

试验旨在探究奶牛乳腺上皮细胞(bovine mammary epithelial cells,BMECs)最佳的冻存液以改善乳腺上皮细胞的冻存质量。BMECs传至第5代后分别加入以下10种不同配方的冻存液。A组:85%DMEM+10%胎牛血清+5%DMSO;B组:80%DMEM+10%胎牛血清+10%DMSO;C组:75%DMEM+10%胎牛血清+15%DMSO;D组:70%DMEM+10%胎牛血清+20%DMSO;E组:85%DMEM+5%胎牛血清+10%DMSO;F组:75%DMEM+15%胎牛血清+10%DMSO;G组:70%DMEM+20%胎牛血清+10%DMSO;H组:80%DMEM+10%胎牛血清+10%甘油;I组:70%DMEM+10%胎牛血清+20%甘油;J组:60%DMEM+10%胎牛血清+30%甘油,冻存前统一调整细胞密度到1×106个/mL冻存。分别对复苏后的细胞进行台盼蓝染色计算存活率和PI/Hoechst33258双染计算凋亡率。结果表明,BMECs经不同冻存剂冻存复苏后,细胞活力、形态学及凋亡率表现有所不同,其中B组和G组的活力和24h贴壁率较其他组高,二者的凋亡率较低,二者之间差异无显著性(P>0.05);传代后B组细胞的生长状况最好。     

不同冻存液对奶牛乳腺上皮细胞冻存质量的影响

试验旨在探究奶牛乳腺上皮细胞（bovine mammary epithelial cells,BMECs）最佳的冻存液以改善乳腺上皮细胞的冻存质量。BMECs传至第5代后分别加入以下10种不同配方的冻存液。A组：85%DMEM＋10%胎牛血清＋5%DMSO;B组：80%DMEM＋10%胎牛血清＋10%DMSO;C组：75%DMEM＋10%胎牛血清＋15%DMSO;D组：70%DMEM＋10%胎牛血清＋20%DMSO;E组：85%DMEM＋5%胎牛血清＋10%DMSO;F组：75%DMEM＋15%胎牛血清＋10%DMSO;G组：70%DMEM＋20%胎牛血清＋10%DMSO;H组：80%DMEM＋10%胎牛血清＋10%甘油;I组：70%DMEM＋10%胎牛血清＋20%甘油;J组：60%DMEM＋10%胎牛血清＋30%甘油,冻存前统一调整细胞密度到1×106个/mL冻存。分别对复苏后的细胞进行台盼蓝染色计算存活率和PI/Hoechst33258双染计算凋亡率。结果表明,BMECs经不同冻存剂冻存复苏后,细胞活力、形态学及凋亡率表现有所不同,其中B组和G组的活力和24h贴壁率较其他组高,二者的凋亡率较低,二者之间差异无显著性（P〉0.05）;传代后B组细胞的生长状况最好。     

不同冻存液对奶牛乳腺上皮细胞冻存质量的影响

试验旨在探究奶牛乳腺上皮细胞(bovine mammary epithelial cells,BMECs)最佳的冻存液以改善乳腺上皮细胞的冻存质量.BMECs传至第5代后分别加入以下10种不同配方的冻存液.A组:85％ DMEM+10％胎牛血清+5％ DMSO;B组:80％ DMEM+ 10％胎牛血清+10％ DMSO;C组:75％ DMEM+10％胎牛血清+15％ DMSO;D组:70％ DMEM+10％胎牛血清+20％ DMSO;E组:85％ DMEM+5％胎牛血清+10％ DMSO;F组:75％ DMEM+15％胎牛血清+10％ DMSO;G组:70％DMEM+20％胎牛血清+10％DMSO;H组:80％DMEM+10％胎牛血清+10％甘油;I组:70％ DMEM+10％胎牛血清+20％甘油;J组:60％ DMEM+10％胎牛血清+30％甘油,冻存前统一调整细胞密度到1×106个/mL冻存.分别对复苏后的细胞进行台盼蓝染色计算存活率和PI/Hoechst 33258双染计算凋亡率.结果表明,BMECs经不同冻存剂冻存复苏后,细胞活力、形态学及凋亡率表现有所不同,其中B组和G组的活力和24h贴壁率较其他组高,二者的凋亡率较低,二者之间差异无显著性(P＞0.05);传代后B组细胞的生长状况最好.     

Effect of suckling period on calf growth and milk yield of Zebu cows

Eighteen multiparous Zebu cows in their third lactation and their calves were randomly allocated to three suckling periods, up to 3, 4 or 5 months of age of the calf. The cows were individually fed natural hay, cottonseed cake and molasses. At 2 months of age, all calves were separated from their mothers, and were offered cottonseed cake mixed with molasses and Mucuna hay individually. The calves stimulated milk ejection by suckling 30 seconds and suckled the residual milk for 45 minutes after milking. The dry matter intake of cows (3.68, 3.29 and 3.31% of body weight) and calves (2.88, 2.80 and 2.55% of body weight) for suckling up 3, 4 and 5 months of age, respectively, was not significantly affected by treatment and neither was the growth rate of the calves (178, 157 and 149 g/d for 3, 4 or 5 months suckling period, respectively). Cows suckling their calves up to 5 months had significantly higher milk yield and higher amount of saleable milk (1.97, 2.93 and 3.69 kg/cow/d for 3, 4 and 5 months suckling period, respectively). The fat content of the milk decreased with increasing length of the suckling period while the protein content was not affected. In conclusion, a suckling period of 5 months resulted in higher total milk production and higher amount of saleable milk but did not seem to have any effect on calf growth when the calves were supplemented.     

影响精液冷冻保存和人工授精效果的因素

本文阐述了精液冷冻保存的细胞反应原理,指出冷冻保护剂甘油对精液保存具有利弊效应,提出精子膜脂组成的差异使得不同品种的精子对冷冻损伤的易感性不同。雌性生殖道解剖结构的品种差异,精子形态,精子运行机制的细微差异,人工授精时间及精子的运行能力,采精方式等因素对人工授精的效果有决定性作用。研究精子质膜的生物学特性可解决低活力精子的问题,然而这并不能解决冷冻后精子质量的个体差异。对精细胞基因组的研究可以找出这些个体的遗传差异。因此,冷冻精子和精原细胞(用于细胞外注射)的差异已经成为完整基因组问题。     

Oocyte and Embryo Quality: Effect of Origin, Culture Conditions and Gene Expression Patterns

In general, the majority of immature bovine oocytes fail to develop to the blastocyst stage following maturation, fertilization and culture in vitro. The evidence suggests that while culture conditions during in vitro embryo production can impact on the developmental potential of the early embryo, the intrinsic quality of the oocyte is the key factor determining the proportion of oocytes developing to the blastocyst stage. In addition, evidence suggests that the period of post‐fertilization embryo culture is the most critical in determining blastocyst quality. This paper reviews the current literature, with emphasis on the bovine model, demonstrating evidence for an effect of oocyte origin and/or in vitro maturation conditions on the developmental capacity and gene expression patterns in the oocyte. Furthermore, the well‐documented effects of post‐fertilization culture environment on embryo gene expression and quality are highlighted.     

天然烟制剂的抗螨效果

大蜂螨(Varroa jacobsoni Oud．)是世界范围内感染西方蜜蜂(Apis mellifera L．)最严重的外寄生虫，蜂螨通过吸取未成年和成年蜜蜂的血淋巴来危害蜜蜂，同时还会传播有害病毒，给养蜂业带来巨大的损失。……     

和牛超数排卵及胚胎移植效果观察

为了研究和牛超数排卵和胚胎移植的效果,试验以和牛作供体,荷斯坦青年母牛作受体,采用PG+FSH方案对7头和牛供体进行超数排卵处理,用非手术法取胚并进行鲜胚移植。结果表明：采用PG+FSH方案头均获得胚胎数为(11.29±4.86)枚,可用胚胎(5.86±3.27)枚(A级和B级);鲜胚移植(A级和B级)妊娠率为51.22%。说明采用该方案对和牛供体进行超数排卵,并进行鲜胚移植的效果较为理想。     

4味中药及其与抗菌药的复方制剂的MIC测定

用改进的试管两倍稀释法测定了黄连、鱼腥草、大青叶和苦参4味中药及其相互配伍（1：1）对标准大肠杆菌的最小抑菌浓度和4味中药与抗菌药配伍（1000：1）对临床分离鸡大肠杆菌的最小抑菌浓度。结果表明，4味中药及其相互配伍对大肠杆菌标准株有一定的抑制作用，其MIC值介于3．91—62．50mg／mL之间，其中大青叶和鱼腥草以1：l配伍对大肠杆菌抑菌效果最好，MIC为3．91mg／mL；对临床分离株，单方药中黄连抑菌效果最好，MIC为31．25mg／mL，24种复方药中，黄连和盐酸多西环素或加替沙星以1000：1配伍的抑菌效果最好，其MIC为3．91mg／mL。     

番茄红素对秦川牛精液冷冻保存及鲜精品质影响

旨在探究番茄红素对秦川牛精液冷冻保存和新鲜精液品质的影响,为番茄红素作为新型精液稀释液添加剂和种公畜日粮添加剂提供依据。本研究采集3头年龄3~5岁健康状况良好秦川种公牛的新鲜精液混合,用0、1、2、3、4 μmol·L^-1浓度的番茄红素稀释液冷冻保存;利用计算机辅助分析仪、低渗肿胀检测法、花生凝集素荧光标记检测法、罗丹明荧光检测法等检测精子的运动性能、顶体完整率、质膜完整性、线粒体活性和抗氧化性能;在秦川种公牛日粮中添加15 mg·kg^-1 BW番茄红素,检测饲喂前后秦川种公牛的新鲜精液品质和血清抗氧化指标。结果表明,在秦川牛精液冷冻保存过程中添加番茄红素可以显著提高冷冻-解冻后精子的活力、顶体完整率、质膜完整性和线粒体活性以及抗氧化酶CAT和GSH-Px酶活性,显著降低精子中MDA含量,且番茄红素的最适添加浓度为2 μmol·L^-1(P<0.05);饲喂15 mg·kg^-1 BW番茄红素后,与饲喂之前相比,秦川种公牛的精子活力显著提高(P<0.05),精子畸形率显著降低(P<0.05),血清中抗氧化酶SOD、CAT、GSH-Px酶活性显著增加(P<0.05),MDA含量显著降低(P<0.05)。综上所述,一定浓度的番茄红素对精子在冷冻过程中造成的精子损伤和秦川牛机体应激造成的繁殖性能降低具有良好的保护作用,这可能与番茄红素通过抗氧化作用减少精子在冷冻-解冻过程中的氧化损伤和提高动物机体的抗氧化能力有关。     

K肥对假俭草草坪绿度和品质的影响

施K对改善假俭草草坪的颜色、延长青绿期、提高品质有一定的作用.适时适量地给假俭草施以K肥,可增加其叶绿素含量,但是施肥量不得超过7.5 g/m2K2O,否则将影响到对其他阳离子尤其是Ca2+、Mg2+的吸收.K肥可延长青绿期2～4 d,尤其是4.5 g/m2K2O分3次施用会使枯黄期推迟4 d,而返青提前6 d,结果使青绿期达313 d.施K肥可以增加草坪的密度,减弱草坪在不良条件下颜色变红泛发等现象,增强草坪抗性,在不同时期都使草坪的品质有所提高.等量肥料分次施用的效果好于一次施用,每次施肥量不得高于2.25 g/m2K2O,年总施肥量不得高于7.5 g/m2K2O.假俭草在不同的生长时期都需要K肥,但在高温和低温条件下,施用的效果较好,这与K可以提高抗逆性有关.     

Research Progress on the Relationship between Cryopreservation of Mammalian Germ Cells and Their DNA Methylation

Cryopreservation of mammalian germ cells and embryos has been widely used in assisted reproduction. It also plays an important role in preservation of species diversity. DNA methylation abnormality induced by freezing in germ cells or embryos has an significant impact on the occurrence of disease and the physiological function of the offspring. In this paper, we summarize the effects of cryopreservation of mammalian germ cells and embryos on their pattern of DNA methylation which in turn make the developmental deficiency in the progeny from these gametes or embryos. Finally, it will provide the reference on the gamete cryopreservation and the development of assisted reproductive technology.     

Effect of different hand-milking techniques on milk production and teat treatment in Zebu dairy cattle

The aim of the study was to test three different hand-milking techniques (“pull down”, “thumb in”, and “full hand grip”) and their effects on milk production and teat treatment. This is important since milk production in many tropical areas still rely on hand-milking. The study was carried out at a peri-urban farm in the Bobo-Dioulasso area in Burkina Faso. Twelve indigenous Zebu cows in early lactation were used in the study. The sequences of the treatments (hand-milking techniques) and the milkers were balanced for carry-over effects between successive periods and days, respectively. The sequences were constructed by using special Latin squares. Yield and composition of saleable milk was not affected by milking technique but there were differences between the milkers in milk yield. There was also a significant interaction for saleable milk yield between milker and milking technique. Hemoglobin in milk was measured as an indicator of teat damage. The hemoglobin content was numerically higher in post-milking strip milk samples than in saleable milk. It was concluded that the amount of milk removed depends mainly on the milker and how well the milking technique works for the individual milker. No effect of milking technique was observed on teat treatment.