猪常温精液稀释剂配方的筛选

选用原精活力在0.80以上的大白、长白和杜洛克种公猪各3头,比较10个稀释剂配方的保存效果,选择保存效果最佳的配方分别添加不同浓度的氨基乙酸和牛血清白蛋白（BSA）进行优化。试验结果表明：5号配方的保存效果较好,保存时间为4.01d;添加1.25 g/L的氨基乙酸和3.00 g/L的BSA均可明显延长常温精液的保存时间（P〈0.05）。     

鸡精液稀释配方对低温保存精子活力和输精效果的影响

为了完善中国南方地区鸡场公鸡精液的保存技术、提高精液的利用率，本试验研究了在低温（4 ℃）保存的条件下，不同的保存时间（0、4、8、24 h）、不同的稀释液配方（原精液、配方Ⅰ和配方Ⅱ）对精液的精子活力变化以及人工输精繁殖效果的影响。选用33周龄黄鸡母鸡192只、公鸡42只，192只母鸡依笼号分为12组（3种处理精液×4个保存时间），每组16只母鸡，公鸡不分组。统一采精后用两种不同稀释液稀释、低温（4 ℃）保存至0、4、8、24 h后观察精子活力，并对母鸡输精，分组收集鸡蛋，对各组受精率、出雏率、健雏率进行比较分析，以原精液低温保存作为对照。结果显示，两种稀释液组的精子低温（4 ℃）保存4、8、24 h，其精子活力极显著高于原精液组（P<0.01），配方Ⅱ组精子的活力高于配方Ⅰ组（P>0.05）；两种配方稀释液组的精液低温（4 ℃）保存4 h，输精受精率高于原精液组（P<0.05）；两种配方稀释液组的精液在低温（4 ℃）保存8 h，输精受精率极显著高于原精液组（P<0.01）。表明这两种精液稀释液更有利于精液保存，经稀释后的精液可以显著提高受精率，可为中国南方鸡场种公鸡精液保存技术的完善提供有力的数据支撑。     

牛血清白蛋白对湖羊精液低温保存效果的影响

为探究牛血清白蛋白(BSA)对湖羊精液低温保存效果的影响,在湖羊精液低温保存的稀释液中添加不同浓度BSA(0、1、2、3、4、5、6 g/L),检测分析BSA对精子在4℃保存条件下的活力、活率、顶体完整率和质膜完整率。结果表明:保存24～96 h时,2～3 g/L BSA组的精子活率显著高于对照组;保存120 h时,3g/LBSA组的精子活率显著高于其他组;保存24、48、120h时,3g/LBSA组的精子活力显著提高;保存24～120 h时,6 g/L BSA组的精子活力低于对照组;保存48～72 h时,1～4 g/L BSA组的精子质膜完整率显著提高;保存48～72 h时,4 g/L BSA组的顶体完整率显著高于对照组;保存96～120 h时,3 g/L BSA组的顶体完整率显著高于对照组。本研究结果表明,在4℃条件下,1～5 g/L BSA可以提高湖羊精子活率、活力和质膜完整率,其中3 g/L BSA的作用效果最佳。     

不同稀释液和不同环境保存公猪精液的效果试验

采取优质长白公猪精液,用已证实较好的5个稀释液配方进行不同环境的保存试验,还将较优配方加与不加牛血清白蛋白(BSA)及与进口稀释液进行比较,配制好的稀释液放置1周后与现配稀释液保存效果的比较。结果表明,不同的保存环境对精子的存活指数影响较大,普通冰箱的保存效果最差,其次为带冰块的泡沫箱,而15℃～17℃恒温箱的保存效果最好;加入BSA对公猪精液的保存有一定效果(P>0.05);较优配方与进口稀释液相比,保存效果差异不显著(P>0.05);现配稀释液优于放置1周的稀释液,精子活率差异显著(P<0.05)。     

猪精液常温保存的试验

设计了4个猪精液稀释液配方,以台湾生产的猪精液常温保存稀释粉为对照,筛选出保存效果较好的配方,再进行不同因素对精液保存的影响试验。结果显示,以含有Tris和半胱胺酸的配方4,并在稀释液中添加庆大霉素(100 IU/mL)和林肯霉素(200 μg/mL)作抗生素,在24-30℃,pH为6．4,稀释倍数为1:1时保存效果最佳。     

不同剂量的稀释液对猪精子活力的影响

家畜精液保存技术在繁殖控制和现代畜牧业生产中日益显出其重要性。而猪新鲜精液常温保存的研究开始于1980年，它要求能保存3d以上，以利于人工授精技术的推广和长距离运输。研究中遇到的主要问题是不同剂量的稀释液在温度为18℃～22℃时对猪精液中精子保存时间长短，精子和精液中的微生物的新陈代谢强度几乎没有下降。因此，既要延长保存时间，又要维持受精力比较困难。1980年，意大利的Gottardi研究出了名为Zorlesco的稀释保存液，由于未添加牛血清白蛋白（BSA）。因此，不能常温保存猪新鲜精液，Zorlesco液的成分不平衡，渗透压低于240毫渗摩尔无助于维持精子活力。     

不同稀释液对绵羊精液4℃保存效果的影响

本实验旨在比较研究3种不同组成成分的绵羊精液低温稀释液对绵羊精子4℃保存效果的影响,3种稀释液分别是以三羟甲基氨基甲烷（Tris）-卵黄-柠檬酸-葡萄糖为基础配方的稀释液1、以脱脂奶粉-大豆卵磷脂-果糖为基础配方的稀释液2和以Tris-卵黄–柠檬酸-果糖为基础配方的稀释液3。通过假阴道方法采集健康成年绵羊精液,用3种不同稀释液按照精液:稀释液=1:8的比例稀释后在4℃环境进行保存,分别在0、24、72、120 h和168 h后检测分析精子活力、运动参数、顶体完整性、线粒体膜完整性、精子总抗氧化能力（T-AOC）、超氧化物歧化酶（SOD）和细胞丙二醛（MDA）浓度。结果表明：精液保存168 h时,稀释液3组的保存效果最好,其总运动精子比例为54.91%、前向运动精子比例为43.27%、快速运动精子比例为19.73%、线粒体完整性为68.03%,均高于稀释液1和稀释液2组（P<0.05）;稀释液3组的顶体完整率为60.55%,高于稀释液2组（P<0.05）。保存120 h时,稀释液3组T-AOC为0.76 mmol/mg prot,高于稀释液2组（P<0.05）;稀释液3...     

甘肃高山细毛羊细管鲜精低温保存效果研究

为了研究甘肃高山细毛羊种公羊细管精液低温保存的效果.用葡萄糖、柠檬酸钠、鲜奶和卵黄为主要成分配制了3种稀释液,10倍稀释精液,并在4℃保存,分别在12h,24 h,36h,48 h,60h和72 h观测精子活力和人工授精受胎率.结果显示,配方Ⅲ稀释精液后,细管封装,4℃保存,72 h后精子活力可达0.49,与配方Ⅰ和Ⅱ差异显著,而且配方Ⅲ稀释精液后,在远距离输精后受胎率可达78.3％.以柠檬酸钠和葡萄糖为基础,添加卵黄和TCM199,在4℃下,能有效保护甘肃高山细毛羊精液,延长精子寿命,且在3d内可以满足远距离人工授精要求,有效利用种公羊价值.     

犬精液低温保存稀释液的研究

试验对苏联红犬精液的低温保存稀释液进行了研究。结果表明,犬精液低温保存的稀释液以Tris-葡萄糖稀释液(Tris3.04%,柠檬酸1.7%,葡萄糖1.25%,卵黄20%)为最佳,在4℃保存3 d后,犬精液的精子活率仍可达0.662 5±0.058 2,精子活率>0.1的存活达5.750 0 d±0.462 9 d。在Tris-葡萄糖稀释液中加入0.5%的甘氨酸或1%的牛血清白蛋白并不能明显提高犬精子的低温保存效果,反而添加BSA后,精子活率在0.1以上的存活时间出现明显缩短(P<0.01)。     

牛血清白蛋白对犬精液冷冻保存效果的影响

为了研究牛血清白蛋白(BSA)对犬精液冷冻保存效果的影响,试验配制含有不同浓度(0、3%、5%、7%)BSA的精液冷冻稀释液用于保存犬精液,测定冷冻-解冻后精子活力、质膜完整率、顶体完整率、总抗氧化能力(T-AOC)、丙二醛(MDA)含量及线粒体膜电位(MMP)等指标。结果表明：3%和5%BSA组冷冻-解冻后精子活力、质膜完整率、顶体完整率和T-AOC均显著高于对照组(P<0.05);3%BSA组MDA含量显著低于其他各组(P<0.05),线粒体膜电位值显著高于其他各组(P<0.05);除精子活力外,7%BSA组和对照组之间各项指标均差异不显著(P>0.05)。说明在犬精液冷冻稀释液中添加BSA能有效改善冷冻-解冻后精液品质,对提高精子抗氧化能力及代谢能力都有显著效果,BSA的最适添加浓度为3%。     

Fertility of ewes inseminated intrauterinally with frozen semen using extender containing bovine serum albumin

The present study was conducted to examine the fertility of ewes inseminated intrauterinally with frozen semen using semen extender containing either egg yolk or bovine serum albumin (BSA). Sixty Suffolk and cross-bred ewes were treated with controlled internal drug release (CIDR) devices during the non-breeding season (July 2006). A CIDR was inserted into the vagina for 12 days and an intramuscular injection of 500 IU equine chorionic gonadotropin was administered one day before its removal. Ejaculates from a suffolk ram were diluted with a Tris-based extender containing either 15% (v/v) egg yolk or 10% (w/v) BSA, and the diluted semen was frozen in 0.25 ml straws. A fixed-time intrauterine artificial insemination (AI) was performed 43-47 h after CIDR removal, regardless of incidence of estrus. There was no significant difference in pregnancy rates at 60 days after AI between the extenders containing egg yolk (66.7%, 20/30 animals) or BSA (65.5%, 19/29 animals). Furthermore, there were no significant difference in the lambing rates (66.7% and 62.1%) and prolificacy (1.25 and 1.56) between the two semen extenders. The present study indicates that a semi-defined semen extender containing 10% BSA produces fertility after intrauterine AI that is similar to that achieved with semen extender containing egg yolk.     

硫脲壳聚糖铜配合物与牛血清白蛋白的相互作用

在模拟动物体生理pH条件下,采用紫外吸收和荧光光谱研究硫脲壳聚糖铜(TUCS-Cu)配合物与牛血清白蛋白(BSA)的结合反应,探讨TUCS-Cu对BSA荧光猝灭过程的猝灭机理。结果表明,TUCS-Cu和BSA结合形成复合物,导致BSA内源性荧光猝灭机理属于静态猝灭。在室温下,TUCS-Cu与BSA的结合常数KA为2.68×106 L/mol。     

Effects of Egg Yolk Source on the Cryopreservation of Stallion Spermatozoa

Three experiments were conducted to determine whether replacement of chicken egg yolk, as a component of freezing extenders, with egg yolk from other avian species would improve the post-thaw motility and percentage of intact acrosomes of stallion spermatozoa. In the first experiment, substitution of chicken egg yolk with chukar egg yolk, as a component of the lactose-ethylenediaminetetraacetic acid extender, improved (P ≤ .05) the post-thaw motility of stallion spermatozoa. These results were not replicated in (IMV Technologies, Maple Grove, MN, USA) a more expansive study comparing 2%, 4%, 6%, or 8% egg yolk combined with INRA 96 when a “slow freeze” method was used, or the same substitution at levels ranging from 13% to 22% when egg yolk was combined with lactose-ethylenediaminetetraacetic acid for diluents used for a “fast freeze” method of cryopreservation. In the third study, egg yolks from regular and high omega-3 chicken eggs as well as from turkey, chukar, and mallard duck eggs were analyzed for lipid content and fatty acid profile. The yolk from the turkey eggs was higher (1,300 mg/100 g) and that from mallard ducks was lower (560 mg/100 g) in cholesterol as compared with the two types of chicken eggs and chukar egg yolk (range, 1,046-1,094 mg/100 g). In addition, the high omega-3 eggs did test higher for fatty acids (4.51 g/100 g) than other types of eggs (range, 0.28-0.73 g/100 g). Substitution of chicken egg yolk with turkey, but not duck, egg yolk resulted in higher post-thaw total motility (P ≤ .05) for spermatozoa obtained from two of the three stallions used in the third experiment.     

Fertility after artificial insemination using a soybean-based semen extender in sheep

The present study aimed to compare the fertility of ewes intrauterinally inseminated with frozen-thawed semen using a soybean-based semen extender (AndroMed) with those of ewes intrauterinally inseminated with frozen-thawed semen using a Tris-based extender containing either egg yolk or BSA. Suffolk ewes (n=104) were treated with an intravaginal sponge containing 40 mg fluoroprogesterone acetate (FGA) for 12 days and an intramuscular injection of 500 IU equine chorionic gonadotropin to induce estrus and ovulation during the non-breeding season (July, 2007). Intrauterine insemination was carried out 40-46 h after removal of the FGA sponge (n=90), regardless of the incidence of estrus. The pregnancy rates were not significantly different among the semen extenders containing egg yolk (64.5%) or BSA (58.6%) and AndroMed extender (56.7%). The lambing rates (64.5, 55.2 and 56.7% for the semen extenders containing egg yolk, BSA and AndroMed, respectively) and prolificacy (1.59 to 1.75) were also not significantly different. The present results indicate that an egg yolk-containing semen extender can be replaced with the non-animal derived extender AndroMed, which could be used for intrauterine insemination using frozen-thawed ram semen without reducing fertility.     

Effect of Different Extenders on In Vitro Characteristics of Feline Epididymal Sperm During Cryopreservation

To evaluate and compare the efficacy of various extenders for the cryopreservation of epididymal cat spermatozoa, two experiments were planned. Bovine and equine commercial extenders in the experiment 1 and TRIS–egg yolk–based extenders in experiment 2 were separately studied since the number of sperm collected per cat is reduced. Epididymal sperm samples were packaged into 0.25‐ml straws and frozen. Vigour, motility, morphology, acrosome status, sperm viability and functional membrane integrity were assessed at collection, after cooling and after thawing, while DNA integrity was evaluated at 0‐ and 6‐h post‐thaw. Experiment 1 compared the effect of three non‐feline commercial extenders – based on TRIS–egg yolk (Triladyl), egg‐yolk‐free medium (AndroMed) and skimmed milk‐egg yolk (Gent) – on the quality of frozen‐thawed epididymal cat sperm. Values for sperm motility and functional membrane integrity in cooled sperm diluted in Triladyl were higher (p < 0.001) than those recorded for Andromed and Gent. Except sperm morphology, the other assessed characteristics showed significant higher values in frozen‐thawed sperm diluted in Triladyl than in Andromed and Gent extenders. Experiment 2 analysed the effects of three TRIS–egg yolk–based extenders, one non‐feline commercial (Triladyl) and the other two prepared using different monosaccharides (glucose and fructose), on freezing‐thawed sperm. Results showed that specifically prepared extenders for cryopreservation of feline spermatozoa performed better than the commercial extender Triladyl, although sperm quality during the freezing‐thawing process did not significantly differ associated with the type of monosaccharide (glucose vs fructose) added to the mentioned extenders. Although TRIS–egg yolk–based extenders prepared in experiment 2 improved sperm cryoprotection, Triladyl remains a good option for practitioners who, for ease of use and availability, prefer to work with commercial extenders.     

多西环素人工抗原的合成与鉴定

将多西环素（doxycycline,DOX）进行化学修饰引入羧基或氨基等活性基团,然后与牛血清白蛋白（BSA）和卵清蛋白（OVA）偶联合成人工免疫原DOX—PABA—BSA、DOX—BSA和包被原DOX—PABA—OVA、DOX～OVA,并用紫外吸收（UV）、凝胶电泳（SDS—PAGE）和EUSA方法对人工抗原进行鉴定;将合成的人工抗原DOX—PABA—BSA、DOX—BSA分别免疫BALB／C小鼠,用间接ELISA方法测定多抗（pAb）效价,用竞争ELISA方法鉴定其敏感性,用交叉反应试验鉴定其特异性。结果表明,二个免疫组6只小鼠血清抗体效价均在1：6400以上,DOXpAb对DOX的50％抑制浓度（IC50）在39．79～53．13μg/L,抗血清与四环素类药物交叉反应很低。本实验为建立多西环素ELISA残留免疫学检测方法和并制备多西环素试剂盒奠定了基础。     

抗牛多杀性巴氏杆菌高免卵黄抗体IgY的制备及间接血凝检测方法的建立

本研究通过自制牛源荚膜血清A型多杀性巴氏杆菌灭活菌苗免疫产蛋鸡,采用卡拉胶结合硫酸铵沉淀法提取卵黄抗体IgY,并采用间接血凝方法检测抗体效价。结果表明,抗原致敏红细胞的最佳浓度是800μg/mL,免疫后第7周抗体效价达到高峰,效价为1：1024,高效价持续5周开始下降,测定提取的IgY浓度为8.258mg/mL,无菌检测及动物安全性实验表明制备的卵黄抗体安全可靠。本研究制备了抗牛多杀性巴氏杆菌卵黄抗体,为防治由荚膜血清A型多杀性巴氏杆菌所致的犊牛肺炎提供了新的手段。     

珍珠鸡蛋与海兰褐鸡蛋蛋品质及营养成分比较分析

珍珠鸡是从国外引进,经人工驯养育成的一个特色品种,其蛋用性能获得了显著进展,为加快蛋产品的开发力度,增加其附加价值,对珍珠鸡蛋和海兰褐鸡蛋蛋品质及营养成分进行比较分析。试验随机抽取新鲜珍珠鸡蛋、海兰褐鸡蛋各100枚,测定其蛋外、内在品质及主要营养成分,采用Duncan新复极差法、皮尔逊相关系数进行比较分析。研究表明,在外部品质方面,海兰褐鸡蛋蛋重高于珍珠鸡蛋15.54g（35.61%）,有极显著差异（P〈0.01）;珍珠鸡蛋蛋壳厚度比海兰褐鸡蛋的蛋壳厚0.14mm（40%）,差异极显著（P〈0.01）;由于蛋重的差异,两种鸡蛋蛋黄比例、蛋白比例、蛋壳比例也均有极显著差异（P〈0.01）。在内在品质方面,两种鸡蛋蛋清pH、蛋黄高度、蛋黄系数均无明显差异;珍珠鸡蛋浓蛋白高度比海兰褐鸡蛋的低3.02mm,有极显著差异（P〈0.01）;浓蛋白系数比海兰褐的高0.87,有显著差异（P〈0.05）;珍珠鸡蛋蛋黄颜色比海兰褐鸡蛋的颜色大一个罗氏比色值,有显著差异（P〈0.05）。在营养成分上,以g/枚为单位,珍珠鸡蛋的主要营养成分除胆固醇高于海兰褐鸡蛋外,其他均低于海兰褐鸡蛋。以g/100g为单位,珍珠鸡蛋水分含量明显低于海兰褐鸡蛋,相差12.62（20.21%）,差异极显著（P〈0.01）,脂肪含量也低于海兰褐鸡蛋,差异显著（P〈0.05）,相差1.34（4.4%）,其它营养成分均高于海兰褐鸡蛋。珍珠鸡蛋是较海兰褐鸡蛋小、壳厚、水分含量低、营养丰富的浓缩型蛋,为珍珠鸡蛋的蛋品开发创造了条件。     

不同培养体系对胎牛成纤维细胞体外培养的影响

研究不同培养体系对胎牛成纤维细胞体外培养的影响及用牛血清白蛋白代替血清培养胎牛成纤维细胞的可行性。利用M199、DMEM、α-MEM、DMEM/F124种培养体系通过组织块贴壁培养对成纤维细胞体外培养液进行筛选,以α-MEM组细胞生长状况较好。分别用含2、4、6、8、10mg/mL BSA的α-MEM培养液对胎牛成纤维细胞进行原代及传代培养,5种浓度的BSA对原代培养时细胞开始游离出组织块的时间影响不明显,均在培养后的48h有成纤维细胞和上皮细胞混合游离出,但在传代培养时,胎牛成纤维细胞在8mg/mL BSA浓度的α-MEM中贴壁率较高。结果表明:培养胎牛成纤维细胞时,可用BSA代替血清,较适宜的培养体系为含8mg/mL BSA的α-MEM培养液。     

Testing Usability of Butylated Hydroxytoluene in Conservation of Goat Semen

The objective of this study was to investigate whether butylated hydroxytoluene (BHT) could be used as a suitable supporter or alternative of egg yolk during preservation of goat spermatozoa. Three in vitro experiments and a fertility test were conducted to evaluate the effect of BHT on viability of chilled‐stored semen as well as motility and kidding rate of frozen‐thawed spermatozoa. In the first two experiments, ejaculates (n = 30/experiment) were collected from 10 bucks, split, diluted with egg yolk‐based and egg yolk‐free extenders supplemented with or without 0.3, 0.6, 2, 5 and 8 mm BHT and stored at 5°C for 168 h. In the third experiment, 30 ejaculates were collected from the above‐mentioned bucks, split and diluted with egg yolk‐free extenders supplemented with or without 0.3, 0.6 and 0.9 mm BHT and egg yolk‐based extenders supplemented with or without 5 mm BHT. Diluted semen was cooled to 5°C over a period of 4 h, frozen and thawed in the form of 0.3‐ml pellets. In the fertility test, 75 ejaculates were collected from two proven fertile bucks, split, diluted with egg yolk‐free extenders containing 0.6 mm BHT and egg yolk‐based extenders supplemented with or without 5 mm BHT, frozen and thawed as described above. An insemination volume of 0.6 ml containing 120–140 × 10⁶ progressively motile spermatozoa was used for a single cervical insemination of cloprostenol‐synchronized does (n = 230). The results showed that addition of 5 mm BHT to egg yolk‐deficient (2.5%) extenders significantly improved viability of chilled‐stored semen together with motility (48.5%) and fertility (62.5%) of frozen‐thawed spermatozoa. Replacement of egg yolk in semen extenders by 0.6 mm BHT could sustain not only viability of chilled‐stored semen but also post‐thaw motility (47.5%) and fertility (53.75%) of frozen‐thawed spermatozoa. In conclusion, supplementation of semen diluents with BHT can ameliorate preservability of goat sperm.