Follicullar Development and Embryo Recovery Following 3 versus 8 FSH Injections in Heifers

Ovarian follicular dynamics and embryo yield were studied during 2 different FSH regimens for superovulation of cattle. Twenty heifers were given intramuscular injections of FSH (total of 35 mg NIH) either once daily for 3 days (Group 3×1) or twice daily for 4 days (Group 4×2). At 72 h after the first FSH injection, each animal was injected with 0.75 mg cloprostenol. Inseminations were performed at 12 h and 24 h after the onset of heat. Transrectal ultrasonography was performed on the day of the first FSH injection, the day of cloprostenol injection, the day of insemination and finally on the day of embryo recovery (day 6 or 7 after heat). The numbers of small (2–4 mm), medium (5–9 mm) and large (>10 mm) size follicles were recorded. The total number of corpora lutea, eggs and transferable embryos were recorded on the day of embryo recovery. No differences were found between the 2 groups in either of the parameters studied (p>0.05). It can be concluded that treatment with this FSH preparation once daily for 3 days gives a folliculogenic and superovulatory response similar to a treatment regimen where it is given twice daily for 4 days.     

PGF2αeCG及hCG 3种生殖激素联用对母犬发情、卵泡发育及胚胎着床的影响

给休情期性成熟母犬肌注 PGF2α(0 .5 m g/ kg) ,2 4 h后肌注 e CG(5 0 IU/ kg) ,再经 12 0 h后肌注 h CG(2 0 U/ kg) ,诱导其发情。将诱导发情的母犬与公犬交配 ,于交配后 16 d,摘取卵巢、子宫 ,观察其卵泡发育和胚胎着床情况。结果表明 ,药物处理可明显促使母犬提前结束休情期而进入发情期 (9/ 10 ) ,促进卵巢内卵泡发育 (P<0 .0 1) ,并使更多的胚胎着床 ,每只母犬子宫胚胎着床点为 (17.4± 2 .2 )个 ,提示 3种生殖激素联合应用可提高母犬繁殖率     

水牛牛巴贝斯虫体外培养的研究：Ⅱ.冷冻血清的应用和筛选

应用冷冻血清对１株采自自然感染的水牛牛巴贝斯虫进行了长达７２ｄ的体外连续培养，共继代２０次，培养７２ｈ红细胞染虫率最高达１４．１％，平均为８％～１０％。培养２０ｄ和３０ｄ的牛巴贝斯虫经液氮保存复苏后，接种于去脾水牛犊均引发了严重的牛巴贝斯虫病，从而说明已建立了水牛牛巴贝斯虫的体外连续培养，且经培养后的牛巴贝斯虫致病力没有改变。本试验利用６头份的冷冻健康水牛血清同时进行培养，结果发现，并非所有的健康水牛血清均适合于体外培养牛巴贝斯虫。这一发现对建立水牛牛巴贝斯虫的体外培养和研究水牛牛巴贝斯虫病均具有重要意义     

鸡促性腺激素释放激素对卵泡膜分泌雌二醇的影响

本实验选用性成熟的京白种蛋鸡,从同一产蛋顺序中取其发育不同阶段的各级卵泡(F_1～F_4),分离膜层,消化为单个的膜细胞,进行短期细胞培养,着重观察鸡促性腺激素释放激素(GnRH)对培养过程中膜细胞雌二醇分泌的影响.用放射免疫法测定细胞培养液中雌二醇的含量,得到以下结果:①未加外源激素处理的对照组细胞,随着卵泡从小到大的发育成熟过程,膜细胞雌二醇的分泌量逐渐降低;②适当剂量的鸡 GnRH-Ⅱ对各级卵泡膜细胞雌二醇的分泌均有促进作用,其中 F_4,F_3,F_2比 F_1更敏感;③加前体物(孕酮或雄烯二酮)之后,再加鸡 GnRH-Ⅱ比单加前体物或单加鸡 GnRH-Ⅱ,膜细胞雌二醇的分泌量增加更明显.实验结果提示,在体外细胞培养的条件下,GnRH-Ⅱ对膜细胞雌二醇的分泌不仅有促进作用,还可能促进雌二醇的合成。     

牛腔前卵泡的简易机械分离方法

采用两种机械法分离牛腔前卵泡。方法一 (M -1) ,皮肤移植刀切割、剪碎、过滤镜下直接捡卵法。方法二 (M -2 ) ,皮质片剪碎、离心、悬浮、过滤镜下捡卵法。M -1平均每卵巢采卵数为 46 9个± 18 2个 ,M -2为 6 8 4个± 12 5个 ;处理时间M -1为 2 5 4min± 6 7min ,M -2为 46 3min± 16 8min ,两种方法采集腔前卵泡数量及处理时间均存在显著差异。平均每小时采集卵泡数分别为 99 5和 87 2个 ,M -1多于M -2。两种方法获得腔前卵泡大小分布规律也有明显差异 ,M -1采集腔前卵泡直径为 6 0～ 15 0 μm ,绝大部分是次级卵泡 ,而M -2获取腔前卵泡则偏小。总的看来 ,M -1处理时间短 ,回收效率高 ,方法简便 ,且分离卵泡直径较大 ,适于体外培养 ,是机械分离牛腔前卵泡的一种理想方法。     

水牛附睾尾精子的体外受精

采用肝素诱导获能 ,比较了 TAL P液和 BO液处理水牛附睾尾精子进行体外受精和细管冷冻精液体外受精的效果。结果表明 ,用 BO液和 TAL P液分别处理水牛附睾尾精子 ,受精后的卵裂率分别为 4 6 .6 7%和 5 3.73% ,发育率分别为 2 1.6 7%和 2 6 .87% ,其受精效果差异不显著。综合 2种方法 ,水牛附睾尾精子的受精率为 5 7.14 % ,受精后的卵裂率为 5 0 .39% ;发育率相对于培养卵为 2 4 .4 1% ,相对于卵裂卵为 4 8.4 4 % ;与细管冷冻精液 (5 6 .0 0 % ,5 4 .31% ,2 6 .72 % ,4 9.2 1% )相比 ,差异不显著。形态学观察还表明 ,用保温干储的方法可获得活率好、存活时间长的附睾尾精子。试验结果说明水牛附睾尾精子用于体外受精可以得到与细管冷冻精液相当的效果。     

水牛精子在胞质内注射后的早期形态变化

应用胞质内精子注射 (intracytoplasmic sperm injection,ICSI)技术探讨了水牛卵母细胞胞质与注射精子间的相互作用以及 ICSI精子的形态变化。ICSI后 13h,5 9.4 %的精子头部已膨大 ,且有 18.8%的精子进入解聚状态。雌雄原核发育不同步 ,雄原核在 ICSI后 16 h和 19h的形成率分别为 3.2 %和 4 0 .0 % ;雌原核在 ICSI后 13h已达到 71.9% ,16 h时提高到 90 .3%。经离子霉素与 6 - DMAP联合激活 ,ICSI后 19h产生的胚胎核型以 2 PN PB1 为主 ,其雌原核形成 (激活 )率为 91.3% ,雄原核形成率为 4 0 .0 % ,5 1.3%为孤雌胚。用 5 mm ol/ L 的 DTT预处理精子 1h,可提高精子的解聚率 (30 .9%比 12 .7% ,P<0 .0 5 ) ,但对雄原核的形成率无显著影响 (33.3%比 32 .7% ,P>0 .0 5 )。结果表明 ,水牛精子 ICSI后的雄原核形成时间晚于卵子雌原核形成时间 ,且其比率低于卵子 ,DTT处理精子能提高其解聚率 ,但对雄原核形成无影响     

Improved milk production through PG–PL system by provision of in‐house shelter management in lactating Murrah buffaloes during winter season

Comprehensive information on the role of β‐casein and plasminogen–plasmin (PG–PL) system in milk secretion of Murrah buffaloes during winter season is lacking, although effects of cold stress can be ameliorated to an extent by altering microclimate at farm level. Hence, this study was aimed to determine the changes in productivity along with PG–PL system of milk, plasma hormones and metabolites of buffaloes during winter (December–January) season under two different management systems. Average minimum temperature and wind chill index during this season were 7.02 and 12.74 °C respectively. Buffaloes were divided in two groups of six animals each: control and treatment, where treatment group animals were placed in‐house with floor bedding of paddy straw and the control group animals in loose housing system without straw bedding. Physiological responses were recorded, and milk and blood samples were collected at weekly intervals for six‐week experimental period. Under in‐house management system, buffaloes experienced better comfort by alleviating environmental stress as their physiological responses such as respiration rate and pulse rate were significantly reduced (p < 0.01) as compared to the control, which subsequently resulted higher milk yield by 9.92% (p < 0.05). Analysis of milk samples revealed higher concentration of plasminogen (10.6 vs. 8.05 μg/ml; p < 0.01) and β‐casein (p < 0.05), and lower plasmin level (0.299 vs. 0.321 μg/ml; p < 0.05) in buffaloes under treatment group. It was also found that plasma cortisol, glucose and non‐esterified fatty acids levels were higher (p < 0.01) in control group as compared to the treatment animals by 13.6%, 8.14% and 12.6% respectively. However, milk composition, growth hormone, epinephrine and norepinephrine level in plasma were similar in both the groups. Hence, it may be concluded that provision of in‐house shelter management with floor bedding of paddy straw during winter was effective to minimize environmental stress and improved milk production through manipulation of PG–PL system in buffaloes.     

Influence of organic phosphorus on reproductive performance and metabolic profiles of anoestrous Farafra ewes in subtropics at the end of breeding season

The effect of organic phosphorus on metabolic, haematological and hormonal status, restoration of ovarian functions and conception rate in anoestrous Farafra ewes in subtropics were evaluated. Anoestrous Farafra ewes (n  = 24; 34.72 ± 0.52 kg body weight) were allocated into two equal groups: control and phosphorus groups. The ewes of phosphorus group were treated with sodium 4‐dimethylamino‐2‐methyl‐phenyl‐phosphonate as an organic bound phosphorous twice a week for successive 3 weeks. Ovarian follicle development and corpora lutea were checked three times a week till occurrence of oestrus using ultrasonography while pregnancy was confirmed at 30 days post‐service. Plasma metabolites, reproductive hormones, thyroid hormones and minerals were detected at weeks ?2, ?1, 0 (mating day) and + 4 weeks post‐oestrus. Phosphorus group had significantly (p  < .05) short interval to oestrous resumption if compared to control ewes (2.1 ± 0.8 weeks vs . 4.6 ± 1.1 weeks). In addition, phosphorous supplementation significantly (p  < .05) increased the number of antral follicles (developed and their sizes in addition to sizes of corpora lutea (8.72 ± 0.3 mm vs . 7.46 ± 0.9 mm) as well. Number of services per conception (2.6 vs . 1.4; p  < .01) was higher in control group than that of phosphorus group. Pregnancy rate (80 vs . 50%) was significantly (p  < .01) higher in phosphorus group when compared to control. White blood cells in treated ewes (10.8 ± 0.44; p  < .05) and monocytes (2.93 ± 0.13; p  < .01) were higher than that of control group (white blood cells; 9.53 ± 0.50 and monocytes; 2.24 ± 0.14). Metabolic parameters did not differ between phosphorus and control groups during different times of treatment. It could be concluded that phosphorous administration to anoestrous Farafra ewes in subtropics could improve reproductive performance and restore ovarian activity at the end of spring and early summer.