中国水牛遗传育种研究进展

我国水牛遗传育种的研究主要集中在染色体核型、蛋白质多态性、水牛胚胎体外生产与胚胎移植技术等方面。本文从中国水牛的类型和分布特征、遗传特性的研究现状、现代生物技术在水牛育种中的应用、中国水牛育种研究前景及展望等四个方面综述水牛的育种进展,认为以现代生物技术为核心的分子育种将成为水牛育种的总趋势,为我国水牛的产业化发展提供了理论依据。     

Effect of body condition score and reuse of progesterone‐releasing intravaginal devices on conception rate following timed artificial insemination in Nelore cows

This study had the aim of investigating the efficiency of timed artificial insemination (TAI) through the progesterone‐releasing intravaginal device (PRID), used in new condition and for the second and third times in Nelore cows. The effects of device reuse and body condition score (BCS) on the conception rate (CR) were evaluated in 1,122 multiparous Nelore cows (mean BCS of 2.7 ± 0.4), which were randomly distributed into three groups that received new (n = 330), once (n = 439) and twice used (n = 353) PRID. Among the 1,122 females that underwent TAI, 573 became pregnant, thus representing an overall CR of 51.06%. Cows with BCS between 2.75 and 4.0 had greater (p < .0001) CR (69.75%) than cows with BSC of 2.0–2.5 (32.98%). It was observed that the CR through using PRID was 60.00%, 51.71% and 41.93% for new, once and twice used PRID, respectively, with difference between all groups (p < .0001). Under tropical conditions, animals with BCS greater than 2.5 had a higher CR, and the CR decreased proportionally with the number of times that the PRID had been used.     

Relationship of the conception rate and the side (left or right) of preovulatory follicle location at artificial insemination in dairy heifers

In this study, we examined the locational effect (left or right ovary) of the preovulatory follicle (PF) on fertility in dairy heifers. In total, 1,111 artificial inseminations (AI) were analyzed. At AI, PF locations were examined using rectal palpation, and heifers were divided into two groups on their PF locations: (i) the PF located in the left ovary (L‐PF); and (ii) the PF located in the right ovary (R‐PF). Pregnancy was diagnosed by rectal palpation 60 days after AI. The conception rate was 50.7% in all heifers. Conception rate was significantly higher in the L‐PF (60.1%) than in the R‐PF (46.2%). The conception rate was significantly lower by sexed semen (48.6%) than conventional semen (59.1%). Conception rates divided by the semen type (sexed: n = 896, conventional: n = 215) were significantly higher in the L‐PF than in the R‐PF for both semen types (sexed; L‐PF vs. R‐PF: 57.3% vs. 44.4%, conventional; L‐PF vs. R‐PF: 72.3% vs. 53.3%). In addition, season, age, AI number, and the number of re‐inseminations at the same estrus did not affect conception rates. In summary, PF development in the left ovary was associated with increased conception rates in dairy heifers.     

Effects of progestagen exposure duration on estrus synchronization and conception rates of crossbreed ewes undergoing fixed time artificial insemination

Synchronization of estrus and ovulation are of paramount importance in modern livestock improvement programs. These methods are critical for assisted reproduction technologies, including artificial insemination and embryo transfer, that can increase productivity. In the current study, subcutaneous implants containing norgestomet were placed for long (14 days), medium (9 days), and short (5 days) periods of time in 70 crossbred ewes undergoing fixed-time artificial insemination. The resulting effects on estrus synchronization and conception rates were subsequently evaluated. Among the synchronized ewes, 85.7% (60/70) underwent estrus over a period of 72 h after progestagen treatment ceased. The shortest mean interval between withdrawal of the device and onset of estrus (34.2 ± 8.9 h) was observed in the G14 days of P4 group (p < 0.05). The conception rate of the G14 days of P4 group was statistically higher than that of the other groups (83.3% vs. 60.9% vs. 47.8%; p < 0.05). In conclusion, 14 days of norgestomet treatment produced higher conception rates and a greater number of pregnancies at the beginning of the breeding season.     

Effects of body condition score in cows peripartum on the onset of postpartum ovarian cyclicity and conception rates after ovulation synchronization/fixed-time artificial insemination

The aim of this study was to examine whether the nutritional state of cows peripartum was associated with the recovery of ovarian function and conception rates after synchronization of ovulation and fixed-time artificial insemination (OVSYNCH/TAI). The effect of the interval in days from calving to the first ovulation on conception rates after OVSYNCH/TAI was also investigated. Conception rates of cows after OVSYNCH/TAI (n=39) were 43.6%. The conception rates of cows with a body condition score (BCS) of 2.75-3.25 at 30 d postpartum and on the day of OVSYNCH treatment were significantly higher than in cows with a BCS < or =2.5 (P<0.05). The percentage of cows establishing ovarian cyclicity before 55 d postpartum in cows with a BCS of 2.75-3.25 at 30 d postpartum and on the day of OVSYNCH treatment were significantly higher than in cows with a BCS < or =2.5 (P<0.05). The conception rates after OVSYNCH/TAI in cows which recovered ovarian cyclicity within 34 d postpartum were significantly higher than in cows with first ovulation > or =56 d (P<0.05). These results indicated that the nutritional state in cows peripartum influenced the conception rates after OVSYNCH/TAI and the postpartum ovarian cyclicity and also suggested that the conception rates after OVSYNCH/TAI decreased in cows with delayed recovery of ovarian cyclicity.     

Growth rate of ovulatory follicles during the first ovulatory oestrus (after seasonal anoestrus) and subsequent oestrous period in Irish Draught mares

It is believed that during the spring transition, the developing follicle tends to grow more slowly, persist longer and grow to a larger diameter prior to ovulation than at subsequent oestrus periods. A general suspicion, that the first ovulation of the year is less fertile than subsequent ovulations could be explained by a slower growth rate of the ovulatory follicle during transition with the consequent production of a subfertile oocyte. By detailed serial examination of the same group of Irish Draught mares over three winter/spring periods, no significant difference was found in either growth rate or pre-ovulatory diameter when compared with subsequent ovulations. Mean growth rates over the ten days prior to ovulation were 2.20 mm/day (range 1.18 to 3.64) and 2.19 mm/day (range 1.25 to 3.41) for first and subsequent ovulations respectively. Mean maximum pre-ovulatory diameters were 44.7 mm (range 35 to 59) and 43.5 mm (range 31 to 57.5) for first and subsequent ovulations respectively. The impression gained by practitioners that the first follicle develops more slowly during the transition to the first ovulation of the season may be due to less frequent examinations and consequently a failure to observe and record that follicles may grow and then regress during this period. The largest follicle observed a few days previously is not necessarily the same large follicle found at a later examination.     

青年奶牛同期发情和同期排卵－定时输精试验

试验旨在研究同期发情和同期排卵－定时输精技术在青年奶牛中的应用。选择1046头澳大利亚进口荷斯坦青年奶牛,随机分为两组:一组自然发情人工授精配种(对照组);另一组采用前列腺素F_2α(PGF_2α)诱导母牛发情或按照同期排卵－定时输精程序(Ovsynch或Ovsynch+CIDR法)处理母牛后人工授精配种(试验组),统计同期发情率、不返情率、第一次人工授精妊娠率和21d妊娠率等繁殖指标。结果表明,试验组青年奶牛人工授精后不返情率和第一次人工授精妊娠率与对照组间无显著差异(P>0.05),但21d妊娠率显著高于对照组(53.1%和35.3%,P<0.05)。试验组中,1次PG法、间隔7d2次PG法和间隔11d2次PG法的同期发情率分别为76.1%、81.7%和84.6%,差异均不显著(P>0.05);同期排卵－定时输精组中,Ovsynch法(GPG)和Ovsynch+CIDR法(GPG+CIDR)的不返情率、第一次人工授精妊娠率和21d妊娠率间无显著差异(P>0.05)。不同输精人员可显著影响青年奶牛的第一次人工授精妊娠率(P<0.05),而不同公牛常规冷冻精液对青年奶牛第一次人工授精妊娠率无显著影响(P>0.05)。同期发情及同期排卵－定时输精技术可使青年奶牛集中发情,提高参配率,从而提高21d妊娠率,有效加快青年奶牛人工授精效率,降低饲养成本。     

Trends in the quality and hygiene parameters of bulk Italian Mediterranean buffalo (Bubalus bubalis) milk: A three year study

Buffalo milk represents an indispensable source of nourishment in many parts of the world and it is the second most consumed milk worldwide. Buffalo milk is actually used for the production of many dairy products such as pasteurized or concentrated milk, butter, yogurt, ice‐cream, dehydrated milk products and cheeses. Due to its high nutritional value and the presence of natural bioactive substances, buffalo milk can also provide health benefits to consumers. In Italy, buffalo milk is used mainly for cheese making, mozzarella PDO (Protected Designation of Origin), which is a highly valued dairy product. This 3‐year study, carried out between 2011 and 2013, was aimed at evaluating the quality of bulk Italian Mediterranean buffalo milk by monitoring physico‐chemical parameters, somatic cell and total bacterial counts. A total of 51 samples of bulk milk were collected from one herd throughout the monitored period. Analysis of variance, carried out to test month, season and year main effects, highlighted remarkable seasonal effects for fat, protein and lactose content, as well as for predicted mozzarella cheese yield, and somatic cell counts. The calculation of simple correlations allowed the identification of positive correlations between estimated cheese yield and fat and protein content.     

Effects of Chemical Activation Treatment on Development of Swamp Buffalo (Bubalus bubalis) Oocytes Matured In Vitro and Fertilized by Intracytoplasmic Sperm Injection

The objective of this study was to optimize the activation protocol for buffalo oocytes after intracytoplasmic sperm injection (ICSI). The release of the second polar body (PB) at 3, 6 and 9 h after ICSI of in‐vitro matured oocytes activated either with 5 μm ionomycin (Io) or with 7% ethanol (EtOH) was preliminary examined. The highest rate of second PB extrusion occurred at 3 h of activation, and the second PB extrusion in EtOH group was significantly higher than that in Io group. Oocytes that extruded the second PB were selected and cultured either with 1.9 mm 6‐dimethylaminopurine (6‐DMAP) for 3 h or with 10 μg/ml cycloheximide (CHX) for 5 h. Significantly higher rate of oocytes formed 2 pronuclei in EtOH combined with CHX (EtOH + CHX) (62%) group compared to those of Io + CHX (42%) and EtOH + 6‐DMAP (48%) groups (p < 0.01) whereas Io + 6‐DMAP group showed intermediate value (58%). Significantly higher blastocyst formation rates were obtained in Io + 6‐DMAP (29%) and EtOH + CHX (24%) groups than in Io + CHX (6%) and EtOH + 6‐DMAP (17%) groups. Our results indicate that buffalo ICSI oocytes are effectively activated by combination treatment of Io with 6‐DMAP and EtOH with CHX resulting in the highest cleavage and blastocyst formation rates.     

Effects of FSH and LH on Steroid Production by Buffalo (Bubalus bubalis) Granulosa Cells Cultured In Vitro Under Serum‐Free Conditions

The objective of this study was to examine the effects of FSH and LH on oestradiol‐17β and progesterone production by buffalo granulosa cells cultured under serum‐free conditions. Granulosa cells (3 × 10⁵) from small (≤5 mm diameter) follicles were cultured for up to 4 days in 48‐well plates coated with 3.3 μg/cm² fibronectin in Dulbecco's modified Eagle's medium (DMEM) : nutrient mixture F‐12 Ham (1 : 1 ratio) supplemented with 10^?7 m androstenedione, 5 μg/ml human apo‐transferrin and 0.1% bovine serum albumin, in the presence or absence of FSH or LH (0, 1, 2, 4, 8, 16, 32 or 64 ng/ml each). Basal oestradiol‐17β production by granulosa cells from small follicles reduced (p < 0.01) from days 1 to 2 of culture and became undetectable by day 3 and basal progesterone production increased (p < 0.05) from day 1 through day 4 of the culture. Although there was no effect of FSH on day 1 of the culture, FSH at 2, 4, 8 and 16 ng/ml increased (p < 0.05) oestradiol‐17β production by granulosa cells from small follicles on day 2. Progesterone secretion was increased (p < 0.05) by all doses of FSH on all days of culture. All doses of LH had no effect on oestradiol‐17β or progesterone production by granulosa cells from small follicles on any day of the culture. The results of this study demonstrate a serum‐free culture system for buffalo granulosa cells and stimulatory effect of FSH but not LH on steroid hormone production by buffalo granulosa cells under these conditions.     

复合添加剂对解冻后奶牛性控冻精质量及受胎率的影响

【目的】试验旨在研究以褪黑素和地诺前列腺素组成的复合添加剂对奶牛性控冻精解冻后精子质量及奶牛人工授精后受胎率的影响。【方法】30份奶牛性控冻精样均分为2组：试验组和对照组,试验组添加由24 ng/mL褪黑素、1.5 mg/mL地诺前列腺素及精液稀释液组成的250μL复合添加剂,对照组添加250μL精液稀释液,试验组和对照组均与解冻后的奶牛性控冻精按照1∶1(V/V)进行混合,室温孵育0、2及4 h,通过免疫荧光染色分析精子活率、顶体完整率、高能线粒体活性精子比率;对216头青年奶牛和82头头胎奶牛输精后通过28 d早孕检测来确定受胎率情况。【结果】与对照组相比,经复合添加剂处理0 h的性控冻精活率、顶体完整率及高能线粒体活性精子比率均无显著变化(P>0.05);经复合添加剂处理2 h的性控冻精活率、顶体完整率、高能线粒体活性精子比率均显著升高(P<0.05);经复合添加剂处理4 h的性控冻精活率和顶体完整率均无显著变化(P>0.05),高能线粒体活性精子比率显著升高(P<0.05)。在输精试验中,试验组青年奶牛和头胎奶牛中的受胎率分别为65.22%和48.21%...     

Effects of AI protocol and time interval from onset of estrus to AI on conception rate in Japanese Black cows

Two different artificial insemination (AI) protocols were investigated to determine the effect of the time interval from the onset of estrus (as detected by a pedometer) to the AI on the conception rate in Japanese Black cows. Seventy‐three cows were divided into two groups that received AI either after the induction of synchronized ovulation (synchronized AI group; n = 26) or at the time of observed estrus (control AI group; n = 47). Each group was further divided into two subgroups, which were artificially inseminated either 0–12 h (early AI group; n = 21) or 12–24 h (late AI group; n = 52) after the onset of estrus. There was no significant difference in the conception rate between the synchronized AI and control AI groups. The AI protocol and the detection of estrus using a threshold of counted steps (as measured by a pedometer) in this study could be effective for planned reproduction management without the observation of standing estrus in Japanese Black cows.     

Effects of Trichostatin A on In Vitro Development and DNA Methylation Level of the Satellite I Region of Swamp Buffalo (Bubalus bubalis) Cloned Embryos

Trichostatin A (TSA), a histone deacetylase inhibitor, has been widely used to improve the cloning efficiency in several species. This brings our attention to investigation of the effects of TSA on developmental potential of swamp buffalo cloned embryos. Swamp buffalo cloned embryos were produced by electrical pulse fusion of male swamp buffalo fibroblasts with swamp buffalo enucleated oocytes. After fusion, reconstructed oocytes were treated with 0, 25 or 50 nM TSA for 10 h. The results showed that there was no significant difference in the rates of fusion (82–85%), cleavage (79–84%) and development to the 8-cell stage (59–65%) among treatment groups. The highest developmental rates to the morula and blastocyst stages of embryos were found in the 25 nM TSA-treated group (42.7 and 30.1%, respectively). We also analyzed the DNA methylation level in the satellite I region of donor cells and in in vitro fertilized (IVF) and cloned embryos using the bisulfite DNA sequencing method. The results indicated that the DNA methylation levels in cloned embryos were significantly higher than those of IVF embryos but approximately similar to those of donor cells. Moreover, there was no significant difference in the methylation level among TSA-treated and untreated cloned embryos. Thus, TSA treatments at 25 nM for 10 h could enhance the in vitro developmental potential of swamp buffalo cloned embryos, but no beneficial effect on the DNA methylation level was observed.     

受体奶牛不同同期发情处理方法对胚胎移植效果的影响研究

本试验选择产后60天以上、空怀、无疾病、体况适宜、年龄2～10岁的182头黑白花奶牛,随机分成第1、第2两组同期发情,发情7天后胚胎移植。第1组为CUEMATETM孕酮栓 PG(n=120),第2组为PG一步法。试验结果表明,第1组牛的黄体不合格率显著高于第2组(P<0.05);产犊率低于第2组,但差异不显著(P>0.05)。从经济实用角度看,本试验初步显示:PG法比CUE-MATETM孕酮栓 PG法好,成本低、操作方便、产犊率较高。     

不同生育期和凋萎时间对多花黑麦草饲用和发酵品质的影响

本研究利用多花黑麦草Musashi品种, 研究孕穗期和开花期饲草在不同凋萎时间的饲用品质动态及青贮品质动态, 旨在选择出适宜不同生育期多花黑麦草凋萎青贮的时间。实验设置了对照、凋萎1, 2, 3, 4和5 h共6个处理, 分别测定了原料的饲用品质相关性状和青贮料的发酵品质性状。结果表明, 原料的非结构碳水化合物(NSC)含量随着凋萎时间的延长持续下降, 但可溶性碳水化合物(WSC)则呈不同的变化趋势:孕穗期多花黑麦草在凋萎过程中的WSC呈不同程度的上升, 在凋萎1 h时含量达到最高, 为10.62%, 之后小幅下降, 但始终高于对照(P<0.05);开花期多花黑麦草在凋萎过程中的WSC则呈先上升后下降的趋势, 在凋萎3 h时含量达到最高, 为14.46%, 之后下降。孕穗期多花黑麦草不同凋萎时间的青贮发酵品质性状中, pH在凋萎3 h时最低, 为4.05, 随后小幅上升;氨态氮(NH₃-N)含量随着凋萎时间的延长持续下降;乳酸(LA)含量先升后降, 凋萎3 h时含量最高, 达8.49%。开花期多花黑麦草不同凋萎时间的青贮发酵品质中, pH随着凋萎时间的延长持续上升;NH₃-N含量随着凋萎时间的延长呈先下降后回升的趋势, 凋萎2 h处理的含量最低, 为2.03 mg/g;LA含量持续下降, 但凋萎3 h以后LA含量大幅降低, 仅为4.56%, 凋萎5 h时LA含量则大幅降至1.38%。适宜的凋萎处理时间, 应该是原料中的WSC含量较高, 干物质含量适宜;青贮料具有较低的pH、NH₃-N含量和较高的LA含量。综合孕穗期多花黑麦草在不同凋萎时间的青贮品质变化趋势, 以凋萎3 h为佳, 凋萎3 h时原料的干物质含量升至26.59%。开花期则以凋萎2 h处理最佳, 凋萎2 h时原料的干物质含量升至28.03%。     

Effects of plane of nutrition on slaughtering traits and meat characteristics in Murrah graded male buffalo (Bubalus bubalis) calves in Nepal

An experiment was conducted using 17 male buffalo calves to assess the effects of plane of nutrition on slaughtering traits and meat characteristics. To attain 250 kg body weight (BW), the calves were allocated into three groups: high (H), low‐high (L‐H) and low (L) corresponding to concentrate levels receiving the concentrate at 1.50% of BW, 0.75% of BW until 190 kg BW and 1.50% thereafter, and 0.75% of BW, respectively. The animals had ad libitum access to urea‐treated rice straw. No significant differences of hot carcass weight, dressing percentage and lean fat–bone yields were observed among the treatment groups. The L group had heavier brisket weight and lower percentage of round weight in the hot carcass than the H and L‐H groups (P < 0.05). The H group had heavier hearts than the L group, and the H and L‐H groups had heavier livers and kidneys than the L group (P < 0.05). There was no significant difference of rib eye area, pH and the contents of moisture, crude protein and fat in loin meat among the groups. The findings indicated that the effects of plane of nutrition affected the weight or percentages of some cut yields in the hot carcasses and internal organs.     

日粮阴阳离子差对奶牛的影响及其在生产实践中的应用研究进展

阴阳离子差(DCAD)是指日粮中每千克或每百克干物质所含的主要阳离子（Na⁺、K⁺、Ca²⁺、Mg²⁺）毫摩尔数与主要阴离子（Cl^-、S^2-、PO^3-₄）毫摩尔数之差。DCAD直接影响奶牛血液的酸碱平衡及酶系统等的功能，可以改善奶牛的健康状况、提高奶牛生产性能。在此，作者综述了国内外关于调节日粮阴阳离子差对奶牛影响的研究结果及现状，探讨了DCAD在奶牛不同阶段生产实践中的应用。     

Effects of plane of nutrition on growth,feed intake,digestibility and nitrogen balance in Murrah graded male buffalo (Bubalus bubalis) calves in Nepal

An experiment was conducted using 17 male buffalo calves to assess the effects of plane of nutrition on dry matter intake (DMI), daily gain (DG), body size measurement, apparent digestibility and nitrogen (N) balance. To attain 250 kg BW, the calves were allocated into three groups: H, L‐H and L, receiving the concentrate at 1.50% of BW, 0.75% of BW until 190 kg BW and 1.50% thereafter and 0.75% of BW, respectively. The animals had ad libitum access to urea‐treated rice straw (UTRS). The DMI of UTRS through the experiment was higher in L and L‐H than H, showing 3.52, 2.90 and 2.62 kg/day, respectively (P < 0.01), but the total DMI did not differ among the treatment groups. The DG throughout the experiment was high in the order of H, L‐H and L, showing 0.72, 0.57 and 0.45 kg, respectively (P < 0.01). The digestibility of DM, organic matter, crude protein, neutral and acid detergent fiber and N retention were higher in H than in L (P < 0.05). The findings of this study thus revealed the greater DG has an advantage of shortening the growing period around 3 months, and consequently increasing benefit in fattening of buffalo calves in Nepal.