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1.
研究旨在建立优化的猪精液冷冻保存方法.实验表明,采用ZORLESCO(ZO)液对猪精液进行预稀释,室温平衡1 h.加入冷冻Ⅰ液后于5℃水浴平衡1.5 h,再加入冷冻Ⅱ液平衡2 h后装入0.25 mL细管,液氮面上3cm处平衡10 min投入液氮,采用37℃水浴解冻30 s,可获得最佳冷冻效果.解冻后的精子活力平均为0.58±0.03,质膜完整率为(63.2±1.2)%,顶体完整率为(51.4±2.6)%;精子畸形率最低,仅为(14.0±3.0)%. 相似文献
2.
芮荣 《农业生物技术学报》2008,16(2)
本研究旨在建立优化的猪精液冷冻保存方法。实验表明，采用ZO液对猪精液进行预稀释，室温平衡1h，加入冷冻I液后于5℃水浴平衡1.5 h，再加入冷冻II液平衡2 h后装入0.25 mL细管，液氮面上3 cm处平衡10 min投入液氮，解冻时采用37℃水浴30s，可获得最佳冷冻效果。采用该优化方案，猪冷冻精液解冻后的精子活力可达0.58。 相似文献
3.
哲罗鲑(Hucho taimen)是分布于额尔齐斯河和黑龙江等水域的冷水性珍稀鱼类,对其精子冷冻保存技术进行研究并建立精子冷冻库,对于哲罗鲑种质保存、人工繁殖发育、苗种培育及种群恢复都具有重要的意义.本研究利用生理盐水、葡萄糖、胎牛血清(fetal bovine serum,FBS)等配制了9种精子稀释液(MPRS,RS,Hanks,ELS,EG1,EG2,Stein,SS-2和D1S),对哲罗鲑精子在稀释液中活力、加入抗冻剂后活力和液氮中冷冻后活力进行检测,发现精子在以上稀释液中均具较高活力(73.00±2.65)％～(96.67±2.08)％,加入二甲基亚砜(dimethyl sulfoxide,DMSO)后活力下降,冷冻后仅在ELS和D15中具有极低活力(1.21±0.00)％和(1.1±0.00)％.进而利用ELS对甲醇、二甲基亚砜浓度进行筛选,发现精子在4％～12％甲醇中平衡20 min后活力仍为(79.00±6.52)％～(88.80±7.89)％,冷冻后仅在6％甲醇中具有相对较高活力(18.33± 10.61)％,但与鲜精活力相比显著下降(P＜0.05).精子在4％～12％ DMSO中平衡5 min后精子活力为(43.75± 11.09)％～(81.67±2.89)％,平衡20 min后精子活力极大下降(3.25±0.30)％～(45.00±5.77)％,冷冻后精子活力极低(1.00±0.00)％～(2.75±1.50)％(P＜0.05).以D15为基础液对葡萄糖浓度进行筛选,发现当葡萄糖浓度为30％(D30)时,冷冻后精子活力相对较高(17.80±2.59)％,但与鲜精相比活力显著下降(P＜0.05).另外对精子在D15、D30、Stein和ELs4种稀释液中短期保存的效果进行比较表明,Stein中精子存活在时间最长(45 h),D30次之(15h).综上所述,分别在ELS和D30中加入6％甲醇冷冻保存哲罗鲑精子具有一定活力,本研究结果为哲罗鲑精子大量冷冻保存和精子库建立等方面研究奠定了基础. 相似文献
4.
额尔齐斯河中蕴藏着丰富的冷水鱼类种质资源,江鳕(Lota lota)是其中之一,建立其精子冷冻保存技术和精子冷冻库,对种质资源保存具有重要意义。本研究利用无机盐、葡萄糖、蔗糖、胎牛血清等配制7种精子稀释液,以5∶1的比例稀释精液,激活后观察统计精子活力。结果显示,江鳕精子仅在SS、D15和CM中具有(13.33±2.89)%~(8.33±2.88)%活力。在此基础上配制22种梯度稀释液(SS-00~SS-10和D15-0~D15-7),以同样的方法稀释和观察精子活力。结果显示,SS-0(56.67±5.77)%、SS-2(63.33±5.77)%、SS-5(53.33±5.77)%和D15-7(63.33±5.77)%中精子活力较高,与鲜精活力(76.67±5.77)%接近(P0.05)。将二甲基亚砜(DMSO)和甲醇(MeOH)以3∶2比例配制成混合抗冻剂(DM),分别以5%~12%浓度加入到精子稀释液SS-5和D15-7中,以5∶1比例稀释精液,冷冻保存精子。结果显示,冷冻前,5%DM的SS-5和D15-7中精子活力(73.33±5.77)%与鲜精无显著差异(P0.05),当DM浓度升高到10%~12%时,精子活力接近于0;冷冻后,在DM浓度为6%的SS-5中,精子活力较高为(26.67±5.77)%(P0.05);在DM浓度为7%的D15-7中,精子活力较高为(26.67±5.77)%(P0.05)。本研究结果为额尔齐斯河江鳕精子冷冻保存提供了一定的理论和技术依据。 相似文献
5.
卢克焕 《广西农业生物科学》1982,(1)
本研究着重对公兔精液冷冻保存所用的防冻剂的选择及其用量、精液的稀释方法、平衡时间、降温速度及解冻温度等方面进行了一系列试验研究,最后进行输精试验,以验证试验效果。公兔采精用假阴道法,采精后立即检查精液一般性状,精子活率达0.7以上始供试验用。稀释液用 Tris 缓冲液为基础液,其成份为:Tris2.52克,葡萄糖1.05克,柠檬酸1.41克,蒸馏水84毫升,卵黄16毫升。稀释液中的防冻剂以 DMSO 和甘油并用效果较好。当 DMSO 最终浓度从0增至9%时,解冻后精子活率亦随之而提高;当甘油最终浓度为1.25、2.5和3.75%时,解冻后精子活率较好,三者间无明显差异(P>0.05),最终浓度为0和5%时,冻后精子活率则比以上三者有明显的下降(P<0.05)。解冻后精子顶体完好率则随着 DMSO 或甘油浓度的增加而下降。以冻后精子活率和顶体完好率这两个因素综合衡量,DMSO 和甘油的最终浓度分别以4.5%和1.25%效果较好。稀释精液时,采用两次稀释法的冷冻效果比一次稀释法的好。但两者差异不显著(P>0.05)。平衡时间以0.5—1小时效果较好,而2小时和3小时的效果较差。冷冻精液时,在-25—-50℃间的降温速度以每分钟下降3—8℃效果较好,如超过11℃,解冻后精子活率则有下降的趋势。颗粒冻精在40—50℃进行干解冻的效果比30℃和60℃的好。输精时,母兔先静注 HCG50—100单位以诱发排卵,输精量为0.5—0.8毫升。输精母兔230头,情期受胎率73.9%,平均每窝产仔数4.4头。 相似文献
6.
带精浆直接稀释法探讨猪精液低温(4℃)保存的可行性 总被引：1，自引：0，他引：1
为研制更为简易、高效的保存稀释液配方及操作程序,探讨4℃条件下稀释保存猪精液的可行性,本实验首次采用带精浆直接稀释法对猪精液的低温保存技术进行了研究.结果表明,在4℃条件下,用Ⅱ液(葡萄糖-卵黄-蔗糖)和Ⅳ液(葡萄糖-卵黄-乳糖)保存猪精液,精子的存活时间和活率分别为144 h、0.517和132 h、0.510,而Ⅰ液(葡萄糖-卵黄-蔗糖-柠檬酸钠)和Ⅲ液(葡萄糖-卵黄-乳糖-柠檬酸钠)精子活率保持0.5以上不到24 h.随着保存时间的延长,四种稀释液中保存的精子活率和顶体完整率均逐步下降,其中Ⅱ液和Ⅳ液均分别在48 h和24 h内下降较快,以后则呈缓慢下降趋势.综上所述,在低温条件下,采用Ⅱ液和Ⅳ液稀释保存猪精液,其精子活率能分别在144 h和132 h内保持0.5以上;所研制配方(Ⅱ液和Ⅳ液)组分简单、配制方便;采用带精浆直接稀释法低温保存猪精液可行,且操作程序简易. 相似文献
7.
为研制更为简易、高效的保存稀释液配方及操作程序,探讨4°C条件下稀释保存猪精液的可行性,本实验首次采用带精浆直接稀释法对猪精液的低温保存技术进行了研究。结果表明,在4°C条件下,用Ⅱ液(葡萄糖-卵黄-蔗糖)和Ⅳ液(葡萄糖-卵黄-乳糖)保存猪精液,精子的存活时间和活率分别为144 h、0.517和132 h、0.510,而Ⅰ液(葡萄糖-卵黄-蔗糖-柠檬酸钠)和Ⅲ液(葡萄糖-卵黄-乳糖-柠檬酸钠)精子活率保持0.5以上不到24 h。随着保存时间的延长,四种稀释液中保存的精子活率和顶体完整率均逐步下降,其中Ⅱ液和Ⅳ液均分别在48 h和24 h内下降较快,以后则呈缓慢下降趋势。综上所述,在低温条件下,采用Ⅱ液和Ⅳ液稀释保存猪精液,其精子活率能分别在144 h和132 h内保持0.5以上;所研制配方(Ⅱ液和Ⅳ液)组分简单、配制方便;采用带精浆直接稀释法低温保存猪精液可行,且操作程序简易。 相似文献
8.
钝吻黄盖鲽(Pseudopleuronectes yokohamae)是鲆鲽鱼类中重要的天然捕捞和养殖对象,但由于生态环境变化、人工捕捞过度等因素,导致其种质资源数量降低,进行精子冷冻保存技术研究,对其种质资源保存具有重要意义.本研究以性成熟的雄性钝吻黄盖鲽为实验材料,对精子稀释液种类及成分、抗冻剂种类、激活精子海水盐度和冷冻精液授精实验等进行筛选,实验数据利用SPSS 19.0软件进行单因素方差分析和Student-Newman-Keuls分析.结果表明,利用MFs-3(8 g/L NaCl+0.65 g/L KCl+15 g/L Glucose)稀释液分别与20％1,2丙二醇(1,2-propylene glycol,PG)和20％乙二醇(ethylene glycol,EG)抗冻剂作为精子抗冻保存液时,冷冻效果最好,其中PG组对应的精子活力、快速运动时间和寿命分别为(95.26±0.39)％、(46.00±1.00)s和(124.33±4.04)s,EG组则为(95.15±0.41)％、(45.67±0.58)s和(124.00±3.00)s.利用盐度为10～50的人工配制的海水激活解冻后的精子,发现当盐度为30时,精子活力高达(95.07±0.69)％,与对照组相比不存在显著性差异.将PG组和EG组冷冻保存的精子解冻后分别与其卵进行授精实验,PG组受精率和孵化率为(80.08±0.68)％和(77.44±1.76)％,EG组则为(80.17±0.45)％和(77.92±1.33)％,与鲜精相比无显著性差异.利用计算机辅助精子分析系统(computer-aided sperm analysis,CASA)测定MFs-3+20％ PG和MFs-3+20％ EG冷冻保存的钝吻黄盖鲽精子的各项运动参数,结果显示,二者的曲线运动速度(curvilinear velocity,VCL)、直线运动速度(straight line velocity,VSL)、平均鞭打频率(beat cross frequency,BCF)、运动的直线性(linearity,LIN)和运动的前向性(straightness,STR)的数值差异性不显著.本研究利用MFs-3+20％ PG和MFs-3+20％ EG成功冷冻了钝吻黄盖鲽的精子,为钝吻黄盖鲽精子冷冻保存技术、人工杂交育种繁殖及种质资源库的建立奠定了基础. 相似文献
9.
以家兔为试验动物,从精液解冻后的精子活率和复苏率2个指标,分别对3种稀释液(1:T ris0.25 m o l/L 蔗糖0.1 m o l/L 柠檬酸钠83 mm o l/L 卵黄10% 二甲基亚砜(DM SO)15%;2:配方1 葡萄糖0.47 mm o l/L;3:T ris 0.29 m o l/L 葡萄糖0.71 mm o l/L 柠檬酸钠67 mm o l/L 卵黄20% 甘油4%)、两种抗冻剂(甘油4%、DM SO 15%)、4°C下平衡时间(30、60、120、180 m in)三个方面进行了比较试验。公兔采精用假阴道法,采精后立即检查精子活率,达0.60以上的供试验用。结果表明:(1)3种稀释液中,用稀释液2稀释的精液,精子解冻后活率和复苏率显著高于稀释液1和稀释液3(精子活率:0.153 vs 0.103、0.109;复苏率:0.205 vs 0.137、0.147,P<0.01),而稀释液1和稀释液3之间差异不显著(P>0.05);(2)在稀释液2中,分别用4%甘油和15%DM SO作防冻剂,两者的精子解冻后活率之间(0.193 vs 0.261,P<0.01),复苏率之间(0.250 vs 0.340,P<0.01)差异极显著;(3)在4°C下平衡30、60、120、180 m in,结果发现,在30~60 m in之间,解冻后精子活率之间(0.261 vs 0.249),复苏率之间(0.340 vs 0.325)差异均不显著(P>0.05),但是随着平衡时间(120~180 m in)的增加,精子活率明显降低,精子几乎死亡。结论:采用稀释液2的配方,在4°C下平衡30~60 m in,可获得较好的兔精液冷冻保存效果。 相似文献
10.
针对星斑川鲽(Platichthys stellatus)养殖群体中雄性个体较雌性小,产精液量低,限制了人工繁殖育苗和杂交育种开展等问题,本研究以性成熟的星斑川鲽为实验材料,对精子稀释液成分、稀释液渗透压、激活海水适宜温度和冷冻精液的适宜稀释比例等进行筛选,并用冷冻精液和鲜精分别与星斑川鲽卵进行受精和孵化实验,实验结果利用SPSS软件中单因素方差分析法和Student-Newman-Keuls进行比较分析。结果表明,利用Ts-2(37.65 g/L蔗糖+10.01 g/L KHCO3+1.21 g/L Tris碱)和SFs-4(60 g/L葡萄糖+10.01 g/L KHCO3+1.21 g/L Tris碱)冷冻保存的精子活力最高,分别为(56.67±5.78)%和(66.67±5.77)%。利用计算机辅助精子分析(computer assisted sperm analysis,CASA)系统对星斑川鲽精子进行分析,测定了用Ts-2和SFs-4保存的冷冻精液中精子运动方式的各项参数。结果显示,测定的各项参数(曲线运动速度(curvilinear velocity,VCL)、直线运动速度(straight line velocity,VSL)、平均鞭打频率(beat cross frequency,BCF)、运动的直线性(linearity,LIN)和运动的前向性(straightness,STR)表明,利用SFs-4保存的精子各项运动参数明显高于Ts-2。利用蔗糖、KHCO3和Tris碱配制不同渗透压稀释液中,在渗透压为319.94 Pa时,精子活力最高,为(61.67±5.00)%,与鲜精相比无显著差异(P0.05)。利用3~18℃海水激活精子,结果显示,当激活海水温度在12℃时精子活力最高,与鲜精相比无显著性差异(P0.05),而其他温度下显著低于鲜精活力(P0.05)。将解冻后的精液稀释10~40倍,与星斑川鲽卵受精,结果显示,在稀释20倍后冷冻精液的受精率最高。本研究利用Ts-2+20%二甲基亚砜(dimethyl sulfoxide,DMSO)和SFs-4+20%DMSO对星斑川鲽精子成功进行冷冻保存,为星斑川鲽精子冷冻保存、种质库的建立和应用提供理论和技术依据。 相似文献
11.
Identification of oxidation products of (-)-epigallocatechin gallate and (-)-epigallocatechin with H(2)O(2) 总被引：4，自引：0，他引：4
Zhu N Huang TC Yu Y LaVoie EJ Yang CS Ho CT 《Journal of agricultural and food chemistry》2000,48(4):979-981
(-)-Epigallocatechin gallate (EGCG) and (-)-epigallocatechin (EGC) are two important antioxidants in tea. They also display some antitumor activities, and these activities are believed to be mainly due to their antioxidative effects. However, the specific mechanisms of antioxidant action of tea catechins remain unclear. In this study are isolated and identified two novel reaction products of EGCG and one product of EGC when they were reacted separately with H(2)O(2). These products are formed by the oxidation and decarboxylation of the A ring in the catechin molecule. This study provides unequivocal proof that the A ring of EGCG and EGC may also be an antioxidant site. This study also indicates an additional reaction pathway for the oxidation chemistry of tea catechins. 相似文献
12.
W R Dantzer J Hopper K Mullin S Hendrich P A Murphy 《Journal of agricultural and food chemistry》1999,47(10):4291-4296
14C-Fumonisin B(1) (FB(1)) was produced by Fusarium proliferatum M-5991 in modified Myro liquid medium and purified to >95% purity with a specific activity of 1.7 mCi/mmol. Nine male and nine female F344/N rats were each dosed by gavage with 0.69 micromol of (14)C-FB(1), (14)C-hydrolyzed FB(1), or (14)C-FB(1)-fructose/kg body weight. Urinary excretion of (14)C-FB(1) and (14)C-FB(1)-fructose was 0.5% and 4.4% of the total dose, respectively, and was similar between male and female rats. Urinary excretion of (14)C-hydrolyzed HFB(1) was significantly greater (P > 0.05) in female rats as compared with male rats (17.3% vs 12.8% of the total dose, respectively). There were no significant (P > 0.05) differences in biliary excretion of the three fumonisin compounds with a mean of 1. 4% of the dose excreted at 4 h after dosing. Lesser amounts continued to be excreted up to 9.25 h after dosing. Although biliary excretion of the (14)C-FB(1), (14)C-hydrolyzed FB(1), and (14)C-FB(1)-fructose was similar, increased urinary excretion of the (14)C-hydrolyzed FB(1) as compared to (14)C-FB(1) and (14)C-FB(1)-fructose indicated a greater absorption of the hydrolyzed form. 相似文献
13.
Roy A 《Journal of agricultural and food chemistry》1999,47(12):5209-5210
Terpinolene oxide, a monoterpene belonging to the p-menthane group, is easily derived from naturally abundant (R)-limonene. It was isomerized with montmorillonite clay catalyst to karahanaenone (2,2, 5-trimethylcyclohept-4-en-1-one) by ring enlargement. The enantiomers of the corresponding alcohol, karahanaenol (2,2, 5-trimethylcyclohept-4-en-1- ol), known for their individual organoleptic properties, were resolved through Pseudomonas cepacia lipase mediated enantiospecific alcoholysis of its acetate derivative. 相似文献
14.
15.
The presence of ethylenediamine-N-(o-hydroxyphenylacetic)-N'-(p-hydroxyphenylacetic) acid (o,p-EDDHA) as the second largest component in commercial EDDHA iron chelates has recently been demonstrated. Here is reported the speciation of o,p-EDDHA by the application of a novel methodology through the determination of the complexing capacity, protonation, and Ca(2+), Mg(2+), Cu(2+), and Fe(3+) stability constants. The pM values and species distribution in solution, hydroponic, and soil conditions were obtained. Due to the para position of one phenol group in o,p-EDDHA, the protonation constants and Ca and Mg stability constants have different values from those of o,o-EDDHA and p,p-EDDHA regioisomers. o,p-EDDHA/Fe(3+) stability constants are higher than those of EDTA/Fe(3+) but lower than those of o,o-EDDHA/Fe(3+). The sequence obtained for pFe is o,o-EDDHA/Fe(3+) >/= o,p-EDDHA/Fe(3+) > EDTA/Fe(3+). o,p-EDDHA/Fe(3+) can be used as an iron chelate in hydroponic conditions. Also, it can be used in soils with limited Cu availability. 相似文献
16.
S. R. Verma J. P. S. Arora J. S. Shankar R. Chand 《Water, air, and soil pollution》1987,36(3-4):247-257
The effect of protein oxovanadium(V) ion concentration and pH on the ratio of diffusion current (id/id0) was studied in vanadium(V) ovalbumin-S and denatured ovalbumin systems. In both the cases marked decrease in diffusion current was observed at the respective pH values, indicating that binding takes place with cationic groups of the proteins. The binding sites (n) were found to be pH dependent. The uniformity of logK and ΔG 0 value at all pH values indicated the involvement of same sites in interaction. Furthermore, the linear scatchard plots in both the systems supported the involvement of single class of independent sites in oxovanadium(V) anion interaction. The difference in binding sites (n) has been attributed to the folded structure of ovalbumin-S while unfolded one of denatured ovalbumin. 相似文献
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The efficiency of As(III) oxidation by MnO2, and retention of oxidation products varies with system pH. Maximum retention by hydrous Mn(IV) oxide occurs at pH < 5, declining at higher pH to about half total As at pH 10. The adsorption capacities of pyrolusite and cryptomelane at pH ～6.5 for As(V) species were 10 and 25 mmol kg?1, respectively. HMO surface saturation (～10 mmol kg?1) was reached with equilibrium As(V) levels of 5 to 8 × 10?6 M but this was supplemented at higher levels by an absorption process where uptake increased linearly with concentration (e.g., 68 mmol kg?1 with 2 × 10?5 M As(V)). Added As(III) was avidly oxidized and most product retained at pH 3. At higher pH increasing amounts of As(III) remained unoxidized due to initial reactions apparently blocking access to internal pores. Added Na+ reduced the amount of As retained by the HMO, with the phosphate salt having a significant effect. Extraction studies confirmed that most As could be released by exposure to reducing agents or chelating agents (EDTA). The environmental significance of the results has been considered. 相似文献