Study On Long-term Extenders For Room And Low Temperature Preservation And Cryopreservation Of Boar Semen

The forms of preserving boar semen have liquid storage (Room and Low temperaturepreservation) and cryopreservation. Room temperature preservation is currently used inproduction practice. The effective survival time of pig sperm preservation at roomtemperature was short, The technology for low temperature and freeze did not mature, itgreatly limits the popularization and application of artificial insemination techniques and pigbreeding, and had became the domestic and foreign scholars had been interested field. Inorder to prolong the effective survival time of pig sperm, to improve the efficiency ofbreeding boars, and promote the development of pig artificial insemination technology andthe production of pig. In the research, the diluents were focused for boar semen preservationat room temperature and low temperature, and on the basis of that, cryopreservation was alsostudied.At the normol temperature(17℃)preservation research, The effects of the base-extenders(TCGI-VI) and the two red cell storage solution (CPDA-1and MAP), the additives(SLX,SSX, adenine and SOD), additives concentration and application together of SLX and SSX onboar sperm effective and total survival time at room temperature were studied. The resultsshowed that:(1) TCG III and TCGI dilution were significantly better than MAP and CPDA-1solution (P<0.01) on sperm effective survival time, The effective survival time of TCG IIIand TCG I dilution were9.30±1.10d and8.30±2.49d separately, MAP and CPDA-1solution were only1.70±0.84d and0.50±0.34d separately. The comparation of six TCGdilution (I-VI) on the total survival time were TCGIII> TCGII> TCGI> TCGIV> TCG V>TCG VI, TCGIII> TCGII> TCG IV> TCG I> TCG V> TCG VI on the effective survivaltime.(2) In TCGIII dilution, SLX could obviously prolong the effective survival time of pigsperm(P<0.05), the effective survival time of SLX group was10.80±2.28d, that of controlgroup was8.20±1.95d, compared to control group, additive SSX, adenine or SOD could notsignificantly prolong the effective survival time (P>0.05). The effective and total survivaltime of different SLX concentration were3.0mg/mL group>1.5mg/mL group>0.75mg/mLgroup>0.4mg/mL group>6.0mg/mL group. The effective survival time and total survivaltime of3.0mg/mL SLX was longest, the effective survival time was12.9±3.58d, totalsurvival time was16.5±1.73d. From SSX concentration, the effective survival time of40 μg/mL and75μg/mL group were significantly longer than that of150μg/mL and300μg/mLgroup(P<0.05),and there were not significant difference between40μg/mL、75μg/mL、150μg/mL and300μg/mL group(P>0.05).75μg/mL>40μg/mL>150μg/mL>300μg/mL group on effective survival time. On total survival time,40μg/mL and75μg/mLgroup were significantly longer than300μg/mL group (P <0.05), there were not obviouslydifference between40μg/mL and75μg/mL group and between300μg/mL and150μg/mLgroup(P>0.05), and40μg/mL>75μg/mL>150μg/mL>300μg/mLgroup.(3) Although theaddition of SLX and SSX in comparision with simple addition of SLX or SSX could notsignificantly prolong the effective survival time and total survival time (P>0.05), but spermmotility of addition of SLX and SSX declined significantly slower at8-13days, and wassimilar with the simple addition of SLX or SSX before8days and13days later.(4) Theacrosome integrity of boar sperm preservated at room temperature at6-12days was alsostudied, The result showed that the acrosome integrity percentage of SLX (1.5mg/mL),SSX(75μg/mL) and SLX+SSX were all above80%.Based at the room temperature research, the effects of the base-extenders (TCG I-VI),the additives (SLX, SSX, adenine and SOD), adding dosage of SSX on boar sperm effectiveand total survival time at low temperature(4℃) were studied. The results showed that:(I)TCG III was the best in six TCG extenders, effective survival time was7.30±1.30d,exceedingly longer than that of TCG V and TCGVI (P<0.01), and significantly longer thanthat of TCGI and TCG IV(P<0.05), TCG III>TCG II> TCG I> TCG IV> TCGVI> TCGVon effective survival time. From the total survival time, TCGI and TCG II were better, its totalsurvival times were separately8.90±0.89d and8.90±1.81d, exceedingly longer than that ofTCG V and TCG VI (P<0.01) and significantly longer than that of TCG IV (P<0.05), TCG I> TCG II> TCG III>TCG IV>TCG VI>TCG V on total survival time.(II) Compared withthe control group (7.30±1.30d,8.5±1.58d), adding SLX, adenine or SOD in TCG IIIextender could not prolonged obviously the effective and total survival time (P>0.05), butonly adding SSX could exceedingly extended the effective survival time (12.90±1.14d) andtotal survival time(14.90±1.95d)(P <0.01). The effect of SSX on effective and total survivaltime of boar sperm at low temperature was related to adding dosage, with the increase of SSXconcentration, the effective survival time and total survival time would be prolonged. Theeffective survival time and total survival time of300μg/mL group were the longest,40μg/mL group were the shortest.In the case of TCG III as basic extender, the effect of glycerol concentration, thawingtemperature and time, and adding SSX on cryopreservation(liquid nitrogen) were also studied.The results showed that:(a) The survival rate of pig sperm after frozed-thawed at4%glycerolconcentration was0.49±0.05, exceedingly higher than those at2%glycerol concentration(P<0.01), significantly higher than at3%glycerol concentration (P<0.05).(b) at the thawingtemperature and time, the survival rate of pig sperm thawed at37℃30s and65℃5s wassignificantly higher than that at4℃60s(P<0.05), the survival rate of pig sperm thawed at37℃30s and65℃5s were0.43±0.06and0.45±0.06separately.(c) Compared with thecontrol group, adding SSX did not significantly improve the survival rate of porcine spermafter freezing-thawing (P>0.05), which showed that SSX could not improve the survival rateof boar semen after freezing-thawing.