水牛发情期唾液生殖激素和结晶与卵泡发育的关系

本研究旨在探明水牛唾液生殖激素、唾液结晶和卵泡发育变化的规律。应用ELISA试剂盒测定水牛唾液和血清中的生殖激素，制作唾液结晶，并分析生殖激素和唾液结晶与卵泡变化的关系。运用唾液结晶法、直肠检查法、唾液结晶+B超法、直肠检查+B超法4种方法在生产中进行验证。结果表明，水牛发情当天唾液雌激素（E₂）的水平为233.51 pg/mL，达到一个峰值，随后开始缓慢降低。水牛唾液中孕激素（P₄）水平在发情前2 d达到16.17 ng/mL，发情当天降到8.91 ng/mL。促卵泡素（FSH）在水牛发情前2 d逐渐下降，从37.91 ng/mL降到34.64 ng/mL，但在发情2 d后逐渐升高，达到61.20 ng/mL。促黄体素（LH）在发情前1 d逐渐下降，从5.15 ng/mL降至发情当天4.12 ng/mL，但在发情1 d后升至5.77 ng/mL。B超监测卵巢卵泡从发情前2 d迅速生长，到发情当天达到20 mm大小，直到破裂排卵卵泡的大小变化不显著。排卵后形成黄体，黄体期一直维持到下次发情前4 d左右，期间在发情后14~17 d的黄体最大，与水牛唾液中LH分泌峰一致。唾液结晶+B超鉴定方法判定的发情率最高（86.67%），与其他几种方法比较差异显著（P<0.05）；妊娠诊断结果也表明，唾液结晶+B超鉴定方法判定的妊娠率（61.53%）显著高于直肠检查法、唾液结晶法和直肠检查+B超法3种方法鉴定的结果（P<0.05）。综上所述，卵巢上卵泡的发育与唾液中生殖激素的浓度显著相关，发情当天唾液结晶呈现典型的树枝状，唾液结晶+B超法的发情鉴定准确率最高。

Relationship Between Salivary Reproductive Hormones and Crystals with Follicular Development in Buffalo During Estrus

The purpose of this study was to investigate the changes of reproductive hormones,salivary crystals and follicular development in buffalo saliva.The reproductive hormones in salivary and serum of water buffalo were measured by ELISA kit,salivary crystals were prepared,and the relationship between reproductive hormones and salivary crystals and follicular changes was analyzed.Four methods,including salivary crystallization,rectal examination,salivary crystallization+B-ultrasound and rectal examination+B-ultrasound were used for verification in production.The results showed that the level of salivary estrogen(E 2)reached a peak of 233.51 pg/mL on the day of estrus,and then began to decrease slowly.The progesterone(P 4)level in buffalo saliva reached 16.17 ng/mL before 2 d of estrus,and decreased to 8.91 ng/mL on the estrus day.The level of follicle-stimulating hormone(FSH)gradually decreased from 37.91 ng/mL to 34.64 ng/mL before 2 d of estrus.However,it gradually increased to 61.20 ng/mL after 2 d of estrus.Luteinizing hormone(LH)gradually decreased from 5.15 ng/mL at 1 d before estrus to 4.12 ng/mL on the estrus day,but increased to 5.77 ng/mL at 1 d after estrus.B-ultrasonography results showed that ovarian follicles grew rapidly from 2 d before estrus to 20 mm on the day of estrus,and the size of follicles was insignificant until ovulation burst.After ovulation,the corpus luteum was formed and maintained until about 4 d before the next estrus,during which the corpus luteum was at its maximum at 14-17 d after estrus,which was consistent with the peak of LH secretion in buffalo saliva.The estrus rate determined by salivary crystallization+B-ultrasound was the highest(86.67%),which was significantly different from other methods(P<0.05).The results of pregnancy diagnosis showed that the pregnancy rate(61.53%)determined by salivary crystallization+B-ultrasound was significantly higher than that by rectal examination,salivary crystallography and rectal examination+B-ultrasound(P<0.05).To sum up,the development of follicles on ovaries was significantly related to the concentration of reproductive hormones in saliva.Saliva crystals on the day of estrus showed typical dendritic shape,and the accuracy of saliva crystals+B ultrasound method for estrus identification was the highest.