酶免疫分析法诊断家畜妊娠研究进展(续上期)

1．2水牛黄右军等（1994）采用EIA法测定了87头杂种水牛，对头摩拉水牛的脱胎乳孕酮含量，初步掌握了水牛发情、配种、妊娠、空怀和流产等不同生殖状态孕酮变化规律。将配种后第21d和27d次取奶样测定孕酮浓度均达到或超过1．7×10~(-9)g／mL者判为妊娠牛，前高后低者（不达者）为空怀牛，对64头配种水牛进行测定，1个月后直检复查，空怀确诊率为97.390（36／37），妊娠确诊率为88．9％（24／27）。与国内外EIA法诊断奶牛妊娠结果基本一致。[11]1．3羊李学斌等（1993）应用ELISA法测定绵羊血清中干扰体液免疫反应的因子，发现在配种后48…     

PAG ELISA早孕检测试剂盒对奶牛繁殖效率的影响

为了及时发现空怀牛并及时实施第2次配种,试验对配种后30~45 d未返情的奶牛采集血样,应用早孕因子(PAG)ELISA早孕检测试剂盒进行早期妊娠诊断。结果表明:应用PAG ELISA早孕检测试剂盒后,奶牛的平均空怀天数下降22.67 d,产犊间隔下降36.82 d。说明使用PAG ELISA早孕检测试剂盒进行奶牛早期妊娠诊断能够及时发现空怀牛并实施第2次配种,能够极显著降低奶牛空怀天数和产犊间隔(P0.01),提高奶牛繁殖效率。     

应用“猪妊娠诊断试剂”早期妊娠诊断

用“猪妊娠诊断试剂”对配种后１８天的１８５头母猪进行早期妊娠诊断试验，以产仔做为判断妊娠的依据，检出妊娠和空怀的准确率分别为９６．７％和１００％，任选其中５６头另用超声波诊断法进行早期妊娠诊断，检出妊娠和空怀的准确率分别为１００％和１８．２％，说明激素法诊断空怀的准确性优于超声波法。上外，激素法未引起母猪流产，并能诱导发情，而且操作简便，因此具有推广应用前景。     

猪早期妊娠诊断的几种方法

养猪生产中，在配种后尽快进行妊娠诊断，及早检出空怀母猪，对于缩短繁殖周期，提高母猪繁殖率和经济效益具有重要意义。 1 外部观察法 配种后经一个发情周期未返情，一般可以认为已经妊娠，准确率为80％。若在40天左右还没有发情，就可断定母猪已经妊娠。     

应用超声扫描仪对家兔早期妊娠诊断的研究

本试验采用超声扫描仪对家兔进行早期妊娠诊断,不仅可及时发观空怀、及时配种,而且还能及时发现死胎。试验结果表明,应用超声扫描仪诊断家兔妊娠的准确率与配种天数呈正相关,在配种后的第9天,准确率达100％。配种后第17天可观察到胎头、胎儿椎体、胎儿的心脏搏动和胎动。此法在诊断准确率上明显优于触诊法和超声多普勒法。     

奶山羊妊娠早期的B超诊断

自从1976年Lindahl首次将B超用于绵羊妊娠诊断以来，国内外学者先后在该领域进行了广泛的探索。他们将B超诊断绵羊早期妊娠的日期提前到配种后18～19d．使B超诊断绵羊妊娠的准确率达到或接近100％，其中对单胎的诊断准确率达84％～97％，对多胎的诊断准确率达95％～100%。     

应用B超探测子叶诊断绵羊早期妊娠的研究

应用B超对26只同期发情配种的美利奴母羊进行了妊娠诊断的研究。选择乳房两旁与膝皱襞之间无毛区域为扫描部位。每只母羊从配种后20天开始,每隔10天检查一次,并于第145天剖腹探查证实结果,以影像显示胎儿及其附属物为妊娠依据。结果,诊断配种后20、30和40天的早期妊娠准确率比较低,分别为18%(3/7)59%(10/17)和88%(15/17);而以扫描妊娠子叶为妊娠阳性诊断依据,相对时期内所得结果比较好,分别为64%(11/17)、94%(16/17)和100%(17/17)。使用两种不同的诊断妊娠指标,以上各阶段所得空怀准确率均为100%(9/9)。剖检妊娠25天的绵羊子宫,肉眼观察子叶的大小和形状已与空怀母羊的子宫肉阜有明显的区别。从而证明绵羊配种后50天以前的早期妊娠诊断可用B超扫描妊娠子叶为妊娠依据,且诊断速度快,效果好,但无法鉴别多胎妊娠。     

PAGmRNA在奶牛血液中表达的研究

本试验采用半定量RT-PCR的方法,检测和分析了牛PAG基因mRNA在妊娠牛及空怀牛血液中的表达规律。结果发现,PAG基因mRNA在妊娠21d的母牛及空怀牛血液中都有表达,而妊娠牛表达量显著高于空怀牛（P〈005）。这说明配种后21d奶牛血液中PAG基因的表达量可以作为奶牛早期妊娠诊断的依据。     

奶牛妊娠判断的方法

简便而有效的早期妊娠诊断,可以及时分辨妊娠牛和空怀牛。对已妊娠母牛要提高饲养管理水平。对空怀牛要查找出未妊娠的原因,在下个情期补配,这样可以减少空怀,缩短产犊间隔,提高繁殖率。1外部观察法配种后的奶牛一般在经过1个发情周期后,未出现发情的,可能已妊娠,而出现发情的,则未妊娠,     

应用B超技术进行绵羊早期妊娠诊断的研究

养羊业是畜牧业的重要组成部分。随着人工授精与胚胎移植等新技术的推广与应用,养羊业逐渐向集约化、产业化发展。绵羊早期妊娠诊断作为胚胎移植技术中的一项配套技术环节,能及时准确地判断并挑选出未妊母羊,对其进行发情控制处理,使空怀母羊及时接受重新配种或胚胎移植,妊娠母     

Effect of prostaglandin F2 alpha administration on early pregnancy in ewes

Two trials were conducted with ewes to determine the effects of prostaglandin F2 alpha (PGF) administration during the first week of gestation. In trial 1, ewes (n = 134) were checked for breeding activity once daily and half of them received 10 mg PGF im at either 0, 24, 48, 72, 96, 120 or 144 h after detection of a breeding mark. The other half served as uninjected controls. In trial 2, ewes (n = 153) were checked for breeding activity twice daily. Two-thirds of the ewes received 10 mg PGF at either 24, 36, 48, 60, 72, 84, 96, 108, 120 and 132 h following detection of a breeding mark. The other one-third of the ewes served as uninjected controls corresponding to treatment times of 24, 48, 72, 96 or 120 h. In trial 1, the percentage of ewes lambing as a result of first service decreased as time of administration of PGF increased. The first-service pregnancy rate was 87.5% for ewes given PGF at 0 h and 0% for ewes given PGF at 144 h. Fewer (P less than .05) ewes given PGF at 96, 120 or 144 h after first mating lambed than control ewes. Similarly in trial 2, fewer (P less than .05) ewes given PGF at 96, 108, 120 or 132 h after first mating lambed than did controls. The total number of ewes lambing as a result of the entire breeding season did not differ (P greater than .05) between treated and control ewes in trial 1 (88.2 vs 87.3%) or trial 2 (85.7 vs 83.3%).(ABSTRACT TRUNCATED AT 250 WORDS)     

Influence of reproductive state on pituitary-adrenal activity in the ewe.

Plasma corticosteroid concentrations are altered in pregnancy, during the reproductive cycle and by ovariectomy in many species. This study was designed to examine basal ACTH and cortisol in ewes of four different reproductive statuses: ovariectomized, nonpregnant cycling, nonpregnant noncycling, and pregnant. Blood samples were drawn every 4 hr for 48 hr from ewes during quiet, undisturbed conditions and analyzed for plasma ACTH, cortisol and progesterone concentrations. There were no significant changes in ACTH, cortisol or progesterone over time. Mean progesterone concentrations were significantly greater in the pregnant ewes than in all other ewes, and were greater in cycling ewes than noncycling or ovariectomized ewes. Mean ACTH was significantly greater in pregnant ewes than noncycling ewes, and mean cortisol was significantly greater in cycling ewes than in nonpregnant cycling or noncycling ewes. Ovariectomized ewes also had significantly greater mean cortisol concentrations than cycling ewes. The results demonstrate that there is an increase in basal ACTH and cortisol in ovine pregnancy.     

Effectiveness of intravaginal progesterone inserts and FSH for inducing synchronized estrus and increasing lambing rate in anestrous ewes

The objectives of this study were to determine whether a new progesterone (P4)-releasing intravaginal insert would induce fertile estrus and whether FSH combined with the insert would increase prolificacy in anestrous ewes introduced to rams. Ewes of mixed breeding on six farms were assigned to four randomized treatments: control (C), n = 73; 12 d P4 (polycapralactone [PCL] insert with 0.82 g P4), (P12), n = 73; 12 d P4 plus i.m. FSH (Folltropin, 55 mg NIH-FSH-P1 equivalent) in propylene glycol, 24 h before insert removal, (P12F), n = 71; and 5 d P4 plus FSH (P5F), n = 77. Growth and ovulation of follicles were observed ultrasonographically in 20 ewes at four farms (five/treatment) at insert removal and 36, 48, 72, and 96 h later. Intact rams (1:15 ewes in multiple-sire groups) were joined at insert removal, and raddle marks were observed every 12 h for 5 d. On d 26 to 30, rams were removed; ewes were examined for pregnancy then and 20 d later. Percentage of ewes marked by rams was greater in P4-treated (66 to 79%) than in C (12%; P < 0.01) ewes and in P5F (79%) than in P12F (66%; P < 0.05). Diameters of largest follicles at insert removal were greater (P < 0.05) in P4-treated (5.5 +/- 0.2) than in C ewes (4.8 +/- 0.2). Progesterone increased numbers of follicles > 3 mm (P < 0.01) or ovulated (P < 0.05; 2.6 +/- 0.6 vs 1.3 +/- 0.6 in C ewes) and FSH increased number of follicles > 3 mm (P < 0.05). In FSH-treated ewes, ovulation rate tended to be greater after treatment with P4 for 5 than for 12 d (P = 0.09, 3.3 +/- 0.6 and 2.2 +/- 0.4, respectively). More P4-treated than C ewes lambed (P < 0.01) to the first (38 to 45 vs 0%) or both (63 to 66 vs 41%) service periods. Prolificacy (first service) did not differ between FSH-treated ewes (P12F + P5F; 1.8 +/- 0.1) and ewes treated with P4 only (P12; 1.6 +/- 0.1). However, FSH increased prolificacy to first service (1.8 +/- 0.1) over prolificacy to second service (C ewes 1.5 +/- 0.1; P < 0.05, and all ewes 1.4 +/- 0.1; P < 0.01). Pregnancy retention did not differ among treatments but was greater (P < 0.01) in ewes that conceived at the first (90.9 +/- 3.7) than at the second (72.5 +/- 3.3) service period. In conclusion, a PCL insert in combination with ram introduction at insert removal was more effective than ram introduction alone to induce synchronized estrus and ovulation and to yield pregnancy after one or two service periods. Treatment with P4 for 5 d was as effective as for 12 d to induce fertile estrus in FSH-treated anestrous ewes.     

Pharmacokinetics of toltrazuril and its metabolites in pregnant and nonpregnant ewes and determination of their concentrations in milk,allantoic fluid,and newborn plasma

The objectives of this study were to determine the pharmacokinetics of toltrazuril and its metabolites in pregnant and nonpregnant ewes following a single oral dose and to determine the plasma concentrations of these compounds in milk, allantoic fluid, and newborn plasma. Eighteen healthy ewes were randomly divided into three groups (n = 6 each): pregnant ewes at 12–13 weeks of gestation (group A), nonpregnant ewes (group B), and pregnant ewes at 1–2 weeks before expected lambing date (group C). Ewes in all groups received a single oral dose of toltrazuril at 20 mg/kg body weight. In groups A and B, blood samples were collected at 1, 3, 5, 7, 9, 12, 15, 18 hr, every 6 hr to day 3, every 12 hr to day 7 and thereafter every 24 hr to day 14 post-toltrazuril administration. In group C, parturition was induced 24–36 hr after toltrazuril administration then milk, allantoic fluid, and newborn plasma samples were collected immediately after birth. Drug metabolites were assayed using ultra high-performance liquid chromatography–ultraviolet detection method (UHPLC-UV). The maximum concentration (C_max), area under the plasma concentration-time curve (AUC_0–t), AUC to 24 and 48 hr (AUC_0–24), and (AUC_0–48) were significantly higher in pregnant ewes. Longer apparent half-life (T_1/2), significantly higher apparent volume of distribution (Vd/F) and total clearance (Cl/F) were observed in nonpregnant ewes. The time to maximum plasma concentration (T_max), mean residence time (MRT) and elimination rate constant (K_el) were similar in both groups. The AUC_0–24 and AUC_0–48 were significantly higher in nonpregnant ewes. The AUC_0–t was significantly higher in pregnant ones. The ratio of plasma toltrazuril concentrations in ewes and toltrazuril concentrations in newborn lambs' plasma, allantoic fluid, and milk were 68%, 2.3%, and 5.3%, respectively. Results of this study showed that toltrazuril is well absorbed after a single oral dose in ewes with widespread distribution in different body tissues.     

TERMINATION OF EARLY PREGNANCY IN EWES BY USE OF A PROSTAGLANDIN ANALOGUE AND SUBSEQUENT FERTILITY

SUMMARY Of 87 Dorset ewes injected at 20 to 60 days of pregnancy with either 125 μg or 250 μg of the prostaglandin F^2α analogue, cloprostenol, 72 (83%) were detected in oestrus by teaser rams within 7 days. A total of 60 ewes mated with fertile rams 14 to 28 days after treatment and 36 of these (60%) subsequently lambed. Thirty-eight ewes mated with fertile rams 29 to 56 days after treatment and 30 of these (79%) subsequently lambed. The difference in fertility between the 2 periods was not significant.
Six additional ewes which did not respond to the cloprostenol lambed normally within 6 weeks. They were more than 100 days pregnant when treated.
In ewes which first exhibited oestrus by 7 days of treatment, plasma progesterone concentrations fell from near 4 ng/ml to 0.6 ng/ml within 48 h of treatment. In ewes not detected in oestrus progesterone concentrations did not decrease to similar low levels (1.4 ng/ml; t-test p < 0.005). Concentrations in the 6 ewes treated near 100 days of pregnancy dropped from 7.4 to 4.4 ng/ml over 48 h. Overall, the progesterone concentrations indicated that 92% of ewes treated at 20 to 60 days of pregnancy experienced rapid luteolysis in response to the cloprostenol.
There were no differences between the 2 doses of cloprostenol in the responses or subsequent fertility of the ewes.     

递增递减光照制对绍鸭产蛋性能及某些生理生化指标的影响

将6月1日出壳的绍鸭雌雏,在自然光照下培育至18周龄,随机分成两组(各50只)。组Ⅰ采用秋冬的自然光照(递减光照组)。组Ⅱ在10月5日自然光照基础上,每周递增15分钟,至31周龄的16小时(递增光照组)。观察两种光照处理对绍鸭产蛋性能、内分泌活动及矿物质代谢等的影响。结果,两种光照对绍鸭产蛋量和蛋品质量均无显著影响。但递增光照显著降低采食量(P<0.01),因而料蛋比显著降低(P<0.05)。两种光照处理对鸭血清孕酮、雌二醇、血清总钙含量无显著影响。据本试验表明,光照对鸡、鸭、鹅等家禽的繁殖季节虽有显著的调节作用,但对繁殖期家禽的生产和繁殖性能的影响不显著。     

Oxytocin increases pregnancy rates after fixed time artificial insemination in Kazak ewes

It is probable that reduced pregnancy rates in ewes after fixed time artificial insemination (FTAI) is attributable, in part, to the reduced number of normal spermatozoa that colonize the oviduct. Administration of oxytocin stimulates both cervical dilation and uterine/oviductal contractility. The hypothesis that oxytocin can enhance sperm transport into the uteri and the oviducts, and thereby increase pregnancy rates, was tested in the present study. Oestrus was synchronized in 199 multiparous Kazak ewes using intravaginal flurogestone-impregnated sponge. The sponge was left in the vagina for 12 days followed with an injection of 330 IU of eCG at sponge removal. Each ewe was intracervically inseminated twice at 50 hr and 62 hr after the removal of sponges using an insemination catheter containing 0.25 ml of diluted semen. Semen was collected from seven Texel rams and all the ejaculates were pooled and diluted in ultra-high temperature-treated commercial skimmed milk without (Control group, 0.05 ml of saline per mL milk, n = 144) or with oxytocin supplement (Oxytocin group, 0.5 U of oxytocin per ml milk, n = 55). Pregnancy status was determined by transabdominal ultrasound examination 45 days after insemination. Lambing performance was recorded at delivery. Significant differences were observed between the Oxytocin group and the Control group in terms of the pregnancy rate and the fecundity rate (85.5% and 92.7% versus 68.8% and 72.9%, respectively). In conclusion, low dose oxytocin supplementation of semen extender significantly increased pregnancy and fecundity rates in oestrus-synchronized Kazak ewes after FTAI.     

Basal and GnRH induced secretion of FSH and LH in ovariectomized ewes treated with bovine follicular fluid

Ovariectomized (OVX) ewes were injected with 5 ml of either bovine serum, charcoalextracted bovine follicular fluid (FF), or whole bovine FF. Five hours after this pretreatment, ewes on each pretreatment were injected with either 0, 1, or 5 μg of GnRH. Ewes that were pretreated with either type of FF had decreased concentrations of FSH regardless of dose of GnRH when compared to ewes pretreated with bovine serum. There was no effect of charcoal extraction. There were no differences among the pretreatment groups in LH response to GnRH. In a second experiment, OVX ewes were pretreated (4 ml) with either bovine serum or bovine FF 5 hr prior to GnRH or with bovine FF 42, 30 and 18 hr prior to GnRH. Ewes were injected with either 0 or 5 μg of GnRH. Pretreatment with FF for 5 or 42 hr prior to GnRH resulted in significantly decreased concentrations of FSH both at the time of GnRH treatment and during the following 2 hr. Concentrations of LH did not differ among pretreatment groups. In a third experiment, OVX ewes were pretreated with either bovine serum or bovine FF 30, 18 and 5 hr prior to GnRH. Ewes were injected with either 0, 5 or 50 μg of GnRH. Pretreatment with FF resulted in decreased concentrations of FSH both at the time of GnRH treatment and during the following 2 hr. Concentrations of LH were also decreased at the time of GnRH treatment.     

Pharmacokinetics of penicillin-G in ewes and cows in late pregnancy and in early lactation

Concentrations of penicillin-G in serum were determined after single intravenous doses of potassium penicillin-G (10 mg/kg body wt) to ewes and cows in late pregnancy and in early lactation. Penicillin-G in serum was analysed by a microbiological method and pharmacokinetic parameters were calculated by model-independent methods. Serum concentrations were lower in early lactation than in late pregnancy in both ewes and cows. The differences were statistically significant at most sampling times in ewes but only during the first hour in cows.
Weight corrected values of clearance and volume of distribution were significantly higher in early lactation than in late pregnancy in both ewes and cows. Mean residence time and elimination-half life were shorter in lactating than in pregnant ewes, however for the latter parameter the difference was not statistically significant. Neither of these parameters changed in cows.
It is concluded that in both ewes and cows pharmacokinetic parameters of penicillin-G are altered from late pregnancy to early lactation but that these changes have little practical impact and do not call for a revised dosage regime of the studied drug.     

Effect of reproductive methods and GnRH administration on long-term protocol in Santa Ines ewes

This study aimed to determine whether reproductive performance of ewes submitted to laparoscopic timed artificial insemination (TAI) would be similar to ante meridiem (AM)/post meridiem (PM) rule and assisted natural mating (NM), and whether GnRH may enhance the pregnancy rate in TAI. In experiment I, 191 non-lactating ewes were synchronized, then TAI was performed either 48 h after progesterone (P4) removal (TAI-48 h) or 12 h after estrus detection (AM/PM); moreover, some ewes were submitted to NM (NM) as control treatment. In experiment II, 247 non-lactating ewes were allocated in five treatments, a control (no-GnRH on protocol) and four treatments arranged in a factorial design 2 × 2. The factors were time and dose of GnRH: ewes that received either 10 μg (TAI-10 μg-36 h) or 25 μg of GnRH (TAI-25 μg-36 h) 36 h after P4 removal and ewes that received either 10 μg (TAI-10 μg-48 h) or 25 μg of GnRH (TAI-25 μg-48 h) at time of insemination, 48 h after P4 removal. In experiment I, pregnancy rate in TAI-48 h was lower (P = 0.03) than AM/PM and NM. Moreover, the probability of pregnancy in TAI-48 h was higher (P = 0.06) in ewes detected in estrus early. In experiment II, the use of GnRH in TAI protocols increased (P < 0.01) pregnancy rate at synchronization, and TAI-25 μ-48 h and TAI-10 μg-36 h treatments increased (P = 0.02) pregnancy rate compered to TAI-10 μg-48 h. We conclude that TAI decreased pregnancy rate compered to NM and AM/PM, which may be improved by GnRH use in TAI to synchronize ovulation.