Effects of GnRH treatment on superovulatory responses of dairy cows

Twenty-eight Holstein cows were randomly divided into two groups. Group received 50 mg of FSH-P intramuscularly in declining doses for five days. Cows in group II received the same treatment as those in Group I but were also administered 250 ug of GnRH on the day of expected estrus. Neither the mean number of corpora lutea palpated nor the mean number of embryos recovered nonsurgically was different (P0.05) between treatments. However, administration of GnRH resulted in a higher fertilization rate and recovery of more developing embryos.     

Temperature responses of dark respiration in relation to leaf sugar concentration

Changes in leaf sugar concentrations are a possible mechanism of short‐term adaptation to temperature changes, with natural fluctuations in sugar concentrations in the field expected to modify the heat sensitivity of respiration. We studied temperature‐response curves of leaf dark respiration in the temperate tree Populus tremula (L.) in relation to leaf sugar concentration (1) under natural conditions or (2) leaves with artificially enhanced sugar concentration. Temperature‐response curves were obtained by increasing the leaf temperature at a rate of 1°C min^?1. We demonstrate that respiration, similarly to chlorophyll fluorescence, has a break‐point at high temperature, where respiration starts to increase with a faster rate. The average break‐point temperature (T_RD) was 48.6 ± 0.7°C at natural sugar concentration. Pulse‐chase experiments with ¹⁴CO₂ demonstrated that substrates of respiration were derived mainly from the products of starch degradation. Starch degradation exhibited a similar temperature‐response curve as respiration with a break‐point at high temperatures. Acceleration of starch breakdown may be one of the reasons for the observed high‐temperature rise in respiration. We also demonstrate that enhanced leaf sugar concentrations or enhanced osmotic potential may protect leaf cells from heat stress, i.e. higher sugar concentrations significantly modify the temperature‐response curve of respiration, abolishing the fast increase of respiration. Sugars or enhanced osmotic potential may non‐specifically protect respiratory membranes or may block the high‐temperature increase in starch degradation and consumption in respiratory processes, thus eliminating the break‐points in temperature curves of respiration in sugar‐fed leaves.     

Follicular development and reproductive endocrinology during and after superovulation in heifers and mature cows displaying contrasting superovulatory responses

To understand the causes for poor response to superovulation in mature cows of high genetic potential, endocrine and follicular events during and after superovulation were compared in heifers (<2 yr old) yielding large numbers of embryos and cows (9 to 13 yr old) known to be poor embryo donors. Follicular development was monitored by daily ultrasonography. Blood samples were taken 2 to 3 times a day for the measurements of P4, E2, FSH and LH by RIA. Intensive blood collections at 15-min intervals for 6 h were also performed during preovulatory and luteal phases. The number of embryos produced in the heifers (15.2 +/- 2; mean +/- SEM) and the cows (0.6 +/- 0.4), was similar to the number of ovulatory follicles derived from ultrasonographic observations in the heifers (16.2 +/- 3.7), but not in the cows (7.8 +/- 2.8). Contrary to that observations in heifers, there was no increase in the number of 4- to 5-mm follicles in cows during superovulation. The number of larger follicles (>5 mm) increased during superovulation in both cattle groups, but it was significantly lower in cows than in heifers. During superovulation, the maximal E2 concentration was greater (P < 0.0001) in heifers than in cows. One cow showed delayed luteolysis during superovulation, while another had abnormally high FSH (>10 ng/ml) and LH (>3 ng/ml) concentrations following superovulation. All the cows had a postovulatory FSH rise which was not detected in the heifers. The results showed that attempts to improve superovulatory response in mature genetically valuable cows are hampered by a number of reproductive disorders that are not predictable from the study of the unstimulated cycle.     

Effects of supplementation of Tinospora cordifolia to crossbred cows peripartum

Guduchi (Tinospora cordifolia), a medicinal plant used in ayurveda, is well documented for its immunomodulatory properties. Since the crossbred periparturient cow is highly susceptible to various diseases that effectively reduces its reproductive performance postpartum we explored the possibility of enhancing the reproductive performance of crossbred cows by guduchi supplementation peripartum. A total of 15 pregnant Karan Fries cows were selected and divided into two groups: treatment group of 8 cows which were supplemented with guduchi at 60 g/day for 45 days prepartum and 120 g/day for 45 days postpartum and unsupplemented control group of 7 cows. Jugular blood samples were collected from all cows during the periparturient period for analysis of endocrine (progesterone, total estrogens and PGFM), immunological and hematological parameters. Incidence of retention of fetal membranes, endometritis, pyometra and calf mortality were higher in control group of cows in comparison to those recorded in treated group. The guduchi supplemented cows exhibited faster uterine involution (28 days vs. 42 days) and early commencement of cyclicity (37 days vs. 58 days; based on plasma progesterone profiles) in comparison to untreated control group of cows. Mean birth weight of calves from treatment group of cows was significantly higher than those from control group however no significant difference was observed in average daily body weight gain of calves in both the groups. A higher total leukocyte, lymphocyte, neutrophil count along with increased neutrophil lymphocyte ratio was recorded in guduchi supplemented cows in comparison to untreated cows although plasma total antioxidant activity was similar between the two groups. Prepartum plasma progesterone concentration was significantly lowered in the treated group however there was no significant change in peripartum plasma total estrogens and PGFM levels due to guduchi supplementation.     

The effect of PMSG-priming on subsequent superovulatory response in dairy cows

Superovulation was induced in 56 dairy cows to evaluate the effect of two different regimens using pregnant mare serum gonadotropin (PMSG). Thirty-two cows (controls) were superovulated between Days 9 and 12 of the estrous cycle with a single dose of PMSG (2 800 IU), while remaining 24 cows (PMSG-primed) received 200 IU of PMSG on Day 4 of the estrous cycle and subsequently a single dose of PMSG (2 800 IU) between Days 8 and 12. The cows in both treatments were each given 0,5 mg of cloprostenol at 48 h after the superovulatory PMSG treatment. They were then artifically inseminated twice, 48 h and 72 h later. Embryos were recovered at sloughter between Days 2 and 5 of the cycle and morphologically evaluated. The number of corpora lutea (CL) in the ovaries of the cows was recorded. The mean number of CL (7.2 vs 17.8) was significantly higher (P 0.01) for PMSG-primed cows. The percentage of recovered ova (60.5 vs 70.2 %) and good embryos (79.3 vs 70.7%) were not significantly different between groups. The percentage of fertilized ova (91.4 vs 83.8%) was significantly (P 0.025) greater for the controls. Results of the study indicate that PMSG-priming increased the ovulation rate in the cows superovulated with PMSG.     

Effect of lameness on estrous behavior in crossbred cows

Lameness, a common disease in crossbred dairy cows, badly affects their fertility. To elucidate, if estrous behavior, comprising of duration and expression of certain estrous behavioral sign(s) (EBS), is involved, an estrous expression score system was applied to objectively compare estrous behavior during 27 and 58 estrous periods, respectively, in 17 normal and 37 lame cows kept in a group of 200 cows under loose housing system in tropical India. The cows were detected in estrus and were closely watched daily for 30 min at 0630, 1030, 1430 and 1830 h from the beginning to the end of estrus. The duration of estrus was similar in the normal and the lame cows (18.7+/-1.2 h versus 17.2+/-0.9 h) and almost similar EBS were shown by two types of cows, albeit with varying frequency. Standing to be mounted, the highest ranking EBS was shown by equal proportions of the cows in both the groups (76.4% versus 73.0%). A lower proportion of lame cows showed head side mounting (35% versus 53%; P<0.05). The frequency of standing to be mounted events was lower in the lame than in the normal cows (2.4+/-0.4 versus 8.0+/-2.5; P<0.05). The lame cows earned non-significantly lower than normal estrous behavior score (1100+/-179 versus 1610+/-357). Further, the presence of more than one cow simultaneously in estrus enhanced the estrous behavior score in the normal (406.4+/-63.0 for one cow in estrus versus 793.0+/-146.0 for three cows simultaneously in estrus; P<0.05), but not in the lame cows. Lameness appeared to have suppressed the playful behavior during estrus probably owing to pain related stress. It is concluded that mildly lame cows appear to have normal physiological estrus. However, some of the EBS remain subdued which prone them to remain undetected for timely breeding.     

Correlations between ovarian follicular blood flow and superovulatory responses in ewes

The primary goal of this study was to employ ultrasonography to examine the ovaries of ewes undergoing superovulatory treatment for correlations between antral follicular blood flow and ovarian responses/embryo yields. Five Santa Inês ewes were subjected to a short- (Days 0–6, Group 1) and five to a long-term progesterone-based protocol (Days 0–12, Group 2) to synchronize estrus and ovulations after the superovulatory treatment. Porcine FSH (pFSH, 200 mg) was administered in 8 decreasing doses over 4 days, starting on Days 4 and 10 in Groups 1 and 2, respectively. After CIDR removal, all ewes were bred by a ram and embryos were recovered surgically 7 days later. Transrectal ovarian ultrasonography was performed the day before and on all 4 days of the superovulatory treatment. Both an arbitrary-scale [(0) non-detectable; (1) small; (2) moderate; (3) intense blood flow] and quantitative analysis of the blood flow area were used to assess the follicular blood flow in color Doppler images. There were no significant correlations between the arbitrary blood flow scores and superovulatory responses in the ewes of the present study. However, there was a positive correlation between the quantitative estimates of follicular blood flow on the final day of the superovulatory treatment, and the number (DA: r = 0.68, P < 0.05; DA/TA × 100%: r = 0.85, P < 0.05) and percentage (DA: r = 0.65, P < 0.05; DA/TA × 100%: r = 0.91, P < 0.001) of unfertilized eggs (DA: Doppler area, TA: total area of the largest ovarian cross section). This experiment presents a commercially practical tool for predicting superovulatory outcomes in ewes and evidence for the existence of follicular blood flow threshold that may impinge negatively on oocyte quality when surpassed during hormonal ovarian superstimulation.     

Induction of synchronized estrus and fertility in anestrous ZebuxTaurus crossbred cows

A major cause for reproductive failures in Zebu x Taurine crossbred cows is postpartum anestrus. Crossbred cows were diagnosed to be in postpartum anestrus by palpation per rectum of nonfunctional ovaries. To induce synchronized estrus, the cows were treated by placement of a progesterone releasing intravaginal device (PRID) for various number of days, together with a single administration of prostaglandin F(2) alpha (PGF, 5 mg intravulvosubmucos), estradiol-17beta (E2, 1 mg i.m.) or pregnant mare serum gonadotrophin (PMSG, 500 IU i.m.). Administration of PRID for 7 d, and of E2 and PMSG on Day 6, significantly improved the degree of behavioral estrus manifestation compared to single PRID for 12 d or PRID for 7 d, and of PGF on Day 6. However, the treatment combination of PRID and PMSG alone was associated with higher (P < 0.01) conception rate to 2 fixed time A.I. at induced estrus. The mean interval from treatment to conception was also shortest (P < 0.01) for this group. These results suggest that administration of PRID for 7 d, and of PMSG on Day 6 is highly effective in achieving synchronized behavioural estrus, a near normal CR to fixed time A.I. and a shorter interval from treatment to conception in anestrus Zebu x Taurine crossbred cows under Indian field conditions.     

Complex vertebral malformation in Holstein cows in Japan and its inheritance to crossbred F1 generation

Complex vertebral malformation (CVM) is an autosomal recessive inherited defect in the Holstein breed. It causes intra-uterine mortality leading to repeat breeding and involuntary culling of cows. This study was carried out to show the prevalence of CVM carriers among Holstein cows in five dairy farms in Hiroshima Prefecture, South Western Japan and whether the defect could be inherited to crossbred F1 generation, when Japanese Black semen was used to inseminate a Holstein cow. Two hundreds Holstein cows were used in the study. Blood samples were collected from all cows in a heparinized tubes and genomic DNA was extracted with a commercial kit. Apart of the bovine solute carrier family 35 member 3 (SLC35A3) gene around the suspected mutation (G to T) was amplified with an allele specific polymerase chain reaction (AS-PCR). For DNA sequencing, PCR products of 522 bp were obtained from the genomic DNA of the cows. Out of 200 examined cows, 26 cows (13.0%) were found to be CVM carriers. Wild type cows showed amplification with the G-specific primer pair but not with the T-specific primer pair, while carrier ones showed amplification of both the G- and T-specific primers, exhibited two DNA bands of 395 bp. Based on the clinical history of the cows, lowered reproductive performance was noticed in carrier ones. Out of four crossbred F1 calves (Holstein x Japanese Black), two were CVM carrier. Crossbred F1 could inherit the CVM defect when Japanese Black semen used to inseminate a carrier Holstein cow. In conclusion, the study reports the occurrence of CVM among Holstein cows in Hiroshima Prefecture, Japan; moreover, it describes the first cases of CVM in crossbred F1 generation.     

Follicular dynamics and superovulatory response in Holstein cows treated with FSH-P in different endocrine states

The effect of follicular and/or endocrine environments on superovulatory response was tested. Eighteen nonlactating Holstein cows were superovulated with 32 mg FSH-P given in decreasing doses at 12-h intervals plus two injections of prostaglandin F2-alpha (25 mg each) on the third day of treatment. Cows were assigned randomly to treatments: T1, superovulatory treatment initiated on estrous cycle Day 10.5; T2, CIDR (intravaginal device containing 1.9 g of progesterone) inserted from Days 3 to 9 and superovulation initiated on Day 6.5; T3, identical to T2 but Buserelin (GnRH agonist) was injected (8 mug, i.m.) on Day 3 at the time of CIDR insertion. Embryos were recovered on Day 7 after the superovulatory estrus. Cows were examined daily by ultrasonography and blood was collected for progesterone and estradiol determinations. Mean diameter of the dominant follicle (frequency of first-wave dominant follicle) at the beginning of FSH injections was 13.7 mm (4 6 ), 11.2 mm (6 6 ) and 8.7 mm (6 6 ) (P<0.01) for T1, T2 and T3, respectively. Following initiation of superovulation, follicles moved into larger follicle classes (Class I, <3 mm; Class II, 3 to 4 mm; Class III, 5 to 9 mm; Class IV >9 mm) earliest in T1 (P<0.01). Cumulative follicular diameter and plasma concentrations of estradiol at Day 4 of superovulation were higher (P<0.01) in T1 (200 mm, 82 pg/ml) compared with T2 (123 mm, 24 pg/ml) and T3 (130 mm, 18 pg/ml). Proportion of cows in estrus prior to 12 h vs 12 to 24 h differed (P<0.05) between groups (T1: 5 vs 1; T2: 2 vs 4; T3: 1 vs 5). Mean number of follicles on the last day of superovulation treatment, number of CL and number of embryos plus unfertilized ova recovered were 17.5, 12.2 and 13.3; 13.8, 10 and 8.2 (P<0.1) and 8.7, 4.5 and 2.3 (P<0.05) for T1, T2 and T3, respectively. The developmental stage of the dominant follicle was associated with not only the number of ovulations, but also the size and periestrous concentrations of plasma estradiol associated with the recruited follicles.     

The effect of estradiol valerate on follicular dynamics and superovulatory response in cows with Syncro-Mate-B implants

Two experiments were designed to evaluate the effect of estradiol valerate on follicular dynamics and superovulatory response in cows with Syncro-Mate-B (SMB)implants. In Experiment 1, 5 mg estradiol valerate (E(2)), injected at the same time as superstimulation treatments were initiated, resulted in fewer corpora lutea (CL), ova/embryos collected and fertilized ova (P<0.05) than if E(2) was administered with the SMB implant 7 days earlier. In Experiment 2, 31 beef cows and 26 Holstein cows were placed in one of four treatment groups. Group I (control) cows were superstimulated on Day 9 (estrus=Day 0). On Day 2, cows in Groups II, III, and IV received SMB and cows in Group III received E(2). On Day 9, cows in Group IV received E(2), and all cows were superstimulated with Folltropin. The number of CL did not differ (P>0.19) among groups. However, there were more follicles < 10 mm and fewer fertilized ova and transferable embryos (P<0.02) in Group IV cows. Ovarian ultrasonography revealed that the diameter of the largest follicle in Group III cows declined from Day 2 to Day 7 and subsequently increased until Day 13. In contrast, Groups I, II and IV were characterized by apparently linear growth between Days 2 and 13. Differences (P<0.05) were detected between Days 5 and 9. Mean diameter of the largest follicle was smaller for cows in Group III than for the remaining groups on Day 9. It was concluded that SMB did not adversely affect superovulatory response and that E(2) administration resulted in atresia of the antral follicles in the cows with SMB implants.     

Decreased superovulatory responses in heifers superovulated in the presence of a dominant follicle

Dairy heifers were superovulated in the presence (dominant group, N = 8) or absence (non-dominant group, N = 6) of a dominant follicle at the start of a a superovulatory treatment on Days 7-12 of the oestrous cycle (Day 0 = oestrus). Daily ultrasonographic observations of ovaries (recorded on videotape) starting on Day 3 were used to assess the presence or absence of a dominant follicle (diameter greater than 9 mm, in a growing phase or at a stable diameter for less than 4 days) and to monitor follicular development before and during treatment. The number of CL estimated by ultrasonography (7.1 +/- 1.8 vs 13.5 +/- 1.4) or by rectal palpation (6.9 +/- 2.0 vs 16.3 +/- 1.6) and mean progesterone concentrations (32.5 +/- 19 vs 80.7 +/- 16 ng/ml) after treatment were lower (P less than 0.01) in the dominant than in the non-dominant group. Based on number of CL, two populations of heifers were identified in the dominant group, i.e. those that had a high (dominant-high, N = 4; greater than 7 CL) or a low (dominant-low, N = 4; less than 7 CL) response to treatment. During treatment, the increases in number of follicles 7-10 mm and greater than 10 mm in diameter occurred sooner and were of higher magnitude in the non-dominant than in the dominant-high or dominant-low groups (P less than 0.01).(ABSTRACT TRUNCATED AT 250 WORDS)     

Cardiac responses in relation to heart size

There is a correlation between heart size and the propensity to develop and maintain fibrillation. Atrial and ventricular fibrillation is more easily induced and sustained in large than in small hearts. But other factors are at work as well, such as the ability to hibernate. The hibernator's heart has an adrenergic innervation which is different in distribution than that in nonhibernators with adrenergic nerves in the ventricles exclusively accompanying the blood vessels, thus leaving the myocardium proper without adrenergic innervation. This indicates that the risk of inhomogeneity of the electrophysiological parameters of the myocardial cells at a high sympathetic tone is less in the heart of a hibernator than in that of a nonhibernator.     

Flowering responses of Lemna perpusilla and L. gibba in relation to nitrate concentration in the culture medium

Flowering responses of Lemna perpusilla strain 6746, a short-dayplant, and L. gibba strain G3, a long-day plant, to nitrateconcentration in Hoagland's type medium with or without EDTA,were compared. Maximum flowering of L. perpusilla under SD occurredat higher nitrate concentrations than did colony proliferation.Even under CL, L. perpusilla grown at sub-optimal nitrate concentrationsfor colony proliferation, flowered irrespective of the presenceof EDTA which reduces flowering. Unlike L. perpusilla, L. gibba failed to flower under SD atany nitrate concentration whether or not EDTA was added. UnderCL, however, L. gibba flowered at almost any nitrate concentrationwith or without EDTA. Double optima for nitrate concentrationwas exhibited in the presence of EDTA; optimal concentrationfor colony proliferation came between the two optima for flowering. We concluded that the nitrogen level of the medium is importantin regulating flowering of duckweeds, and that the effect ofEDTA, if any, may primarily be on colony proliferation and onlysecondarily or antagonistically on flowering. ¹ Present address: Institute for Agricultural Research, TohokuUniversity, Sendai 980, Japan. (Received September 25, 1971; )     

Endocrine and superovulatory responses in heifers pretreated with FSH or bovine follicular fluid

This study examined the effects of altered serum FSH concentration on subsequent ovarian response to superovulation. Synchronized heifers were assigned randomly on Day 1 of the cycle (estrus = Day 0) to three pretreatment groups that consisted of 6-d of saline (7ml, s.c., b.i.d.; Group I), FSH-P (0.5 mg, i.m., b.i.d.; Group II) or charcoal-extracted bovine follicular fluid (BFF; 7 ml, s.c., b.i.d.; Group III) injections. Superovulation was initiated on Day 7 and consisted of FSH-P in decreasing dosages over 4 d (4,3,2,1 mg; i.m., b.i.d.), with cloprostenol (500 mug) on the morning of the third day. A second replicate with 14 heifers was conducted using the same protocol but twice the pretreatment dosage of FSH-P (1 mg) and BFF (14 ml). Endogenous plasma FSH decreased during BFF and FSH-P pretreatments compared to controls (P < 0.02). Endogenous FSH concentrations in both primed groups (II and III) were similar to control values (Group I) 12 h after the start of superovulation. Basal LH concentrations were not different between pretreatment groups. The interval from cloprostenol treatment to the preovulatory LH surge in Group III was 21.3 and 23.9 h longer (P < 0.0001) than it was in Groups I and II. The postovulation progesterone rise was delayed in Group III. The number of corpora lutea (CL) was lowest in the BFF-primed group (4.2 +/- 0.8) compared with the FSH-primed (7.4 +/- 1.3) and the control (12.0 +/- 1.8; P < 0.003) groups. In the FSH-primed group (0.68 +/- 0.06 cm(3)), CL volumes were larger than in the control group (0.45 +/- 0.03 cm(3)), whereas in the BFF-primed group (0.27 +/- 0.02 cm(3)) CL volumes were smaller compared with the control group (P < 0.0001). Mean FSH concentrations for 48 h preceding superovulation and the number of CL per cow were positively correlated (r = 0.55; P < 0.004; n = 26). We concluded that both FSH-P and BFF pretreatments decreased the superovulatory response of heifers to FSH-P. The mechanism for this would appear to be associated with reduced endogenous FSH prior to the start of superovulation.     

Hormone responses to exogenous GnRH pulses in post-partum dairy cows

Nine Friesian dairy cows were treated with 2.5 micrograms GnRH i.v. at 2-h intervals for 48 h commencing between Days 3 and 8 post partum. Hormone concentrations were measured in jugular venous plasma. An episodic pattern of LH release was induced in all animals and there was no significant change in amplitude during treatment. However, cows treated between Days 7 and 8 ('late') showed higher LH episode peaks than did those treated between Days 3 and 6 ('early'). Plasma FSH concentrations showed a less clear episodic pattern in response to GnRH injection. The mean height of FSH responses to GnRH tended to be higher in the 'early' group than in the 'late' group, as did mean FSH concentrations during the pretreatment sampling period. Although clear episodic changes were not observed, GnRH treatment induced a rapid sustained rise in plasma oestradiol-17 beta concentrations, indicating the responsiveness of ovarian follicles to gonadotrophin stimulation early in the post-partum period. There was no difference in oestradiol-17 beta concentrations between the 'early' and 'late' groups during the treatment period. Only one cow exhibited preovulatory-type LH, FSH and oestradiol-17 beta surges during the 96-h post-treatment sampling period. It is concluded that: (1) responsiveness to GnRH pulses increases significantly and FSH responsiveness tends to decrease with time post partum, (2) ovarian follicles are able to secrete oestradiol-17 beta in response to GnRH-induced LH and FSH release during the early post-partum period and there is no time-dependent change in responsiveness; and (3) the lack of preovulatory surges, except in one cow, may reflect a temporary defect in the positive-feedback mechanism by which high concentrations of oestradiol-17 beta induce preovulatory gonadotrophin release.