Comparison of reproductive management programs for submission of Holstein heifers for first insemination with conventional or sexed semen based on expression of estrus, pregnancy outcomes, and cost per pregnancy.

Our objective was to evaluate reproductive management programs for submission of Holstein heifers for first insemination with conventional or sexed semen. In experiment 1, nulliparous Holstein heifers (n = 462) were submitted to a 5-d progesterone-releasing intravaginal device (PRID)-Synch protocol [d 0, GnRH + PRID; d 5, PGF2α - PRID; d 6, PGF2α; d 8, GnRH + TAI] and were randomly assigned for PRID removal on d 5 or 6 of the protocol followed by timed artificial insemination (TAI) with conventional semen. Delaying PRID removal decreased early expression of estrus before scheduled TAI (0.9 vs. 12.2%), and pregnancies per AI (P/AI) did not differ between treatments. In experiment 2, nulliparous Holstein heifers (n = 736) from 3 commercial farms were randomized within farm to 1 of 3 treatments for first AI with sexed semen: (1) CIDR5 [d -6, GnRH + controlled internal drug release (CIDR); d -1, PGF2α - CIDR; d 0, PGF2α; d 2, GnRH + TAI]; (2) CIDR6 (d -6, GnRH + CIDR; d -1, PGF2α; d 0, PGF2α - CIDR; d 2, GnRH + TAI); and (3) EDAI (PGF2α on d 0 followed by once-daily estrous detection and AI). Delaying CIDR removal decreased early expression of estrus before scheduled TAI (0.004 vs. 27.8%); however, CIDR5 heifers tended to have more P/AI at 35 (53 vs. 45 vs. 46%) and 64 (52 vs. 45 vs. 45%) days after AI than CIDR6 and EDAI heifers, respectively. Overall, CIDR5 and CIDR6 heifers had fewer days to first AI and pregnancy than EDAI heifers which resulted in less feed costs than EDAI heifers due to fewer days on feed until pregnancy. Despite greater hormonal treatment costs for CIDR5 heifers, costs per pregnancy were $16.66 less for CIDR5 than for EDAI heifers. In conclusion, delaying PRID removal by 24 h within a 5-d PRID-Synch protocol in experiment 1 suppressed early expression of estrus before TAI, and P/AI for heifers inseminated with conventional semen did not differ between treatments. By contrast, although delaying CIDR removal by 24 h within a 5-CIDR-Synch protocol in experiment 2 suppressed early expression of estrus before TAI, delaying CIDR removal by 24 h tended to decrease P/AI for heifers inseminated with sexed semen. Further, submission of heifers to a 5-d CIDR-Synch protocol for first AI tended to increase P/AI and decrease the cost per pregnancy compared with EDAI heifers.

Short communication: Effect of timing of induction of ovulation relative to timed artificial insemination using sexed semen on pregnancy outcomes in primiparous Holstein cows.

Our objective was to determine the effect of increasing the interval from induction of ovulation to timed artificial insemination (TAI) on fertility by decreasing the interval from TAI to ovulation using sexed semen within a synchronized breeding program. Our hypothesis was that induction of ovulation earlier relative to TAI would increase pregnancies per artificial insemination (P/AI). Primiparous Holstein cows from 3 commercial dairy farms in the United States were submitted to a Double-Ovsynch protocol for first service as follows: Pre-Ovsynch (GnRH; 7 d, PGF2α; 3 d, GnRH), followed 7 d later by Breeding-Ovsynch [GnRH (G1); 7 d, PGF2α; 24 h, PGF2α], followed by the last GnRH treatment (G2), which varied between treatments, and TAI. To vary the interval between G2 and TAI, cows were randomized to 2 treatments to receive G2 either 16 (G2-16; n = 373) or 24 (G2-24; n = 357) h before TAI, which was fixed at 48 h after the second PGF2α treatment of the Breeding-Ovsynch portion of the protocol. All cows were inseminated with sexed semen, and each herd used sires of their choosing, which were randomly allocated between treatments. Pregnancy diagnosis was conducted by herd veterinarians using transrectal ultrasonography. In disagreement with our hypothesis, G2-24 cows had fewer P/AI than G2-16 cows at 34 ± 3 d (44 vs. 50%) and 80 ± 17 d (41 vs. 48%) after TAI. Pregnancy loss (5 vs. 6%) and fetal sex ratio (92:8 vs. 90:10, female:male) did not differ between treatments for G2-16 and G2-24 cows, respectively. Thus, we reject our hypothesis and conclude that induction of ovulation earlier relative to TAI with sexed semen for first service after a Double-Ovsynch protocol decreased P/AI in primiparous Holstein cows.

Factors affecting the first service conception rate of cows in smallholder dairy farms in Bangladesh.

The successful outcome of an insemination is a combination of both male and female fertility-linked factors. We investigated the first service conception rate of cows at artificial insemination (AI) in the smallholder dairy farms in Bangladesh. Frozen straws were prepared from ejaculates of Bos indicus (n = 7) and Bos indicus × Bos taurus (n = 7) AI bulls. Fertility was determined from 6101 first services in cows that were performed by 18 technicians in four regions between April 2004 and March 2005. Pregnancy was diagnosed by rectal palpation between 60 and 90 days post-insemination. The Asian version of Artificial Insemination Database Application (AIDA ASIA) was used for bulls-, cows- and AI-related data recording, and later retrieved for analysis. The mean ± SD number of inseminations performed from individual bulls and their conception rates were 436.0 ± 21.6 and 50.7 ± 1.9%, respectively. Logistic regression demonstrated body condition scores (BCS), heat detection signs, months of AI and their interactions had greatest effects (odds ratios: 1.24-16.65, p < 0.04-0.001) on first service conception rate in cows. Fertility differed (p < 0.02-0.001) between the regions, previous calving months, months of AI, BCS, parity and heat detection signs of cows. Inseminations based on mounting activity (n = 2352), genital discharge (n = 3263) and restlessness and/or other signs (n = 486) yielded a conception rate of 53.6%, 48.8% and 50.1%, respectively (p < 0.05). Conception rate between technicians ranged between 43.4% and 58.6% (p < 0.05). The days interval from calving to first service (overall mean ± SD = 153.4 ± 80.6) had relationship (p < 0.001) with BCS, months of previous calving and parity of the cows. Fertility at AI in smallholder farms can be improved by training farmers on nutrition and reproductive management of the cows.

Disparate consequences of heat stress exposure during meiotic maturation: embryo development after chemical activation vs fertilization of bovine oocytes.

Consequences of heat stress exposure during the first 12 h of meiotic maturation differed depending on how and when bovine oocytes were activated. If heat-stressed oocytes underwent IVF at ~24 h, blastocyst development was less than for respective controls and similar to that obtained for nonheat-stressed oocytes undergoing IVF at 30 h (i.e. slightly aged). In contrast, if heat-stressed oocytes underwent chemical activation with ionomycin/6-dimethylaminopurine at 24 h, blastocyst development was not only higher than respective controls, but also equivalent to development obtained after activation of nonheat-stressed oocytes at 30 h. Developmental differences in chemically activated vs IVF-derived embryos were not related to fertilization failure or gross alterations in cytoskeletal components. Rather, ionomycin-induced calcium release and MAP kinase activity were less in heat-stressed oocytes. While underlying mechanisms are multifactorial, ability to obtain equivalent or higher development after parthenogenetic activation demonstrates that oocytes experiencing heat stress during the first 12 h of meiotic maturation have the necessary components to develop to the blastocyst stage, but fail to do so after fertilization.

Positive effect of FSH but not LH on early development of the dominant follicle in mares.

The effects of FSH, LH or both on follicular growth and intrafollicular free insulin-like growth factor (IGF)-1 and oestradiol were investigated in mares after the beginning of deviation (largest follicle >/= 20 mm; Hour 0). A single treatment with a gonadotropin-releasing hormone antagonist (acyline) was given at Hour 3 to suppress the concentrations of FSH and LH. Five groups (n = 5 mares per group) were evaluated in the present study: (1) control; (2) acyline treated; (3) acyline + recombinant equine (re) FSH treated; (4) acyline + reLH treated; and (5) combined acyline + reFSH + reLH treated. Beginning at Hour 3, reFSH and reLH were given at 6-h intervals in eight decreasing or increasing doses, respectively. The reFSH and reLH prevented the acyline-induced decreases in FSH and LH, respectively. Diameters and concentrations of intrafollicular free IGF-1 and oestradiol of the two largest follicles at Hour 48 did not differ significantly between the control and acyline + FSH groups, but were reduced (P < 0.05) similarly in the acyline and acyline + LH groups. The combination of reFSH and reLH was no more effective than reFSH alone. The results demonstrate a role for FSH but not LH in the growth of the largest follicle and intrafollicular concentrations of free IGF-1 and oestradiol during the 48 h after the beginning of deviation in mares.

Short communication: Validation of in vitro fertility genes in a Holstein bull population.

Previously, we constructed an in vitro fertilization system for the identification of genes affecting fertility traits in dairy cattle. The efficiency of this system has been demonstrated by the identification of several genes affecting fertilization rate and early embryonic survival. However, to employ these genetic markers in marker- and gene-assisted selection programs, there is a need to validate in vitro results in phenotypic data sets collected in vivo. Thus, the objective of this study was to validate, in a population of Holstein bulls, the fertility trait genes we previously identified in an in vitro system. Estimated relative conception rate (ERCR) data from 222 Holstein bulls were obtained from 5 different artificial insemination companies in the United States. Bulls were genotyped for the genes FGF2, POU1F1, PRL, PRLR, GH, GHR, STAT5A, OPN, and UTMP, and the data were analyzed for association with ERCR using a mixed effects sire model. A stepwise model selection procedure revealed evidence of association with ERCR for FGF2 and STAT5A polymorphisms. The in vivo validation suggests that these genes can be used in gene-assisted selection programs for reproductive performance in dairy cattle. The genotypes found to be associated with low bull fertility in this study have been reported to be associated with high milk composition in previous studies. These findings provide molecular evidence for the antagonistic relationship between milk production and fertility observed for many years in different breeds of dairy cattle.

Effect of suppression of FSH with a GnRH antagonist (acyline) before and during follicle deviation in the mare.

A GnRH antagonist (Acyline) was used to study the role of FSH in early development of a follicular wave in 61 mares. In Experiment 1, a single dose of 3 mg per mare, compared with 0 and 1 mg, suppressed both the FSH and follicle responses to exogenous GnRH. In Experiment 2, high concentrations of FSH were induced by two successive ablations of all follicles >/= 6 mm on days 10 and 13 (day 0 = ovulation). A single treatment with Acyline resulted in significantly greater suppression of plasma concentrations of FSH than a single treatment with charcoal-extracted follicular fluid (source of inhibin) or oestradiol. Suppression of FSH was not significantly different between the group treated with Acyline alone and a group treated with a combination of Acyline, inhibin and oestradiol. In Experiment 3, all follicles were ablated on day 10 to induce an FSH surge and a new follicular wave. Acyline treatment on day 10 resulted in an immediate decrease in FSH, without a significant effect on day of emergence of a new wave or growth of follicles from 7 to 11 mm on days 11-13. Treatment on day 15, a day before expected follicle deviation and after the peak of the wave-stimulating FSH surge, resulted in an immediate decrease in FSH and cessation of follicle growth. Results indicated that growth of follicles for about 2 days after wave emergence was independent of FSH. In contrast, during the decline in the wave-stimulating FSH surge and before follicle deviation, growth of follicles was dependent on FSH.

Quantification of bovine sperm separation by a swim-up method. Relationship to sperm motility, integrity of acrosomes, sperm migration in polyacrylamide gel and fertility.

The number of bovine spermatozoa separated in a swim-up procedure was quantified using an electronic cell counter. In an initial test of the swim-up procedure, non-frozen sperm samples with different ratios of live to dead cells were prepared and tested for the number of spermatozoa counted by the swim-up procedure. In ejaculates from six bulls, the number of spermatozoa swimming up was related to the number of live cells present (R2 = 0.97). Next, sperm quality of frozen-thawed semen immediately after thawing was measured at 37 C by swim-up sperm count, sperm motility, spermatozoa with an intact acrosome and migration in polyacrylamide gel and then compared with the fertility of the semen used for artificial insemination. Twenty-nine ejaculates of frozen-thawed semen from 11 bulls were evaluated. Correlations with fertility were highest on an ejaculate basis for motility (r = 0.41, P = 0.05) and for swim-up sperm count (r = 0.35, P = 0.06). On a bull basis, swim-up sperm count had the highest correlation with fertility (r = 0.59, P = 0.06). In a multiple regression model to predict male fertility that included all described measures of semen quality, a R2 value of 0.69 was obtained. This is the first report showing that the ability of spermatozoa to swim out of a more dense medium (whole milk-glycerol extender) into culture media is quantitatively related to in vivo fertility.

Measurement of bovine sperm nuclear shape using Fourier harmonic amplitudes.

An objective method for measuring bovine sperm nuclear shape was developed. Digital images of bovine sperm stained with propidium iodide were collected and Fourier functions used to describe the perimeters of individual sperm nuclei. Harmonic amplitudes from Fourier functions were first shown to be independent of sperm orientation during digitization. Sperm from 12 different bulls were used, and 6 harmonic amplitudes per sperm were found to adequately describe sperm nuclear shape. Based on harmonic amplitudes 0 to 5, cluster analysis was used to generate 20 different groups. Sperm within groups had similar morphologies and groups were distinguished by statistically unique shape characteristics. Harmonic amplitudes 0 to 5 can be used to distinguish previously reported abnormalities such as tapered, pyriform, macrocephalic, and microcephalic, as well as gradations in between. Furthermore, differences were detected among bull harmonic amplitude centroids (P < .05), indicating that bulls differ in mean sperm nuclear shape.

Relationship of bull fertility to sperm nuclear shape.

The relationship between sperm nuclear shape and bull fertility was determined. Two groups of bulls, 3 per group, were selected. Bulls differed in fertility based on lifetime nonreturn rates. Digital images of propidium iodide-stained sperm from each bull were collected and shape-evaluated by Fourier harmonic amplitudes 0 to 5. A discriminant function (P < .05) was constructed based on harmonic amplitudes and the 2 fertility groups. When individual sperm were classified as being of high or lower fertility, the percentage of each bull's sperm placed in the high-fertility group had a linear relationship (r = .89, P < .05) with fertility. To construct a plot of mean sperm shapes, a novel technique to automatically orient and identify the anterior tip of the sperm head was developed. The mean nuclear shape of high-fertility sperm was more elongated and tapered than those of lower fertility. A discriminant function (P < .05) was also constructed that separated the 6 bulls into 2 groups based only on the harmonic amplitudes or sperm nuclear shape. The bulls were correctly classified into the 2 fertility groups. A comparison of sperm chromatin structure analysis (SCSA) and harmonic amplitudes found that overall size variance, anterior roundness, and posterior taperedness of sperm nuclei were related to chromatin stability (P < .05). Some of the differences observed in sperm nuclear shape between the high- and lower-fertility bulls may be explained by varying levels of chromatin stability. However, sperm nuclear shape appears to contain additional information from chromatin stability alone. In this particular study, with 6 bulls, all with good chromatin quality, sperm nuclear shape was a better predictor of bull fertility.

Current concepts of cell-cycle regulation and its relationship to oocyte maturation, fertilization and embryo development.

Genetic and biochemical approaches have contributed to an explosion of literature on cell-cycle control. Regulation of the cell-cycle is controlled by a series of kinases and phosphatases. Key control points are during the G(1)-S and G(2)-M transitions. During both transitions, cyclins interact with a specific kinase to allow a cell to pass through that phase. The meiotic maturation of oocytes, fertilization and embryo development are all events influenced by cell-cycle regulation. Understanding cell-cycle control should provide new ways for gamete and embryo biologists to approach culture and development problems.

In vitro capacitation of bovine spermatozoa: role of intracellular calcium.

The development of successful methods of in vitro fertilization for bovine oocytes has advanced the bovine as a model for reproductive technology. The discovery of heparin as a capacitating agent has made it possible for investigators to have an inexpensive, readily available supply of bovine gametes for experimentation in reproductive biotechnologies such as gene transfer and cloning. The central event that mammalian sperm must undergo before being able to fertilize an oocyte is capacitation. Although we have methods which lead to efficient in vitro fertilization, we still lack understanding about the molecular mechanisms of capacitation. While numerous events occur during capacitation, it appears that regulation of intracellular Ca2+ (Ca(i)) is one of the most important. We found that the influx of Ca2+ into sperm during the first 2 hours of incubation is critical to heparin-induced capacitation. This is a period during capacitation when Ca(i) has not yet increased. We propose that during capacitation, the initial influx of Ca2+ into sperm is used to fill an intracellular Ca2+ store located in the acrosome. We found that thapsigargin, an inhibitor of an acrosomal Ca2+-ATPase, can stimulate capacitated sperm to acrosome react, trigger the opening of a store-operated calcium channel in the plasma membrane and has greater effects on capacitated sperm compared to noncapacitated sperm. An increase in intracellular Ca2+ was also detected in the anterior sperm head during capacitation, suggesting the loading of the acrosome with Ca2+. These observations may be important in the development of new methods for capacitation and understanding the death of sperm after cryopreservation.

Effect of bovine sperm separation by either swim-up or Percoll method on success of in vitro fertilization and early embryonic development.

The objectives of these experiments were to characterize separation of frozen-thawed bovine spermatozoa on a Percoll gradient and then to compare sperm separation by either a swim-up or Percoll gradient procedure for the ability of spermatozoa to fertilize oocytes in vitro. The Percoll gradient was a 45 and 90% discontinuous gradient. Initial experiments found that centrifugation of semen on the Percoll gradient for 15 min at 700 g was sufficient to obtain optimal recovery of motile spermatozoa. Most of the nonmotile spermatozoa were recovered at the interface of the 45 and 90% Percoll layers, while the motile spermatozoa were primarily in the sperm pellet at the bottom of the gradient. When frozen-thawed semen from each of 7 bulls was separated by swimup, a mean +/- SEM of 9% +/- 1 of the motile spermatozoa were recovered after the procedure. In contrast, more spermatozoa were recovered after Percoll gradient separation (P < 0.05), with 40% +/- 4 of the motile spermatozoa recovered. The effect of separation procedure on in vitro fertilization found swim-up separated spermatozoa penetrated a mean +/- SEM of 74% +/- 5 of the oocytes, while fewer oocytes were penetrated by Percoll separated spermatozoa at 52% +/- 8 (P < 0.05). There was no effect of the separation procedure on the rates of polyspermy as measured by sperm/penetrated ova, with a mean +/- SEM of 1.25 +/-.09 for swim-up separated spermatozoa and 1.14 +/-.07 for Percoll separated spermatozoa (P>0.05). A carry over of Percoll into the fertilization medium with the Percoll separated spermatozoa was found not the cause for the decreased penetration of oocytes by these spermatozoa. In 2 of 3 bulls tested, the decreased penetration of oocytes by Percoll separated spermatozoa could be overcome by increasing the sperm concentration during fertilization from 1 x 10(6) to 5 x 10(6)/ml. When development of embryos fertilized by either swim-up or Percoll separated spermatozoa was compared for the semen from 2 bulls, a difference in cleavage rate was found in favor of swim-up separated spermatozoa (P < 0.05), but there was no effect of separation procedure on development (Day 7) to the morula + blastocyst or blastocyst stage (P>0.05). The disadvantages of the Percoll procedure could easily be overcome and the procedure was faster and yielded a six-fold greater recovery of motile spermatozoa than the swim-up method.

Effect of heparin and chondroitin sulfate on the acrosome reaction and fertility of bovine sperm in vitro.

Glycosaminoglycans (GAGs) were reported to induce acrosome reactions (AR) in epididymal and ejaculated bovine sperm (4,5). The GAGs chondroitin sulfate A (CS-A) and heparin were tested on ejaculated bovine sperm for their ability to increase in vitro fertilization (IVF) frequencies. Regardless of treatment, a sperm-egg incubation time of 18 hr was sufficient to achieve maximal rates of fertilization. The IVF frequency of sperm incubated 6 hr with 10 mug/ml heparin (116 173 , 67%) was increased (P<0.05) above control levels (56 181 , 31%); however, 10 mug/ml CS-A (56 164 , 34%) was without effect (P>0.05). In contrast to previous reports, CS-A did not (P>0.05) induce AR in ejaculated (9.5-hr incubation) or epididymal sperm (22.5-hr incubation). Linear increases in fertilization frequency (40% to 81%; P=0.001) and AR (9% to 32%; P

Selection of follicles, preculture oocyte evaluation, and duration of culture for in vitro maturation of equine oocytes.

Equine oocytes (n = 537) were collected from slaughterhouse ovaries (n = 118 mares) by scraping the internal follicular wall. Preculture record was made of the appearance of oocyte investments (no cumulus, corona radiata only, compact cumulus, expanded cumulus), appearance of cytoplasm (homogeneous, condensed heterogeneous/fragmented), and nuclear maturation stages (germinal vesicle, germinal-vesicle breakdown, metaphase I, metaphase II, degenerated). There was no difference between follicles > 30 mm and follicles < or = 30 mm in the preculture frequency distribution among the 5 nuclear stages; 96% were at either the germinal vesicle or germinal-vesicle breakdown stages. Oocytes from follicles 5 to 30 mm were cultured in modified TCM-199 for 18, 24, 36 and 48 h. Postculture nuclear maturation classifications were immature (germinal vesicle, germinal-vesicle breakdown, and metaphase I), mature (metaphase II or secondary oocyte), and degenerated. The frequency distribution of oocytes among the 3 postculture maturation classifications changed (P < 0.05) at 18 h (15% mature oocytes), changed (P < 0.05) further at 24 h (55% mature oocytes), with no additional change for 36 or 48 h. The only preculture cytoplasm group that affected the postculture results was the heterogeneous/fragmentation group which had a high proportion of postculture degenerated oocytes (67%); however, only 4% of oocytes were in this group. Luteal status of the mare had an effect (P < 0.05) on the frequencies of the maturation classifications, but not enough to be useful in selecting oocytes. Consistency of the follicle and the type of oocyte investment did not alter significantly the maturation frequencies. The frequency of degenerated oocytes after culture was high under the following conditions: 1) diameter of the follicle from which the oocyte was selected was 5 to 10 mm (44% degenerated oocytes), 2) the largest follicle per pair of ovaries was < or = 10 mm (63%), and 3) the mare was pregnant (66%). These results were probably related to the reported high frequency of atretic follicles in the 5- to 10-mm population. In summary, oocytes from individual follicles < or = 10 mm or from follicles in which the largest follicle per mare was < or = 10 mm were the poorest candidates for in vitro maturation.

Bovine in vitro fertilization with frozen-thawed semen.

A procedure to obtain high and repeatable fertilization frequencies for bovine in vitro fertilization (IVF) with frozen-thawed sperm was developed. IVF frequency of in vitro matured oocytes was increased by a swimup sperm separation procedure (P=0.01) or treatment of sperm with the glycosaminoglycan heparin (P=0.0001), but the two factors did not interact (P=0.23). Heparin was the most important factor in increasing IVF frequencies. The fertilization frequency was not affected by the batch of oocytes used (P=0.38), but bull effects were present (P<0.05). Within a bull, the IVF system was highly repeatable and varied between trials no more than +/- 12% in fertilization frequency with an overall fertilization frequency of 299 379 (79%) on four trials over four bulls. In vivo matured oocytes fertilized in vitro were transferred to ewe or heifer oviducts. Morulae or blastocysts were recovered from ewes after four to five days, while conceptuses were present in the bovine after 25 days (diagnosed by ultrasound). Embryonic development from the IVF system either pre- or postimplantation was normal.

Female traits, ovary and follicle characteristics, and the conditional probability of normal oocyte development after superovulation of beef cows.

The proportion of transferable beef embryos obtained after superovulation, follicle aspiration, and in vitro maturation and fertilization has been small. To seek possible explanations, cows on different planes of nutrition were treated with exogenous gonadotropin and oocytes were isolated from their ovaries. The record for each oocyte included characteristics of the follicle, ovary, and cow from which it was obtained and the response to in vitro maturation, fertilization, and development. The sample was used to obtain estimates of the relationships among the variables. The logistic function with the probability of normal development as the dependent variable was the basic equation of the statistical model. When an explanatory variable was itself a result of the biological system, an equation explaining variation therein was added to the model. Had equations representing endogenous regressors not been added to the model a simple, single equation would have represented oocyte development response; given an oocyte at aspiration only one variable, cumulus quantity, was found to condition the probability of normal development directly. However, the complete model included four additional equations: 1) the probability that an oocyte was recovered at aspiration was conditional on the plane of nutritional treatment and progesterone concentration in follicular fluid; 2) cumulus quantity was conditional on the presence on a corpus luteum, follicle size, and progesterone concentration; 3) progesterone concentration was dependent on plane of nutrition; and 4) corpus luteum was conditional on plane of nutrition. The estimated model provided some insight into the complexity of oocyte development response and the role nutrition may play.

The foetal thyroid of the dromedary: its relation to body weight and dimensions and to the weight of the Dam's thyroid.

Growth and iodine content of the thyroid glands in relation to body weight, length [total (BCVRT), crown-rump (CR), and curved crown-rump (CVR)], chest circumference (CC), and age were investigated in 83 camel foetuses from 70 to 355 days of gestation (mean gestation length 390 days). Weight and iodine content of the dam's thyroid were also studied. Highly significant (P less than 0.01) correlations of 0.883, 0.797, 0.798, 0.792, and 0.813 were obtained between foetal thyroid weights on the one hand and body weight, BCVRT, CR, CVR, and CC on the other hand. A significant (P less than 0.05) correlation of 0.345 was obtained between weights of the foetal thyroid and dam's thyroid. The relative weight of the foetal thyroid to body weight was 0.045% at 70-99 days of gestation, but decreased to 0.028 (0.023 to 0.036%) in the subsequent stages. Iodine trapping by the foetal thyroid was first detected at 100-129 days of gestation. Total iodine then increased until 310 days, but decreased thereafter. The foetal thyroid has 4 to 25 times higher avidity to iodine accumulation than that of the dam.

Functional relationships among intrafollicular insulin-like growth factor 1, circulatory gonadotropins, and development of the dominant follicle in mares.

The functional relationships among intrafollicular free insulin-like growth factor 1 (IGF1), circulatory gonadotropins, and development of the dominant follicle were studied in 40 mares in two experiments. A GnRH antagonist (Acyline) was given i.m. at the expected beginning of follicular deviation (largest follicle or F1> or =20mm; Day 0) alone (Acyline group) or in combination with intrafollicular treatment of F1 with rhIGF1 (Acyline/IGF1 group). In Experiment 1, blood samples, follicular-fluid samples, and diameter of F1 were taken on Days 1 and 2. In Experiment 2, daily follicular diameter and blood samples were taken from Day 0 to ovulation. The GnRH antagonist induced a 50% decrease in circulatory FSH concentrations for 1 d and in LH for 2 d. In Experiment 1, control and Acyline/IGF1 groups had greater intrafollicular free IGF1 (P<0.05) and inhibin-A concentrations (P<0.08) than the Acyline group. The intrafollicular concentration of estradiol on Day 2 was greater (P<0.05) in the control group than in the Acyline and the Acyline/IGF1 groups. In Experiment 2, a decrease in diameter of F1 in the Acyline group was followed by a new follicular wave. All IGF-treated follicles grew and ovulated. Results indicated that the increase in intrafollicular free IGF1 observed in F1 in association with deviation is gonadotropin dependent. During the period of lesser gonadotropin concentrations from Acyline treatment, intrafollicular IGF1 stimulated follicular growth and inhibin concentrations, but not intrafollicular estradiol production.