Estrous Cycle Synchronization Research Articles

Synchronization of the bovine estrous cycle: Current and emerging opportunities

Controlling the bovine estrous cycle is a necessity if performing fixed-time artificial insemination or embryo transfer. Addition­ally, irrespective of the genetic objectives driving the decision to use reproductive technologies, the use of synchronization protocols often has beneficial effects on the overall reproduc­tive performance and commercial production output of a beef cattle operation. This proceedings paper provides a general review of the conceptual goals and challenges associated with synchronization of estrus, and directs readers to resources on current protocols and emerging opportunities in the field.

Follicular dynamics in dairy sheep within and out of the breeding season employing high or low progesterone concentration in estrous cycle synchronization

Resumo A dinâmica folicular em ovelhas leiteiras é pouco compreendida. Neste trabalho foi avaliado a dinâmica folicular de ovelhas leiteiras, dentro e fora da estação reprodutiva observando o efeito de duas concentrações de progesterona (60 e 120mg) utilizadas em pessários vaginais na pré-sincronização. O experimento foi realizado nos meses de abril/maio e outubro/novembro, utilizando 40 ovelhas multíparas leiteiras hígidas, predominantemente da raça Lacaune, que foram submetidas a exames ultrassonográficos diários por via transretal. Na estação reprodutiva foram avaliados o número de ovulações (1,35 vs 1,05), o diâmetro(mm) do folículo ovulatório (5,97 vs 5,05), a duração da fase lútea (11 vs 9,14 dias) e a duração do ciclo estral (16,83 vs 16,6 dias) dentro da estação em comparação com fora da estação reprodutiva respectivamente. Fora da estação as ovelhas não apresentaram corpo lúteo no momento da inserção do pessário vaginal, caracterizando anestro. Mesmo com menor duração da fase folicular, o folículo ovulatório apresentou maior diâmetro na estação reprodutiva, sugerindo maior viabilidade e qualidade do oócito ovulado. A aplicação de progesterona exógena amplia a fase lútea na estação reprodutiva (11 vs 9,14 dias). O anestro observado fora de época sugere a necessidade de maior concentração de progesterona nos protocolos reprodutivos. Desta forma, é possível adaptar protocolos reprodutivos para diferentes épocas, sugerindo o uso de pessários vaginais com maior concentração de progesterona (120 mg) fora da estação reprodutiva. Os dados obtidos permitiram caracterizar a dinâmica folicular em ovelhas leiteiras, demonstrando suas diferenças em cada estação a partir do uso de diferentes doses de progesterona na pré-sincronização

Human-relevant exposure to di-n-butyl phthalate tampers with the ovarian insulin-like growth factor 1 system and disrupts folliculogenesis in young adult mice.

Phthalates are compounds used in consumer and medical products worldwide. Phthalate exposure in women has been demonstrated by detection of phthalate metabolites in their urine and ovarian follicular fluid. High urinary phthalate burden has been associated with reduced ovarian reserve and oocyte retrieval in women undergoing assisted reproduction. Unfortunately, no mechanistic explanation for these associations is available. In short term in vivo and in vitro animal studies modeling human-relevant exposures to di-n-butyl phthalate (DBP), we have identified ovarian folliculogenesis as a target for phthalate exposures. In the present study, we investigated whether DBP exposure negatively influences insulin-like growth factor 1 (IGF1) signaling in the ovary and disrupts ovarian folliculogenesis. CD-1 female mice were exposed to corn oil (vehicle) or DBP (10 µg/kg/day, 100 µg/kg/day, or 1000 mg/kg/day) for 20-32 days. Ovaries were collected as animals reached the proestrus stage to achieve estrous cycle synchronization. Levels of mRNAs encoding IGF1 and 2 (Igf1 and Igf2), IGF1 receptor (Igf1r), and IGF-binding proteins 1-6 (Ifgbp1-6) were measured in whole ovary homogenates. Ovarian follicle counts and immunostaining for phosphorylated IGF1R protein (pIGF1R) were used to evaluate folliculogenesis and IGF1R activation, respectively. DBP exposure, at a realistic dose that some women may experience (100 µg/kg/day for 20-32 days), reduced ovarian Igf1 and Igf1r mRNA expression and reduced small ovarian follicle numbers and primary follicle pIGF1R positivity in DBP-treated mice. These findings reveal that DBP tampers with the ovarian IGF1 system and provide molecular insight into how phthalates could influence the ovarian reserve in females.

Effect of Progestin based Estrus Synchronization Protocol on Estrus Parameters, Hormonal Profile and Vaginal Cytology in Surti Goats

This investigation was conducted to study the effect of progestin and prostaglandin-based estrous synchronization protocol on estrus parameters, hormonal profile and vaginal cytology in Surti goats. Ten non-pregnant Surti goats were selected and subjected to estrus synchronization with intra-vaginal progesterone sponges (60 mg Medroxyprogesterone acetate, MAP) for 11 days followed by PGF2α injection (125 μgCloprostenol). Whole blood was collected on day 0 (before treatment), day 5, 11 (after treatment), at proestrus, estrus and diestrus for analyzing progesterone as well as estrogen hormone concentration. The vaginal exfoliative cytology was studied at proestrus, estrus and diestrus. Estrus parameters such as induction rate was 100%, time of onset of estrus from PG injection was 29.70 ± 1.06 h and duration was 30.74 ± 0.19 h. Significantly higher population of parabasal cells and superficial cells were observed during proestrus, intermediate cells in diestrus and cornified cells during estrus. At estrus, level of progesterone was lowest and estrogen was highest. Combination of progestin and prostaglandin-based protocol is successful in synchronization of estrous cycle with 100% estrus induction rate in Surti goats. Further in addition to other signs, vaginal exfoliative cytology can also be used for determining different phases of estrous cycle.

Attainment of Sexual Maturity and Gonadotropin Priming in Gilts Determine Follicular Development, Endocrine Milieu and Response to Ovulatory Triggers.

The routine procedure of estrous cycle synchronization in pigs allows for the use of gonadotropins to stimulate ovarian activity. The applied protocols of eCG and hFSH priming similarly affected development of ovarian follicles in two classes 3–6 mm and >6 mm of diameter, however, the number of small follicles (<3 mm) was 2-fold higher in hFSH- than in eCG-primed prepubertal gilts. The attainment of sexual maturity increased concentration of estradiol, testosterone and androstenedione in the follicular fluid of hFSH/eCG-primed gilts, however, prostaglandin E2 and F2α metabolite increased in mature hFSH- and eCG-primed gilts, respectively. The maturity increased mRNA and/or protein expression of key steroidogenic enzymes, prostaglandin synthases or luteinizing hormone receptors in follicular walls. Both hormonal primers played a moderate role in affecting expression of steroidogenic enzymes in follicular walls. In vitro studies showed higher estradiol production in r-hLH (p = 0.04)- and r-hCG (p = 0.049)-stimulated follicular walls of mature gilts than in prepubertal hFSH-primed gilts. Both ovulatory triggers decreased the abundance of LHCG/FSH mRNA receptors in follicular walls, which mimic downregulation of these receptors by a preovulatory LH surge, confirmed in vivo. These data revealed the importance of sexual maturity in the protection of the estrogenic environment, and the selective, moderate role of eCG and FSH in the activation of steroidogenic enzymes in preovulatory follicles.

Influence of hormonal synchronization protocols in heifers-recipients on their pregnancy rate with the use of frozen-thawed embryos

At the moment, there are a great number of protocols enabling to prepare recipients for embryo transfer. However, because of their effectiveness variety on cattle breeding farms the issue of searching of practical schemes that allow successful reproductive cycle synchronization in recipients with the achievement of their highest pregnancy rate becomes relevant. Based on the information above, the aim of the study was to determine the most effective estrous cycle synchronization protocol in recipient heifers before embryo transfer which provide the high level of their pregnancy rate with the use of frozen-thawed embryos. The research was carried out on the beef cattle in Samara region of Russia. In the course of the study, the heifers (n=107) were selected as the potential recipients and then divided into three groups depending on the applied protocol for the estrous cycle synchronization. All the recipients were transferred with frozen-thawed embryos obtained in vivo from Aberdeen Angus donor cows. The doses of the drug products indicated below were administrated per one animal. During the study, it was found that in group I where we used a protocol included a double administration of 2 ml of the drug "Oestrophan" (Czech Republic) with an interval of 11 days estrous cycles were synchronized in 72.73±0.80 percent of animals. The pregnancy rate in group I was 29.17±0.97 percent. In group II where we applied a protocol included a double administration of "Surfagon" (Russia) at a dose of 10 ml and Oestrophan with an interval of 11 days estrous cycles were synchronized in 50.0±0.86 percent of animals. The pregnancy rate in that group was 61.11±1.22 percent. In group III that included the application of intravaginal progesterone implant CIDR with a simultaneous administration of Surfagon at a dose of 2 ml and at the time of CIDR extraction the administration of 2 ml of Oestrophan, estrous cycles were synchronized in 63.16±0.80 percent of animals. The pregnancy rate in that group was 66.67±1.00 percent. Thus, the group III was characterized by the highest pregnancy rate that was achieved by the application of the protocol included intravaginal progesterone implant CIDR in combination with both medical products Surfagon and Oestrophan.

Coal Dust Exposures Change the Spiral Artery Remodeling and Natural Killer Cells Counts in the Uterus of Pregnant Rats

BACKGROUND: Indonesia has numerous coal resources, but the effects of environmental pollution by coal dust to human health, especially the reproductive system, are still less investigated. Chronic coal exposures during pregnancy might cause systemic and uterine inflammation that induces disturbances of spiral artery remodeling. AIM: This research was conducted to analyze the effect of coal dust exposures to uterine spiral artery remodeling and natural killer (NK) cell counts. METHODS: There were 42 female adult Rattus norvegicus rats which had been simultaneously mated after synchronization of estrous cycle used as animal subjects. The rats then divided into seven groups, those were K0 (without exposure); K1-1, K1-2, K1-3; and K2-1, K2-2, K2-3 which were exposed to 6.25 mg/m3, 12.5 mg/m3, and 25 mg/m3 dose for 1 h (K1) and 2 h (K2) per day, respectively. The rats were exposed by a dust exposure machine (NKBS-1-2010- 0.5) every morning starting from the day-1 to day-19 post-mating and were sacrificed on day-19 afternoon. All uterus lobes of each pregnant rat were taken and histologically processed by HE staining. Five histological slides were randomly taken as samples representing each pregnant rat and were examined for ten visual fields per slide for the measurements of diameter and wall thickness of spiral arteries. NK cells were observed immunobiological. Data analyses used Kruskal–Wallis. RESULTS: Result showed that there were significant differences of diameters and wall thickness of spiral arteries and uterine NK cells in all exposed groups compared to control (P &lt; 0.05). Diameter and wall thickness of spiral arteries were thicker in coal dust exposure than normal pregnant rats. NK cell number was more in coal dust exposure than normal pregnant. CONCLUSION: Coal dust exposures caused the thickening of wall and narrowing of lumen of spiral arteries and NK cell counts that might influence pregnancy.

Leptin Signaling in the Ovary of Diet-Induced Obese Mice Regulates Activation of NOD-Like Receptor Protein 3 Inflammasome.

Obesity leads to ovarian dysfunction and the establishment of local leptin resistance. The aim of our study was to characterize the levels of NOD-like receptor protein 3 (NLRP3) inflammasome activation in ovaries and liver of mice during obesity progression. Furthermore, we tested the putative role of leptin on NLRP3 regulation in those organs. C57BL/6J female mice were treated with equine chorionic gonadotropin (eCG) or human chorionic gonadotropin (hCG) for estrous cycle synchronization and ovary collection. In diet-induced obesity (DIO) protocol, mice were fed chow diet (CD) or high-fat diet (HFD) for 4 or 16 weeks, whereas in the hyperleptinemic model (LEPT), mice were injected with leptin for 16 days (16 L) or saline (16 C). Finally, the genetic obese leptin-deficient ob/ob (+/? and −/−) mice were fed CD for 4 week. Either ovaries and liver were collected, as well as cumulus cells (CCs) after superovulation from DIO and LEPT. The estrus cycle synchronization protocol showed increased protein levels of NLRP3 and interleukin (IL)-18 in diestrus, with this stage used for further sample collections. In DIO, protein expression of NLRP3 inflammasome components was increased in 4 week HFD, but decreased in 16 week HFD. Moreover, NLRP3 and IL-1β were upregulated in 16 L and downregulated in ob/ob. Transcriptome analysis of CC showed common genes between LEPT and 4 week HFD modulating NLRP3 inflammasome. Liver analysis showed NLRP3 protein upregulation after 16 week HFD in DIO, but also its downregulation in ob/ob−/−. We showed the link between leptin signaling and NLRP3 inflammasome activation in the ovary throughout obesity progression in mice, elucidating the molecular mechanisms underpinning ovarian failure in maternal obesity.

Endocrine and molecular milieus of ovarian follicles are diversely affected by human chorionic gonadotropin and gonadotropin-releasing hormone in prepubertal and mature gilts

Different strategies are used to meet optimal reproductive performance or manage reproductive health. Although exogenous human chorionic gonadotropin (hCG) and gonadotropin-releasing hormone (GnRH) agonists (A) are commonly used to trigger ovulation in estrous cycle synchronization, little is known about their effect on the ovarian follicle. Here, we explored whether hCG- and GnRH-A-induced native luteinizing hormone (LH) can affect the endocrine and molecular milieus of ovarian preovulatory follicles in pigs at different stages of sexual development. We collected ovaries 30 h after hCG/GnRH-A administration from altrenogest and pregnant mare serum gonadotropin (eCG)-primed prepubertal and sexually mature gilts. Several endocrine and molecular alternations were indicated, including broad hormonal trigger-induced changes in follicular fluid steroid hormones and prostaglandin levels. However, sexual maturity affected only estradiol levels. Trigger- and/or maturity-dependent changes in the abundance of hormone receptors (FSHR and LHCGR) and proteins associated with lipid metabolism and steroidogenesis (e.g., STAR, HSD3B1, and CYP11A1), prostaglandin synthesis (PTGS2 and PTGFS), extracellular matrix remodeling (MMP1 and TIMP1), protein folding (HSPs), molecular transport (TF), and cell function and survival (e.g., VIM) were observed. These data revealed different endocrine properties of exogenous and endogenous gonadotropins, with a potent progestational/androgenic role of hCG and estrogenic/pro-developmental function of LH.

The effects of the anti-Müllerian hormone on folliculogenesis in rats: light and electron microscopic evaluation

ABSTRACT In this study, we evaluated the effects of anti-Müllerian hormone on follicle development and oocyte quality with light and electron microscopy. Twenty-four adult female rats were divided into four groups. After estrous cycle synchronization, on the first day, control group rats were injected with 0.5 ml saline, 2nd, 3rd, and 4th groups were injected 1 µgr, 2 µgr, and 5 µgr anti-Müllerian hormone, respectively. On the third day, intracardiac blood samples were taken for follicle-stimulating hormone, luteinizing hormone, estradiol, and progesterone serum level measurements. Ovaries were obtained for light and electron microscopic examinations. Secondary (antral) follicles were decreased while atretic follicles were increased in number parallel with an increased dose of anti-Müllerian hormone injection. Atresia of the follicles was demonstrated with apoptosis of granulosa cells characterized by apoptotic bodies and with paraptosis characterized by the vacuole formation in the cytoplasm, enlargement of granular endoplasmic reticulum cisternae and perinuclear cisternae in granulosa cells. Premature luteinization characterized by increased lipid droplets, mitochondria with tubular cristae, and smooth-surfaced endoplasmic reticulum in the cytoplasm of granulosa cells were detected in some growing follicles. In the anti-Müllerian hormone injected experimental groups, cystic follicles characterized by a large antrum, attenuated granulosa cell layer, and flattened granulosa cells that face the antrum were observed. Corpus luteum and stroma were similar in all groups. It was concluded that increasing doses of anti-Müllerian hormone caused increased atresia in developing follicles, premature luteinization of granulosa cells in some follicles, and cystic follicle formation in the further developing follicles.

The Effectiveness of Pharmacological Synchronization of the Estrous Cycle in Hinds (Cervus elaphus L.): A Pilot Field Trial.

Simple SummaryReproductive biotechnics of Cevidae species may increase valuable traits for the agricultural sector, and improve genetic diversity in relatively small farmed populations. The first step of biotechnics in terms of suitability for embryo transfer or insemination is preparation of females for proper phase of the estrous cycle. Based on available synchronization protocols applicated in red deer and domestic ruminant females we estimated the protocol for hinds in view of future routine use. Progestin analogues in the form of an insert (CIDR) or a sponge (Chronogest) and injection of equine chorionic gonadotropin (eCG; Folligon) were used on the appropriate day for inducing the estrus. The effect of synchronization was checked by measurement of progesterone (P4) and 17-beta estradiol (E2) concentration by RIA, pregnancy-associated glycoproteins (PAG)s concentration by EIA and insemination effectiveness—pregnancy rate. The better method of the estrous cycle synchronization found down to be progestin sponge and eCG based protocol in hinds considering hormone responsiveness and pregnancy rate.The aim was to estimate the effective pharmacological method of the estrous cycle synchronization by checking the effects of synchronization by measurement of progesterone (P4) and 17-beta estradiol (E2) concentration by RIA and artificial insemination. The experiment was performed at the red deer farm in Rudzie (North-East Poland; 3 year’s old). The herd (N = 14) was kept away from bulls and was divided in two groups of seven animals. In the Group I, CIDR insert (0.3 g of P4) was applicated intravaginally for 12 days; a second insert replaced the first one for the next 12 days, and next 200 IU of equine chorionic gonadotropin (eCG) was injected intramuscularly (Folligon). Estrus was expected 48 h after eCG injection. In the Group II, Chronogest sponge (20 mg of flugestone acetate) was applicated intravaginally and after 7 days replaced with second chronogest sponge for 7 days. After removing the sponge, on the same day eCG was injected and estrus was expected after 48 h. Artificial insemination was provided with frozen-thawed semen twice: 12 and 24 h after expected estrus. The peripheral blood from the jugular vein was collected each time when the inserts or sponge were applicated and 40 days after insemination. The concentration of P4 and E2 in plasma was measured by RIA. The effectiveness of insemination was monitored by pregnancy-associated glycoproteins determination and observed by the number of calves born. Two pregnancies were confirmed in Group I and five in Group II based on PAG concentration. One newborn was observed in Group I and five in Group II. Both methods of synchronization are effective in hinds based on the received profile of steroids. Although the sponge shape in case of chronogest is better comparing with CIDR, which was not completely deposited in the vagina of hind, potentially leads to bacteria inflammation, and it disturbs the rightful endocrine regulation. Moreover, pregnancy rate and hormone responsiveness were better in Group II.

Follicular dynamic and reproductive performance of gilts submitted to estrous cycle synchronization using two different progestogen sources

This study evaluated reproductive indicators of gilts treated with altrenogest or an intravaginal device (IVD) containing medroxyprogesterone acetate (MPA) for estrous cycle synchronization, starting the protocol on different days of the estrous cycle or replacing the IVD in the middle of treatment. In Experiment 1, 126 gilts were assigned, according to the day of treatment onset (Day 5 or 10 of the estrous cycle), to the following treatments: Control-5 (no hormone); Control-10 (no hormone); IVD-5 (IVD with MPA); IVD-10 (IVD with MPA); ALT-5 (altrenogest); or ALT-10 (altrenogest). The first day of the previous estrus was considered as Day 0 of the estrous cycle, and progestogen groups were treated for 14 d. In Experiment 2, 63 gilts were assigned to Control, ALT, or IVD groups. Progestogen treatment started on Day 10 of the estrous cycle, and the IVD was replaced after 7 d of treatment. In both experiments, no gilts expressed estrus during progestogen administration. In Experiment 1, the interval hormonal withdrawal-to-estrus (IHE) tended to be shorter when treatment started on Day 10 than on Day 5 (3.6 vs. 4.1 d, respectively; P = 0.09). The percentage of gilts expressing estrus after hormone withdrawal was lower for IVD-gilts (76.3%) compared to ALT (100%) and Control-gilts (92.9%; P ≤ 0.07). The percentage of persistent follicles (PFOL) was greater in IVD-10 (60.0%) and ALT-10 (33.3%) than CONT-10 (0.0%; P ≤ 0.06). The adjusted farrowing rate (AFR) was lower in IVD (65.5%) and ALT (80.5%) compared with CONT (97.4%; P ≤ 0.08). In Experiment 2, the IHE was longer for ALT than IVD (4.9 vs. 3.9 d, respectively; P < 0.01). No difference among groups was observed in the percentage of gilts expressing estrus (overall 86.4%), but the occurrence of PFOL was higher in IVD (61.5%) compared to ALT (5.3%), and Control groups (10.5%; P < 0.01). The AFR was lower in IVD (53.8%) than in ALT (88.2%) and Control (94.7%; P ≤ 0.05). The total number of piglets born was not affected by hormonal treatments in either experiment. Estrous expression was delayed in gilts treated with altrenogest or IVD–MPA. However, the reproductive performance of IVD-gilts was compromised, which was not circumvented by IVD replacement in the middle of treatment. Therefore, further studies are necessary to understand MPA pharmacodynamics and investigate alternative devices for a steady release of progestogens in gilts.

Поведенческие и гормонально-метаболические характеристики самок крыс, селекционированных по длительности пассивно-оборонительной реакции

The study was conducted on the female GC (Genetic Catatonia) rats selectively bred for the duration of the passive defensive response to the weak stimuli. Only male rats of this line were used in the previous studies, therefore the aim of this study was to characterize the behavioral, hormonal and metabolic parameters of the female GC rats, and to elucidate the specific properties of their estrous cycle synchronization. We determined the estrous cycle phases using the vaginal smear classification. Other parameters were recorded in the running wheel: the turn and rocking counts, the overall motor activity (amount of runs, grooming, vertical postures, and occurrences on the central axis of the running wheel), and the excitability level in response to an auditory stimulus. We also assayed the progesterone, estradiol, cholesterol, and triglycerides blood levels. Based on the obtained results, we confirm the passive response type and elevated anxiety in female GC rats. The GS rats demonstrated lower fertility and lower body weight in comparison with the Wistar rats. The progesterone blood level was elevated in the proestrus phase in GC rats. The female GC rats demonstrated similarly low triglyceride blood level to the male rats of the same line. The progesterone levels in the synchronized phase were higher than in the unsynchronized one in both GC and Wistar rats. These data show that the selective breeding for the passive defensive response in GC female rats is associated with the moderate stress reactions, changes in the hormonal and metabolic profiles, and decreased fertility.

Recent Technical Breakthroughs for ARTs in Mice

Abstract: Assisted reproductive technologies (ARTs) in mice were recently advanced when two long-existing technical barriers were overcome. The first barrier was the limited number of mature oocytes after conventional superovulation, especially in inbred strains of mice. A combination of estrous cycle synchronization and antiinhibin serum treatments increased the number of collected oocytes from female mice by approximately 3–4 times in many strains. The second barrier was the low fertilization rate after in vitro fertilization (IVF) using frozen-thawed spermatozoa. The addition of reduced glutathione in the fertilization medium dramatically increased the IVF yields, even in cryopreserved/warmed spermatozoa from the C57BL/6J strain, which is one of the strains most sensitive to cryoinjury. This result encouraged the use of cryopreserved spermatozoa in mouse strains worldwide for the preservation and transportation of their genetic characteristics. The final yield to produce offspring from one female was i...

Estrous cycle synchronization in the Persian onager (Equus hemionus onager).

The endangered Persian onager (Equus hemionus onager) has experienced significant population declines over the past century due to poaching, habitat destruction, and resource competition. Remaining animals in zoos and the wild are regionally isolated. Artificial insemination (AI) may be particularly useful as a means of aiding in global genetic management of these isolated populations. The first successful AI in onagers was performed in 2009 utilizing urinary hormone analyses and regular transrectal ultrasound examinations that required specialized handling devices. A method for estrous synchronization in this species would alleviate the need for daily handling and provide a more feasible approach to AI. This study tested long-acting controlled-release preparations of estradiol and progesterone, followed by a single injection of prostaglandin 10 days later, in six adult female Persian onagers to determine whether ovulation would occur within a narrow window of time. Serial transrectal ultrasound exams were performed to determine the day of ovulation following hormone treatment. Means and standard deviations were determined for the lengths of follicular and luteal phases, follicle sizes, and time to ovulation, and compared to historical data in this species. All six onagers ovulated between Days 18 and 22, with three females ovulating on Day 19, as determined by the presence of a corpus luteum. This is an apparently safe and effective method for the synchronization of estrous cycles in the Persian onager, and may be used to develop a timed AI protocol for use at institutions that do not have specialized handling facilities to enable regular transrectal ultrasound.

Possibility of Diagnosing Uterine Function in Cows

Declining in reproductive efficiency is a worldwide problem, although remarkable progress has been made in reproductive techniques including artificial insemination programs, estrous cycle synchronization, and effective embryo production. One means of solving this problem is to select breeding cows based on highly precise diagnosis for high reproductive ability, including ovarian and uterine function. Many recent studies have documented the regulation of uterine function, with the results suggesting that said functions might be an effective diagnostic index for reproductive performance. Therefore, the production of oxytocin-induced prostaglandin F2α and epidermal growth factor in bovine endometrium, such as the physiological reaction in vivo, may be useful in detecting repeat breeder cows. Although the diagnosis is possible using relatively simple techniques such as biopsy, culture, and hormone assay, it is difficult to apply such diagnosis to the field conditions. However, further improvements in the exhaustive gene expression analysis of uterine function in repeat breeder or low fertility cows, so as to determine the genes associated with physiological reactions, could lead to new diagnostic indexes resulting in improved reproductive efficiency. Discipline: Animal Industry Additional key words: endometrial biopsy, epidermal growth factor, oxytocin, prostaglandin F2α

High-Yield Superovulation in Adult Mice by Anti-Inhibin Serum Treatment Combined with Estrous Cycle Synchronization.

Producing many mature oocytes is of great importance for assisted reproductive technologies. In mice, superovulation by consecutive injections of equine chorionic gonadotropin (eCG) and human chorionic gonadotropin (hCG) has been the gold standard for oocyte collection. However, the yield of mature oocytes by this regimen can fluctuate according to the stage of the estrous cycle, strain, and age. Therefore, our objective was to develop a high-yield superovulation protocol to collect higher numbers of oocytes from adult female mice of different strains and ages. First, we aimed to synchronize the estrous cycle using C57BL/6 (B6) female mice. Most (93%) were synchronized to metestrus after two daily injections of progesterone. Second, we found that with the injection of anti-inhibin serum (AIS) instead of eCG, the mean number of ovulated oocytes almost doubled (21 vs. 41 per mouse). Third, by combining estrous cycle synchronization with two AIS injections, we obtained 62 oocytes per mouse, about three times that with the eCG-hCG protocol. Importantly, this approach increased the proportion of mice that ovulated >25 oocytes from about 40% (eCG-hCG) to 90%. The same protocol was also effective in other inbred (BALB/cA), outbred (ICR), and hybrid (B6D2F1) strains. In addition, B6 female mice aged over 1 yr ovulated 1.8-fold more oocytes by this protocol. Thus, estrous cycle synchronization followed by AIS-hCG yielded a broadly applicable, highly efficient superovulation. This protocol should promote the effective use of invaluable female mouse strains and decrease the numbers of animals euthanized.

Synchronization of the Estrous Cycle against the Background of Increased Excitability in Rats Selected for Catatonic Type of Reaction.

Selection for the elevation of catatonic type of reaction in female GC rats ("genetic" and "catatonia") was followed by an increase in the number of intense running episodes in response to acoustic stimulation. The observed changes are typical of increased general excitability in these animals. The phenomenon of estrous cycle synchronization was confirmed by variations in the concentrations of estradiol and progesterone in groups of Wistar and GC rats differing by the degree of synchronization. Some differences were found in the concentration of main sex hormones in the blood from female GC and Wistar rats.

Timed artificial insemination programs during the summer in lactating dairy cows: Comparison of the 5-d Cosynch protocol with an estrogen/progesterone-based protocol

The objective of this study was to compare a GnRH-based to an estrogen/progesterone (E2/P4)-based protocol for estrous cycle synchronization and fixed timed artificial insemination (TAI), both designed for synchronization of ovulation and to reduce the period from follicular emergence until ovulation in cows with a synchronized follicular wave. A total of 1,190 lactating Holstein cows (primiparous: n=685 and multiparous: n=505) yielding 26.5±0.30kg of milk/d at 177±5.02d in milk were randomly assigned to receive one of the following programs: 5-d Cosynch protocol [d −8: controlled internal drug release (CIDR) + GnRH; d −3: CIDR removal + PGF2α; d −2: PGF2α; d 0: TAI + GnRH] or E2/P4 protocol (d −10: CIDR + estradiol benzoate; d −3: PGF2α; d −2: CIDR removal + estradiol cypionate; d 0: TAI). Rectal temperature and circulating progesterone (P4) were measured on d −3, −2, 0 (TAI), and 7. The estrous cycle was considered to be synchronized when P4 was ≥1.0ng/mL on d 7 in cows that had luteolysis (P4 ≤0.4ng/mL on d 0). To evaluate the effects of heat stress, cows were classified by number of heat stress events: 0, 1, and 2-or-more measurements of elevated body temperature (≥39.1°C). Pregnancy success (pregnancy per artificial insemination, P/AI) was determined at d 32 and 60 after TAI. The cows in the 5-d Cosynch protocol had increased circulating P4 at the time of PGF2α injection (2.66±0.13 vs. 1.66±0.13ng/mL). The cows in the E2/P4 protocol were more likely to be detected in estrus (62.8 vs. 43.4%) compared with the cows in the 5-d Cosynch protocol, and expression of estrus improved P/AI in both treatments. The cows in the 5-d Cosynch protocol had greater percentage of synchronized estrous cycle (78.2%), compared with cows in the E2/P4 protocol (70.7%). On d 60, the E2/P4 protocol tended to improve P/AI (20.7 vs. 16.7%) and reduced pregnancy loss from 32 to 60d (11.0 vs. 19.6%), compared with the 5-d Cosynch protocol. In cows withtheir estrous cycle synchronized, the E2/P4 protocol had greater P/AI (25.6 vs. 17.7%) on d 60 and lower pregnancy loss from 32 to 60d (6.7 vs. 21.7%) compared with cows in the 5-d Cosynch protocol. Follicle diameter affected pregnancy loss from 32 to 60d only in the cows in the 5-d Cosynch protocol, with smaller follicles resulting in greater pregnancy loss. Pregnancy per AI at d 60 was different between protocols in the cows with 2 or more measurements of heat stress (5-d Cosynch=12.2% vs. E2/P4=22.8%), but not in the cows without or with 1 heat stress measurement. In conclusion, the 5-d Cosynch protocol apparently produced better estrous cycle synchronization than the E2/P4 protocol but did not improve P/AI. The potential explanation for these results is that increased E2 concentrations during the periovulatory period can improve pregnancy success and pregnancy maintenance, and this effect appears to be greatest in heat-stressed cows when circulating E2 may be reduced.

Producer survey of herd-level risk factors for nursing beef calf respiratory disease

To identify herd-level risk factors for bovine respiratory disease (BRD) in nursing beef calves. Population-based cross-sectional survey. 2,600 US cow-calf producers in 3 Eastern and 3 Plains states. The associations of herd characteristics with BRD detection in calves and cumulative BRD treatment incidence were determined. 459 (177%) surveys were returned and met the inclusion criteria; 48% and 52% of these surveys were completed by producers in Plains and Eastern states, respectively. Mean (95% confidence interval) number of animals in herds in Plains and Eastern states were 102 (77 to 126) and 48 (40 to 56), respectively. Bovine respiratory disease had been detected in ≥ 1 calf in 21% of operations; ≥ 1 calf was treated for BRD and ≥ 1 calf died because of BRD in 89.2% and 46.4% of operations in which calf BRD was detected, respectively. Detection of BRD in calves was significantly associated with large herd size, detection of BRD in cows, and diarrhea in calves. Calving season length was associated with BRD in calves in Plains states but not Eastern states. Cumulative incidence of BRD treatment was negatively associated with large herd size and examination of cows to detect pregnancy and positively associated with calving during the winter, introduction of calves from an outside source, offering supplemental feed to calves, and use of an estrous cycle synchronization program for cows. Results of this study indicated factors associated with calf BRD risk; modification of these factors could potentially decrease the incidence of BRD in nursing calves.