Investigation of vaginal mucus parameters: Development of models for staging the oestrous cycle of the Bunaji cow

Changes in some vaginal mucus parameters were studied in order to generate predictive models capable of enhancing oestrous cycle staging, using equal groups (unsynchronized-USC [no treatment] and synchronized-SC [Synchromate^® i/m on d0, d11]) of Bunaji cows (n = 48) aged 3–4 years. Vaginal mucus was collected (starting d11 in SC) daily over 26 days using standard procedures. Physical (viscosity, elasticity, density, resistivity) and biochemical (pH, glucose, cholesterol, total protein, calcium, magnesium, sodium, potassium) parameters were evaluated using standard procedures. Data were analysed using chi-square and multinomial logit regression modelling. Models generated using oestrus as reference categories were ascertained for accuracies. Chi-square values for viscosity, elasticity and density were significant (p < .01) in USC and SC across stages of the cycle. Results for USC showed that pH and cholesterol were predictive (p < .01) for pro-oestrus, metoestrus and dioestrus, while total protein was predictive (p < .01) for dioestrus only. Similarly, magnesium was predictive (p < .05) for pro-oestrus. For SC, pH, magnesium and cholesterol were predictive (p < .01) for pro-oestrus, metoestrus and dioestrus, while total protein was predictive (p < .01) for pro-oestrus and dioestrus. Potassium and total protein were also predictive for metoestrus at 10% and 5% significance levels, respectively. Though findings suggest the usefulness of magnesium in staging the oestrous cycle only in synchronized cows, pH, total protein and cholesterol appeared to be the more important vaginal mucus parameters in Bunaji cows, regardless synchronization. Furthermore, the models developed showed high accuracy levels for staging the oestrous cycle in USC (100%) and SC (89%).     

Influence of a bovine respiratory disease vaccine with a temperature‐sensitive modified live or killed infectious bovine rhinotracheitis component on oestrous cycle parameters and anti‐Müllerian hormone concentration in nulliparous heifers

The objective of this study was to examine the impact of a bovine respiratory disease complex (BRDC) vaccine with a temperature‐sensitive modified live vaccine (MLV) infectious bovine rhinotracheitis (IBR) component on oestrous cycle parameters and the follicular pool. Twenty‐four Holstein heifers (12.4 ± 0.5 months) previously calfhood vaccinated with an IBR MLV component were enrolled in two replicates (Spring; n = 10 and Fall; n = 14) and were blocked by pre‐vaccination bovine viral diarrhoea (BVD) serum neutralizing (SN) titres. Upon enrolment, heifers were oestrous synchronized with sampling beginning at detected oestrus. At their second heat, heifers were vaccinated with a BRDC calfhood vaccine with a MLV (MLV; n = 12) or killed (K; n = 12) IBR component and sampled for two additional cycles. Serum samples for oestrogen (E2) and progesterone (P4) as well as ultrasound data of ovarian structures were collected every other day. Serum samples for anti‐Müllerian hormone (AMH) were collected at oestrus and mid‐cycle for each cycle, and serum for titres was collected prior to and following vaccination. Data were analysed with the PROC MIXED and GLM procedures of SAS. There was no difference in pre‐ or post‐vaccination titres between MLV and K heifers (p > .5). Vaccination had no impact on P4 concentrations, P4 area under the curve, luteal tissue area, peak E2 production or oestrous cycle length (p > .05). Cycle number did impact AMH concentration (p < .05). In MLV heifers, AMH concentration was highest in cycle 1 (p < .05) while cycles 2 and 3 did not differ (p > .05). This was also true for the K heifers in the Fall replicate (p < .05). Within cycle 2, AMH concentrations were numerically lower between vaccine types (K = 308.22 ± 33.3 pg/ml, MLV = 181.13 ± 32.9 pg/ml; p > .05). Although no differences were seen in overall cycle parameters, differences in AMH concentrations may indicate a reduction of the follicular pool following vaccination and requires further investigation.     

Plasma Inhibin Levels in Relation to Steroids and Gonadotrophins during Oestrous Cycle in Buffalo

The study was conducted on six Murrah buffalo synchronized and induced to oestrus. An indwelling catheter was placed in the jugular vein of each buffalo 4 days before the expected onset of the oestrus following the induced oestrus and blood samples were collected at 8 h intervals from each animal throughout the oestrous cycle. Plasma immunoreactive inhibin, follicle-stimulating hormone (FSH), luteinizing hormone (LH), estradiol-17β and progesterone were estimated by radioimmunoassay to study the variations in the peripheral levels of these hormones and their inter-relationships in order to elucidate the feedback systems controlling them during the oestrous cycle of buffalo. Plasma inhibin levels ranged between 391.25 and 631.97 pg/ml during various phases of the oestrous cycle and were found to be higher than reported in cows. Peak LH and FSH levels during oestrus were 38.40 ± 9.21 and 24.04 ± 4.75 ng/ml, respectively and estradiol-17β and progesterone were 19.50 ± 5.51 pg/ml and 0.61 ± 0.25 ng/ml, respectively. The mean plasma inhibin concentration on the day of oestrus was 562.5 ± 18.9 pg/ml. Levels of FSH in the plasma showed three mid-cycle elevations which corresponded to comparatively lower inhibin and elevated estradiol-17β levels during the same period. From this observation it was deduced that both inhibin and estradiol-17β have a feed-back regulatory effect on FSH secretion in buffalo.     

The post-ovulatory rise in progesterone is lower and the persistence of oestrous behaviour longer during the first compared with the second cycle of the breeding season in mares

Mares are seasonally polyoestrous breeders. Therefore, the first ovulation of the season, following winter anoestrus, is the only cycle in which mares ovulate without the presence of an old CL from the previous cycle. The objective of this study was to compare the length of oestrous behaviour, and plasma progesterone concentrations during the early post-ovulatory period between mares after the first and second ovulation of the breeding season. Overall, 38 mares and 167 oestrous periods were used in the study. From those, 11 mares were used during the first and subsequent oestrous period to measure and compare the post-ovulatory rise in progesterone concentration, whereas all the mares were used to compare the length of the post-ovulatory oestrous behaviour between the first and subsequent cycles of the breeding season. The persistence of the post-ovulatory oestrus was longer (p < .001) following the first ovulation of the year (median of 52 h) compared with the subsequent ovulations (median of 36 h for second and later ovulations groups; n = 38 mares). The progesterone concentration at any of the four 8 h-intervals analysed (28, 36, 76 and 84 h post-ovulation) was lower (p < .01) following the first versus the second ovulation of the year. By 36 h post-ovulation the progesterone concentration of mares at the second ovulation of the year had passed the threshold of 2 ng/ml (2.1 ± 0.33 ng/ml), whereas in the first cycle it was 1.2 ± 0.13 ng/ml. In conclusion, mares had lower progesterone concentrations in their peripheral circulation and longer persistence of oestrous behaviour following the first ovulation of the year compared with the second and subsequent ovulatory periods of the breeding season.     

The effects of luteinizing hormone as a suppression factor for apoptosis in bovine luteal cells in vitro

The fate of the corpus luteum, a transient endocrine gland formed and degraded during an oestrous cycle, is decided by various physiological factors, such as luteinizing hormone (LH). As a stimulator of progesterone, LH is known to maintain corpus luteum functional and structural integrity by inhibiting apoptosis, a programmed cell death. Therefore, we aim to investigate its action during the mid-luteal phase hypothesized that LH suppresses the death mechanism of bovine luteal steroidogenic cells (LSC) by analysing the expression of proteins involved. Cultured bovine LSC obtained from corpus luteum were treated for 24 hr with recombinant TNF and IFNG in the presence or absence of LH. The result showed that LH proved to have a protective effect by increased cell viability (p < .05) and prevented DNA fragmentation (p < .05), as demonstrated by the WST-1 colorimetric assay and TUNEL assay. Expression analysis of mRNA and protein levels showed that LH altered the expression of BCL2 (p < .05), CASP3 (p < .05), FAS (p < .05), and BAX (p < .05) to support cell survival. In conclusion, our study suggests that LH prolongs the corpus luteum life span through the anti-apoptotic mechanism by increasing cell viability and suppressing apoptosis-related genes and protein expression.     

Expression and localization of angiopoietin family in corpus luteum during different stages of oestrous cycle and modulatory role of angiopoietins on steroidogenesis,angiogenesis and survivability of cultured buffalo luteal cells

The objective of this study was to document the expression and localization of angiopoietin (ANGPT) family members comprising of angiopoietin (ANGPT1 and ANGPT2), and their receptors (Tie1 and Tie2) in buffalo corpus luteum (CL) obtained from different stages of the oestrous cycle, and the modulatory role of ANGPT1 and ANGPT2 alone or in combinations on progesterone (P₄) secretion and mRNA expression of phosphotidylinositide‐3kinase‐protein kinase B (PI3K‐AKT), phosphoinositide‐dependent kinase (PDK), protein kinase B (AKT), Bcl2 associated death promoter (BAD), caspase 3 and von willebrand factor (vWF) in luteal cells obtained from midluteal phase (MLP) of oestrous cycle in buffalo. Real‐time RT‐PCR (qPCR), Western blot and immunohistochemistry were applied to investigate mRNA expression, protein expression and localization of examined factors whereas, the P₄ secretion was assessed by RIA. The mRNA and protein expression of ANGPT1 and Tie2 was maximum (p < .05) in mid luteal phase (MLP) of oestrous cycle. The ANGPT2 mRNA and protein expression was maximum (p < .05) in early luteal phase, decreased in MLP and again increased in late luteal phase of oestrous cycle. ANGPT family members were localized in luteal cells and endothelial cells with a stage specific immunoreactivity. P₄ secretion was highest (p < .05) with 100 ng/ml at 72 hr when luteal cells were treated with either protein alone. The mRNA expression of PDK, AKT and vWF was highest (p < .05) and BAD along with caspase 3 were lowest (p < .05) at 100 ng/ml at 72 hr of incubation period, when cultured luteal cells were treated with either protein alone or in combination. To conclude, our study explores the steroidogenic potential of angiopoietins to promote P₄ secretion, luteal cell survival and angiogenesis through an autocrine and paracrine actions in buffalo CL.     

Luteal Blood Flow and progesterone concentration during first and second postpartum estrous cycle in lactating dairy cows

The aim of the present study was to determine the differences in corpus luteum (CL) functionality between the first postpartum estrous cycle and the following cycle in lactating dairy cows. Luteal blood flow (LBF), luteal size and blood progesterone (P4) concentration were monitored during the first and second postpartum estrous cycle. During the first and second postpartum estrous cycle, the mean LBF value increased (p < .05) from early to late dioestrus, while it decreased rapidly in proestrus, resulting statistically lower (p < .05) than those registered in all previous phases. Statistically significant differences were not observed between overall LBF during first and second postpartum estrous cycle (p > .05). During the first postpartum estrous cycle, P4 blood concentrations showed a significant reduction (p < .05) from dioestrus to proestrus. A different trend of P4 concentrations was observed during the second postpartum estrous cycle, where mean P4 value registered in proestrus resulted statistically lower than those registered in the previous cycle phases (p < .05). The mean P4 concentration registered over the first postpartum estrous cycle resulted statistically lower (p < .05) than that registered during the second one. A significant correlation between P4 concentrations and LBF was registered only during the second postpartum estrous cycle. Results indicate that during the first postpartum estrous cycle, P4 concentration was independent of luteal blood flow and luteal size.     

Metabolic blood profile of beef heifers during oestrous and non‐oestrous states

Haematological metabolic profiles in heifers could contribute to the development of proxies for oestrous detection and provide clues to further characterize biological changes during oestrus. One hundred and seven beef heifers were observed for oestrous behaviour twice daily for 124 days. Feed intake and productive performance (body weight and composition) traits were measured, and feed efficiency was determined using residual feed intake (kg DM/day). Blood plasma samples were collected when signs of oestrus were observed and every 30 ± 2 days. Heifers were considered in oestrus (n = 71) when plasma progesterone concentrations were <0.6 ng/ml. Least square means of blood metabolic parameters were compared between oestrous and non‐oestrous states and within oestrous groups according to performance traits and age. Heifers in oestrus exhibited higher concentrations of alkaline phosphatase, aspartate aminotransferase (AST), beta‐hydroxybutyric acid, creatine kinase (CK) and triiodothyronine (T3) than heifers in non‐oestrus. Heifers in oestrus revealed lower osmolality and concentrations of calcium, sodium and total protein than during non‐oestrus. Younger (and smaller) heifers had greater concentrations of CK, gamma‐glutamyl transferase (GGT), glucose and sodium than the older heifers. Heifers with lower fatness had increased osmolality and concentrations of cholesterol, CK, phosphorus, sodium and reduced T3 levels. Feed efficient heifers had greater levels of AST, cholesterol and GGT than inefficient heifers. Blood plasma parameters may be complementary to oestrous detection upon further validation; effects of age, feed efficiency, body size and body composition should be considered to optimize this haematological assessment.     

Effect of melatonin on regulation of apoptosis and steroidogenesis in cultured buffalo granulosa cells

This study was aimed to address melatonin receptor expression, mRNA level of hypothalamus and hypophysis hormone receptors (GnRHR, FSHR, and LHR), steroidogenesis, cell cycle, apoptosis, and their regulatory factors after addition of melatonin for 24 hr in cultured buffalo granulosa cells (GCs). The results revealed that direct addition of different concentrations of melatonin (100 pM, 1 nM, and 100 nM) resulted in significant upregulation (p < 0.05) of mRNA level of melatonin receptor 1a (MT1) without affecting melatonin receptor 1b (MT2). Melatonin treatment significantly downregulated (p < 0.05) mRNA level of FSH and GnRH receptors, whereas 100 nM dose of melatonin significantly increased mRNA level of LH receptor. Treatment with 100 nM of melatonin significantly decreased the basal progesterone production with significant decrease (p < 0.05) in mRNA levels of StAR and p450ssc, and lower mRNA level of genes (Insig1, Lipe, and Scrab1) that affect cholesterol availability. Melatonin supplementation suppressed apoptosis (100 nM, p < 0.05) and enhanced G2/M phase (1 nM, 100 nM, p < 0.05) of cell cycle progression which was further corroborated by decrease in protein expression of caspase‐3, p21, and p27 and increase in bcl2. Our results demonstrate that melatonin regulates gonadotrophin receptors and ovarian steroidogenesis through MT1. Furthermore, the notion of its incorporation in apoptosis and proliferation of buffalo GCs extends its role in buffalo ovaries.     

AQP8 participates in oestrogen-mediated buffalo follicular development by regulating apoptosis of granulosa cells

Aquaporins (AQPs), a family of small membrane-spanning proteins, are involved in fluid transport, cell signalling and reproduction. Regulating AQP8 expression influences apoptosis of granulosa cells (GCs), ovarian folliculogenesis, oogenesis and early embryonic development in mice, but its role has never been investigated in other species. The aim of the present study was to characterize the AQP8 function in buffalo follicular development. The expression pattern of AQP8 in buffalo follicle was analysed by immunohistochemistry method. 17β-Estradiol (E2) or oestrogen receptor antagonist ICI182780 was used to treat GCs cultured in vitro, and the expression of AQP8 was detected using qRT-PCR. Its roles in apoptosis of buffalo GCs were investigated by shRNA technology. AQP8 was found to be expressed higher in secondary follicles (p < .05), and its mRNA level in GCs was upregulated by E2 via receptor-mediated mechanism in a dose-dependent manner. A 732-bp buffalo AQP8 coding region was obtained, which was highly conserved at the amino acid level among different species. AQP8-shRNA2 had more effective inhibition on target gene than AQP8-shRNA1 (66.49% vs. 58.31%) (p < .05). Knockdown of AQP8 induced GCs arrested at G2/M stage and occurred apoptosis. Compared with the control group, higher Caspase9 expression were observed in AQP8-shRNA2 lentivirus infected GCs (p < .05), while Bcl-2 and Bax expression levels had no obvious change (p > .05). Altogether, the above results indicate that AQP8 is involved in oestrogen-mediated regulation of buffalo follicular development by regulating cell cycle progression and apoptosis of GCs.     

Association of interservice interval with conception rate in Japanese Black cattle

This study aimed to clarify the association of interservice interval (ISI) with conception rate (CR) and to evaluate factors affecting ISI in Japanese Black cattle. Data used in this study covered 32,639 artificial insemination (AI) records on 972 farms. The mean ± SEM of ISI and CR was 57.0 ± 0.3 days and 44.6 ± 0.3%, respectively. The relative frequency of ISI differed significantly between parity groups (p < .05). For parity 0, the proportion of ISI at an 18‐ to 24‐day interval was 43.6%, whereas the proportions in the other parity groups ranged from 27.2% to 29.1%. The CR was associated significantly with ISI, parity and AI number (p < .05), but not with season. Cows reinseminated at an 18‐ to 24‐day interval had the highest CR (46.8%; p < .05). Cows reinseminated at a 39‐ to 45‐day interval had a similar CR to those reinseminated at 25‐ 38‐day and 46‐ to 59‐day intervals. Cows reinseminated at 11–17, 60–66 and 67 days or longer intervals had the lowest CR (p < .05). Thus, cows returning to oestrus in a normal cycle tended to have a higher probability of conceiving compared with those that returned to oestrus after a normal cycle for any degree of parity and at any number of AI.     

Expression of Frizzled 2 in the mouse ovary during oestrous cycle

The Wnt/β‐catenin pathway is a conserved signalling pathway that regulates gene expression and controls diverse developmental processes such as cell fate specification, cell proliferation and cell differentiation. To our knowledge, the potential role of this pathway in the adult ovary has been poorly addressed. To this end, we investigated the expression pattern of Frizzled 2 in the mouse ovary during oestrous cycle by real‐time Q‐PCR, in situ hybridization, Western blotting and immunohistochemistry. In this study, we used uterine wet weight and H3.2 mRNA level as two markers for classification of mouse oestrous cycle. We found that both uterine wet weight and H3.2 mRNA are the highest in the proestrus and decrease from oestrus to diestrus. During oestrous cycle, Frizzled 2 mRNA and protein exhibited the highest level in the proestrus stage and rapidly reduced from oestrus to diestrus. In situ hybridization results showed that the positive signals for Frizzled 2 are highly detected in the oocyte and stroma in proestrus stage, while moderate or weak Frizzled 2 mRNAs are localized in the oocyte, granulosa cells, stroma and corpus luteum from oestrus to diestrus. The localization pattern of Frizzled 2 protein is mostly consistent with its mRNA, but stronger Frizzled 2 proteins are present in the granulosa cells and membrane of oocyte in oestrous cycle. Our data suggest that Frizzled 2 may be involved in regulating follicle growth, oocyte maturation and luteinization during oestrous cycle.     

Ion,Protein, Phospholipid and Energy Substrate Content of Oviduct Fluid During the Oestrous Cycle of Buffalo (Bubalus bubalis)

The aim of this research was to analyse the composition of oviduct fluid (ODF) in buffalo cows at different oestrous cycle phases to fulfil the requirements of buffalo embryos in vitro. ODF was collected by chronic cannulation from three cows that were synchronized by administering a synthetic prostaglandin. Based on hormonal profiles, the pre‐ovulatory, ovulatory, post‐ovulatory and luteal phases of the oestrous cycle were defined. The volume of ODF produced (ml/24 h) was influenced by the oestrous cycle, with values (mean ± SE) around ovulation (1.0 ± 0.2) greater (p < 0.05) than in both the luteal (0.4 ± 0.1) and the post‐ovulatory phases (0.5 ± 0.1), but not different from the intermediate values in the pre‐ovulatory phase (0.8 ± 0.2). Among cycle phases, no differences were found in sodium, potassium, calcium and magnesium concentrations (130.0 ± 1.1, 5.1 ± 0.3, 2.8 ± 0.1 and 0.59 ± 0.04 mmol/l respectively). Interestingly, the chloride secretion (μm /24 h) was higher (p < 0.05) at ovulation (150.2 ± 16.5) than during both the luteal (73.7 ± 22.0) and the post‐ovulatory phases (63.7 ± 11.2), with intermediate values in the pre‐ovulatory phase (113.4 ± 23.5). Glucose concentration (mmol/l) was higher (p = 0.056) in the pre‐ovulatory phase (0.06 ± 0.02) than in the luteal (0.02 ± 0.01) and post‐ovulatory (0.02 ± 0.01) phases but not different from values in the ovulatory phase (0.04 ± 0.02). Concentrations of pyruvate and lactate among oestrous cycle phases were similar (0.08 ± 0.01 and 1.0 ± 0.1 mmol/l respectively). The total quantity of phospholipids (μmol/24 h) was greater (p < 0.05) at ovulation (0.21 ± 0.02) compared with the luteal, pre‐ovulatory and post‐ovulatory phases of the cycle (0.09 ± 0.02, 0.13 ± 0.02 and 0.09 ± 0.01 respectively). No differences were found in either the protein concentration (1.8 ± 0.3 mg/ml) or the quantity of proteins secreted in 24 h (1.8 ± 0.4 mg) among oestrous cycle phases. In conclusion, this study provides the first characterization of buffalo ODF during the oestrous cycle, showing species‐specific differences that may be useful for developing suitable media for buffalo in vitro embryo production.     

Immunoexpression of cytokine tumour necrosis factor-α suggesting its role in formation and regression of corpus luteum in Indian buffalo

Tumour necrosis factor-α (TNF-α) is a cytokine that plays multiple important roles in corpus luteum (CL). Immunolocalization of expression of TNF-α in CL of buffalo was studied in different stages of its development and regression. Corpus luteum of healthy buffaloes (24) was collected from local slaughterhouses and categorized into early (stage I, 1–5 days, n = 6), mid (stage II, 6–11 days, n = 6), late luteal (stage III, 12–16 days, n = 6) and regressing phase (stage IV, 17–20 days, n = 6). In earliest phase of cyclic CL, per cent immunoexpression of TNF-α was significantly (p < .05) lower as compared to all phases with its expression being restricted to few developing luteal cells, usually in neutrophils. A significantly (p < .05) higher number of neutrophils with TNF-α immunoexpression were observed as compared to mid-luteal phase that indicated its role in initiation of angiogenesis at this stage. TNF-α immunoexpression almost doubled in mid-luteal phase, but the number of neutrophils exhibiting TNF-α was significantly (p < .05) lower with respect to all phases of CL. Immunoexpression percentage in late luteal phase increased sharply being significantly (p < .05) higher than earlier two phases of CL. In regressing phase, per cent immunostaining was maximum with highly significant (p < .05) difference as compared to all other stages, observed in all degrading luteal cells, abundant immune cells, that is neutrophils and macrophages which finally led to apoptosis and phagocytosis. Immunoexpression of TNF-α in early luteal phases served its role in initiation of angiogenesis, and its intense expression in regressing phase of CL suggested a shift in its role to apoptosis and structural luteal regression signifying both luteotropic and luteolytic roles in buffalo. This is probably the first study of its kind in buffaloes.     

A longitudinal study of oestrous characteristics and conception in tie-stalled and loose-housed Swedish dairy cows

The presence and importance of visual oestrous symptoms and the relation to conception, breed, housing system and milk yield were studied. Data were collected from a Swedish research herd during a 16-year period. Local, secondary and primary oestrous symptoms and an overall oestrus intensity score (OI) were studied. Decreasing trends in oestrous expression and conception rates (CR), and an increase in CR with more intensive oestrus, from 24% at OI score one (very week) to 54% at OI score five (very strong) were observed. The mean interval from first to last observed oestrus symptom was long with a large variation (54?±?SD?=?32?h). Generally, irrespectively of housing system, Swedish Red cows had higher CR, more intensive oestruses and longer oestrus durations compared to Swedish Holstein cows. Local oestrous symptoms, with CR of 24–28%, were most frequently detected, which is why more attention should be paid to local oestrous symptoms.     

Changes in Plasma Concentrations of LH, FSH, Estradiol 17-β and Progesterone During Oestrus in Mithun (Bos frontalis)

The objective of the present study was to establish the changes in plasma concentrations of LH, FSH, estradiol 17-beta (E2) and progesterone (P4), as well as to understand their temporal relationships during oestrus in mithun (Bos frontalis). The experiment was conducted on 11 mithuns during third or fourth postpartum oestrous cycle. Since oestrus onset the jugular vein blood samples were collected every 2 h for 72 and 96 h, respectively from the animals without and with standing heat. The LH, FSH, E2 and P4 concentrations were estimated in plasma. The P4 concentration was fluctuated throughout the oestrus period and the average P4 concentration was found significantly (p<0.05) lower on the day of oestrus onset. The multiple rises in LH and FSH concentrations above the basal level in spike like fashion were observed throughout the oestrus period irrespective of the occurrence of standing heat. A significant (p<0.01) gradual increase in the average daily E2 concentration was observed till day 2 following oestrus onset irrespective of the occurrence of standing heat. A significant (p<0.05) simultaneous increase in LH, FSH and E2 concentrations and a transient increase in P4 concentration at approximately the time of standing heat onset were observed. During investigation a definite temporal coupling between LH and FSH rises was absent throughout the oestrus period. The results suggest that (1) the multiple short-duration low-amplitude LH and FSH surges during oestrus may be crucial for the final maturation of ovulatory follicle and subsequent ovulation in mithun; (2) a differential mechanism for controlling LH and FSH secretions probably exists in mithun.     

In vitro Release of Ovarian Progesterone is Decreased During the Oestrous Cycle and Pregnancy of Swine with Obesity/Leptin Resistance

Previous studies indicate that reproductive prolificacy of obese swine breeds is markedly influenced by embryo losses in early pregnancy. In such period, adequate secretion of progesterone (P4) by the ovary is essential for pregnancy success. This study analyses the luteal functionality during the oestrous cycle and early pregnancy of Iberian sows and Large White x Landrace females, in terms of P4 secretion after in vitro culture of luteal tissue stimulated or not with luteinizing hormone (LH). The secretion of progesterone (expressed in ng/mg of luteal tissue or ng/mgLT) of the corpora lutea of obese Iberian swine was always hampered when compared to lean genotypes, either during early oestrous cycle (110.7 ± 37.8 vs 259.7 ± 10.2 ng/mgLT; p < 0.0001), late oestrous cycle (49.0 ± 3.5 vs 75.92 ± 7.14 ng/mgLT; p < 0.0001) or early pregnancy (38.4 ± 2.1 vs 70.7 ± 5.3 ng/mgLT; p < 0.0001). The differences in basal P4 secretion remained after stimulation with LH. Finally, P4 secretion during early pregnancy of Iberian sows decreased with age and, hence, with obesity features (46.6 ± 4.2 vs 65.5 ± 4.8 ng/mgLT; p < 0.001). In conclusion, the results of the present study provide convincing evidence of a reduced luteal function during oestrous cycle and early pregnancy of sows with obesity/leptin resistance like Iberian sows, which may contribute to the low reproductive efficiency reported in this breed.     

Plasma progesterone, oestradiol-17β and total oestrogen profiles in relation to oestrous behaviour during induced ovulation in Murrah buffalo heifers

The objectives of this study were to establish the characteristics of oestrous behaviour in Ovsynch (induction of ovulation through administration of GnRH-PGF2-GnRH in a systemic manner on 0, seventh and ninth day respectively) and Ovsynch plus Norprolac (Quinagolide hydrochloride – an inhibitor of prolactin secretion) treated Murrah buffalo heifers and to determine the relationships between this behaviour and the plasma concentrations of oestradiol-17β (E2), total oestrogen, and progesterone. Oestrus was detected by visual observations of oestrus signs, per rectal examination of genitalia and bull parading thrice a day during treatment period. Among all the symptoms, it was observed that bull mounting of heifers in oestrus was highest. Examination of genital tracts per rectum revealed that the cervix was relaxed, uterus was turgid and ovaries had palpable follicle in animals with oestrus. The peak concentrations of E2 (10.81 ± 0.62 pg/ml) and total oestrogen (17.11 ± 1.21 pg/ml) occurred at 9.45 ± 0.85 and 9.64 ± 0.93 h after second GnRH administration, respectively, in Ovsynch treated animals. However, the peak levels of E2 (20.02 ± 2.87 pg/ml) and total oestrogen (32.71 ± 3.15 pg/ml) occurred at 10.18 ± 0.50 and 10.36 ± 0.75 h after second GnRH administration, respectively, in Ovsynch plus Norprolac treated animals. Plasma progesterone concentration was basal (0.20 ± 0.001 ng/ml) during the peri-oestrus period. The plasma progesterone concentration was the lowest on the day of oestrus and increased to register a peak on day 13 ± 2 of the cycle. Oestrous behaviour was positively correlated with the peak concentration of E2 (p < 0.001) and total oestrogen (p < 0.001) during the peri-oestrus period. Inhibition of prolactin by Norprolac administration significantly increased the concentration of E2 and total oestrogen during oestrus in buffaloes in comparison to those recorded in animals subjected to Ovsynch protocol alone. In conclusion, our results suggest that the peak concentrations of E2 and total oestrogen and mean level of E2 and total oestrogen during the peri-oestrus period are the important factors contributing the behavioural manifestation of oestrus in buffalo cows.     

Effect of the side of ovulation on the uterine morphometry,blood flow,progesterone, oestradiol and nitric oxide during spontaneous and induced oestrus in lactating dairy cows

This study investigates the changes in endometrial thickness between ipsilateral and contralateral uterine horns, diameter, blood flow area and hormonal concentrations in cows during natural and induced oestrus. In the induced group, six cows received a controlled internal device insert for 7 days. In the normal and induced oestrous group (n = 12), Doppler was performed day after day from day 5 till day 15. Oestradiol, progesterone and nitric oxide metabolites (NOMs) were measured. Results showed that dorsal, ventral and cross-section diameter of the ipsilateral horn increased during the induced oestrous group to reach a maximum on day 2 than during normal one. The total coloured area in both horns in a normal and induced group was also affected (p < .001), as the total coloured area in the ipsilateral horns, increased in the induced group, reach a maximum on day 2 after ovulation, while in normal it reaches a maximum on day of ovulation. Oestradiol increased during the induced cycle than during the normal one, but progesterone increased during days after ovulation reaches a higher value on day 10 in both groups. Nitric oxide showed two peaks, one on the day 3 and another peak on day 5. Conclusion: The ipsilateral uterine horn different diameters and thickness increased more rapidly in the induced oestrous group than the natural oestrus group. A complete analysis of variations in P4 and E2 concentrations and endometrial thickness suggested that decreases in P4 concentrations were related to an increase in endometrial thickness and that increases in E2 concentrations enhanced these endometrial changes.