Onset Of Proestrus Research Articles

Luteal phase decrease in packed cell volume in healthy non-pregnant and pregnant bitches.

To establish packed cell volume (PCV) ranges for non-pregnant, pregnant and post-partum bitches from day 10 of proestrus, investigating any relationship with parity and litter size. This prospective cohort study used 37 healthy breeding bitches to examine PCV counts from routine blood samples collected every 4weeks, from day 10 of proestrus, as part of routine PCV monitoring. For pregnant (n=19) and non-pregnant (n=18) bitches, PCV decreased until week 8 (corresponding to 8.5±1.1days before whelping for pregnant bitches) and recovered by 16-20weeks after the initial sample; bitches that whelped average and large litters showed greater declines. PCV began to recover sooner for bitches that had previously whelped one or two litters compared to bitches that had previously whelped three or more litters. There was a significant three-way interaction between time after the onset of proestrus, litter size and the number of previous litters which demonstrated that the large decrease in PCV for bitches that had previously whelped three or more litters only occurred in bitches that were expecting an average or large sized litter. Chronological variation in PCV for pregnant and non-pregnant bitches was established during the reproductive cycle. There was no evidence to suggest that routine PCV measurement for normal, healthy bitches would be beneficial. However, knowledge from this study may be useful when deciding whether to prospectively monitor a bitch where there is a history of previous pregnancy-related anaemia, when performing a caesarean section due to the anticipated blood loss during surgery, or when examining blood profiles for post-litter bitches.

Simultaneously administration of cabergoline and PMSG reduces the duration of estrus induction in anestrous bitches.

This study designed a protocol that would combine pregnant mare serum gonadotrophin (PMSG) and cabergoline (CAB) to induce estrus in bitches. Twenty clinically healthy adult and anestrous female dogs were randomly assigned into four groups. The first group was treated with 5.00 μg kg-1 CAB until the onset of proestrus or for 25 days. The second group was treated with 20.00 IU kg-1 PMSG for 5 days and 500 IU human chorionic gonadotropin (hCG) on the 5th day. The third group was treated with 5.00 μg kg-1 CAB for 10 days in combination with 20.00 IU kg-1 PMSG for 5 days and 500 IU hCG on the 10th day. The control group received 1.00 mL of normal saline. Ovarian changes were evaluated ultrasonographically, and the estrus cycle phase was examined by vaginal cytology. Respectively, three, three and four bitches showed clinical signs of proestrus in each treatment group. The intervals between treatment and proestrus for each group were 30.00 ± 3.05, 7.67 ± 1.20 and 13.00 ± 1.20 days, respectively. Two weeks after estrus, the progesterone mean was 14.51 ± 6.24, 19.96 ± 17.16 and 19.12 ± 9.26 ng mL-1 for each group, respectively. In ultrasonography examination, the largest follicle was identified at 15.66 ± 1.33, 11.66 ± 2.40 and 8.75 ± 2.17 days after the onset of proestrus and the largest follicle's size was measured 6.50 ± 0.55, 4.83 ± 1.64 and 7.07 ± 1.49 mm for each group, respectively. Although the combined use of CAB and PMSG reduced the duration of treatment, alteration of the duration or PMSG dosage can be helpful to improve the results.

Evaluation of Bovine Progesterone Semi-Quantitative Test Kit (Rapid p4) for Estimation of Blood Serum Progesterone Levels in Follicular Phase of Bitch

BACKGROUND: Detection of ovulation time using progesterone measurement is important to inseminate or breeding during the optimum time. OBJECTIVES: The aim of present study was evaluation of bovine progesterone semi-quantitative test kit for estimation of blood serum progesterone levels in bitch. METHODS: Five healthy intact anestrus bitches were used in the present study. Dogs were treated with cabergolin until onset of proestrus. Vaginal cytology, ultrasonography and blood sampling were performed from the 7th day of proestrus until two days after ovulation. Blood serum samples were divided as follows: one part to measure progesterone level by using RIA, and the other part was investigated by bovine progesterone semi-quantitative test kit. Ovarian changes were evaluated by ultrasonography and assessment of the estrus cycle was performed by vaginal cytology examination. RESULTS: Following Spearman analysis, significant positive correlation was observed between the semi-quantitative test kit and RIA results (r=0.916; p =0.00). The results of ROC and Kappa analysis suggested that the highest diagnostic accuracy of progesterone semi-quantitative test kit was observed in the blood serum progesterone levels of 2 and 5 ng/ml. The sensitivity and specificity of the used kit at the level of 5 ng/ml of progesterone were 88.5 and 93.7 %, respectively. The sensitivity and specificity of the used kit at the level of 10 ng/ml of progesterone were 82.4 and 96 % and at the level of 10 ng/ml were 82.4 and 96%, respectively. The observed scores during the late proestrus were 2-3 and just one dog showed score 4. During the estrus phase, in two dogs scores 4-5 and in three dogs scores 3-5 were revealed. CONCLUSIONS: Bovine progesterone semi-quantitative test kit is useful for estimation of estrus cycles in bitch. Scores of 4 and 5 of semi-quantitative test kit indicate ovulation time and proper breeding time.

Effects of Two Commercial Diets on Several Reproductive Parameters in Bitches: Note One-From Estrous Cycle to Parturition.

Simple SummaryReproductive efficiency is a key aspect of all breeding species. Several factors, such as infective diseases, hormonal, and nutritional status could affect female fertility. As demonstrated by high numbers of newborns per litter and proper fetal development, nutritional status at mating, and correct nutritional management during pregnancy are fundamental. In this study, two diets with different ingredients (protein and lipid sources) and different macro and micronutrients concentrations (crude protein, fatty acids profile, vitamins, and mineral concentration) were administered from two months before the expected onset of proestrus to parturition to 18 bitches divided into two groups. The experimental diet, richer in protein, essential and polyunsaturated fatty acids and vitamins, when compared to control diet, seems able to guarantee better clinical presentation of estrus and embryonic development in the first third of pregnancy with a lower incidence of a fetal resorption.The close link between nutrition management and reproductive efficiency is well known, but there is very little data available concerning this topic in canine species. The present study aimed to compare the effect of two different diets upon the follicular period and gestation in bitches. Eighteen pluriparus medium and large size bitches were recruited and divided into control (CTR) and experimental (EX) groups and fed, respectively, with a commercial kibble diet and a specially formulated diet from two months before the expected onset of proestrus up to the end of the trial. It was possible to observe how the EX group had a better clinical presentation of the estrous phase, a higher number of ovarian follicles (p < 0.05), a lower percentage of fetal resorption (p < 0.05), and lower oxidative status, expressed by d-ROMs (p < 0.01), at the moment of pregnancy diagnosis compared to CTR group. Moreover, the EX group showed a lower fetal resorption rate and higher litter size (p < 0.05). These results highlight how a diet characterized by high protein and fat content and richer in essential fatty acids can improve reproductive performance in dogs.

Effect of aglepristone (RU534) administration during follicular phase on progesterone, estradiol-17β and LH serum concentrations in bitches.

Aglepristone was administered in bitches during the follicular phase to evaluate its effects on progesterone, estradiol-17β and LH serum concentrations. Ten German Shepherds were divided into two groups (treated n=5; control n=5). Treated bitches received 10mg/kg BW of aglepristone subcutaneously during the early follicular phase, 24hr after and then 7days later. The control group was injected, at the same time periods, with saline solution (0.3ml/kg BW). For the steroid evaluations, blood was collected daily from the onset of proestrus until the first day of cytological dioestrus. For LH base-line serum determination, blood was also collected every 20min for 2hr at the onset of proestrus. For LH surge identification, blood was collected daily (every 6hr) starting from the day of the first administration of aglepristone or saline solution until the first day of dioestrus. All animals ovulated but the treated group presented longer ovulation-dioestrus intervals than the control group (5.2±2.2days p<.05). Serum concentrations of the evaluated hormones were similar between experimental animals except for serum LH. Indeed, no LH peaks were detected in the treated group while LH surges were clearly observed in the control group (9±1days after the beginning of proestrus. In particular, the area under the curve for LH was significantly lower in treated than control animals (12±4ng/ml x Day; p=.01). In conclusion, administrations of aglepristone during the follicular phase of the bitch does not affect the steroid hormone patterns but does prevent the occurrence of a LH surge. This work raises significant questions and opens perspectives concerning the mechanisms of ovulation in bitches.

Impact of hormonal modulation at proestrus on ovarian responses and uterine gene expression of suckled anestrous beef cows

BackgroundThis study evaluated the impact of hormonal modulation at the onset of proestrus on ovarian response and uterine gene expression of beef cows.MethodsA total of 172 anestrous beef cows were assigned to one of four groups according to the treatment with estradiol cypionate (ECP) and/or equine chorionic gonadotropin (eCG) [CON (n = 43), ECP (n = 43), eCG (n = 44) and ECP + eCG (n = 42)].ResultsECP-treated cows (ECP and ECP + eCG groups) presented greater occurrence of estrus (44.6% vs. 65.4%; P = 0.01) and pregnancy per AI [47.1% vs. 33.3%; P = 0.07], but similar progesterone (P4) concentration at subsequent diestrus than cows not treated with ECP (CON and eCG groups). Nonetheless, eCG-treated cows (eCG and ECP + eCG groups) presented larger follicle at timed AI (12.6 ± 0.3 vs. 13.5 ± 0.3 mm; P = 0.03), greater ovulation rate (96.5% vs. 82.6%; P = 0.008) and greater P4 concentration at d 6 (3.9 ± 0.2 vs. 4.8 ± 0.2 ng/mL; P = 0.001) than cows not treated with eCG (CON and ECP groups). Next, cows with a new corpus luteum 6 d after TAI were submitted to uterine biopsy procedure. Uterine fragments [CON (n = 6), ECP (n = 6)] were analyzed by RNA-Seq and a total of 135 transcripts were differentially expressed between groups (73 genes up-regulated by ECP treatment). Subsequently, uterine samples were analyzed by qPCR (genes associated with cell proliferation). ECP treatment induced greater abundance of PTCH2 (P = 0.07) and COL4A1 (P = 0.02), whereas suppressed EGFR (P = 0.09) expression. Conversely, eCG treatment increased abundance of HB-EGF (P = 0.06), ESR2 (P = 0.09), and ITGB3 (P = 0.05), whereas it reduced transcription of ESR1 (P = 0.05). Collectively, supplementation with ECP or eCG at the onset of proestrous of anestrous beef cows influenced ovarian responses, global and specific endometrial gene expression. ConclusionProestrus estradiol regulate the endometrial transcriptome, particularly stimulating proliferative activity in the endometrium.

148 EXPRESSION OF OVIDUCT-SPECIFIC GLYCOPROTEIN IN THE CANINE OVIDUCT DURING THE PERIOVULATORY PERIOD

In the canine species, the oocyte is ovulated at the immature germinal vesicle (GV) stage and will reach metaphase II stage after 3 to 4 days spent in the oviduct. Fertilization and embryonic development to the blastocyst stage also take place in the oviduct. In a previous study (Tahir et al. 2012 Reprod. Domest. Anim. 47, 487), we reported the expression of oviductin (oviduct-specific glycoprotein) mRNA in the oviduct. The present study aimed to describe the oviductin protein expression (immunolocalization and Western blot quantification) and the effect of the oviducal region and the ovarian cycle. Beagle bitches were ovariectomized at 6 stages (6 bitches/stage): anestrus, after the onset of proestrus and before the LH peak (Pre-LH), after the LH peak and before ovulation (Pre-ov), 1 day (Day 1), 4 days (Day 4), and 7 days (Day 7) after ovulation. Three oviducal regions were collected [i.e. ampulla, isthmus, and ampulla-isthmus junction (AIJ)]. Ampulla and isthmus were fixed in paraformaldehyde, embedded in paraffin, and 7-μm sections were used for immuno-histochemistry using a goat polyclonal anti-human oviductin (N20; Santa Cruz Biotechnology, Santa Cruz, CA, USA) and the ImmPress kit (Vector Laboratories, Burlingame, CA, USA). Total protein from the AIJ was extracted and used for Western Blot using a mouse monoclonal anti-mouse antibody (H8; Santa Cruz Biotechnology). The expression of oviductin in AIJ was quantified in duplicate on blots using ImageJ software and normalized with actin levels. Relative amounts of oviductin were compared between stages by ANOVA followed by a Tukey test. Immuno-histochemistry revealed that oviductin was specifically expressed in the nonciliated cells of the oviducal epithelium from Pre-LH to Day 7, with a stronger staining in the isthmus than in the ampulla at all stages. Furthermore, the expression of oviduct-specific glycoprotein, detected by Western Blot, varied significantly with the stage (P &lt; 0.0001). The oviductin protein expression was at its lowest level at anestrus, then increased significantly at Pre-LH and Pre-ov (35- and 41-fold higher levels than anestrus, respectively), reached a maximal level at Day 1 (66-fold higher than anestrus), then decreased at Days 4 and 7 (47- and 20-fold higher than anestrus, respectively). In conclusion, this is the first report of oviductin protein expression in the canine oviduct. The region-specific higher expression of oviductin at Day 1 post-ovulation suggests a potential role of this glycoprotein in gamete maturation and fertilization in the bitch.

The relationship between vaginal mucous electric resistance and plasma progesterone concentration for optimal mating time in Beagle bitches

The objective of this experiment was to assess the relationship between electrical resistance of the vaginal mucosa and plasma progesterone for optimal mating time in the bitch. Eight mature beagle bitches were examined, and we observed eight times of estrus. Vaginal electric resistance was recorded weekly using a Draminski ovulation detector in anestrus, and daily in estrus. Plasma progesterone concentration was estimated by radioimmunoassay. In the bitch, incline in vaginal electric resistance (376.20 ± 105.63 units) showed a closely association with the onset of proestrus. Ovulation day was determined as the first day when plasma progesterone concentration increased above 5.0 ng/ml (Day 0). On Day 0, vaginal mucous electric resistance was 438 ± 132 units. Vaginal mucous electric resistance showed a slight decrease or was maintained until Day 0. However, it showed an explosive increase, and peaked on Day 1~3, which was above 600 units. Two of eight cases peaked on Day 1, three of eight cases were revealed on Day 2, and others were revealed on Day 3. After Day 4, resistance showed a rapid drop to below 600 units and reached 200 units on Day 8. The optimal mating time was determined when vaginal mucous electric resistance was above 600 units.

Immunolocalization of Progesterone Receptors in the Canine Oviduct around Ovulation

In the bitch, oocyte maturation, sperm storage, fertilization and early embryo development take place within the oviducts under high and increasing circulating progesterone concentrations. To investigate the potential effects of progesterone on the canine oviduct, nuclear progesterone receptors (PR) were localized. Oviducts were collected by ovariectomy from adult Beagle bitches during anestrus, after the onset of proestrus but prior to the Luteinizing Hormone (LH) peak (Pre-LH), after the LH peak but before ovulation (Pre-ov) and on Days 1, 4 and 7 after ovulation (n = 3 bitches per stage). The cellular distribution of PR was studied by immunohistochemistry (IHC) in the ampulla, isthmus and tubal part of the utero-tubal junction (UTJ). Plasma progesterone and 17β-oestradiol were assayed on the day of surgery. PR were specifically expressed in the nuclei of epithelial, stromal and muscular cells in the ampulla, isthmus and UTJ. The IHC scores did not vary from one oviductal region to another. However, the epithelium displayed higher scores than the stroma at anestrus, Pre-ov, Days 4 and 7, and also higher scores than muscle at Days 4 and 7 (p < 0.05). Immunohistochemistry scores in the stroma and muscle decreased at Days 4 and 7 compared with previous stages (p < 0.05). Furthermore, muscular IHC scores were positively correlated with circulating 17β-oestradiol concentrations and negatively correlated with circulating progesterone concentrations (p < 0.05). In conclusion, PR were identified in the canine oviduct, with differences in expression between tissues and times around ovulation, suggesting that progesterone may regulate tubal functions and reproductive events in this species.

Comparison Between Vestibular and Subcutaneous Insertion of Deslorelin Implants for Oestrus Induction in Bitches

Investigations using sustained-release deslorelin implants at various insertion sites have shown that this method consistently induces oestrus in anoestus bitches. However, fertility comparisons between implant insertion sites have not been performed. Anestrous beagle bitches received one 2.1 mg deslorelin implant beneath the vestibular mucosa (VM group; n = 6) or in the subcutaneous tissue between the shoulder blades (SubQ group; n = 8). Vestibular implants were removed when serum progesterone concentrations first exceeded 1.5 ng/ml. Vaginal cytologies and blood samples were collected daily and bitches were inseminated during oestrus. Serum progesterone and deslorelin concentrations were measured and pregnancy status was determined using ultrasonography. There were no differences between groups in the intervals between implant administration and the onset of proestrus, the time of the luteinizing hormone surge and the onset of cytologic diestrus. There were also no differences in the number of corpora lutea or foetuses. However, conception rate was significantly lower in the SubQ group. The pregnancy rate did not differ significantly between the VM and SubQ groups [4 out of 6 (66.7%) and 3 out of 8 (37.5%), respectively]. One bitch (16.7%) in the VM group and three bitches (37.5%) in the SubQ group suffered distinct, premature declines in serum progesterone concentrations starting 1-4 weeks after cytologic diestrus. Serum progesterone concentrations did not recover (premature luteal failure), resulting in abortion. Bitches with premature luteal failure in the SubQ group still had serum deslorelin concentrations >100 pg/ml 20 days after implant insertion, suggesting a possible association between prolonged deslorelin release and luteal failure.

33 REPRODUCTIVE CHARACTERISTICS OF CLONED DOGS DERIVED FROM SOMATIC CELL NUCLEAR TRANSFER

Several cloned dogs have been successfully produced by somatic cell nuclear transfer. However, there is no investigation of its reproductive characteristics including changes of hormone and ovarian follicles in cloned dogs. Thus, this study was to examine the onset of puberty, follicular dynamics, and reproductive hormone profiles in cloned beagle dogs. Two female cloned beagles, derived from fetal fibroblasts, were compared to 2 individual age- and weight-matched female beagles produced by natural breeding. All dogs were examined twice weekly from 6 months for the presence of swelling of the vulva and serosanguineous vaginal discharge, which were used as markers of the onset of proestrus. From the first day of proestrus, jugular venous blood samples were collected twice a day for hormonal analysis. Also, from the day when progesterone concentration was &gt;1 ng mL–1 until 5 days after ovulation, blood sampling was carried out 3 times a day. Ultrasound examination of ovaries and uterus was performed daily from the onset of proestrous until 5th day post-ovulation. Cloned female dogs (332.5 ± 1.5 days) reached puberty later than controls (300 ± 8 days). The mean number of ovarian follicles was 4.5 ± 0.5 v. 6.5 ± 0.5 for clones and controls, respectively. The largest size of ovarian follicles in clones and controls detected by ultrasound were 0.6 ± 0.03 v. 0.69 ± 0.07 cm, respectively. It took 8 days from the initiation of vulvar bleeding to the LH peak in all beagles. Moreover, there were no differences in the profile of hormonal changes (LH, FSH, estradiol, and progesterone) in cloned and control beagles. These results demonstrate that cloned dogs have normal development of reproduction. This study was financially supported by the Korean MEST, through KOSEF (grant # M10625030005-08N250300510) and the BK21 program for Veterinary Science.

Ultrastructure of canine oocytes during in vivo maturation

The aim of the present study was to describe the canine oocyte ultrastructural modifications during in vivo maturation, with precise reference to the timing of the LH surge and of ovulation. Twenty-five bitches were ovariectomized at specific stages between the onset of proestrus and the fifth day post-ovulation: 65 oocytes were observed by transmission electron microscopy (TEM), either before the LH surge (n = 10), between the LH surge and ovulation (n = 12) or after ovulation (n = 43). Prior to the LH surge, the oocyte nucleus had already begun its displacement to the vicinity of the oolemma and reticulated nucleoli were infrequent. The cytoplasm showed signs of immaturity (few organelles preferentially located in the cortical zone, "mitochondrial cloud", scarce cortical granules). The LH surge was immediately followed by cumulus expansion but the ovulation occurred 2 days later. Retraction of the transzonal projections and the meiotic resumption occurred after another 3 days (5 days after the LH peak). The ovulation was then followed by gradual cytoplasmic modifications. Nucleoli re-assumed a reticulated aspect around 24 hr post-ovulation. From 48 hr post-ovulation mitochondria and SER were very numerous and evenly distributed. In conclusion canine oocyte maturation began prior to the LH surge and no cytoplasmic or nuclear modifications followed immediately the LH surge and ovulation. This study suggests that two distinct signals are needed for the final in vivo maturation: one prior to the LH surge (to induce maturation) and another one, around 3 days post-ovulation (to induce meiotic resumption).

Plasma progesterone and prolactin concentrations in overtly pseudopregnant bitches: A clinical study

Plasma concentrations of progesterone (P 4) and prolactin (PRL) were measured in 35 bitches presented at veterinary clinics for symptoms of overt pseudopregnancy (PSP) between 50 and 95 days after the onset of proestrus. Results were compared to those from samples collected from 35 control bitches at comparable stages of the ovarian cycle (expressed as days after the onset of observed signs of proestrus). In the PSP bitches at 71.4 ± 1.6 (mean ± S.E.M.) days of the cycle, P 4 (1.5 ± 0.2 ng/mL) was lower ( P < 0.01) and PRL (16.0 ± 1.9 ng/mL) was higher ( P < 0.01), compared to P 4 (2.7 ± 0.4 ng/mL) and PRL (2.9 ± 0.6 ng/mL) in control bitches at 70.6 ± 1.5 days of the cycle. Low P 4 was not a prerequisite for elevated PRL. Although elevated (≥10 ng/mL) PRL (20.9 ± 2.0 ng/mL) occurred more often with low (<2 ng/mL) P 4 (20 of 24 cases) it also occurred with P 4 above 3 ng/mL in two affected bitches and in two control bitches. Whether the occurrence of relatively low PRL concentrations (<4 ng/mL) in samples obtained from 4 of the 35 pseudopregnant bitches reflected variable and often elevated PRL secretion or increased sensitivity to PRL in the absence of elevated prolactin in those animals was not determined. We inferred that elevated plasma PRL was often involved in the etiology of overt PSP; furthermore, a premature decline in circulating P 4 concentrations may be a factor in some instances.

The effects of a low dose of cabergoline on induction of estrus and pregnancy rates in anestrous bitches

This is the first report of successful induction of normal estrus and ovulation in breeder bitches with as a low dose as 0.6 μg/kg/day of cabergoline formulation marketed for use in women. Sixty-one pure breed bitches from various breeds were used in the study at their already determined periods of anestrus. Twenty-four dogs formed the control group, while 37 bitches were administered with two different doses of cabergoline (recommended dose group, n = 10, 5 μg/kg/day and low dose group, n = 27, 0.6 μg/kg/day). Induced estrus rates and mean treatment and proestrus durations of dogs in these two dose groups were compared. At the second phase of the study, the effects of 500 IU human chorionic gonadotropin (hCG) administered on days 1 and 3 of estrus induced by the low dose of cabergoline, on the duration of behavioral estrus, ovulation rates, pregnancy rates and the number of offspring were investigated. For this purpose, the dogs with signs of proestrus (22/27) following the treatment in the low dose group were assigned into two subgroups. Five hundred IU of hCG (Pregnyl, Organon, Turkey) was intramuscularly administered to eight of these dogs [low dose (hCG+) group] on days 1 and 3 of estrus. The remaining 14 dogs were not treated with hCG [low dose (hCG−) group]. An aqueous solution of cabergoline (Dostinex, Pharmacia, Italy) was orally administered until 2 day after the onset of proestrus or for a maximum of 42 days. Blood samples were taken daily from all treatment and 11 control bitches during the first five days of behavioral estrus to measure progesterone concentrations. In the recommended dose and low dose groups, estrus was induced between days 8–45 and 4–48 (mean: 23.63 ± 14.33 and 24.41 ± 14.31 days), in the ratio of 80.0 and 81.5%, respectively ( p > 0.05). In both dose groups, post-treatment interestrous intervals were significantly shorter than both those of the control group and their own pre-treatment interestrous intervals ( p < 0.05). Ovulation rates, pregnancy rates and mean number of offspring delivered by the dogs in the recommended dose, low dose (hCG−), low dose (hCG+) and control groups were found to be similar ( p > 0.05). However, the mean duration of behavioral estrus of the dogs in the low dose (hCG+) group was found to be significantly longer compared to dogs in all other groups ( p < 0.05). In both dose groups, no correlation could be found between the anestrus stages and treatment durations ( p > 0.05). Shortly, it has been concluded from the study that (1) normal and fertile estrus can be induced more economically in bitches during different stages of anestrus using as a low dose of 0.6 μg/kg of cabergoline formulation marketed for use in women, and that (2) hCG injections on days 1 and 3 of the estrus induced by this method has no positive effects on the ovulation rates, pregnancy rates and the number of offspring per pregnancy.

Polygonum hydropiper crude root extract mimics estrogenic properties in females: Evidence of uterine protein profiles studied by sodium dodecyl sulfate polyacrylamide gel electrophoresis.

Polygonum hydropiper is a widely grown weed in the north-eastern states of India. In the present study, estrogenic effects of the crude root extract (CRE) of Polygonum hydropiper on uterine protein was tested in ovary-intact and ovariectomized (OVX) female albino rats. The methanolic crude extract of Polygonum hydropiper was given to adult ovary-intact and OVX female albino rat in a dose of 1000mg/kg bodyweight per day. Oral administration of the CRE was carried out for a period of 12days from the onset of proestrus and for a similar period of time for the OVX female. To study the estrogenic effect, OVX females were subcutaneously injected with 0.1µg of estradiol-17β (E2). E2 was injected at the interval of 24h for a total period of 72h. The uterine protein of ovary-intact and OVX females was studied by single dimensional sodium dodecyl sulfate polyacrylamide gel electrophoresis. The results showed similar protein bands in OVX females treated with E2 and CRE. In addition, treatment with CRE stimulated expression of more proteins in the uterus. The results showed that CRE of Polygonum hydropiper mimics the effect of estradiol-17β in the uterine protein profiles of adult female albino rats. (Reprod Med Biol 2006; 5: 155-160).

Polygonum hydropiper crude root extract mimics estrogenic properties in females: Evidence of uterine protein profiles studied by sodium dodecyl sulfate polyacrylamide gel electrophoresis

Polygonum hydropiper is a widely grown weed in the north‐eastern states of India. In the present study, estrogenic effects of the crude root extract (CRE) of Polygonum hydropiper on uterine protein was tested in ovary‐intact and ovariectomized (OVX) female albino rats. The methanolic crude extract of Polygonum hydropiper was given to adult ovary‐intact and OVX female albino rat in a dose of 1000 mg/kg bodyweight per day. Oral administration of the CRE was carried out for a period of 12 days from the onset of proestrus and for a similar period of time for the OVX female. To study the estrogenic effect, OVX females were subcutaneously injected with 0.1 µg of estradiol‐17β (E2). E2 was injected at the interval of 24 h for a total period of 72 h. The uterine protein of ovary‐intact and OVX females was studied by single dimensional sodium dodecyl sulfate polyacrylamide gel electrophoresis. The results showed similar protein bands in OVX females treated with E2 and CRE. In addition, treatment with CRE stimulated expression of more proteins in the uterus. The results showed that CRE of Polygonum hydropiper mimics the effect of estradiol‐17β in the uterine protein profiles of adult female albino rats. (Reprod Med Biol 2006; 5: 155–160)

169 PREDICTION OF THE OPTIMAL DAY OF MATING IN THE BITCH USING A BOVINE PLASMA PROGESTERONE ELISA KIT

The present study was conducted to predict the optimal day of mating by measurement of progesterone concentration in peripheral blood of bitches using a bovine plasma progesterone ELISA kit (enzyme-immunoassay practice P kit; Kawasaki Mitaka K.K., Mitaka-shi, Tokyo, Japan). In the first experiment, blood samples were collected from 20 mature beagle bitches once a day from 1 to 13 days after the onset of proestrus (Day = 0). All bitches were mated with male beagle dogs twice on Day 9 and Day 12 after the onset of proestrus and then allowed to undergo parturition. In the second experiment, blood samples were collected from 16 mature and 7 prepubertal beagle bitches; the blood collections were randomly started between Day 4 and Day 6 and performed three times after the initiation of blood collection (the first, third, and fourth days). The bitches with progesterone concentration of &amp;lt;10 ng mL-1 at the end of progesterone analysis were mated with male beagle dogs twice at 3-day intervals between Day 9 and Day 15 according to the hormonal profile. Data were compared by chi-square analysis. In the first experiment, a total of 20 bitches were mated with male dogs irrespective of hormonal profiles, of which 17 bitches became pregnant and delivered offspring. Two of the non-pregnant bitches did not show an increase of progesterone concentration during the progesterone analysis period, and the other one showed a high progesterone concentration (&amp;gt;10 ng mL-1) before the mating. In the second experiment, 6 of 23 bitches were excluded from the mating with male dogs because their progesterone concentrations had reached &amp;gt;10 ng mL-1 by the second day of progesterone analysis. The percentage of prepubertal bitches with high progesterone concentration before the mating (4/7, 57%) was significantly higher (P &amp;lt; 0.05) than that of mature bitches (2/16, 13%). A total of 17 bitches were mated with male dogs, of which 15 bitches became pregnant and delivered offspring. These results indicate that the bovine plasma progesterone ELISA kit is useful for the prediction of the optimal day of mating with male dogs, and progesterone analysis may be effective for exclusion of bitches that have ovulated before mating.

Shortening of Interestrous Intervals With Cabergoline in Bitches: A Clinical Trial

Two consecutive interestrous intervals (n=46) were recorded in 23 bitches of different breeds. At varying times after day 100 from the onset of the second proestrus, cabergoline (5 microg/kg per os q 24 hours) was administered from early (n=11), mid- (n=10), and late (n=2) anestrus until 2 days after the beginning of the following proestrus. Interestrous intervals (IEI) were significantly shorter in the cabergoline-treated time periods when compared to the nontreated IEI (184+/-4.5 days versus 239+/-4.5 days; P<0.01). The mean number of days of cabergoline treatment until the onset of proestrus was 21.4+/-2.9 (least square means and standard error of the mean [LSM+/-SEM]). Mean cabergoline treatment durations beginning in early, mid-, and late anestrus were 27.4+/-3.7, 17.6+/-3.8, and 5+/-3 days (LSM+/-SEM), respectively. A significant correlation was found between the stage of anestrus in which the treatments began and the duration of the treatments required to induce estrus (0.51, P=0.01).

Use of cabergoline to treat primary and secondary anestrus in dogs.

To determine whether cabergoline would be safe and effective for induction of estrus in dogs with primary or secondary anestrus. Prospective case series. 6 privately owned otherwise healthy pure-bred dogs with primary or secondary anestrus. Dogs were treated with cabergoline (5 microg/kg [2.3 microg/lb], p.o., q 24 h) until 2 days after the onset of proestrus. Follicular development was assessed by means of cytologic examination of vaginal smears; ovulation was assessed by measuring serum progesterone concentration 3 weeks after the onset of estrus. Five bitches were mated during behavioral estrus. All dogs had normal estrus periods, and all 5 dogs that were mated whelped normal litters. Mean duration of cabergoline treatment was 16 days. None of the dogs had any adverse effects associated with cabergoline administration. Results suggest that administration of cabergoline is safe and effective for treatment for primary and secondary anestrus in dogs.

Assessment of optimal mating time in the bitch

THE bitch is described as being monoestrus, which means that a long period of anoestrus occurs between successive periods of oestrus, whether she becomes pregnant or not. Correct monitoring and...