Comparison of progesterone assay by chemiluminescence or radioimmunoassay for clinical decision-making in canine reproduction.

The Coat-A-Count® radioimmunoassay has been long and widely used to determine the concentration of progesterone in serum or plasma of bitches (progRIA), but was discontinued in 2014. The Immulite® 1000 LKPG1 chemiluminescence immunoassay has gained prominence since 2003 to determine the concentration of progesterone in serum of bitches, but the assay changed in 2012 (Immulite® 1000 LKPW1). This study assessed the feasibility of using Immulite® 1000 LKPW1 (progImm) to estimate the time of clinically relevant events during oestrus and compared progRIA and progImm 2 and 3 days after the first or only day of the luteinizing hormone surge (LH1). ProgImm first exceeded 5.1 nmol/L on the same day that progRIA first exceeded 6 nmol/L, a proxy for the occurrence of the LH surge, or the day before in 28 of 31 (90%) of oestrous periods. ProgImm first exceeded 13.6 nmol/L on the same day that progRIA first exceeded 16 nmol/L (a proxy for the day of ovulation) or the day before in 34 of 35 (97%) oestrous periods. ProgImm first exceeded 5.4 nmol/L on LH1 or the day before in 24 of 25 (95%) of oestrous periods. The median of progImm 2 days after LH1 was 1.2 nmol/L lower than the 10.7 nmol/L of progRIA (p = 0.001). The mean of progImm 3 days after LH1 was 2.2 nmol/L lower than the 19.0 nmol/L of progRIA (p 0.001). In conclusion, the days on which progImm first exceeded 5.1 nmol/L, 13.6 nmol/L and 5.4 nmol/L effectively estimate the days on which progRIA reached 6 nmol/L or 16 nmol/L or LH1.

An objective volumetric method for assessment of ovarian follicular and luteal vascular flow using colour Doppler ultrasonography.

Our goal was to develop an objective computer-assisted volumetric method of assessing vascular flow from colour Doppler ultrasound data of ovarian structures recorded by free-hand movement. We hypothesized that a vascularity index (ratio of the region of blood flood to the region of ovarian structure) obtained from the three-dimensional volumetric analysis would be more precise (less variable) than conventional two-dimensional analysis of single images in estimating the functional status of the preovulatory follicles and corpus luteum. Doppler ultrasound cineloops of water buffaloes (Bubalus bubalis; n = 22) ovaries were recorded daily from 12 h before GnRH treatment to four days after ovulation. Cineloops were processed using Fiji and Imaris software packages for segmenting the area (two-dimensional analysis) and the volume (three-dimensional analysis) occupied by the blood-flow and associated tissue to calculate the vascularity index. For volumetric measurement, all images in a cineloop were used (i.e., no a-priori selection of images) while for two-dimensional analysis, three images from the region with apparent maximum vascularity were selected. The volumetric method was verified with theoretical ellipsoidal volume of the follicle (r = 0.96 P < 0.01) or corpus luteum (r = 0.58 P = 0.02). The variability in the follicular vascularity index among animals was lower using the volumetric method than two-dimensional analysis (0.018 ±â€¯0.002 vs 0.030 ±â€¯0.005, P < 0.01), while the variability for CL vascularity was similar between methods (P = 0.23). An increase in the follicular vascularity index was detected at 12 h after GnRH treatment using both methods (two-dimensional: 0.030 ±â€¯0.008, P < 0.01; three-dimensional: 0.016 ±â€¯0.006, P < 0.02). Buffaloes that ovulated tended to have a greater increase in 3D vascularity index than non-responding buffaloes (P = 0.06); the two-dimensional method was not able to detect these changes. Using the three-dimensional method, a moderate positive correlation (r = 0.59; P = 0.02) was evident between the follicular vascularity index at 14-16 h after GnRH treatment and follicular diameter. In conclusion, an objective volumetric method for assessing relative ovarian blood flow changes was developed using Doppler ultrasound cineloops recorded by free-hand movement. The 3-dimensional method eliminates the need for a-priori selection of images and is more precise as a result of decreased technical variability.

Use of injectable progesterone and hCG for fixed-time artificial insemination during the non-breeding season in goats.

The objective of this study was to determine the estrous response and the moment of ovulation and fertility after a progesterone (P4) priming plus human chorionic gonadotropin (hCG) administration in multiparous and nulliparous goats. Therefore, two experiments were conducted during non-breeding season (April and May, 26° N) and all the animals received a single injection of 20 mg of P4 and 100 IU of hCG 24 h later. In Experiment 1, 13 multiparous and 9 nulliparous goats were subjected to estrus detection twice a day from P4 administration, and their ovaries were scanned by transrectal ultrasonography every 12 h from the onset of estrus to determine ovulation. The proportion of goats in estrus did not differ between multiparous and nulliparous females. The onset of estrus (60.5 ±â€¯12.4 h vs. 52.0 ±â€¯5.2 h after hCG administration) and the moment of ovulation (91.5 ±â€¯10.3 h vs. 85.3 ±â€¯11.5 h) were also similar in multiparous and nulliparous goats. In Experiment 2, a total of 299 multiparous and nulliparous goats managed under intensive (n = 112 and 41 goats, respectively) or extensive (n = 85 and 61 goats, respectively) production systems were divided to receive a fixed-time artificial insemination (FTAI) with fresh semen at 60 or 72 h after hCG administration. The pregnancy rate did not differ significantly between multiparous and nulliparous goats in both production systems. Nonetheless, in the intensive system pregnancy rate was affected by the moment of insemination (P < 0.05). In this system, the pregnancy rate was higher in goats inseminated at 60 h than in 72 h (55.6%, 44/79 vs. 35.1%, 26/74, respectively; P < 0.05). On the contrary, in the extensive system the pregnancy rate was not affected by the time of insemination (29.4%, 23/78 vs. 22.0%, 15/68). To conclude, both the ovulatory response and the pregnancy rate after a single P4 injection plus hCG was similar between multiparous and nulliparous goats during anovulatory season. Although the pregnancy rate was not affected by the time of insemination in the extensive production system, under intensive conditions FTAI should be performed at 60 h after hCG treatment.

Canine ovulation timing: A survey on methodology and an assessment on reliability of vaginal cytology.

Ovulation timing in bitches is a routine procedure in small animal practice around the world. It is most frequently used for supporting breeding management, high-risk pregnancy monitoring and determination of the time of parturition. To learn more about how and for what reasons veterinarians interested in small animal reproduction perform canine ovulation timing, an online survey was conducted in 2017. The link to the survey was distributed via the mailing list cafereprod-l@list.cornell.edu and the website, the Twitter account and the Facebook page of EVSSAR. All respondents recommended using quantitative progesterone measurement for ovulation timing alone or in combination with other diagnostic tests. Vaginal cytology was also a commonly used technique. The 63 respondents followed different protocols for sampling and staining vaginal epithelial cells. Interestingly, 50 respondents used vaginal cytology routinely, but only two consider it as a very reliable and another nine as a somewhat reliable test if used alone. In a second project, delegates attending the EVSSAR congress held in Vienna, Austria, in June 2017 had the opportunity to examine seven stained vaginal smear slides under optical microscopy in a blinded approach. The results showed a marked heterogeneity in the identification of vaginal cells and assessment of the time in relation to ovulation. This heterogeneity supports the opinion of survey participants that vaginal cytology alone is not a valid tool for determining the day of ovulation. Results of both projects suggest that there is no standard procedure for the examination of vaginal smears in dogs. It is not recommended to determine the optimal time for mating based on the examination of a single vaginal smear only. According to some comments of survey participants, it is more useful to assess vaginal samples repeatedly and to use the findings to determine when to start progesterone measurement.

Dynamics of putative sex pheromone components during heat periods in estrus-induced cows.

Determination of the optimal insemination time in dairy cows is vital for fertilization success and is a challenging task due to silent or weak signs of estrus shown by some cows. This can be overcome by combining several estrus detection methods, leading to higher detection rates. However, an efficient, noninvasive method for detecting estrus in cows is still needed. Chemical cues released by the cow during estrus have been proposed to have pheromonal properties and signal readiness to mate to the bull. Such cues could be used in an industrial setting to detect cows in estrus. However, no conclusive published data show temporal changes in putative sex pheromone levels during estrus. The goal of this study was to determine the temporal pattern of putative sex pheromone components during estrus and to assess the reproducibility of changes in pheromone concentration with respect to ovulation time. Two injections of the hormone PGF2α were administered over a 2-wk interval to induce and synchronize the estrous cycles of 6 Holstein cows. The precise time of ovulation was determined by means of an ultrasound technique, and estrus was determined by visual observation. Using solid-phase microextraction gas chromatography-mass spectrometry techniques, we showed that acetic and propionic acids, which have been proposed to be putative sex pheromone components in cows, were present in the headspaces of all estrous and diestrous fecal samples, whereas 1-iodoundecane was not detected by solid-phase microextraction or by solvent extraction with diethyl ether. Low levels of acids were observed until 1 d before ovulation, at which point their concentrations increased, peaking around 0.5 d before ovulation. The application of labeled synthetic standards revealed that during the peak of release, 36 ± 8 ng (average ± SD) of acetic acid and 10 ± 3 ng of propionic acid were present in 0.5-g samples of estrous-phase fecal matter compared with 19 ± 5 and 2.3 ± 1 ng of acetic and propionic acids, respectively, in the control diestrous samples. After the peak, the amounts of the compounds decreased sharply to match those of the control samples and afterward returned to the baseline readings. This decrease in the amounts of putative pheromone components was registered about 12 h before ovulation, indicating that acetic and propionic acids could be used as biomarkers for the electronic detection of ovulation.

Reproductive analysis of male and female captive jaguars (Panthera onca) in a Colombian zoological park.

A reproductive analysis of a captive group of jaguars (Panthera onca; n = 6) at the Santacruz Zoological Foundation in Cundinamarca, Colombia, was conducted by performing a longitudinal, noninvasive, hormonal analysis of estradiol and progestogens in females and of androgens in males. During four seasons, female jaguars confined in solitary were evaluated for ovarian activity and spontaneous ovulation, male jaguars for testicular activity. A second hormonal follow-up was conducted in the females after administration of gonadotropins. Hormones were extracted from fecal samples of three females (n = 3) and two males (n = 2). Estradiol measurements were obtained by RIA and progestogens by enzyme immunoassay. The linear mixed-effect regression showed that there was a significant effect of seasons in the concentrations of estradiol (chi square = 15.97, degrees of freedom = 3, P < 0.01). Posthoc comparisons of all pairs of seasonal means were conducted according to Tukey's honest significant difference, revealing significant differences between seasons: Dry 1 versus Rains 2 (P < 0.01), Rains 1 versus Rains 2 (P < 0.05), and Dry 2 versus Rains 2 (P < 0.05). Elevations of progestogens compatible with spontaneous ovulation occurred in three jaguars, and the linear mixed-effect regression showed that there was also a significant effect of seasons (chi square = 28.56, degrees of freedom = 3, P < 0.01). Posthoc comparisons showed significant differences only between seasons: Dry 2 versus Rains 2 (P < 0.01). The season with the lowest average concentration was Rains 2 (October, November, and December). During this season, periods of anestrous were registered that lasted between 31 and 83 days. The three females presented estradiol peaks after the administration of eCG. A noninvasive longitudinal analysis for androgens was also made (males 1 and 2) over the course of 1 year, and no significant differences were found between the different seasons. A seminal analysis of three adult male jaguars (Panthera onca; n = 3) was also performed after electroejaculation under general anesthesia (male 1 and 2) and by a postmortem epididymal wash (male 3). The mean concentration of spermatozoids was 5.7 × 106 ± 1.1 × 106 spermatozoa/mL. The progressive motility + standard deviation averaged 80%. The percentage of normal spermatozoids obtained by electroejaculation was 80 ± 2.8%, and the abnormalities found more frequently were head defects (7 ± 1.4%). The seminal fluid obtained by epididymal flush contained 35 ± 1.4% normal spermatozoids, and the most frequent abnormalities found corresponded to distal cytoplasmic droplets (39 ± 11.3%).

Synchronization of ovulation in cattle with an aromatase inhibitor-based protocol.

A study was designed to determine the effect of stage of the estrous cycle on the proportion of animals that ovulated and the synchrony of ovulation of heifers treated with an aromatase inhibitor-based protocol. Forty-eight heifers were treated intramuscularly with 500 µg of cloprostenol (PGF) followed by 100 µg of GnRH 24 hours later to serve as control data for comparison of the ovulatory response to a subsequent aromatase inhibitor protocol. Daily ultrasound examinations were done to determine the incidence of and interval to ovulation. At the time of ovulation (Day 0), heifers were assigned randomly to five day-groups (n = 8-11/group) and given an intravaginal device containing 3 g of letrozole for 4 days starting on Day 0, 4, 8, 12, or 16. At the time of device removal, heifers were given PGF followed by GnRH 24 hours later. Ultrasound examinations were done daily from 2 days before device insertion to 9 days after the posttreatment ovulation. The preovulatory follicle diameter after letrozole treatment was larger in the Day 4 group compared to the Day 0 and 16 groups and intermediate in the Day 8 and 12 groups (P < 0.001). Compared to control data, the percentage of heifers that ovulated after letrozole treatment was greater (87.1% vs. 69.4%, respectively; P < 0.05) as was the synchrony of ovulation (residuals: 0.24 ± 0.07 vs. 0.68 ± 0.13; P < 0.01). The day on which letrozole treatment was initiated did not affect the proportion of heifers that ovulated or the interval to ovulation. Plasma estradiol concentrations at the time of removal of the letrozole device in the Day 0 and 4 groups was lower (P < 0.05) than in the corresponding controls. Estradiol concentrations in the Day 8 and 12 groups did not differ from already low concentrations in the respective controls. Corpus luteum diameter profiles and progesterone production were not affected by day-group although reduced luteal lifespan after letrozole treatment was observed and requires further investigation. In summary, a protocol involving a letrozole-impregnated intravaginal device for 4 days, PGF treatment at device removal, and GnRH 24 later resulted in a greater ovulation rate and greater synchrony of ovulation than in heifers not given letrozole. Results suggest that the protocol may be initiated effectively at random stages of the estrous cycle and may provide impetus for further studies to assess the efficacy of a letrozole-based synchronization protocol for fixed-time insemination.

Variations in the vulvar temperature of sows during proestrus and estrus as determined by infrared thermography and its relation to ovulation.

The prediction of ovulation time is one of the most important and yet difficult processes in pig production, and it has a considerable impact on the fertility of the herd and litter size. The objective of this study was to assess the vulvar skin temperature of sows during proestrus and estrus using infrared thermography and to establish a possible relationship between the variations in vulvar temperature and ovulation. The experimental group comprised 36 crossbred Large White × Landrace females, of which 6 were gilts and 30 were multiparous sows. Estrus was detected twice daily and the temperature was obtained every 6 hours from the vulvar area and from two control points in the gluteal area (Gluteal skin temperature [GST]). A third variable, vulvar-gluteal temperature (VGT) was obtained from the difference between the vulvar skin temperature and the GST values. The animals were divided into two subgroups: group A consisting of 11 animals with estrus detected at 6:00 AM, Day 4 postweaning, and group B comprising seven animals with estrus detected at 6:00 AM, Day 5 post-weaning. Both groups showed a similar trend in the VGT. The VGT increased during the proestrus, reaching a peak 24 hours before estrus in group A and 48 hours before estrus in group B. The VGT then decreased markedly reaching the lowest value in groups A and B, respectively, 12 and 6 hours after estrus. Although the time of ovulation was only estimated on the basis of a literature review, the matching between the temporal variations of the VGT values and the predicted time of the peak of estradiol secretion that ultimately leads to the ovulation processes suggests that the VGT values represent a potential predictive marker of the ovulatory events.

Seasonal fluctuations in the response of Italian Mediterranean buffaloes to synchronization of ovulation and timed artificial insemination.

A comprehensive study of the efficiency of synchronization of ovulation and timed artificial insemination (TAI) was undertaken in a large group of Italian Mediterranean buffaloes at a commercial dairy. A total of 2791 synchronization protocols were carried out on 857 animals over 3 years. Of these protocols, 823 (29.5%) did not proceed beyond Day 7 (due to the absence of a vascularized CL) and 620 (22.2%) were discontinued on Day 10 (due to the absence of follicles >1.0 cm and tonic uteri); hence, 1443 (51.7%) protocols did not progress to TAI. Data were analyzed for four periods: P1, transition to spring (from breeding season to low breeding season); P2, low breeding season; P3, transition to fall (low breeding season to breeding season); and P4, breeding season. No differences were found among the four periods in terms of the proportion of protocols that did not result in TAI. Of the 857 buffaloes, 660 (77%) conceived and delivered a calf. The average number of TAI per pregnancy was 2.1 and ranged from 1.9 to 2.3 across years. Logistic regression analysis showed that buffaloes that calved during P3 had a higher odds ratio for pregnancy (1.380; P < 0.05) than buffaloes that calved in other periods. Pregnancy was also influenced by the calving to service period (odds ratio = 0.977; P < 0.01) and the pregnancy per AI (P/AI) at successive TAI (odds ratio = 1.480; P < 0.01). The pregnancy per AI at the first TAI (424/857, 49.5%) was greater (P < 0.01) than in subsequent TAI. The occurrence of late embryonic mortality (between Days 27 and 45 after TAI) was similar among the four periods. These findings indicated that there are distinct seasonal differences in the response of Italian Mediterranean buffaloes to synchronization and TAI.

Monitoring ovarian cycle activity via progestagens in urine and feces of female mountain gorillas: A comparison of EIA and LC-MS measurements.

Understanding the reproductive biology of endangered mountain gorillas (Gorilla beringei beringei) is essential for optimizing conservation strategies, determining any demographic impact of socioecological changes, and providing information for comparative studies of primates. Non-invasive techniques have been used to assess the reproductive function of many primates and the importance of validating the measurements of hormones metabolites is widely recognized because they may vary even within closely related species. To determine if it is possible to non-invasively monitor ovarian activity in wild mountain gorillas, we used enzyme immunoassays (EIA) to quantify both urinary and fecal excretion of immunoreactive pregnanediol-3-glucuronide (iPdG), defined as all metabolites detected by a pregnanediol-3-glucuronide immunoassay (PdG EIA). Simultaneously, we performed the liquid chromatography mass spectrometry (LC-MS) to quantify the excretion of pregnanediol in urine and feces. Samples were analyzed over nine cycles of five females from the habituated gorillas monitored by Karisoke Research Center, Rwanda. As an additional indicator for ovulation timing, estrone conjugates (E1C) were measured in a subset of urine samples. The concentrations of iPdG and pregnanediol measured in the same samples were significantly correlated. Urinary concentrations of iPdG and pregnanediol fluctuated over the menstrual cycle but did not reveal any cyclic pattern, whereas a typical preovulatory urinary E1C surge and postovulatory increases of fecal iPdG and pregnanediol were detected. The luteal peaks of iPdG and pregnanediol levels in feces were on average 2.8 and 7.6 times higher, respectively, than averaged levels in the corresponding follicular phase. The relative number of days with observed matings was higher within the presumed fertile window than in the preceding period. Overall, the results indicate that fecal analysis of iPdG and pregnanediol is suitable for detecting ovulation in female mountain gorillas. Urinary measurements using both EIA and LC-MS appeared to be uninformative for monitoring ovarian activity in this primate.

Determination of ovulation time in sows based on skin temperature and genital electrical resistance changes.

Different physical and physiological parameters may be used to determine ovulation time in sows. In the present study, we analysed the ear and vulvar skin temperature fluctuations, and the changes in genital electrical resistance, at a distance of 4, 8 and 12 cm from the vulva during oestrus in order to predict the time of ovulation. Multiparous sows were checked by transrectal real-time ultrasonography and luteinising hormone (LH) plasma concentration was determined. Temperature was measured using a thermoprecision infrared thermometer, and the electrical resistance was measured with a commercial resistance probe. All measurements were carried out every 12 hours from one day after the weaning to three days after oestrus onset. Skin temperature showed significant difference around periovulatory period. The electrical resistance at 4 cm from the vulva showed marked changes during oestrus, which were different from those described at 8 and 12 cm from the vulva. At 12 hours before ovulation time, skin temperature decreased significantly, and negative correlation (P<0.05) was found between vulvar skin temperature and vaginal resistance. There was no relationship between skin temperature, electrical resistance and LH plasma concentration. The measurement of several physiological traits may provide more accurate predictions of the moment of ovulation.

Changes in blood flow in ovine follicles and serum concentration of estradiol 17 beta (E2) and nitric oxide (NO) around the time of ovulation in Ossimi ewes.

The aim of the present study was to examine the relation between follicular blood flow of the ovulatory follicle and the levels of serum E2 and nitric oxide (NO) in Ossimi ewe. Seven cyclic ewes were synchronized with a double injection PGF2α. The follicular wave was examined daily until ovulation (disappearance of the large dominant follicle ultrasonographically) with transrectal color Doppler ultrasonography (8-10MHz linear array transducer). The number of recruited follicles was 4.8±0.9 (3-8 follicles) with diameter of 2.8±0.1mm. The interval from PGF2α injection to follicle deviation was 2.35±0.07 days. The diameter of the first largest follicle (LF1) at recruitment day was 4±0.3mm while the diameter of the second largest follicle (LF2) was 3.7±0.1mm. The diameter of LF1 at the day of deviation was 5.1±0.5mm while the diameter of the LF2 was 4±0.7mm. The diameter of the ovulatory follicle was 6.1±0.5at day of ovulation. We detected the blood flow area of the ovulatory follicle at D2. At ovulation, the blood flow area and blood flow area percent increased significantly to be 11.9±0.6mm(2) and 44±3.4% respectively. The results showed a positive correlation between E2 and NO (r=0.85, P<0.009). Both increased concomitantly with the diameter of the ovulatory follicle. Besides, NO and E2 reached a maximum level at ovulation (12.1±1.8ng/ml and 16.4±1.7pg/ml respectively).

Follicular-phase concentrations of progesterone, estradiol-17ß, LH, FSH, and a PGF2α metabolite and daily clustering of prolactin pulses, based on hourly blood sampling and hourly detection of ovulation in heifers.

Circulating concentrations of hormones were determined each hour in 13 heifers from the end of the luteolytic period to ovulation (follicular phase, 3.5 days). Diameter of the preovulatory follicle was determined every 8 hours, and the time of ovulation was determined hourly. The diameter of the preovulatory follicle decreased 0.8 ± 0.1 mm/h in heifers when there was 1 to 3 hours between the last two diameter measurements before ovulation. The concentration of progesterone (P4) after the end of the luteolytic period (P4 < 1 ng/mL) changed (P < 0.0001), as shown by a continued decrease until Hour -57 (Hour 0 = ovulation), then was maintained at approximately 0.2 ng/mL until 2 hours before the peak of the LH surge at Hour -26, and then a decrease to 0.1 ng/mL along with a decrease in estradiol-17ß. Concentrations of LH gradually increased (P < 0.007) and concentrations of FSH gradually decreased (P < 0.0001) after the end of luteolysis until the beginning nadirs of the respective preovulatory surges. A cluster of prolactin (PRL) pulses occurred (P < 0.0001) each day with approximately 24 hours between the maximum value of successive clusters. Hourly concentrations of a PGF2α metabolite decreased (P < 0.007) until Hour -40, but did not differ among hours thereafter. Novel observations included the gradual increase in LH and decrease in FSH until the beginning of the preovulatory surges and follicle diameter decrease a few hours before ovulation. Results supported the following hypotheses: (1) change in the low circulating P4 concentrations during the follicular phase are temporally associated with change in LH concentrations; and (2) PRL pulses occur in a cluster each day during the follicular phase of the estrous cycle.

Chronology of early embryonic development and embryo uterine migration in alpacas.

The objectives were to: (1) describe the chronology of early embryonic development from ovulation to entry into the uterus; and (2) to determine the timing of embryo migration to the left uterine horn when ovulation occurred from the right ovary. The experiment was conducted in Peru. Females (n = 132) were randomly assigned to 15 experimental groups. All females were mated to an intact male, given 50 µg GnRH im (Cystorelin) and ovulation time determined by transrectal ultrasonography, conducted every 6 hours, starting 24 hours postmating. Animals were slaughtered at a specific intervals postovulation and reproductive tracts were recovered and subjected to oviductal and uterine flushing for females slaughtered between 1 and 6 days postovulation (dpo; Day 0 = ovulation) and uterine flushing for females slaughtered from 7 to 15 dpo for recovery of oocytes/embryos. Season of mating did not influence the interval from mating to ovulation (winter: 29 ± 6 hours vs. summer: 30 ± 6 hours; P = 0.49). Ovulation rates for females mated during winter and summer were 92% versus 100%, respectively (P = 0.05). Fertilization rates for winter and summer mated females were 72% and 82% (P = 0.29). Unfertilized ova were not retained in the uterine tube. All embryos collected were in the uterine tube ipsilateral to the side of ovulation between 1 and 5 dpo. Embryos reached the uterus on 6 dpo. Embryos began to elongate on 9 dpo; at this time, 83% of embryos derived from right-ovary ovulations were collected from the left uterine horn. Embryos occupied the entire uterine cavity by 10 dpo. In conclusion, we characterized early embryo development and location of embryo during its early developmental stages in alpaca. This was apparently the first report regarding chronology of embryo development and migration to the left horn in alpaca which merits further investigation regarding its role in maternal recognition of pregnancy.

[Sonographical visibility of the corpus luteum in dairy cows]. / Sonographische Darstellbarkeit des Corpus luteum bei der Milchkuh.

OBJECTIVE: The study aimed to evaluate the day from which a corpus haemorrhagicum in the dairy cow after spontaneous or induced ovulation can be visualized using a mobile ultrasound device. MATERIAL AND METHODS: During three uninfluenced control cycles and three cycles in which six cows received a simultaneous treatment with GnRH and PGF2a, either on day 7, 14 or 17 post ovulation, the ovaries were scanned sonographically. In every control cycle a daily ultrasound examination was conducted starting from the 18th day of a cycle, in the treatment cycles beginning from the day of the hormonal treatment. Thus the formation of a new corpus luteum was followed sonographically from the day of ovulation onwards. Concentrations of progesterone were determined on day 1 of any cycle, on the day of hormonal treatment and on the 5 days following the treatment to support the sonographical results through endocrine values. RESULTS: A corpus luteum was visible in 100% of the cases on day 5 of the cycle, regardless of whether it had been a spontaneous or induced ovulation. In the treatment cycles, a sonographically visible luteolysis induced by the hormonal treatment began between 1.3 and 1.8 days after the hormone injection and lasted on average 3.7 days. The day of the cycle on which hormonal treatment was performed had no influence on the occurrence of luteolysis. A central fluid-filled cavity was visible in 33% of the corpora lutea, becoming smaller during luteolysis and disappearing before luteolysis was completed. CONCLUSION AND CLINICAL RELEVANCE: A corpus luteum is sonographically visible on day 5 after ovulation. Ultrasound examinations can be used to confirm induced luteolysis. The formation of a new corpus luteum can be visualised to confirm the success of an induced ovulation.

Crystallisation pattern of vestibular mucus and its relation to vestibular electrical resistance in cycling sow.

Changes in the genital mucus around the oestrus are used by different diagnostic methods to determine optimal fertilisation time. In the current study, the authors evaluated the different arborisation patterns found in vestibular mucus, and also established its relationship with vestibular resistance changes during oestrus. Thirty multiparous sows were checked by transrectal ultrasonography to determine ovulation time every 12 hours. Vestibular resistance was measured with a commercial resistance probe, and vestibular mucus ferning was also evaluated every 12 hours during the oestrus. Significant changes (P < 0.05) in vestibular resistance were detected, registering high variation among individuals. Maximum resistance data was reached between 12 and 24 hours after ovulation time in 83 per cent of the sows. Crystallisation samples were classified into three different patterns according to the fern-like crystal degree. Arborisation peak occurred from 48 to 36 hours before the moment of ovulation, when vestibular resistance values increased gradually. In the optimal insemination moment, vestibular resistance increased significantly (P < 0.05) and vestibular mucus showed a low crystallisation pattern (P < 0.05). Combining several methods to measure genital mucus changes may predict the ovulation time and the best insemination moment.

Field investigation of whether corpus luteum formation during weeks 3-5 postpartum is related to subsequent reproductive performance of dairy cows.

This study analyzed the effect of corpus luteum (CL) formation during weeks 3-5 postpartum on the subsequent reproductive performance of dairy cows. Factors contributing to CL formation during the postpartum period were also determined. Data were collected from 1524 Holstein dairy cows on 18 farms using a single ultrasonographic examination to determine the presence or absence of a CL during weeks 3-5 postpartum. The dates of calving, AI, conception and cow parity were also collected. Data were acquired for a subset of 475 cows on five farms related to peripartum reproductive events and the body condition score (BCS) during weeks 3-5 postpartum. The hazard of first postpartum insemination by 150 days in milk (DIM) was higher for cows with a CL compared with herd mates without a CL during week 3 (hazard ratio [HR]: 1.40, P=0.007), week 4 (HR: 1.28, P=0.004) and week 5 postpartum (HR: 1.43, P<0.0001). Furthermore, the pregnancy hazard was also higher by 210 DIM for cows with a CL compared with cows without a CL during week 3 (HR: 1.56, P=0.0009), week 4 (HR: 1.28, P=0.006) and week 5 postpartum (HR: 1.20, P=0.04). Cows calved during autumn were more likely to have a CL than cows calved during spring (odds ratio [OR] =2.32, P=0.003). Primiparous cows were less likely to have a CL than multiparous cows (OR=0.63, P=0.03). Cows with a BCS < 3.00 were less likely to have a CL than cows with a BCS ≥ 3.00 (OR=0.51, P=0.0013). In conclusion, CL formation during weeks 3-5 postpartum was related to subsequent improved reproductive performance when compared with herd mates without a CL.

Relationship between dose of cloprostenol and age of corpus luteum on the luteolytic response of early dioestrous mares: a field study.

The objective of this study was to establish and characterize the relationship between the dose of cloprostenol (37.5, 250, 500 and 750 µg) and the age of the early corpus luteum (CL) (80, 88, 96, 104 and 112 h) on the luteolytic response of mares. Behavioural oestrus and ultrasonographic signs of return to oestrus were considered as the occurrence of full luteolysis. A total of 298 mares were divided into groups according to dose of cloprostenol and CL age. There was an effect of dose of cloprostenol (p < 0.001) and age of the CL at the time of treatment (p < 0.001) on the percentage of mares with full luteolysis. The efficacy of 37.5 µg of d-cloprostenol was similar to that of 250 µg of d,l-cloprostenol (p > 0.05); and that of 500 similar to that of 750 µg (p > 0.05). The higher dose groups (500 and 750 µg) induced full luteolysis more frequently than the lower dose groups (37.5 and 250 µg) 96-104 h post-ovulation. There was no effect of CL age or cloprostenol dose on the interovulatory interval (p > 0.05). In conclusion, the effect of cloprostenol on the percentage of mares undergoing full luteolysis is dose-dependent. However, this effect is only evident in mares with CLs aged between 96 and 104 h. There is no advantage of administering more than 500 µg of d,l-cloprostenol (Estrumate(®)), to obtain a higher percentage of mares with full luteolysis in mares with CLs aged 80-112 h.

Hormonal monitoring of reproductive status in monogamous wild female owl monkeys (Aotus azarai) of the Argentinean Chaco.

The Neotropical owl monkeys (Aotus spp.) are a good model for evaluating the hypothesis that monogamy may arise if female reproductive cycles limit the mating potential of males. To evaluate this hypothesis, we first needed to assess the feasibility of using fecal sampling for monitoring the reproductive status of females. We collected fecal samples (n = 242, from 7 females) from wild adult Aotus azarai females in the Gran Chaco forests of Argentina during 3 years. Fecal estrone-1-glucuronide (E(1)C) and pregnenadiol-3-glucuronide (PdG) tended to rise in parallel during the luteal phase. The average cycle length was 22 ± 3 days (n = 5 females, 10 cycles). We identified 2 conceptive cycles and characterized the E(1)C and PdG profiles of 2 pregnancies. This report is the first of its kind on wild female owl monkeys. Despite the difficulties in sample collection and processing in the field and providing a species-specific validation in the laboratory, we show that fecal samples from A. azarai can be used for monitoring female reproductive status and function.