Determination of expression patterns of miR-26a, and preimplantation factor levels for early pregnancy detection in nulliparous and multiparous cows.

For maximum productivity in a dairy farm, the earliest and the most accurate detection of pregnancy is essential. The aim of this study was to determine the efficacy of expression patterns of miR-26a, and serum Preimplantation Factor (PIF) levels for pregnancy diagnosis during the early pregnancy in nulliparous and multiparous cows. A total of 60 cows (30 nulliparous and 30 multiparous Holstein cows) were enrolled in the study. Blood samples were collected for miR-26a on days 8 and 16 (D8 and D16), and for the PIF on days 10 and 20 (D10 and D20) following insemination (D0). Pregnancies were determined by ultrasonography on the 28th day after insemination. Expression levels of miR-26a determined by qPCR. PIF levels were assessed by using commercial ELISA kits. All data were analyzed by using the MIXED procedure of SPSS. The expression levels of miR-26a were 6.64 folds higher on D16 in pregnant compared to non-pregnant multiparous cows (p < .05). On D8 and D16, miR-26a expression levels were found higher 13 folds in pregnant compared to non-pregnant nulliparous cows (p < .05). Additionally, miR-26a expressions were higher 5.42 folds (p < .05) on D8, 7.19 folds higher (p < .01) on D16 in pregnant nulliparous and multiparous cows, and were 6.30 folds higher (p < .001) on D8 and D16 according to non-pregnant animals. PIF levels were greater in pregnant animals (p < .05). Analyzing miR-26a on D8 might be considered as sufficient in nulliparous cows. Pregnancy detection in multiparous cows can be made on the 16th day with this method. Furthermore, PIF evaluations may be sufficient on D10 in multiparous cows. Besides, PIF levels and miR-26a expression levels might be used safely in field conditions and clinical applications.

Association between metabolic parameters and embryo production in superovulated dairy cattle.

This study aims to investigate the relationship between metabolic parameters and the number of embryos produced in superovulated cows with high genetic characteristics in milk yield. Eighteen Holstein donors were treated with classic superovulation protocols, AI and flushing. During superovulation, decreasing doses of FSH (follicle-stimulating hormone) were administered at 12-h intervals for 4 days. Plasma insulin-like growth factor (IGF1), glucose (GLU), beta-hydroxybutyric acid (BHB), non-esterified fatty acid (NEFA), blood urea nitrogen (BUN) and total protein (TP) levels were determined by using an autoanalyzer. The mixed model analysis of variance was used for statistical analysis. As a result, plasma IGF1, BHB and BUN had significant interactions with both groups and days (p < .05). Additionally, plasma TP-days interactions were significant (p < .05). Furthermore, there was a negative correlation between the number of embryos and plasma BHB levels (p < .05). In conclusion, under appropriate environmental conditions, metabolic profile control of donors can contribute to the embryo production process and to the studies on the metabolic infrastructure.

Semen collection by trans-rectal digital stimulation and insemination campaign in goat.

The overall purpose of this study was to describe a method of semen collection via trans-rectal digital massage (TDM) and to carry out a related fertility trial in Angora goat. Sixteen Angora bucks (ranging 1-4 years) and 28 nulliparous does (1-2 years) were used in this study. Semen samples were collected via trans-rectal massage from 85.71% of the bucks in multiple attempts (18/21). The mean values of volume, pH, mass motility, total motility, concentration, viability, abnormal spermatozoa rate and ejaculation time were 0.64 ± 0.09 ml, 6.3 ± 0.21, 2.7 ± 0.34, 58.18 ± 5.1%, 3.68 ± 0.31 × 109 /ml, 71.38 ± 7.12%, 18.22 ± 2.48% and 3.4 ± 0.33 min respectively. Oestrus was detected with teaser buck and confirmed by using infrared thermography and ultrasonography (US). The success rate of synchronisation was found as 71.4% (20/28). On Day 21, pregnancy diagnosis was performed trans-rectally with US and the pregnancy rate was determined as 78.57% (11/14). TDM method of semen collection seems to be easily applicable to the buck and it could be a good alternative to collect semen as well as its use in artificial insemination campaign. Thermal monitoring is found to be a valuable tool to monitor the response to hormonal driven ovulatory synchronisation in Angora does during timed artificial insemination.

Ultrasonographic patterns of different regions of uterus according to pregnancy in cows and heifers.

This study aimed to detect pregnancy within the first 20 days after artificial insemination by evaluating the ultrasonographic patterns of the different regions of uterus in Holstein heifers and cows. Animals were divided into subgroups according to pregnancy on 28th day as pregnant (heifer, n: 15; cow, n: 15) and non-pregnant (heifer, n: 15; cow, n: 15). Images were taken from the ovulation-side cornu uteri (OSC), non-ovulation-side cornu uteri (NOSC), and the corpus uteri (CU) on alternate days from D0 to D20. The images were evaluated by ImageJ software in terms of mean gray value (MGV), homogeneity (HOM), and contrast (CON) and endometrium thickness (ET). The mean MGV and G*T and P*T interactions, the mean CON and G*T and G*P interactions, the mean HOM and G*P interactions, and the mean ET and G*P interactions were statistically significant (p < 0.05). In receiver operating characteristic analyses, D2-D6 for CON and D2, D6, D8, D16, and D20 for HOM of OSC in cows and D8 and D10 MGV and D18 and D20 ET of OSC in heifers had high relationship with positive pregnancy (p < 0.05). The use of echogenicity evaluations and endometrium thickness measurements in reproductive management seems to be suitable for the prediction of pregnancy in cows and heifers.

The thermographic monitoring in early pregnancy detection in Holstein cows and heifers.

This study aimed to conduct a comprehensive investigation to demonstrate early pregnancy detection using thermography in heifers and cows. A total of 60 heifers (n: 30) and cows (n: 30) were divided into two groups as pregnant (n: 15 heifers, n: 15 cows) and non-pregnant (n: 15 heifers, n: 15 cows) according to the day 28 of gestation. Thermographic images were taken from the vulvar and anal regions on alternate days from D0 to D20. Blood samples were collected to determine estrogen and progesterone concentrations. The mean temperature difference between the anal and vulvar regions (ΔT °C) was used in the statistical analyses. Based on the hormonal profiles, no abnormalities were observed for follicular waves or luteal profiles in heifers and cows. The ΔT °C values between heifers and cows and between days were statistically significant (P < 0.001). In thermographic analyses, the differences observed in other main effects and interactions of the group, sampling time, and pregnancy were not statistically significant (P > 0.05). In receiver operating characteristic (ROC) analyses, it was concluded that the ΔT °C value of ≤ 2.9 °C (100% Se - 61.9% Sp) was highly correlated with pregnancy diagnosis in cows on day six after artificial insemination (AI) (P < 0.001). In conclusion, it was determined that the clinical application of thermography can be used for the detection of pregnancy on day six after AI in cows. However, further studies are needed to determine heifers' thermographic characteristics and profiles.

Fertility comparison of frozen bull semen stored in cryogenic deep freezer (-152 °C) and LN2 container.

The present study aimed to use cryogenic deep freezers that could be a feasible alternative for cryopreserved semen storage. A total of 284 straws from three Simmental bulls and 272 Simmental cows were used. The experimental group consisted of 151 semen straws that were stored at -152 °C for a week. Moreover, the control group consisted of 133 semen straws that were stored at -196 °C. Firstly, two samples per bull (n = 6) were examined in terms of sperm kinetic parameters by CASA. Furthermore, plasma membrane, acrosome integrity (PMAI) and high mitochondrial membrane potential (HMMP) were analyzed by flow cytometry. Then, artificial inseminations were performed on Simmental cows with 272 straws belonging to two groups. Then, 56th-day Non-return Rate (NRR56) was determined. All spermatological data were subjected to a linear mixed model. Chi-Square test was performed to NRR56 between storage temperature groups. Also, logistic regression analysis was used to examine the effect of bull, storage temperature and age of cows on pregnancy status. While age of cows was included in the final logistic regression model, effect of bull x storage temperature was not included because it was found as non-significant. The post-thaw PMOT and STR of cryopreserved bull semen, which was stored at -152 °C, had lower and statistically significant values (p < 0.05). However, frozen bull semen, which were stored at -152 °C, kept its fertility ability as which stored at -196 °C. Besides, NRR56 of semen stored at -152 °C and -196 °C were detected as 57.24% (83/145) and 55.91% (71/127), respectively (p > 0.05). Nevertheless, these results should be supplemented with more pre-freezing and post-thaw sperm quality analyses and more fertility data for increasing the accuracy of the method.

Vulvar thermal pattern following synchronization of estrus is linked to fertility after timed artificial insemination in goat.

Aim of this study was to study the vulvar thermal pattern variation during the timed artificial insemination protocol in Angora goat and identify the relationship with the successful rate. Does (36 adult healthy females) were synchronized using PGF2α at the day 0, 11 days of progesterone impregnated sponges intra-vaginally, PMSG 48 h before sponges withdraw (day 11) and the intra-cervical inseminations were carried out 48 h later (Timed Artificial Insemination: TAI) with chilled semen. Vulvar (VST) and perivulvar (PST) areas were considered to evaluate the thermal pattern during the protocol at the day 0 and at the TAI using a thermo camera (E60, FLIR System). Differences of temperature (ΔT) between the surfaces were calculated for each time. The does were monitored for pregnancy, delivery time and prolificacy. Pregnant (P) and non-pregnant (NP) does were compared in terms of VST, PST and ΔT using two ways ANOVA considering time and pregnancy as sources of variability. VST was lower than PST in all the monitored does (P < 0.05) (34.79 ± 0.14 vs 36.58 ± 0.14 °C) and without differences between P and NP at day 0 (35 ± 0.18 vs 36.39 ± 0.22 °C). Significant difference (P < 0.05) between P and NP does was recorded at TAI in terms of VST (33.89 ± 0.31 vs 35.40 ± 0.24 °C) and ΔT (-3.16 ± 0.34 vs -1.62 ± 0.26 °C). In conclusion thermal emission by glabrous surfaces in goat may be used to identify the right response induced by hormonal treatments and to optimize the application of assisted reproductive techniques at the field level.