Day Of Artificial Insemination Research Articles

The Potential of Infrared Thermography for Early Pregnancy Diagnosis in Nili-Ravi Buffaloes

This study was designed to explore the potential of infrared thermography (IRT) as an alternate approach for early pregnancy diagnosis in buffaloes. The surface temperature (ST) of different regions (eyes, muzzle, flanks, and vulva) was determined in 27 buffaloes using IRT from the day of artificial insemination (AI; Day 0), and measurement was repeated every fourth day until Day 24 post-AI. From all regions, the ST in each thermograph was recorded at three temperature values (maximum, average, minimum). Pregnancy status was confirmed through ultrasonography on Day 30, and animals were retrospectively grouped as pregnant or non-pregnant for analysis of thermographic data. In pregnant buffaloes, all three values of ST were significantly greater (p ≤ 0.05) for the left flank, while, in the left eye and vulva, only the maximum and average values were significantly greater. By contrast, the maximum ST of the muzzle was significantly lower (p ≤ 0.05) in pregnant buffaloes compared to non-pregnant buffaloes. However, the ST of the right eye and right flank did not show significant temperature variation at any value. These findings suggest that IRT has the potential to identify thermal changes associated with pregnancy in buffaloes at an early stage.

Early prognostic markers to predict unsuccessful pregnancy in dairy cattle.

This study aimed to investigate maternal serum levels of some angiogenic factors and certain proteins in dairy cattle for (1) early prediction of unsuccessful fertilization and (2) early detection of possible pregnancy failures (early EM) after positive insemination Serum samples were collected from the same cattle at three distinct time points: 30 days before artificial insemination (B-AI), on the day of artificial insemination (AI), and 30 days after artificial insemination (A-AI). As a result of the pregnancy examination, the cows were divided into two main groups according to whether they were pregnant. The results showed that leucyl/cystinyl aminopeptidase (LNPEP) concentration was significantly decreased B-AI and Secreted frizzled-related proteins (SFRP-3), Vascular Endothelial Growth Factor (VEGF) and LNPEP levels were significantly decreased on day of AI, while PRL level was increased, and these data have prognostic significance as early indicator of the risk of potentially failed pregnancy. Additionally, a significant decrease in LNPEP, SFRP3, and VEGF levels, along with an increase in PRL levels was also observed in A-AI. These results suggest that these biomarkers can be used as a screening test to monitor the course of pregnancy. There were no significant differences in serum levels of Insulin-Like Growth Factor 2 (IGF-2), Tissue inhibitors of metalloproteinases (TIMP-1), angiopoietin (ANG), Endoglin (ENG), Fibroblast growth factor (FGF), Inhibine-A (INH-A) and Transforming growth factors-β1 (TGF-β1) between the evaluated periods neither unsuccessful nor the successful pregnancy groups. This is the first study reporting that the maternal serum levels of LNPEP, SFRP3, VEGF, and PRL have important roles in pregnancy success and may indicate whether insemination outcome will be successful B-AI and predict the risk of unsuccessful pregnancy after AI in dairy cattle. The increase in such studies will allow the development of more specific, practical, and applicable markers.

99 Diversity of vaginal bacteria in lactating dairy cattle on pregnancy establishment

Abstract Establishment and maintenance of pregnancy has a substantial economic impact within the dairy industry each year. Recent research has indicated a potential role of the reproductive microbiome on impacting pregnancy status as a result of hormone concentrations and breeding strategies. However, the association between alpha (within sample metric) and beta (between samples metric) bacterial diversity with pregnancy establishment has not been extensively evaluated in dairy cattle. Thereby, the objective of this study was to analyze vaginal bacterial diversity in lactating dairy cows 1) between day of artificial insemination (AI; day 0) and maternal recognition of pregnancy (MRP; d 18), and 2) between resulting Open and Pregnant females. Lactating Holstein dairy cows (n = 53) were sampled of day of AI and day of MRP. Pregnancy determination occurred on day 32 via transrectal ultrasonography. Sterile swabs were inserted into the vagina, rotated eight times against the vaginal wall, and stored in sterile microcentrifuge tubes at -80°C. Swab samples were utilized for subsequent bacterial diversity analyses targeting the V4 hypervariable region of the 16S rRNA gene. Cows underwent transrectal ultrasonography on gestational day 32 for pregnancy determination. Sample quality was determined by FastQC before being processed with the qiime2 pipeline for taxonomic classification. Alpha diversity metrics were analyzed with Kruskal-Wallis and beta diversity was measured utilizing PERMANOVA in the qiime2 pipeline. At day 32 of gestation, the pregnancy rate was 47.2%. According to Faith’s Phylogenetic Diversity metric for alpha diversity, there was no significant difference in diversity richness (i.e., number of different bacterial species within a sample) between day of AI and MRP or between cows diagnosed as Open or Pregnant (P >0.05). Pielou’s Evenness metric (i.e., distribution or balance of bacterial species within a sample) for alpha diversity indicated greater diversity within samples at day of AI compared with day of MRP (P = 0.02). Shannon’s diversity index metric (i.e., richness and evenness within a sample) for alpha diversity also indicated greater diversity at day of AI compared with day of MRP (P = 0.04). However, there was no difference between resulting Open and Pregnant females for Pielou’s or Shannon’s metrics (P > 0.05). Distinctions between samples (beta diversity) were compared utilizing pairwise comparisons of the unweighted and weighted Unifrac distance metrics. Specifically, day of AI indicated lower diversity compared with day of MRP for both metrics (P < 0.01). However, upon comparisons of Open and Pregnant status, there was no difference between samples for unweighted or weighted Unifrac distances (P > 0.05). Based on these results, bacterial diversity within and between vaginal samples appear to shift between day of AI and day of MRP. Additional studies are needed to elucidate the mechanisms altering bacterial diversity during early gestation in lactating dairy cows.

288 Awardee Talk: Vaginal Microbiomes of Lactating Dairy Cattle Bred Utilizing Timed Artificial Insemination Or Estrus Expression

Abstract Within the cattle industry, implementation of timed artificial insemination (TAI) or estrus detection protocols aid in determining the optimal breeding date. However, previous studies utilizing both breeding strategies have found differences between pregnancy rates, further emphasizing pregnancy establishment as a prominent issue in cattle production systems. Recent research has begun to evaluate the role of the female reproductive microbiome as it pertains to pregnancy establishment in cattle. It has been demonstrated that shifts in the female reproductive bacterial communities may alter pregnancy outcomes in cattle. Therefore, the objective of this work was to evaluate vaginal microbiomes on day of artificial insemination (AI) in dairy cattle bred utilizing either TAI or estrus detection and the impacts on subsequent pregnancy outcomes. Lactating Holstein cows (104 days in milk) were classified into two groups based on breeding protocols: 1) TAI (n = 44), and 2) Heat (n = 9). On day of AI, sterile swabs were inserted into the vagina, rotated eight times, and subsequently stored at -80°C in sterile collection tubes for microbiome analysis. For microbiome analysis, the V4 hypervariable region of the 16S rRNA bacterial gene was targeted. Statistical analysis for microbiome data were conducted using PROC TTEST in SAS where equality of variances was performed. Pregnancy rates by TAI versus Heat were 56.8% and 33.3%, respectively. The phylum Firmicutes had greater relative abundance in cows bred by Heat (64.1 ± 2.3% vs. 53.9 ± 2.5%, respectively; P < 0.01) compared with phyla Proteobacteria (7.1 ± 0.8% vs. 13.7 ± 1.7%), Fusobacteria (0.9 ± 0.3% vs. 5.8 ± 1.5%), and Actinobacteria (1.8 ± 0.3% vs. 3.2 ± 0.5%) which had a greater relative abundance in cows bred by TAI (P < 0.01). Within Firmicutes, genera Ruminococcus (13.0 ± 1.0% vs. 8.5 ± 0.6%), Blautia (10.3 ± 0.7% vs. 8.0 ± 0.5%), Clostridium (5.8 ± 0.2% vs. 4.9 ± 0.3%), Oscillospira (2.2 ± 0.2% vs. 1.7 ± 0.1%), and Caloramator (2.1 ± 0.1% vs. 1.7 ± 0.1%) were all greater in relative abundance in cows bred by Heat compared with cows bred by TAI (P < 0.05). Interestingly, within Fusobacteria, genera Sneathia (0.6 ± 0.3% vs. 3.5 ± 1.0%) and Fusobacterium (0.2 ± 0.1% vs. 1.5 ± 0.5%) were greater in relative abundance in cows bred by TAI (P < 0.01). Within phyla Proteobacteria and Bacteroidetes, genera Gallibacterium and Porphyromonas respectively, were both greater in relative abundance in cows bred by TAI (P < 0.05). Collectively these results indicate that the vaginal microbiomes of lactating dairy cows bred via TAI or estrus expression differs. Additionally, a shift in the vaginal microbiome to a prevalence of the phylum Firmicutes on day of AI appears to be negatively impacting pregnancy outcomes.

Effect of Folic Acid Supplements on Progesterone Profile and Blood Metabolites of Heat-Stressed Holstein Cows during the Early Stage of Pregnancy.

Simple SummaryThe fertility of a dairy cow can be described as the female’s ability to conceive and maintain pregnancy when insemination is performed at the proper time relative to ovulation. In this context, poor estrous detection and embryonic or fetal losses are common causes for reduced reproductive performance in dairy cows. Furthermore, heat stress severely deteriorates conception and pregnancy rates in dairy farms. The aim of this study was to see how oral folic acid (FA) supplements affect progesterone levels, blood metabolites, endocrine patterns, and blood biochemical concentrations in heat-stressed pregnant cows. Oral FA supplementation (10 μg kg−1) in the first month of gestation improved the progesterone profile, as well as blood folates, pregnancy associated glycoprotein (PAG), growth hormone (GH), and Insulin-like growth factor-1 (IGF-1) concentrations in heat-stressed Holstein cows.The aim was to elucidate the impact of oral folic acid (FA) supplements on progesterone profile, blood metabolites and biochemical indices of heat-stressed Holstein cows during the early stage of pregnancy. The study lasted from the day of artificial insemination through the end of the fourth week of pregnancy. The first group (CON, n = 17) received 0 μg of FA/kg BW as a control. The second and third groups received oral FA doses of 5 (FA5, n = 19) and 10 (FA10, n = 20) μg kg−1 BW, respectively. At the 2nd and 3rd weeks of pregnancy, the FA10 group had greater progesterone levels than the CON group (p < 0.05). The FA10 group had a greater progesterone level than the FA5 and CON groups at the fourth week of pregnancy (p < 0.01). The FA10 group had higher folate levels than CON group during the first three weeks of pregnancy (p < 0.01). Both FA-supplemented groups had significantly greater serum folates than the CON group by the end of the fourth week of pregnancy (p < 0.01). At the 2nd and 4th weeks of pregnancy, the FA10 group had greater levels of serum glucose and globulin than the CON group (p = 0.028 and 0.049, respectively). Both FA-supplemented groups had greater serum growth hormone (GH) levels at the 4th week of pregnancy (p = 0.020). Additionally, the FA10 group showed significantly higher levels of IGF-1 at the 2nd and 4th week of gestation (p = 0.040 and 0.001, respectively). FA supplementation decreased the levels of non-esterified fatty acid (NEFA) at the 2nd and 4th week of gestation (p = 0.020 and 0.035, respectively). Additionally, the FA10 group showed significantly higher pregnancy-associated glycoprotein (PAG) levels at the 2nd and 4th week of gestation (p = 0.005 and 0.001, respectively). In conclusion, oral FA supplementation (10 mcg kg−1) in the first month of gestation improved the progesterone profile, as well as blood folates, PAG, GH, and IGF-1 concentrations in heat-stressed Holstein cows. These findings could be useful in developing practical strategies to keep dairy cows’ regular reproductive patterns under heat stress conditions.

Telomere length in bovine sperm is related to the production of reactive oxygen species, but not to reproductive performance

Over the last decades, selection in cattle has mainly been based on milk production rather than on reproductive efficiency. While, when applied, focus on reproduction has involved females, attention has barely been paid to males and, if so, it has only looked at classical sperm quality parameters. In effect, variables such as telomere length have been missed, despite the fact that longer telomeres have been suggested to be linked to male fertility in humans. For this reason, the present study aimed to determine the length of telomeres in bovine sperm and their relationship with a) sperm quality evaluated through the conventional spermiogram and flow cytometry, and b) bull reproductive performance. For this purpose, 29 bulls were involved in this study. Sperm telomere length was evaluated through quantitative Fluorescent In Situ Hybridization (qFISH), and sperm quality was determined at 0 h and 4 h post-thaw. Bull fertility was assessed as non-return to estrus rates after 90 days of artificial insemination. Although the mean telomere length in bovine sperm was 12.06 ± 2.75 kb, the intra-individual variability in length led us to observe three different groups of telomeres in each sperm cell: short telomeres (7.14% ± 5.79% of telomeres; 8.29 ± 2.34 kb), medium telomeres (31.03% ± 12.92% of telomeres; 16.00 ± 2.72 kb) and long telomeres (61.93% ± 18.11% of telomeres; 30.13 ± 11.35 kb). Moreover, whereas reactive oxygen species (ROS) were found to be correlated to sperm telomere length (Rs = −0.492; P= 0.007), no correlation with other sperm quality parameters was found (P > 0.05). Reproductive performance after artificial insemination was not seen to be correlated to sperm telomere length (Rs = 0.123; P= 0.520). In conclusion, this study determined, for the first time, the mean telomere length in bovine sperm and also reported that there is a high variability within each sperm cell. Yet, while telomere length was found to be correlated to ROS generation, it was not related to bull reproductive performance.

Comparative effectiveness of the use of various gonadoliberin analogues and the effect of the morphobiochemical composition of blood on the fertilization of cows

Synchronization schemes of sexual cyclicity are widely used in dairy cattle breeding to optimize reproductive function in cows and heifers. A comparative evaluation of the use of three synthetic analogues of gonadotropin-releasing hormones (GnRH) in a complex synchronization scheme was carried out, as well as the influence of the morphobiochemical composition of blood on fertilization was determined. For experimental work, 3 groups of full-aged cows were formed, 10 in each. The first group of animals was intramuscularly administered surfagon at a dose of 10 ml, the second – fertagil 2.5 ml, the third - fertagon 5 ml. In cows treated with surfagon fertilization after the first insemination was 70%, fertagil - 20%, fertagon - 50%. According to the results of 3 months of observations, after treatment with surfagon and fertagon, fertilization was 80%, with a multiplicity of inseminations of 1.13 and 1.38 times, respectively, whereas after treatment with fertagil, fertilization was 60% with a multiplicity of 1.67. The morphological picture of blood in infertile cows on the day of artificial insemination was characterized by a lower content of erythrocytes (by 10.7%; P&lt;0.05), lower hematocrit (by 6.3%; P&lt;0.05), with a higher content and concentration of hemoglobin in the erythrocyte (by 7.0%; P&lt;0.05 and 2.9%; P&lt;0.05, respectively). It was also found that infertile cows had lower progesterone levels on the day of insemination. Thus, in the group treated with surfagon, the hormone concentration was 2.7 times lower compared to those fertilized, in animals treated with fertagil 4.2 times (P&lt;0.05), fertagon 2.0 times (P&lt;0.05), which is apparently due to the different functional activity of the pituitary-gonadal system.

Evaluation of the relationship between serum paraoxonase-1 activity and superovulation response/embryo yield in Holstein cows.

In this study, the effect of serum paraoxonase-1 (PON-1) activity on superovulation response and embryo yield was evaluated. The study material comprised 50 Holstein cows aged 3–4 years on postpartum day 90–120 with a body condition score of 3–3.25. A progesterone-based estrus synchronization protocol was initially administered to the selected donors. For this purpose, progesterone source was inserted intravaginally (day 0) and gonadotropin-releasing hormone injection was performed (day 6). Seven days after the insertion of progesterone device, follicle-stimulating hormone injections (total dose of 500 µg in decreasing doses for 4 days) were administered for superovulation. On the morning of the ninth day, prostaglandin (PG) F2α was administered, and the progesterone device was removed from the vagina in the evening on the same day. Two days after PGF2α administration, fixed-time artificial insemination was performed in the morning and in the evening. On the day of artificial insemination, blood samples were taken from the donors to determine the serum PON-1 activity. Uterine flushing was performed seven days after insemination. The results revealed that the serum PON-1 activity (mean ± SD, 562.71 ± 140.23 U/l) of the cows that responded to superovulation (donors with total corpus luteum count of ≥3 in both ovaries) was higher than those (389.91 ± 80.51 U/l) that did not (P<0.05). On the day of insemination, a positive correlation was determined between serum PON-1 activity and the counts of total corpus luteum (r=0.398), total oocyte/embryo (r=0.468), transferable embryo (r=0.453), and Code I embryos (r=0.315, P<0.05). Unlike the Code I embryos, there was no significant correlation between serum PON-1 activity and the number of Code III embryos. Moreover, no significant difference in the number of Code III embryos between the two PON-1 groups was observed. However, embryo yield and quality were found to have increased with increased PON-1 activity. Therefore, it was concluded that serum PON-1 activity may be associated with superovulation response, embryo yield and quality in donor cows.

PSVIII-16 Interferon-TAU in the pathogenesis and prevention of intrauterine growth restriction and embryonic death in dairy cows

Abstract The aim of the research was to assess the state of the hormonal and cytokine blood profiles of Holstein dairy cows during the early period of gestation with physiological formation of the embryo, its intrauterine growth restriction and death, clarify the pathogenetic mechanisms of these embryopathologies development and identify the efficacy of their prevention using recombinant bovine INFT. The dynamics of serum concentration of P4, cortisol (F) and cytokines: INFT, interleukin IL-2, tumor necrosis factor (TNFα), interferon-γ (INF-γ) at the physiological formation of the embryo (n = 9), its growth restriction (n = 9) and death (n = 8) were studied. The blood samples were obtained on the day of artificial insemination, during early blastogenesis (days 8–9), implantation of the embryo (days 15–16), the formation of primary genital organs (days 32–33), and at the end of the embryonic period of development (days 60–65). In cows with intrauterine growth restriction and embryonic death, serum concentration of P4, compared with the cows demonstrating their physiological formation, was lower by 14.0–30.1%, F - by 11.2–35.5%, INFT - by 21.0–37.7% when the content of IL-2 was exceeded by 46.5–310.5%, TNFα - by 11.7–36.2%, INF-γ - by 58.0–296.0%. A statistically significant positive correlative relationship between the content of INFT, P4, F and a negative one with the indices of the concentration of proinflammatory cytokines was found. Violation of the hormonal-cytokine balance in the organisms of fertilized animals is a universal pathogenetic mechanism for the development of early embryopathies. Injections of recombinant bovine INFT to inseminated cows during the pre- and implantation periods increased the level of progesterone production and reduced the synthesis of proinflammatory cytokines. The efficacy of insemination of cows increased by 1.96 times, the manifestation of fetal/embryonic intrauterine growth restriction syndrome decreased by 2.35 times in comparison with the control animals.

Relationship between total antioxidant/oxidant status, and oxidative stress index and superovulation response in donor cows

• TAS, TOS, and OSI affect superovulation response in donor cows. • Positive correlation was determined between TAS and transferable embryo count, and embryo quality. • TOS and OSI were negatively correlated with the count of transferable embryo. • Positive correlation was found between unfertilized oocyte count and TOS, and OSI. The aim of the present study was to determine the effects of serum total antioxidant status (TAS), total oxidant status (TOS), and oxidative stress index (OSI) on counts of corpus luteum, transferable embryo, and embryo quality in cows treated with superovulation protocol. A total of 60 Simmental cows (postpartum 90 to 120 days) were used. The donors were treated with a 9 day-progesterone based estrus synchronization protocol. On the 7th day of the progesterone treatment, donors were treated with FSH twice daily as decreasing doses for 4 days. On day 9, PGF 2α injection was performed in the morning and progesterone source was removed in the evening. The cows were inseminated twice artificially on day 11 at 12 hours interval. On the day of artificial insemination, blood samples were collected to determine serum TAS, TOS and OSI concentrations. Uterine flushing was performed on day 7 after artificial insemination. Serum TAS, TOS and OSI concentrations of cows were 1.62 μmol/L, 5.44 μmol/L and 0.45 AU, respectively. There was a positive correlation between TAS concentration and the counts of transferable embryo, Code I and II quality embryo ( P <0.05). TOS and OSI concentrations were negatively correlated with the counts of transferable embryo and Code I embryo, and positively correlated with unfertilized oocyte counts ( P <0.05). As a result, it was concluded that TAS, TOS, and OSI affected the counts of transferable embryo, embryo quality, and unfertilized oocyte in donor cows.

Rewiring of gene expression in circulating white blood cells is associated with pregnancy outcome in heifers (Bos taurus)

Infertility is a challenging phenomenon in cattle that reduces the sustainability of beef production worldwide. Here, we tested the hypothesis that gene expression profiles of protein-coding genes expressed in peripheral white blood cells (PWBCs), and circulating micro RNAs in plasma, are associated with female fertility, measured by pregnancy outcome. We drew blood samples from 17 heifers on the day of artificial insemination and analyzed transcript abundance for 10,496 genes in PWBCs and 290 circulating micro RNAs. The females were later classified as pregnant to artificial insemination, pregnant to natural breeding or not pregnant. We identified 1860 genes producing significant differential coexpression (eFDR < 0.002) based on pregnancy outcome. Additionally, 237 micro RNAs and 2274 genes in PWBCs presented differential coexpression based on pregnancy outcome. Furthermore, using a machine learning prediction algorithm we detected a subset of genes whose abundance could be used for blind categorization of pregnancy outcome. Our results provide strong evidence that transcript abundance in circulating white blood cells is associated with fertility in heifers.

Implantation associated changes in expression profile of indoleamine-2, 3-dioxygenase 1, Th1-Th2 cytokines and interferon-stimulated genes on neutrophils and peripheral blood mononuclear cells of crossbred cows

Effective bidirectional communication between the embryo and dam improves the reproductive efficiency of dairy cows. Possible role of immunosuppressive indolamine-2, 3-dioxygenase 1 (IDO1) enzyme in the regulation of maternal systemic cytokine balance/shift during early pregnancy establishment along with various interferon-stimulated genes (ISGs) expression in neutrophils and peripheral blood mononuclear cell (PBMCs) were investigated in crossbred cows. Blood was collected on days 0 i.e. day of Artificial Insemination (AI), 10, 18 and 36 post-AI followed by isolation of neutrophils and PBMCs for gene expression study of IDO1, anti-inflammatory cytokines (IL-4, IL-10 and TGFβ1), pro-inflammatory cytokines (IFNγ and TNFα) and ISGs (ISG15, MX1, MX2, OAS1) in pregnant and non-pregnant cows. Cows were grouped as pregnant and non-pregnant after pregnancy confirmation by non-return to heat, ultrasonography, per rectal examination along with progesterone and IFNτ assay. Significantly (P < 0.05) higher relative mRNA expression of IDO1 and anti-inflammatory cytokines on days 10 and 18 post-AI were observed in both neutrophils and PBMCs of pregnant cows. Pregnant cows showed significantly (P < 0.05) higher mRNA transcripts of IFNγ and TNFα genes on days 18 post-AI in both neutrophils and PBMCs. Expression of ISGs was higher (P < 0.05) on day 10th and 18th post AI in both the neutrophils and PBMCs of pregnant cows. The study indicates that systemic immune regulation by IDO1 (through cytokine shift) and ISGs in peripheral immune cells are essential for the establishment of pregnancy and may be targeted in future as biomarkers for pregnancy diagnosis.

A comparative study on various immunological parameters influencing embryo survivability in crossbred dairy cows

Since long embryonic mortality has remained an area of concern affecting the reproduction, production, and profitability of dairy cows. We investigated the possible interaction between interleukins, hormones, and neutrophil associated CD markers during the implantation window in Karan Fries (KF) cows naturally coming to heat. Blood collection was done on days 0 i.e. day of Artificial Insemination (AI), 10, 18, 21, 30 and on day 40 post-AI. Total leucocyte count (TLC) and neutrophil to lymphocyte (N:L) ratio were recorded. Blood neutrophils were isolated and their number, phagocytic activity (PA), myeloperoxidase (MPO) concentration and relative mRNA expression of cell adhesion molecules (CD-11b, CD-31, CD-44, CD-62L) as well as progesterone-inducing-blocking-factor (PIBF) and glucocorticoid receptor alpha (GRα) were examined. Plasma progesterone, cortisol, IL-2, IL-8, IL-6, and IL-10 were also measured. Pregnancy was confirmed by non-return to heat, ultrasonography and per rectal examination along with progesterone assay. Cows were further divided into pregnant (P), early embryonic mortality (EEM) and late embryonic mortality (LEM) groups. Embryonic losses cows showed lower plasma concentration of IL-10 (<100 pg/ml) and a higher concentration of IL-2 (>500 pg/ml). Also, a 4 fold increase in the relative mRNA expression of CD-11b and 2.5 fold changes in CD-44 expression were observed in embryonic mortality. We observed a 1.5 fold increase in the relative mRNA expression of PIBF and a 0.5 fold increase in GRα expression in pregnant cows compared to EEM (on day 21) and LEM (on days 30 and 40) cows. Our results depicted that the hyperimmune status of the dam which could be due to multifactorial events that led to the pregnancy failure. The above basic values may be used for checking the immune status and thus timely management strategies can be taken to prevent embryonic losses.

Effect of BMI on blood value of patients on HCG day with IUI treatment

BackgroundThis study aims to analyze the effect of the body mass index (BMI) on E2, P and LH values in females who received intrauterine insemination (IUI) treatment on human chorionic gonadotropin (HCG) day.MethodsA total of 2319 cycles of IUI-assisted pregnancy treatment were selected in our hospital. Based on the BMI, female infertility patients are divided into three groups: normal weight group, overweight and obese group.ResultsFor patients with natural cycles and ≤ 35 years old, there were 440, 178 and 197 cases in the three groups, respectively. For patients with natural cycles and > 35 years old, there were 90, 83 and 81 cycles in the three groups, respectively. For patients with induced ovulation cycle and ≤ 35 years old, there were 425, 203 and 516 cases in the three groups, respectively. For patients with induced ovulation cycle and > 35 years old, there were 26, 26 and 54 cases in the three groups, respectively.ConclusionWhen a patient is ≤35 years old, the BMI affects the E2, LH and P values on the day of artificial insemination. However, the BMI is negatively correlated with E2, LH and P in IUI on HCG day. After controlling for age and assisted pregnancy, the correlation analysis revealed that the BMI is negatively correlated with hormone E2 and LH. The higher the BMI was, the lower the levels of hormones E2, LH and P became. However, in the present study, the BMI did not significantly improve the clinical pregnancy rate of patients who received IUI.

Administration of aromatase inhibitor MPV-2213ad to blue fox vixens (Vulpes lagopus) as a model for contraception in female dogs

The interest in non-surgical approaches to contraception and fertility control in female dogs has increased in recent years. In this study the effect of an aromatase inhibitor (finrozole) was evaluated in fur production animals, farmed blue fox vixens, as a model for contraception in bitches. A total of 80 vixens were divided into 4 groups, receiving orally placebo (A) or finrozole 0.5 mg/kg (B), 3.5 mg/kg (C) or 24.5 mg/kg (D) for 21 consecutive days beginning in the pre-ovulatory period of heat. Monitoring of the vixens included clinical signs of heat, measurement of vaginal electrical resistance (VER) as well as oestradiol and progesterone concentrations in plasma. The approximate relation of the start of treatment to ovulation varied from 11 days before to one day after ovulation provided that the LH peak occurred 0.5–2 days before the VER peak and ovulation was then estimated to occur 2 days after the LH peak. Seventy vixens were artificially inseminated within 8 h after a 50 Ω decline in vaginal electrical resistance was detected. Ten vixens were not inseminated. Pregnancy was confirmed by transabdominal ultrasound examination and birth of cubs was recorded. The pregnancy rates in the groups were 89.5% (A), 81.3% (B), 55.6% (C) and 52.9% (D). The average number of live born pups in the four groups was 9.4 (A), 7.0 (B), 5.8 (C), and 3.8 (D), respectively. No deleterious effects (for instance malformations) of finrozole on pups could be verified. The administration of finrozole did not have a significant effect on oestradiol parameters and VER values in vixens. Progesterone values were significantly higher in treatment groups compared with the placebo group. The results indicate that pregnancy could be avoided by finrozole provided that doses of ≥3.5 mg/kg were used and the treatment was initiated at least four days before the day of artificial insemination. This corresponds with two to six days before ovulation provided that the LH peak occurred 0.5–2 days before the VER peak and that ovulation then occurred in average 2 days after the LH peak.

Effects of human chorionic gonadotropin and intravaginal progesterone device treatment after artificial inseminations on the reproductive performance of normal and repeat breeder lactating dairy cows.

We examined the effects of human chorionic gonadotropin (hCG) treatment on Day 5 (Day 0 = day of artificial insemination: AI) and intravaginal progesterone device (IVPD) treatment from Day 5 to 19 on the conception and detection rates of return to estrus (re-estrus) in lactating dairy cows. A total of 306 cows from a commercial dairy farm were divided into the following three groups on Day 5: non-treatment group (n = 128), untreated; hCG group (n = 71), 3,000 IU hCG was administered (intramuscularly); IVPD group (n = 107), IVPD was inserted into the vagina from Day 5 to 19. Re-estrus detection was performed up to Day 25. Pregnancy was diagnosed by rectal palpation between Day 50 and 60. There was an interaction between treatment and AI number (P < 0.01) on the conception rate of first-AI. For cows with more than three AIs, the IVPD treatment (66.7%) was more effective than the non-treatment (23.1%) (P < 0.05). The re-estrus detection rate was significantly (P < 0.05) higher in the IVPD group (60.7%) than that in the non-treatment group (41.4%) and tended (P < 0.1) to be higher than that in the hCG group (37.8%). Our results suggested that the conception rate can be improved by IVPD treatment, especially in cows with more than three AIs. In addition, IVPD treatment can induce higher estrus expression up to 25 days after AI in non-pregnant cows.

DAILY DYNAMICS OF SEXUAL HUNTING SIGNS IN COWS AND HEIFERS OF MATING AGE OF DAIRY HERD WITH LOOSE HOUSING

The article reveals the feasibility of organizing and conducting the selection of cows and heifers by sexual activity for artificial insemination day and night. Revealed for three months in females (n=717): in 600 signs of sexual activity in 0,70 %, 700-19,94 %; 800- 19,25 %; 900 – 9,21 %; from 1000-1200-14.78 %; from 1300-1400- 4.32 %; from 1500-1600-15.2 %; from 1700-1900 – 9,76 %; 2000-6,83 %. It was revealed that most animals had sexual activity in the morning (49.10 %). In August, 213 heads were studied, of which the signs of sexual activity in 600 had 2.35 %; in 700-14.08 %; in 800 - 11.74 %; in 900 - 8.45; in 1000 decreased to 3.29 %, in 1400 - an increase to 11.74 %; in 1600 - 12.68 %; in 1800 - 13.15% and 2000 - 22.54 %. Round-the-clock monitoring revealed a state of libido, for a period of 0000-400 hours morning 25,00 %, 400-800 – 18,27 %, 800-1200 – 17,34 %, 1200-1600 – 20,19 %, 1600-2000 – 4,81 %, 2000-2400 - 14,42 %. Of the 104 cows identified in the hunt, 62.5 % showed sexual activity in the period from 2000 to 800, and only 37.5 % of cows in the hunt were detected during the working hours of the artificial insemination operator. Also, from 22 heifers, 40.9 % showed signs of sexual activity in the period from 2000 to 800, and 59.09 % showed signs of sexual activity during working hours. Therefore, given the acceleration of physiological processes in the warm season, there are prerequisites for reducing the efficiency of the operator. In September (an hour later), it was revealed in females (n=270): in 700 - 18.89 %, in 800 - 21.85 %; before 900 there was a decrease to 1000 %; and by 1000 hours - a rapid increase to 27.04 %; by 1400, a decrease to 1.85 %; from 1600 to 2000 - 12.96, 7.04 and 0.37 %, respectively. In October, out of 234 animals in 700, 26.50 % of females showed signs of sexual activity, which is 7.60 more than in September and 12.42 % more than in August; in 800 - 23.08 %; in 900-8.97 %; in 1000, a slight increase to 11.11 % was observed, by 1400 - a decrease to 0.43 % (similar to the previous month); in 1600, 20.09 % were detected, in 1800 - 9.83 %. So, during the daylight hours, cows and heifers show signs of sexual activity not evenly: the first wave of excitement begins with a sunrise and lasts 4-6 hours, the second begins at 1400-1600 and continues until sunset. The data obtained indicate that it is advisable to identify cow’s sexual activity both during the day and at night. Such studies should be carried out depending on the time of year: the length of daylight and the temperature of the environment. The conducted research can serve as a basis for compiling the multiplicity of detection of cow’s sexual activity during the day and the mode of their detection libido during working shifts by artificial insemination operators, their assistants and other workers.

Study on the effect of insemination time on pregnancy rate of Bangladeshi buffalo in intensive farming

The study was designed to evaluate the insemination time and pregnancy rate of Bangladeshi buffalo at Lal Teer Livestock Breeding and Research Farm, Bhaluka, Mymensingh, Bangladesh during January 2017 to June 2018. A total of 30 cyclic buffaloes were included in this study. The animals were inseminated under four different times, i.e. inseminated i) between 0-6 hours after seeing first sign of estrous, ii) between 6-12 hours after seeing first sign of estrous, iii) between 12-18 hours after seeing first sign of estrous and v) between 18-24 hours after seeing first sign of estrous. The pregnancies (positive or negative) were confirmed by rectal palpation and transrectal ultrasonography after 60 days of artificial insemination (AI). The total pregnant animals in this study were 40%. The highest pregnancy rate (60%) was observed in insemination between 12-18 hours after seeing first sign of estrous and the lowest pregnancy rate (0%) was observed in insemination between 0-6 hours after seeing first sign of estrous. The findings of the study suggest that, the conception rate of buffalo depend on the time of AI, detection of proper estrus symptoms and site of semen deposition.&#x0D; SAARC J. Agri., 16(2): 143-152 (2018)

Follicular fluid composition of ovulatory follicles in repeat breeder Holstein dairy cows

Objective: To examine differences in the metabolite, steroid and lipopolysaccharide of follicular fluid collected from the ovulatory follicle of Holstein repeat breeding cows, lactating cows, and virgin fertile heifers. Methods: Estrus was induced in animals possessing functional corpus luteum by intramuscular administration of prostaglandin F2 α. Six to twelve hours after detection of the standing estrus, cervical samples were collected to detect subclinical endometritis via counting neutrophils on stained smears of cervical swabs. Then, follicular fluid of ovulatory follicles and serum samples were collected from repeat breeding cows (n=11), lactating cows (n=8) and virgin fertile heifers (n=10). Sodium and potassium were measured with a flame photometer method. Urea, total protein, glucose, cholesterol and β -hydroxybutyric acid were assayed with commercial spectrophotometry kits. Chloride concentration was also measured with titration of samples against silver nitrate. Progesterone, estradiol-17 β and lipopolysaccharide concentrations were measured using enzyme-linked immuno sorbent assay kits. Results: All analysis of follicular fluid samples showed that repeat breeding and lactating cows had a mean higher lipopolysaccharide concentration than that of the virgin fertile heifers (P Conclusions: High follicular fluid concentration of lipopolysaccharide in ovulatory follicles results in the occurrence of repeat breeding syndrome in dairy cows. Further, a lower serum estradiol-17 β concentration and a higher percentage of neutrophil in the cervical secretion on the day of artificial insemination may have resulted in the occurrence of repeat breeding syndrome in dairy cows.

Influence of Urea Level in the Milk on the Day of Artificial Insemination in Holstein Cows

Influence of Urea Level in the Milk on the Day of Artificial Insemination in Holstein Cows