Cyclic Cows Research Articles

104 Non-invasive pregnancy diagnosis in Holstein dairy cows using computer vision analysis of thermal images

Abstract Thermal visualization of Holstein dairy cattle skin can be indicative of the pregnancy state (cyclic or pregnant) of the cow by observing the size and shape of skin thermal patterns. However, visual assessment of skin thermal patterns can be subjective and laborious. Further, infrared thermography implementation on-farm requires automated analysis to be feasible for dairy producers. The objectives of this study were to implement a machine learning convolutional neural network (CNN) to detect features in thermal images associated with pregnant (Pregnant) and non-pregnant (Cyclic) and to evaluate the accuracy of pregnancy diagnosis in dairy cows. A total of 938 images from 18 Pregnant cows and 982 images from 18 Cyclic were used to build the CNN model, and 120 images from Pregnant (60 images) and Cyclic (60 images) dairy cows were used to validate the model. Pregnant cows were confirmed to be pregnant 90 d after insemination via transrectal ultrasonography. Cyclic cows were confirmed to be non-pregnant using follicular dynamics and corpora lutea presence via transrectal ultrasonography. Images were recorded using a T620s infrared camera at 640 × 480 (307,200) to train the CNN model and images from an E40 FLIR camera at 320 x 240 (76,800 pixels) were used for the validation test. The CNN model was created using TensorFlow in Google Colab to binary classify images into Pregnant or Cyclic using a single output neuron (1: Pregnant and 0: Cyclic) with different Epochs (10, 25, and 100). The diagnosis accuracy (acc) was 0.72 with a validation accuracy (val_acc) of 0.41 for 10 Epochs, acc 0.76 with val_acc 0.37 for 25 Epochs, and acc 0.80 with val_acc 0.63. The acc and val_acc of the training model and validation test were optimized when using images from the T620s exclusively (Epochs 10: acc = 0.70 and val_acc = 0.64; Epoch 25: acc = 0.74 and val_acc = 0.70; Epoch 100: acc = 0.81 and val_acc = 0.80). Greater resolution of thermal images was proven to increase the accuracy of pregnancy diagnosis (307,200 compared with 76,800 pixels) and the greater number of Epoch used to train the model. Machine learning algorithms can diagnose pregnancy using thermal images with an acceptable evaluation accuracy (> 0.70).

Effect of Heat Stress on Subsequent Estrous Cycles Induced by PGF2α in Cross-Bred Holstein Dairy Cows.

This study aimed to determine the effect of heat stress (HS) on reproductive parameters (calving to first service (CTFS) and the first-service conception rate (FSCR)) and general physiological responses (rectal temperature (RT) and respiratory rate (RR)) in tropical cross-bred Holstein dairy cows raised in Ratchaburi province, Thailand. HS was determined using the temperature-humidity index (THI), calculated from temperature and humidity inside the barns, and was classified as moderate HS (THI: 80.67 ± 0.79) and mild HS (THI: 77.81 ± 1.09) in this study. Cows with detected corpus luteum were defined as cyclic cows and were injected with PGF2α at the beginning of the experimental period. Reproductive and physiological parameters were recorded. Cows showed significantly lower RT and RR in the mild HS group (38.47 ± 0.21 °C and 41.04 ± 4.55 bpm, respectively) than in the moderate HS group (38.87 ± 0.15 °C and 51.17 ± 10.52 bpm). The percentage of cows that ovulated after being induced by PGF2α and showed estrus signs was higher in the mild than the moderate HS groups (54.55% vs. 18.18%). Furthermore, the FSCR of cows under mild HS tended to be higher than that in the moderate HS group (42.11% and 15%, respectively) (p = 0.06), while the average CTFS interval was significantly shorter under mild HS than moderate HS (69.47 ± 18.18 and 84.60 ± 27.68 days, respectively). These results indicate that moderate HS impairs reproductive performance in crossbred Holstein cows, compared to mild HS conditions.

Study the Prevalence of Anestus in Relation to Some Threats in Heifer and Postpartum Cows

The objectives of current study were to assess the prevalence and risk factors associated with postpartum anestrus in lactating dairy cows in Rajshahi, Bangladesh. A total of 160 postpartum cows of different breed, genotype and age were selected randomly from different dairy farms in studied area. Structured questionnaire was used to collect data from farm owner on prevalence and risk factors for postpartum anestrus. Observed heat within 60 days after parturition was considered as normal cyclic cows and others were considered as postpartum anestrus. The prevalence of postpartum anestrus was 54.42%. The study shows that older cows take relatively more time for ovarian resumption. The highest percentage (54.64%) of cows showed anestrus in crossbred cows that native in Rajshahi. When compared with genotype the highest percentage anestrus was observed 100.00 in Local x Jersey than 66.66% in Local x Sahiwal genotype to 53.07% in Local x Friesian and lowest was 50.00% in indigenous cattle. The age of cattle also relation to anestrus and the highest, lowest and average percentage of anestrus were showed 60.00%, 39.39% & 50.49% in < 4 year, > 6 years and 4 to < 6 years age group of cattle at Rajshahi area of Bangladesh.

Performance and Cost-Efficiency of Single Hormonal Treatment Protocols in Tropical Anestrous Dairy Cows.

This retrospective study aimed to evaluate the performance of hormone treatment protocols, determine the factors associated with pregnancy success after hormone treatment, and compare the cost-efficiencies of two types of hormone treatment among cyclic and noncyclic anestrous dairy cows. The clinical records of 279 anestrous cows that received hormone treatment for artificial insemination (AI) from 64 herds in the western region of Thailand were obtained from Kasetsart University Veterinary Teaching Hospital from January to August 2017. The performance of the hormone treatment protocols, fixed-time AI (TAI) and estrus detection before AI (EAI), showed that the pregnancy risk for the TAI protocol was higher than that for the EAI protocol, but pregnancy per AI did not differ significantly between the two protocols in cyclic and noncyclic cows. Multivariate logistic regression analysis showed that cows receiving the TAI protocol were more likely to be pregnant compared to those treated with the EAI protocol. Cows with a 3.00 body condition score (BCS) < 3.75 after treatment and loose-housed cows were more likely to become pregnant. Treatment during winter showed higher pregnancy success than that in the summer and rainy seasons. The cost-efficiency analysis showed that the TAI protocol was the most cost-efficient option for noncyclic cows, whereas the EAI protocol was the most cost-efficient option for cyclic cows.

Randomized clinical trial of ketoprofen or ceftiofur for treatment of metritis in dairy cows

Our objectives were to compare the efficacy of ketoprofen or ceftiofur for treatment of metritis in dairy cows considering subsequent health, production, and reproduction. Cows from 2 commercial dairy farms in Ontario, Canada were examined with a Metricheck device 3 times per week from 2 to 14 d in milk (DIM). Cows with metritis (fetid vaginal discharge; n = 193) were blocked by parity and fever (rectal temperature ≥39.5°C or <39.5°C) and within each block per farm, randomly assigned to receive 3 mg/kg BW of ketoprofen (KET) or 2.2 mg/kg of ceftiofur hydrochloride (CEF), once a day for 3 d. Day of enrollment was considered study d 0. Rectal temperature and attitude were evaluated in cows with metritis on study d 0, 3, 4, 7, 10, and 13, and vaginal discharge was evaluated on study d 4, 7, 10, and 13. Body condition was scored at enrollment and 35 DIM, and serum concentration of haptoglobin was measured at d 0, 2, 4, and 7. Cows with rectal temperature ≥39.5°C or a depressed attitude on d 3 were classified as clinical failure and received treatment with ceftiofur for 3 d (KET), or 2 additional days (CEF), to a maximum of 5 d of treatment with ceftiofur. At 35 ± 3 DIM cows were examined for uterine involution by transrectal palpation, purulent vaginal discharge (PVD) by Metricheck, and endometritis by endometrial cytology. Time to onset of cyclicity was assessed by serum progesterone (P4) measurements at 28, 42, and 56 DIM. Contemporary cows from the same farms without metritis (NOMET; n = 1,043) were used for comparison. Data were analyzed with mixed linear or logistic regression or Cox's proportional hazard models, including herd as a random effect. The proportion of clinical resolution of metritis on d 3 (96% vs. 92%), of cows with fever (from d 3 to d 13 after enrollment) or fetid discharge (from d 4 to d 13 after enrollment), and the number of medical treatments (3.1 vs. 3.3) were not different between CEF and KET, respectively. Cows in KET received fewer antibiotic treatments than cows in CEF (0.3 vs. 3.1). Uterine involution, the prevalence of PVD (50% vs. 47%) and subclinical endometritis (6.6% vs. 4.3%), and the proportion of cyclic cows (82% vs. 86%) did not differ between CEF and KET. Cows in KET had greater serum haptoglobin concentration from d 2 to 7 after enrollment. The incidence of mastitis, lameness, or displaced abomasum to 60 DIM and subclinical ketosis to 21 DIM did not differ among CEF, KET, and NOMET. There were no differences in median days to first AI (CEF = 68 d; 95% CI: 65-70; KET = 69 d; 95% CI: 68-72; NOMET = 69 d; 95% CI: 68-70), and median days to pregnancy (CEF = 118 d; 95% CI: 92-145; KET = 113 d; 95% CI: 90-135; NOMET = 105 d; 95% CI: 101-109), pregnancy at first AI at 33 d after insemination (CEF = 42%; KET = 41%; NOMET = 41%), pregnancy loss after first AI (CEF = 8%; KET = 11%; NOMET = 8%), hazard of pregnancy or hazard of culling up to 300 DIM. Milk yield was not different between CEF and KET during the first 10 weeks, but lesser in KET at wk 2 and 4 and CEF at wk 2, 4, and 6 than in NOMET. In this pilot-scale study, given early detection, we did not detect differences in subsequent health, milk yield, or reproductive performance in cows with metritis initially treated for 3 d with CEF or KET. Additional, larger studies are warranted.

MicroRNA-148b secreted by bovine oviductal extracellular vesicles enhance embryo quality through BPM/TGF-beta pathway

BackgroundExtracellular vesicles (EVs) and their cargoes, including MicroRNAs (miRNAs) play a crucial role in cell-to-cell communication. We previously demonstrated the upregulation of bta-mir-148b in EVs from oviductal fluid of cyclic cows. This miRNA is linked to the TGF-β pathway in the cell proliferation. Our aim was to verify whether miR-148b is taken up by embryos through gymnosis, validate its target genes, and investigate the effect of miR-148b supplementation on early embryo development and quality.MethodsZygotes were cultured in SOF + 0.3% BSA (Control) or supplemented with: 1 µM miR-148b mimics during: D1-D7 (miR148b) or D1-D4 (miR148b-OV: representing miRNA effect in the oviduct) or D4-D7 (miR148b-UT: representing miRNA effect in the uterus) or 1 µM control mimics was used during: D1-D7 (CMimic). Embryos at ≥ 16-cells and D7 blastocysts (BD7) were collected to examine the mRNA abundance of transcripts linked to the TGF-β pathway (TGFBR2, SMAD1, SMAD2, SMAD3, SMAD5, BMPR2, RPS6KB1, POU5F1, NANOG), total cell number (TC), trophectoderm (TE), and inner cell mass (ICM) were also evaluated. One-way ANOVA was used for all analyses.ResultsWe demonstrated that miR-148b can be taken up in both 16-cell embryos and BD7 by gymnosis, and we observed a decrease in SMAD5 mRNA, suggesting it’s a potential target of miR-148b. Cleavage and blastocysts rates were not affected in any groups; however, supplementation of miR-148b mimics had a positive effect on TC, TE and ICM, with values of 136.4 ± 1.6, 92.5 ± 0.9, 43.9 ± 1.3 for miR148b and 135.3 ± 1.5, 92.6 ± 1.2, 42.7 ± 0.8, for miR148b-OV group. Furthermore, mRNA transcripts of SMAD1 and SMAD5 were decreased (P ≤ 0.001) in 16-cell embryos and BD7 from miR148b and miR148b-OV groups, while POU5F1 and NANOG were upregulated (P ≤ 0.001) in BD7 and TGFBR2 was only downregulated in 16-cell embryos. pSMAD1/5 levels were higher in the miR148b and miR148b-OV groups.ConclusionsOur findings suggest that supplementation of bta-miR-148b mimics during the entire culture period (D1 - D7) or from D1 - D4 improves embryo quality and influences the TGF-β signaling pathway by altering the transcription of genes associated with cellular differentiation and proliferation. This highlights the importance of miR-148b on embryo quality and development.

Temporal alteration in ovarian follicle fate with hormonal and hemodynamic perspectives during receptive phase of estrous cycle in endometritic cattle.

The present research was carried out to assess the serum progesterone (P4) concentration and uterine hemodynamics at estrus till ovulation in cyclic cows (N = 130) with healthy or diseased uterus. At estrus, 85 cows were diagnosed with clinical endometritis (CE; n = 44) and sub-clinical endometritis (SCE; n = 41); whereas, 45 cows being served as control namely no endometritis (NE; n = 45) were included in the study. Serum progesterone estimation at 12 - 14 and 40 hr after the onset of estrus and Doppler sonography of both middle uterine arteries were done to envisage the uterine hemodynamics and ovulation. The serum progesterone concentration was significantly higher at 12 - 14 hr after onset of estrus in CE and SCE cows. At 12 - 14 hr after onset of estrus, a cut-off value of ≥ 0.48 ng mL-1 P4 was obtained, above which 22.72% CE, 26.82% SCE and only 8.88% NE cows failed to ovulate within 36 - 40 hr of estrus onset. Among the Doppler indices, pulsatility and resistance indices were significantly higher; whereas, volume and velocity indices were significantly lower in NE cows. In cows diagnosed with CE and SCE, a higher supra-basal P4 concentration, and velocity and volume of blood flow to uterus at estrus negatively affected the duration to ovulation.

Exploring Prevalence and Contributing Risk Factors of Repeat Breeding Syndrome of Crossbred Dairy Cows in Bangladesh

The dairy sector suffers significant financial losses due to repeat breeding syndrome (RBS), a significant reproductive disease in dairy cattle. For a very long time, repeat breeding has been regarded as one of the most critical reproductive issues in cattle. The objective of this study is to investigate the frequency and identify the risk factors related to repeat breeding in dairy cows in the Bogura district of Bangladesh. Additionally, the study aims to assess the influence of repeat breeding on reproductive performance in this population. This study collected indigenous and crossbred 179 cow's data using a pre-structured questionnaire. The cows that needed more than three consecutive inseminations to conceive were considered repeat breeders, and those that needed ≤3 inseminations were regular cyclic cows. The incidence rate of RBS in this study was 33.50%. In BCS 2.5 to 3.0, the incidence rate was significantly higher (58.30%) in repeat breeders than in others. Cows with &lt; 5-litre average daily milk production and reared in intensive farming systems with moderate to poor levels of hygienic management have a significant (p&lt;0.005) effect of repeat breeding syndrome. Different reproductive diseases have a significant (p&lt;0.001) effect on the conception rate of cows. Stillbirth (77.87%) followed by fetal death (76.47%) were recorded as major reproductive obstacles in repeat breeding, whereas pyometra (71.43%), followed by abortion (66.67%), dystocia (70.00%) and endometritis (57.14%) were the major recorded complications in repeat breeders. The rate of conception would go up if cows had optimal BCS, good hygiene and management practices were used, and farmers were trained on how to manage cattle reproduction. This would lower the repeat breeding syndrome.

COMPARATIVE ANALYSIS OF ANTIOXIDANT ACTIVITY AND REPRODUCTIVE HORMONES IN POSTPARTUM ANESTRUS COWS DIAGNOSED BY ULTRASOUND IN BASRAH PROVINCE

This study was conducted in Basrah province from September 2022 till December 2022 to evaluate the ratio of enzymatic antioxidants; catalase (CAT), superoxide dismutase (SOD), glutathione peroxidase (GSH-px), glutathione (GSH), and the concentration of malondialdehyde level (MDA) in the serum of 10 cows that suffered from postpartum anestrus compared five cyclic cows as a control group. In addition, a hormonal evaluation (Estrogen E2, progesterone P4, and cortisol) in postpartum anestrus cows compared regular control group. Postpartum anestrus and cyclic cows are diagnosed ultrasonographically by examining the genital tract per rectum using ultrasound. The results revealed a significant decrease (P&lt;0.05) in all antioxidant enzymes (SOD, CAT, GSH-px) in postpartum anestrus cows in compared with to regular cyclic cows. The results of GSH analysis revealed that nonsignificant elevation occurred in postpartum anestrus cows compared to normal cyclic cows, and a significant increase (P&lt;0.05) in the MDA level occurred in anestrus cows compared control group. The hormonal analysis showed a significant decrease (P&lt;0.05) in both estrogen and progesterone with a significant elevation (P&lt;0.05) for cortisol in anestrus cows compared to cyclic cows. The study concluded the postpartum anestrus cows were under oxidative stress, which plays an essential role in anestrus, especially after parturition

Efficacy of an automated technology at detecting early postpartum estrus events: Can we detect resumption of cyclicity?

The objective of this observational study was to evaluate the efficacy of a neck-mounted automated activity monitor (AAM) at detecting early postpartum resumed ovarian cyclicity. A total of 192 lactating cows (primiparous = 73 and multiparous = 119) were enrolled in this study. Cows were continuously monitored by a neck-mounted AAM early postpartum (7 to 30 d in milk; DIM). Calving was classified as assisted (forced extraction of a calf) or unassisted (normal calving). Retained fetal membrane, metritis, hyperketonemia, clinical mastitis, and milk production were recorded. Cows were classified as healthy (i.e., no disease events) or sick (i.e., any disease event). Estrus events were alerted by the AAM using a proprietary algorithm set by the AAM company. Blood samples, from the coccygeal vein, were collected at 15, 18, 21, 24, 28, and 30 DIM for progesterone (P4) analysis. Resumption of cyclicity was considered when P4 concentration was ≥1 ng/mL on any collection day. Cows were considered anovular when P4 concentration was <1 ng/mL on all collection days. Cows were classified as true positive: P4 ≥ 1 ng/mL and at least one estrus alert; false positive: P4 < 1 ng/mL and at least one estrus alert; true negative: P4 < 1 ng/mL and no estrus alerts; and false negative: P4 ≥ 1 ng/mL and no estrus alerts. Statistical analyses were performed by frequency distribution and mixed effects logistic regression procedures on SAS (SAS Institute Inc.). The specificity, sensitivity, accuracy, and positive predictive value of the sensor to detect cows that had resumed cyclicity were 84.0%, 34.1%, 52.1%, and 79.2%, respectively. Out of the 192 cows, 35.9% (69/192) were anovulatory and 37.5% (72/192) had no estrus events between 7 to 30 DIM. Healthy cows were more likely to resume cyclicity in early lactation compared with cows that were sick (78.3 ± 1.9 vs. 32.8 ± 3.1%, respectively) independent of parity. In conclusion, the sensor had a high specificity for detecting anovular cows, but it had lower sensitivity, and thus was not effective at detecting cyclic cows, perhaps due to silent ovulation early postpartum.

PSII-10 Effect of Equine Chorionic Gonadotrophin-Like Glycoprotein Treatment on Fertility in Angus Cows Subjected to a 7-Day Co-Synch Protocol

Abstract This study evaluated the effect of administering a glycoprotein with equine chorionic gonadotropin like activity produced by recombinant DNA technology (eCG-like) associated with a 7-d Co-Synch protocol on estrus rate (ER) and pregnancy per timed AI (P/TAI). A total of 1,344 suckled Angus cows (primiparous, n = 381; multiparous, n = 963) from 11 herds were scanned at initiation of synchronization protocol by transrectal ultrasonography and the presence of corpus luteum (CL) and uterine tone was used to classify them by cycling. On day -10, cows received 100 μg GnRH analogue and a progesterone intravaginal device (IVD; 1.0g progesterone) for 7 days. At IVD removal, cows were administered 500 μg of cloprostenol and randomly assigned to receive 400 IU of eCG-like glycoprotein (Syntex S.A., Argentina, PCT/EP2019/073277; eCG-like, n = 665) or to remain as untreated (Control, n = 679). Also on day -3, paint was applied on the sacrum to the tail head of each cow. Finally, 100 μg GnRH analog were injected concurrently with TAI on Day 0 (66h after IVD removal) and the ER was evaluated by the degree of paint loss. Pregnancy to TAI was determined on day 30 by transrectal ultrasonography when an embryo and fluid were detected in the uterus. Body condition score (BCS; scale = 1-9) was evaluated on days -10 and 30. Optimal BCS was considered ≥ 5 and BCS balance between days -10 and 30 was classified as negative (lost BCS) or positive (maintained or gained BCS). The associations among treatment, parity, cyclicity at initiation of protocol, optimal BCS on day -10, and their interactions and ER and P/TAI were analyzed by GLIMMIX (SAS 9.3). Estrus expression and BCS balance were also included as effects in the model of pregnancy per TAI. The effect of herd was included as a random effect. The model specifications included an option to retrieve the mean, SEM, and an adjustment for multiple comparisons (Tukey-Kramer). The percentage of acyclic cows was 31.3%. The ER in cyclic cows was 64.9% and the eCG-like treatment increased ER only in anestrus cows (55.8 vs. 44.3%; Treatment x cyclicity interaction; P = 0.03). The percentage of cows with negative BCS balance was 27.2%. The P/TAI in cows with positive BCS balance was 48.4% and the eCG-like treatment increased P/TAI only in cows with negative BCS balance (44.7% vs. 30.5%; Treatment x BCS balance interaction; P = 0.01). In conclusion, the addition of a recombinant eCG-like glycoprotein to a 7-day Co-Synch protocol increased estrus expression in anestrus Angus cows and P/TAI in those with negative body condition score balance. However, this addition did not influence the estrus expression in cycling cows or on P/TAI in those with positive BCS balance.

Comparative study on plasma mineral profiles of normal cyclic and pregnant cows with infertile cows of North-Western Himalayan region of India

The present study was undertaken to study the effect of different minerals on infertility in cows (n=885) belonging to different Northern Western Himalayan regions of the state. Cows suffering from different types of infertility such as endometritis, anestrus, silent estrus and cows showing metestrual bleeding were evaluated and compared with normal cyclic and pregnant cows. Minerals evaluated were calcium, phosphorous, magnesium, copper, zinc, sodium, and potassium. The levels of calcium were statistically low in anestrus and silent estrus cows when compared with normal cyclic and pregnant animals. The levels of phosphorus were significantly low in cows suffering from endometritis and in cows showing abnormal metestrual bleeding in comparison to normal cyclic cows. Similarly, magnesium levels were high in pregnant cows than other categories. Copper was statistically high in normal cyclicand pregnant cows. Similarly, sodium and potassium were statistically high in normal cyclic cows than infertile and repeat breeder cows.

Total protein, albumin, and globulin levels of blood serum in repeat breeder Holstein Friesian cows

This study aimed to determine the relationship between total protein, albumin, and globulin in the blood with the incidence of repeat breeders in dairy cows. Ten normal cyclic cows and ten repeat breeders cows were taken randomly based on the reproductive records of the Tani Wilis Village Cooperative, Sendang District, Tulungagung Regency. Questionnaire data in the form of farmers' knowledge about dairy cows' reproductive systems and feed treatment is displayed descriptively. Dairy cows blood samples were taken from the jugular vein 6 mL to measure total protein, albumin, and globulin levels. Data analyzed using the Statistical Product and Service Solution software at a significance level of 5%. The results showed that the total protein, albumin, and globulin levels in normal dairy cows and dairy cows with repeat breeders were not significantly different (p&gt;0.05). However, in normal cyclic cows, most of the total protein, albumin, and globulin levels (80-100% of normal cyclic cows samples) are within the normal range. Meanwhile, the majority of repeat breeder cows (60-70% of the repeat breeder cows) had total protein, albumin, and globulin levels lower than the normal range (p&lt;0.05). It could be concluded that there was a relationship between the incidence of repeat breeder and total protein, albumin, and globulin levels in the blood serum of dairy cows with repeat breeder. Therefore, it is suggested to raise awareness of smallholder farmers about proper feeding and feeding management to reduce repeat breeder cases.

Total protein, albumin, and globulin levels of blood serum in repeat breeder Holstein Friesian cows

This study aimed to determine the relationship between total protein, albumin, and globulin in the blood with the incidence of repeat breeders in dairy cows. Ten normal cyclic cows and ten repeat breeders cows were taken randomly based on the reproductive records of the Tani Wilis Village Cooperative, Sendang District, Tulungagung Regency. Questionnaire data in the form of farmers' knowledge about dairy cows' reproductive systems and feed treatment is displayed descriptively. Dairy cows blood samples were taken from the jugular vein 6 mL to measure total protein, albumin, and globulin levels. Data analyzed using the Statistical Product and Service Solution software at a significance level of 5%. The results showed that the total protein, albumin, and globulin levels in normal dairy cows and dairy cows with repeat breeders were not significantly different (p&gt;0.05). However, in normal cyclic cows, most of the total protein, albumin, and globulin levels (80-100% of normal cyclic cows samples) are within the normal range. Meanwhile, the majority of repeat breeder cows (60-70% of the repeat breeder cows) had total protein, albumin, and globulin levels lower than the normal range (p&lt;0.05). It could be concluded that there was a relationship between the incidence of repeat breeder and total protein, albumin, and globulin levels in the blood serum of dairy cows with repeat breeder. Therefore, it is suggested to raise awareness of smallholder farmers about proper feeding and feeding management to reduce repeat breeder cases.

Intensity of estrus expression – valuable obvious determinant of fertility in Bos indicus cows

This study aimed to characterize estrus response and to establish relationships between intensity of estrus, preovulatory follicle (POF) size and estradiol (E2) concentrations on day of AI, luteal profiles and pregnancy outcome in lactating Hariana breed of cows. 200 cyclic cows were subjected to Ovsynch (n = 54) and Pre-OV treatment (n = 146). Ovsynch: Buserelin acetate (BA; 10 µg), Cloprostenol (500 µg) and BA (10 µg) were injected i.m. on day 0, 7 and 9, respectively, irrespective of treatment. Pre-OV: BA (10 µg) and Cloprostenol (500 µg) was also injected i.m. simultaneously 7 days prior to initiate Ovsynch. On the basis of estrus behavior, the cows were classified into three groups: weak, moderate and intense. Artificial insemination performed at 18–24 hours after 2nd BA of Ovsynch in both treatments. The average duration of estrus did not differ (p > 0.05) between Ovsynch and Pre-OV treatment. A positive correlation was observed between estrus response and POF size, concentration of E2 on day of AI and luteal profiles on day 12 post-AI. First service conception rate was higher in cows exhibited intense (45.46%) and moderate (42.56%) estrus response than weak (28.57%) estrus response. In conclusion, intensity of estrus expression could be considered as important determinant for deciding pregnancy outcomes in Bos indicus cows.

Exploring molecular dynamic indicators associated with reproductive performance of Bos indicus cattle in blood plasma samples through data-independent acquisition mass spectrometry

Improving reproductive performance of cattle is of paramount importance for sustainable dairy farming. Poor reproduction performance (RP) hinders the genetic improvement of important Bos indicus cattle breeds. It is well known that incorporation of molecular information along with conventional breeding method is far better than use of conventional method alone for the genetic improvement of reproductive performance traits in cattle. Therefore, the present study sought to investigate the plasma proteome of the Deoni cows in cyclical (n = 6) and pregnant (n = 6) reproductive phases with varying reproductive performance (high and low). High-throughput data independent acquisition (DIA) based proteomics was performed to understand corresponding proteome. We identified a total of 430 plasma proteins. Among cyclic cows, twenty proteins were differentially regulated in low RP as compared to high RP. BARD1 and AFP proteins were observed upregulated in cyclical cows whose upregulation reported to affect reproductive performance in cattle. Among the pregnant cows, thirty-five proteins were differentially regulated, including the downregulation of FGL2 and ZNFX1 that modulates the maternal immune response mechanism which is required for successful implantation of the embryo. Also, proteins such as AHSG, CLU and SERPINA6 were upregulated in the pregnant cows whose upregulation reported to reduced reproductive performance. The results of this study will be helpful in establishing a framework for future research on the aspect of improving reproductive performance in Bos indicus cattle breeds. SignificanceThe Indian subcontinent is the center of domestication for Bos indicus cattle breeds and they are known for their disease resistance, heat tolerance, ability to survive in low input regime and harsh climatic conditions. In recent times, population of many important Bos indicus breeds including Deoni cattle is declining due to various factors, especially due to reproductive performance. Traditional breeding methods are not sufficient enough to understand and improve the reproductive performance traits in important Bos indicus cattle breeds. Proteomics approach is a promising technology to understand the complex biological factors which leads to poor reproductive performance in cattle. The present study utilized DIA based LC- MS/MS analysis to identify the plasma proteins associated with reproductive performance in cyclical and pregnant cows. This study if improved further, can be used to develop potential protein markers associated with reproductive performance which is useful for the selection and genetic improvement of important Bos indicus breeds.

Effect of repeated low doses of GnRH analogue (buserelin) on fertility performance of dairy cows with anovulation type I.

The aim of this study was to evaluate the fertility response of dairy cows with anovulation type I on repeated low doses of GnRH agonist buserelin. The study was conducted on 83 anovulatory and 60 cyclic Polish Holstein Friesian cows. Anovulation type I was defined as small ovaries with follicles of ≤ 5 mm in diameter and without corpus luteum on two examinations in a 7-10 day interval between 50-60 days after parturition. Cows from the experimental group (n=58) received 0.4 μg of buserelin i.m. once a day for 5 consecutive days. Cows from the negative control group (n = 25) received saline. Sixty cyclic cows receiving no treatment served as positive controls. Intervals from calving to estrus and from calving to conception, pregnancy rate 30-35 days and 260 days after AI, and pregnancy loss were calculated. The anovulatory cows had a substantially prolonged calving to conception interval, decreased pregnancy rate and increased pregnancy loss and culling rate compared to cyclic herd mates. The average calving to conception interval was significantly (p⟨0.05) shorter in treated cows compared to non-treated anovulatory cows (153.7 days vs 209.3 days). In conclusion, repeated low doses of GnRH analogue buserelin led to a significant shortening of calving to conception interval. More clinical trials are needed to determine the practical usefulness of this method for the treatment of anovulation type I in dairy cows.

Comparison of the 7-day CO-Synch and 8-day estradiol-based protocols for estrus synchronization and timed artificial insemination in suckled Bos taurus cows

Two experiments were conducted to compare, follicle diameter (FD) on Day −1, corpus luteum (CL) area on Day 7, progesterone (P4) concentration on Day 7 and 18, pregnancy per timed artificial insemination (TAI) on Day 30, and pregnancy loss (PL) between Days 30 and 60 after TAI (TAI, Day 0) using two different synchronization protocols. In Experiment 1, Angus cows (n = 1148) were randomly assigned to either 7-d progesterone CO-Synch (7-d CO-Synch) or 8-d progesterone + estradiol (8-d P + ES) synchronization protocols for TAI. On Day −10, cows in the 7-d CO-Synch treatment group (n = 574) received a progesterone-releasing intravaginal device (PIVD; 0.5 g P4) and GnRH (0.105 mg), on Day −3 the PIVD was removed and cows received cloprostenol (0.150 mg), then, on Day 0 (64 h after PIVD removal), cows received GnRH (0.105 mg) and were TAI. On Day −10, cows in the 8-d P + ES treatment group (n = 574) received a PIVD (0.5 g P4) and estradiol benzoate (2.0 mg), on Day −2 the PIVD was removed, and cows received cloprostenol (0.150 mg) and estradiol cypionate (0.5 mg), then, on Day 0 (48 h after PIVD removal), cows were TAI. Pregnancy per TAI was determined on Days 30 and 60. In a subset of cows (7-d CO-Synch, n = 41; 8-d P + ES, n = 40), serum P4 concentration was evaluated on Day 18. In Experiment 2, anestrus (n = 34) and cyclic (n = 34) suckled beef cows were selected and submitted at random on Day −10, to either 7-d CO-Synch or 8-d P + ES treatment groups. Follicle diameter on Day −1, CL area, and serum P4 concentration on Day 7 were determined. In Experiment 1, pregnancy per TAI on Day 30 did not differ (7-d CO-Synch = 48.9%; 8-d P + ES = 45.6%) between treatments but it was greater for cows with BCS ≥5 (P < 0.01). Pregnancy loss between Days 30 and 60 did not differ between treatment groups but tended to be greater in cows with BCS <5.0 (P < 0.1). In a subset of cows, serum P4 concentration on Day 18 did not differ between treatment groups but tended to be lower (P < 0.1) in cows that had PL between Days 30 and 60 compared to cows that had no PL. In Experiment 2, FD tended to be greater (P < 0.1) and CL area was greater (P = 0.05) in anestrus cows from 7-d CO-Synch treatment. In cyclic cows, the treatment did not affect the FD or CL area. In conclusion, there was no difference in pregnancy per TAI on Day 30 and PL between Days 30 and 60 between cows using 7-d CO-Synch + PIVD or 8-d estradiol-based + PIVD protocols for estrus synchronization and TAI.

The immunolocalization of cadherins and beta-catenin in the cervix and vagina of cycling cows.

The adherens junctions (AJs) maintain the epithelial cell layers' structural integrity and barrier function. AJs also play a vital role in various biological and pathological processes. AJs perform these functions through the cadherin-catenin adhesion complex. This study investigated the presence, cell-specific localization, and temporal distribution of AJ components such as classical type I cadherins and beta-catenin in the cow cervix and vagina during the estrous cycle. Immunohistochemistry and Western blot analysis results demonstrated that beta-catenin and epithelial (E)-, neural (N)-, and placental (P)-cadherins are expressed in the cow cervix and vagina during the estrous cycle. These adhesion molecules were localized in the membrane and cytoplasm of the ciliated and non-ciliated cervical cells and the stratified vaginal epithelial cells. Positive immunostaining for P-, N-cadherin, and beta-catenin was also observed in the vascular endothelial cells of the cervical and vaginal stroma. Quantitative immunohistochemistry examinations revealed that in the cervical and vaginal epithelia, P-cadherin's optical density values (ODv) were the highest; in contrast, the N-cadherin ODv were the lowest. The ODv of P-cadherin and beta-catenin in the cervical epithelium and E-cadherin in the vagina were significantly higher in the luteal phase versus the follicular phase of the estrous cycle. Furthermore, the ODv of P-cadherin, N-cadherin, and beta-catenin in the cervix's central and peripheral epithelial regions were different during the estrous cycle. These findings indicate that classical cadherins and beta-catenin in the cervix and vagina exhibit cell- and tissue-specific expression patterns under the influence of estrogen and progesterone hormones during the estrous cycle.

Salivary crystallization pattern: a possible unconventional tool for timing of insemination and early pregnancy diagnosis in zebu cows.

The present study assessed if salivary crystallization pattern (ferning pattern formed as a result of the higher levels of salt content in the dried sample) could be used for estrus detection and for diagnosis of pregnancy/non-pregnancy in dairy cows. Saliva and blood samples were collected from non-pregnant cycling cows (Sahiwal breed; n = 20) on alternate days from the day of estrus till next estrus. Then, all the cows were inseminated and saliva and blood sampling were continued further for a period of 22 d post-insemination. Pregnancy diagnosis was carried out on day 45 post-insemination and eight cows were found to be pregnant. The salivary crystallization pattern and estradiol:progesterone ratio during estrous cycle and during pregnancy were compared among these cows. Six types of salivary crystallization patterns were discerned; distinct patterns such as branch-like, fern-like, fir-like and combinations of these. Fern-like pattern was observed in all the cows on the day of estrus (first measurement day) and furthermore, all of the cows that subsequently became pregnant had fern-like salivary crystallization pattern at the time of insemination. Saliva of all the pregnant cows showed branch-fir type of crystallization pattern on day 16 post-breeding while only 50% of non-pregnant cows showed this pattern on day 16 of estrous cycle. The appearance of fern-like pattern was positively and significantly related to estradiol:progesterone ratio (r = 0.86; P < 0.001). The findings were validated on a separate group of cycling cows (n = 32). We can conclude that salivary crystallization pattern might serve as a non-invasive and cost effective and easy-to-use cow-side tool for estrus detection and early pregnancy/non-pregnancy diagnosis in cows upon validation on a larger sample size.