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Theses (M.Sc.)

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2019 - 201942020 - 202321
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Type: Thesis × Subject: Veterinary Gynaecology and Obstetrics ×
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    ThesisItemOpen Access
    EFFECT OF DIETARY CURCUMIN SUPPLEMENTATION ON SEMINAL ATTRIBUTES OF CROSSBRED BULLS DURING INDUCED HEAT STRESS
    (ICAR-NDRI, KARNAL, 2023) MANISHA YADAV; NISHANT KUMAR
    An imbalance between heat production in the body and its dissipation is called as state of heat stress. When environmental temperature is high, it challenges the animal’s capacity to maintain energy, thermal, water, hormonal and mineral balance ultimately resulting in heat stress. Since bull fertility is considered to be adversely affected by hot humid condition, the current increase in global temperature as a result of climate change may have profound implications for future livestock production. Any strategy which could reduce the adverse effect of heat stress on bull fertility would be a welcome innovation. The present study was conducted with objectives (i) to study the effect of dietary curcumin supplementation on seminal and biochemical attributes in crossbred bulls and (ii) to evaluate the effect of dietary curcumin supplementation on physiological and sexual behavior of crossbred bulls. Heat stress was induced by placing the animals in psychrometric chamber. Both control and supplemented group were exposed to a temperature and relative humidity 37±20C and 50% respectively for 8 hours (9 AM to 5 PM) per day for 15 days. Pre and post heat exposure semen samples were evaluated during the experiment. The results indicated that the ejaculate volume was not significantly altered by induced heat stress. Sperm concentration, mass activity, motility, viability, plasma membrane integrity and sperm morphology at fresh and post thaw stage were significantly (P<0.05) altered by heat stress. However, these adverse effects were significantly (P<0.05) reduced in Curcumin supplemented groups. There was also a significant (P<0.05) difference in respiratory rate, rectal temperature and skin surface temperature between control and supplemented groups. There were significant (P<0.05) difference in the serum cortisol and testosterone concentration in both groups. Heat stress deteriorates semen quality from 2nd week and recovery occurs after 5th-6th week in supplementedgroup and after 8th week in control group and alters some of the physiological parameters. It can be concluded that supplementation with Curcumin have been found to ameliorate adverse effects of heat stress in cross bred bulls. The nutritional supplements like curcumin to ameliorate the adverse effects of heat stress in animals could be a beneficial and novel strategy particularly for animals during heat stress.
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    ThesisItemOpen Access
    EFFECT OF EXPOSURE OF URINE AND URINARY PHEROMONES OF BULLS ON EARLY RESUMPTION OF CYCLICITY IN POSTPARTUM SAHIWAL COWS
    (ICAR-NDRI, KARNAL, 2023) SANCHI KAMAL; NISHANT KUMAR
    Reproductive performance is a major predictor of profitability in dairy farms. Resumption of ovarian cyclicity is linked to better reproductive performance. Animals emit pheromones that communicate information between members of the same species. Objective of present study was identification of urinary pheromones of Sahiwal bulls and evaluation of effect of urine and urinary pheromones of Sahiwal bulls on reproductive performance and hormonal profile of Sahiwal cows. In first objective, 6 urinary samples were collected from 6 bulls for identification of chemical metabolites by Gas Chromatography Mass Spectrometry (GC-MS). Results indicated that 95 volatile compounds were identified with constituents belonging to alkanes, alkenes, alcohols, amides, aldehydes, carboxylic acids, ether, ketones, and phosphates. Compounds possessing pheromonal property were found to be undecane, eicosane, dimethyl sulfone, 1,2 benzene dicarboxylic acid, nonane, and octacosane. For second objective, a total of 18 Sahiwal cows were selected and divided equally in 3 groups. T0 group was exposed to NSS, while T1 group was exposed to bull urine and T2 group was exposed to synthetic pheromonal compounds after 20 days postpartum. Results indicated that interval from calving to resumption of cyclicity was significantly (P≤0.05) lower in T1(55.33±5.36days) as compared to T0(88.33±7.68days) and T2(82.33±12.37days), first behavioural estrus was also significantly (P≤0.05) lower in T1(60.17±5.45days) compared to T0(92.83±7.63days) and T2(85.83±12.57days). Interval from exposure to first insemination was observed to be lower in T1(100.5 ± 20.42 days) than T0(145±16.26days) and T2(113.33±11.29days). The amount of progesterone concentration in T1 group was significantly (P≤0.05) higher as compared to T0 and T2 group. Concentration of E2 on the day of estrus in T0, T1 and T2 was 20.39±1.43, 37.90±1.15 and 22.48±0.79 pg/ml respectively. Overall mean estrogen level was significantly (P≤0.05) higher in T1 group as compared to T0 and T2 group. The mean concentration of LH on the day of estrus in T0, T1 and T2 was 8.46±0.31, 11.63±0.56 and 9.27±0.34 ng/ml respectively and peak LH was found to be 11.18±1.61, 17.04±1.61 and 12.63±1.52 ng/ml respectively with significant difference (P<0.05) in T1 as compared to T0 and T2. Average leptin concentration at the start of experiment was 4.50±0.15, 4.60±0.17 and 4.59±0.19 ng/ml in T0, T1 and T2 respectively which had no significant difference. As time after parturition progress, level of leptin hormone had shown significant (p<0.05) difference in T1 group as compared to T0 and T2 group. The levels of BHBA during all fortnights were within normal physiological ranges indicating that animals were in positive energy balance. Size of largest follicle was found to be 12.26±0.34 mm, 14.98±0.38 mm and 12.68±0.33 mm in T0, T1 and T2 respectively, which was significantly (p<0.05) higher in T1 group as compared to T0 and T2. Overall score of estrus intensity in T0, T1 and T2 group was weak to moderate (66.67%, 33.33%), moderate to intense (66.67%, 33.33%) and weak to moderate (50%, 50%), respectively. The total duration of standing estrus (hours) was significantly (P<0.05) higher in T1(10.94±0.42 hours) as compared to T0(6.24±0.33 hours) and T2(6.51±0.32 hours). It can be concluded that bull urine exposed group resumed ovarian cyclicity 27-33 days earlier possibly indicating a rapid response of cows to male stimuli. It resulted in improved reproductive performance, hormonal profile, expression and intensity of estrus behaviour as compared to control and synthetic pheromones treated group.
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    ThesisItemOpen Access
    EFFECT OF LETROZOLE: A NON-STEROIDAL AROMATASE INHIBITOR ON OVARIAN FUNCTION AND ESTRUS SYNCHRONIZATION IN DAIRY CATTLE
    (ICAR-NDRI, BENGALURU, 2023) SAKHARE ABHIJEET CHANDRAKANT; S. JEYAKUMAR
    Estrus synchronization is becoming a necessity in the period of contemporary reproductive biotechnology for the augmentation of bovine fertility. There is demand for the development of short-term, cost-effective estrus synchronization protocols for adaptation at the field level. Letrozole, a non-steroidal aromatase inhibitor, can be an alternative approach for estrus synchronization in cattle. Letrozole reversibly inactivates the aromatase enzyme responsible for the synthesis of estrogen, delays the preovulatory LH surge, and prolongs the life span or induces growth of the dominant follicle by increasing LH pulse frequency. To date, most of the studies on the use of Letrozole for estrus synchronization have been conducted in beef cattle, and a perusal of the literature revealed a lack of studies on dairy cattle and also in indigenous cattle. With this background, the objectives of the present study were to determine the effect of Letrozole on ovarian function in dairy cattle and to evaluate the efficiency of Letrozole on estrus synchronicity and ovulation rate. In the present study, a total of 18 Deoni cows were selected and grouped as a blank vaginal sponge group (n = 6), a blank vaginal sponge plus PGF2α plus GnRH group (n = 6), and a Letrozole vaginal sponge plus PGF2α plus GnRH group (n = 6). The ovaries of all the animals were observed ultrasonographically before and after treatment to monitor follicular growth, ovulation, and corpus luteum development. Animals that came to heat in each group were inseminated with Deoni bull semen. Results of the present study revealed that treatment with Letrozole intravaginal sponges for four days at random stages of the estrous cycle resulted in synchronization of the estrus. The diameter of the dominant and pre-ovulatory follicles significantly (p < 0.05) increased in the Letrozole treatment group as compared to the control group. The diameter of the resultant corpus luteum (CL) formed after ovulation was significantly larger (p < 0.05) in the Letrozole treatment group. Similarly, the CL volume and area were significantly larger (p < 0.05) in the Letrozole treatment group, indicating the luteotropic property of Letrozole. There was no significant difference in CL blood flow area between the Letrozole treatment and control groups. Ovulation was more synchronous in the Letrozole treatment group. The present study demonstrates that the Letrozole+PGF2α+GnRH protocol improved the ovulation rate, increased the growth of the dominant follicle, the size of the preovulatory follicle, and the resultant CL. It is suggested that the Letrozole-based protocol can effectively be used for estrus synchronization programmes in Bos indicus cattle.
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    ThesisItemOpen Access
    OXIDATIVE STRESS INDUCED ALTERATIONS IN ZEBU BULL SPERM TRANSCRIPTOMIC PROFILE
    (ICAR-NDRI, BENGALURU, 2023) THAKOR HIRALBEN PRABHATSINH; A. KUMARESAN
    At physiological concentrations, Reactive Oxygen Species (ROS) play an important role in sperm functions and fertility; on the other hand, excessive production of ROS (oxidative stress), results in lipid peroxidation, DNA damage and protein oxidation affecting sperm fertility. Although there is a consensus among researchers that oxidative stress is associated with male infertility, the mechanism by which it affects fertility is not fully understood. Oxidative stress induced phenotypic/functional alterations in sperm are known to some extent, but the alterations induced by oxidative stress in the composition of sperm biomolecules important for fertilization and subsequent embryonic development are not known. The current study aimed to investigate the alterations in the global transcriptomic profile of zebu bull spermatozoa induced by oxidative stress. Oxidative stress was induced in freshly ejaculated spermatozoa from Deoni bulls (zebu; Bos indicus) by incubating them with sperm TALP containing 50 μM H2O2 for 1 h at 37 oC. Spermatozoa incubated in sperm TALP only under similar conditions served as the control. Then, the spermatozoa from the control and treatment groups were subjected to global transcriptomic analysis using high throughput next generation sequencing method. A total of 16,972 and 18,131 transcripts were detected in the control and treatment groups, respectively. It was found that 537 transcripts were unique to the control group while 1,696 transcripts were unique to the oxidative stress induced group. Among the 2737 dysregulated transcripts in the treatment group, 121 were upregulated while 176 were downregulated (p < 0.05) as compared to the control group. The top 10 upregulated transcripts in oxidative stress-induced spermatozoa included PEPB1, CYCS, PRC1, NDUFS3, LST1, SDHB, CDK4, EIF5B, CSNK2A1 and CCDC107 while the top 10 downregulated transcripts in oxidative stress-induced spermatozoa included LMNB1, FUT6, HDGF, DGKA, TMEM270, BTG1, KCTD4, GLB1L3, RAB8A and CNPPD1. All those differentially regulated transcripts were found to be involved in spermatogenesis, sperm motility, capacitation and acrosome reaction, which are important for fertilization. Further, it was identified that H2AFZ, PRDX2, TMED9, ADAMST2 and MRPL53 were detected uniquely in oxidative stress-induced spermatozoa, suggesting that these transcripts hold the potential to serve as indicators of oxidative stress in bull spermatozoa.
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    ThesisItemOpen Access
    DEVELOPMENT OF GOLD-NANOPARTICLE BASED COLOUR REACTION FOR DETECTION OF ESTRUS CANDIDATE PROTEINS IN SALIVA OF BUFFALOES
    (ICAR-NDRI, KARNAL, 2023) APARNA RAJ; RUBINA K. BAITHALU
    The domestic water buffalo (Bubalus bubalis) is an important triple purpose livestock species contributes immensely to the Indian dairy industry. However, one of the major constraints in the exploitation of production potential of buffalo is its inherently poor reproductive efficiency and factors which are contributing includes delayed sexual maturity, poor expression of estrus signs, seasonal expression of estrus signs leading to higher incidence of silent estrus, wrong time insemination and conception failure. Estrus detection is a major problem in buffaloes because of its shy breeding nature and poor expression of estrus signs that leads to wrong time insemination. Therefore, developing an easy and reliable test for estrus detection in buffaloes is essential. In this regard several studies carried out in our laboratory and by others to identify estrus biomarker proteins in easily accessible body fluids like saliva by employing proteomics approaches. But till date no on-spot test is available for accurate estrus identification in buffaloes. Therefore, present study was carried out to develop a gold-nanoparticle based colour test using earlier identified estrus candidate proteins in saliva of buffaloes. For this study a total of 26 estrous cycles from 8 heifers and 6 pluriparous buffaloes were included during breeding season (November-February). The standing to be mounted behaviour by teaser bull was most reliable sign of estrus and 92.3 % and 90.81 % of heifers and pluriparous buffaloes, respectively has shown this sign during estrus. Estrus stage was further confirmed by per-rectal examination, ultrasonography, cervical mucus ferning pattern and hormone estimation (progesterone and estradiol) in blood serum. Gold nanoparticles were prepared by citrate reduction method and the average size of prepared AuNPs was 21.26 nm with zeta potential of -24.3 mV as determined by DLS and characteristic plasmon resonance peak observed at 520 nm. Antibody concentration was optimized by considering different concentrations of VMO1, HSPA1A, CYP11A1 and ENO3 antibodies, respectively for making stable conjugate with AuNPs. Further, AuNP based colour assay was standardized for detection of estrus candidate peptides viz., VMO1, HSPA1A, CYP11A1 and ENO3. Dilution of saliva samples was optimized @ 1:15 dilution and BSA concentration was optimized @ 10% and 0.1% for initial and final blocking. AuNP-based colour assay was developed for detection of estrus candidate proteins viz., VMO1, HSPA1A, CYP11A1 and ENO3 in saliva of buffaloes. Sensitivity, specificity and accuracy of AuNP-based colour assay using HSPA1A antibody was found to be highest i.e., 91%, 77.2%, & 82%, respectively for differentiating estrus from other non-estrus stages. Several protocols were tried for optimization of conjugation of antibody with AuNPs using different buffer at different pH and antibody concentration for development of competitive lateral flow assay (cLFA). For finding the optimum amount of peptide at test line for the competitive interaction of AuNP-antibody conjugate and peptide, various dilutions of peptide were made. Saliva samples from proestrus, estrus, metestrus and diestrus stages of estrous cycle were taken for checking the efficiency of developed LFA strip against HSPA1A protein. The test line was observed with higher intensity during proestrus, metestrus and diestrus stages but not for estrus stage depicting higher presence of HSPA1A protein in saliva samples during estrus. Altogether this study developed a proof of concept for developing a method for accurate estrus identification in buffaloes.
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    ThesisItemOpen Access
    ELUCIDATING THE EFFICACY OF SODIUM ALGINATE AS AN ANTIBACTERIAL ALTERNATIVE IN CRYOPRESERVATION OF SAHIWAL BULL SEMEN
    (ICAR-NDRI, KARNAL, 2022) MOHIT KUMAR; NISHANT KUMAR
    The present study was conducted to investigate the efficacy of sodium alginate (S.A.) as an antibacterial alternative in cryopreservation of Sahiwal bull semen. In 1st objective, each of the pre-dominant Gram –ve (Proteus spp.) & Gram +ve (Bacillus spp.) bacterial isolates from Sahiwal bulls were collected and the antibacterial potency of sodium alginate was determined. Minimum inhibitory concentration (MIC) was determined by the broth microdilution method, in which two-fold serial dilutions of S.A. were done from 10 to 0.078 mg/ml concentration. It was found that the MIC of S.A. against Proteus spp. & Bacillus spp. was 0.625 and 1.25 mg/ml respectively. Similarly, the minimum bactericidal concentration (MBC) of S.A. against Proteus spp. & Bacillus spp. was found to be 0.625 and 2.5 mg/ml respectively. In 2nd objective, effect of S.A. as an antibacterial alternative on microbial load and freezability of Sahiwal bull semen was studied. Dose tolerance test in semen, as a pilot study was done by using six different S.A. concentrations (without streptopenicillin) based on MIC doses keeping one group as negative control (without streptopenicillin). Among these treated groups, three best doses (i.e. 1.25, 0.625 & 0.313 mg/ml) were selected on the basis of progressive motility & CFU/ml count for the standardization experiment. In standardization experiment, extended ejaculates were split into 3 treatment groups (1.25 mg/ml T1; 0.625 mg/ml T2; and 0.353 mg/ml T3- no antibiotics) and one control (Penicillin-1000IU/ml; Streptomycin-1mg/ml) group. Post-dilution, pre-freeze, post-thaw seminal parameters & bacterial load (CFU/ml) were evaluated, and T2 (0.625 mg/ml S.A.) was selected as the best dose for the final experiment. In the final experiment, ejaculates were split into 3 groups; one was kept as control (Penicillin-1000IU/ml; Streptomycin-1mg/ml), T1 (0.625 mg/ml S.A., no antibiotics) and T2 (0.625 mg/ml S.A. + antibiotics) groups. It was found that progressive motility, viability, HOST and acrosomal integrity of spermatozoa in the post-thaw stages was found to be significantly higher (p<0.05) in the T2 group as compared to the control & T1 group. Bacterial load (CFU/ml) at all the cryopreservation stages was found to be significantly lower (p<0.05) in the T2 group as compared to the control. Cryo-capacitation were significantly (p<0.05) reduced in the T2 group as compared to the control and T1 group at the post-thaw stages. No significant difference (p<0.05) was recorded in the protamine deficiency of the spermatozoa between the control, T1 & T2 groups at post-thaw stages. In the CASA study, total motility (TM), and progressive motility (PM) was found to be significantly higher (p<0.05) in T2 as compared to the control and T1 group. While there was no significant difference recorded in curvilinear velocity (VCL), straight linear velocity (VSL), average path velocity (VAP), and beat cross frequency (BCF) in the control and T2 groups. It can be concluded that S.A. has limited anti-bacterial activity as compared to strepto-penicillin, however it potentiates the antibacterial activity of strepto-penicillin if used in the supplementation. Also, S.A. in supplementation with antibiotics, enhances progressive motility, viability, plasma membrane integrity and acrosomal integrity of the sperm, as it protects the sperm from the adverse effects of cryopreservation.
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    ThesisItemOpen Access
    Early pregnancy diagnosis using color doppler ultrasonographic monitoring of luteal blood flow and growth dynamics in cattle
    (ICAR-SRS-NDRI, KARNAL, 2022) Majumder, Kaushik; S. JEYAKUMAR
    Early pregnancy diagnosis is a fundamental aspect of reducing the calving interval by identifying open animals early and rebreeding them as soon as possible. The Corpus luteum (CL) is a highly vascularized transitory endocrine gland that produces progesterone to support pregnancy. Luteal blood flow is a more accurate predictor of the CL function and serum P4 concentration than the CL size, particularly during the CL's regression phase. In this context, the evaluation of luteal blood flow by color Doppler ultrasonography can be a reliable parameter for diagnosing pregnancy. The aim of this study was to evaluate the CL growth dynamics and luteal blood flow changes in relation to early pregnancy. Multiparous (n=18) Deoni cows between 3-5 parity were selected for this study. Ultrasonographic examination and serum progesterone estimation was performed on days 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 33, 40, and 47 after estrus or until exhibition of next estrus, with or without insemination. Grayscale (B-mode) wasused for examining CL morphometry like diameter, tissue area, volume, and color-flow Doppler was used for assessing luteal blow area. Deoni cows were retrospectively allocated as being pregnant (n=6) or non-pregnant (interoestrus interval 18-21 days; n=6) by per rectal pregnancy diagnosis method on day 47, and were compared with nonbred cyclic control Deoni cows (n=6). The results revealed that the CL morphometry and blood flow area were progressively increasing from day 5 to 47 of the gestation period in pregnant cows, whereas it decreased from day 19 in inseminated non-pregnant and cyclic control cows. The CL diameter, volume and tissue area of pregnant cows was significantly higher than non-pregnant, and cyclic control (P<0.05) on day 19. Similarly, corpus luteal blood flow area was significantly higher (P<0.05) on day 19 in pregnant than in non-pregnant, or cyclic control cows. Though the CL vascularity index showed a similar trend, there was no significant difference between the groups. The serum P4 concentration declined from day 17 in non-pregnant and cyclic control cows and on day 19 it was significantly higher (P<0.05) in pregnant cows. In the present study, the CL morphometry and luteal blood flow on day 19 was significantly higher in pregnant than non-pregnant cows and are reflective of serum P4 concentration. Therefore, it was concluded that CL morphometry and CL blood flow area could be potential non-invasive predictors for early pregnancy diagnosis on day 19 in Deoni cows.
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    ThesisItemOpen Access
    EFFECT OF Murraya koenigii AND MELATONIN ON MODULATION OF BUFFALO GRANULOSA CELL FUNCTION IN HEAT STRESS CONDITION
    (ICAR-NDRI, KARNAL, 2022) CHIRAG PRUTHI; RUBINA K. BAITHALU
    This study aimed to establish the buffalo granulosa cells (GCs) culture model mimicking the intra-follicular environment in order to access the effect of heat stress on cell viability and functional characteristics of buffalo GCs and its amelioration by treatment of melatonin and Murraya koengii plant extracts. Buffalo GCs isolated from small antral follicles were cultured for 6 days in 3D-like by using gelatine pre-coated plates and a 3D culture model by using the hanging drop (HD) method. Three different cells seeding densities (2, 3, and 4X105/mL) were compared and found that seeding density of 4X105/mL was optimum to develop GCs of preovulatory phenotype after 96 hrs of culture based on morphology and functional attributes (CYP19, FSHR, RUNX, and PCNA) in both the culture models. Based on structural and functional characteristics, a 3D culture model using the hanging drop method was selected to access the effect of heat stress by exposing GCs to higher temperatures (40.5˚C & 41.5˚C) and the functional attributes of cells were studied. Exposure to acute heat stress for 2-24 hrs after attaining preovulatory phenotype showed a significant difference in morphology, growth characteristics, and functional attributes. Chronic heat stress exposure for 24-120 hrs after 24 hrs of culture affected the GCs growth and cells didn’t form compact spheroids and started to lose their phenotype earlier (after 72 hrs) than in the control group (120 hrs). Further, heat stress exposure affected the function of GCs by reducing the expression of PCNA, CYP19, FSHR, and RUNX and increasing the expression of HSP70 after 72 hrs. Subsequently, the GCs in the 3D culture model were treated with melatonin and Murraya koenigii plant extracts to study their ameliorating effect against heat stress on GCs functional attributes. Total phenolics (TP), Total flavonoid content (TFC), and reducing power of Murraya koenigii leaves were 459.67±9.1 μg GAE/mg, 788.83±5.49 μg QE/mg, and 16556.81±1445.2 μmol TE/mg in methanolic extract respectively and 417±69.9 μg GAE/mg, 52.21±14. μg QE/mg and 20042.95±1866.32 μmol TE/mg in aqueous extract respectively. Cytotoxicity assay revealed maximum cell proliferation at concentrations of 0.390 mg/mL, 3.125 mg/mL, and 300 ng/mL for methanolic extract, aqueous extract, and melatonin, respectively. Melatonin & plant extracts (aqueous and methanolic) treatment to GCs has shown an ameliorative response against heat stress by the maintenance of their structural & functional phenotype evidenced by significantly higher expression (p<0.05) of PCNA, CYP19, FSHR, antioxidant genes (CAT and SOD2) & reduced expression (p<0.05) of HSP70 & BAX transcript under acute & chronic heat stress condition. The estradiol-17-β (E2) analysis in the spent culture medium revealed reduced basal production of estradiol in heat stress conditions (acute- 6.48 pg/μg of cellular protein; chronic- 6.76 pg/μg of cellular protein) compared to the control (acute-7.52 pg/μg of cellular protein and chronic-8.26 pg/μg of cellular protein) whereas, melatonin treatment improved basal production of estradiol under control (acute-9.06 pg/μg of cellular protein and chronic-8.07 pg/μg of cellular protein) and heat stress condition (acute-8.48 pg/μg of cellular protein and chronic-7.27 pg/μg of cellular protein). It could be concluded that the hanging drop method with a seeding density of 4X105 cells/ mL is one of the best strategies to culture buffalo GCs mimicking the inta-follicular environment. Heat stress exposure to GCs hampers the growth, differentiation, and steroidogenesis which can be ameliorated by the use of Murraya koenigii leaves extracts and melatonin.
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    ThesisItemOpen Access
    PROTEOMIC PROFILING OF OVARIAN FOLLICULAR FLUID FROM NORMAL AND PROLONGED FOLLICULAR DOMINANCE IN DEONI COWS
    (ICAR-SRS-NDRI, KARNAL, 2022) NIRIBILI RAJBANGSHI; S. JEYAKUMAR
    Dairying is an integral part of animal husbandry, and efficient reproduction in dairy cattle makes a distinct contribution to the socio-economic growth of the country. Prolongation of follicular dominance is one of the conditions associated with disconcerted follicular dynamics that result in substantial economic losses through low reproductive efficiency. Despite the fact that there is a lot of research available on the mechanisms of prolonged follicular dominance and oocyte viability, the proteomic alterations in the microenvironment of the prolonged follicle remain unknown. In this regard, we investigated the ultrasonographic characteristics of follicles as well as the proteomic alterations in follicular fluid from normal (NDFF) and prolonged dominant follicles (PDFF). Prolonged dominance was induced in Deoni cows (n = 6) by placing CIDR (previously used for 7 days) intravaginally from day 4 to 8 of estrus, with a luteolytic dose of PGF2 on day-6 and day-7resulting in subluteal progesterone concentration. Ultrasonographic examination revealed no significant difference in the size and growth pattern of normal and prolonged dominant follicles. The concentration of serum progesterone in the prolonged follicular dominance group was significantly higher (P<0.05) than in the normal follicular dominance group on the day of transvaginal follicular aspiration (day-8). Furthermore, global proteomic analysis of follicular fluid detected 217 proteins in the Deoni cow, with the majority of the identified proteins involved in 21 pathways, 42 molecular functions, and 106 biological processes. Complement and coagulation cascades (22.8%) and cholesterol metabolism (4.68%) were the major pathways in which identified proteins were involved. In comparison to NDFF, 26 proteins were dysregulated in PDFF. Among them, 15 proteins showed upregulation, and 11 proteins showed downregulation. Proteins involved in complement and coagulation cascades, and vitamin digestion and absorption were found to be dysregulated. In this study, the ovarian follicular characteristics were not different in cows experiencing prolonged follicular dominance from those of normal follicular dominance. However, the expression of proteins involved in inflammation, oocyte metabolism, and ovulation of follicles was dysregulated in the follicular fluid of cows with prolonged follicular dominance, which might be responsible for delayed ovulation.