64 COULD FROZEN–THAWED BOAR-SEMEN FERTILITY BE INCREASED WITH SEMINAL PLASMA ADDITION?

2015 ◽  
Vol 27 (1) ◽  
pp. 125
Author(s):  
M. A. Torres ◽  
G. M. Ravagnani ◽  
M. de Lima Oliveira ◽  
D. F. Leal ◽  
G. Amorim de Campos ◽  
...  
Keyword(s):  
Seminal Plasma ◽  
Mixed Models ◽  
Intrauterine Insemination ◽  
Final Concentration ◽  
Fertility Rate ◽  
Experimental Conditions ◽  
Orthogonal Contrasts ◽  
Binary Distribution ◽  
Boar Semen ◽  
C 30

Post-thawed addition of seminal plasma (SP) in equine cryopreserved semen increased the integrity of plasma and acrosomal membranes (Andrade et al. 2011 Reprod. Dom. Anim. 46, 682–686) and these possibly affect sperm lifespan, improving fertility (Garcia et al. 2010 Anim. Reprod. Sci. 119, 160–165). This study was conducted to verify the pregnancy (PR) and fertility rate (FR) of addition of homologous SP in thawed boar semen. First, SP collections were made with polled sperm rich-fraction. Semen was centrifuged (500 × g for 10 min) and supernatant was removed, centrifuged one more time (2500 × g for 30 min), vacuum filtered through membranes (0.22 μm), and stored at –80°C for future use. Samples collected to frozen were pooled and divided in 2 aliquots, control (cryopreserved with SP; CON) and cryopreserved without SP (WSP; SP was removed and discarded after centrifugation – 500 × g for 10 min). Samples were extended in freezing extender (Botupharma®) to a final concentration of 300 × 106 spermatozoa per milliliter, packaged in 0.5-mL straws, and frozen in an automatic system (TK Tecnologia em Congelação®) using a rate of –0.5°C min–1 until 5°C, –20°C min–1 until –120°C, and then immersed in LN (–196°C). Ten straws, from each treatment, were thawed in water bath (37°C/30 s) and extended in Beltsville thawing solution to obtain 1.5 × 109 sperm in 40 mL. The other 10 straws of WSP were thawed and extended in Beltsville thawing solution plus 10% (v:v) of SP to originate treatment TSP (thawed added of seminal plasma). Thirty-three (11 per treatment) gilts had synchronized ovulation with altrenogest (25 mg/5 mL, Regumate®) administration per 18 days. Following day after withdrawal the altrenogest was administered with an intramuscular injection of 600 IU of eCG (Novormon®) and 2.5 mg of pLH (Lutropin®-V) after 72 h; a single deep intrauterine insemination was made 36 h after. Five days after, females were slaughtered and embryos were collected (by oviducts flushed) and evaluated by esteromicroscopia. Fertility rate and PR were analysed by SAS program (SAS Institute Inc., 2010, Cary, NC, USA). Fertility rate was analysed by Mixed models, and treatment effects were analysed by orthogonal contrasts (C1: effect without SP = CW v. NC; C2: effect of post-thawed addition of SP = CP v. CW), and PR was evaluated by binary distribution with PROC GLIMMIX test. Fertility rate was not affected (P > 0.05) by cryopreservation of boar semen in presence or absence of SP nor by its addition in Beltsville thawing solution (10.58 ± 3.92, 9.57 ± 4.92, 21.29 ± 7.37 for CON, WSP, and TSP, respectively). Treatments did not influence (P > 0.05) PR (50.00, 36.36, 72.73 for CON, WSP, and TSP, respectively). Thus, neither SP addition in thawed boar semen nor cryopreservation with or without SP can be beneficial to PR and FR, in our experimental conditions. However, a numeric large difference was observed. Therefore, these results lead us to believe that SP have a potential to increase the fertility and pregnancy rate, and that can be verified in further studies, with more repetitions.Research was supported by Agroceres Multimix, Botupharma and FAPESP process 2013/08070-8 and 2011/23484-8.

scholarly journals A practical solution to pseudoreplication bias in single-cell studies

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Kip D. Zimmerman ◽  
Mark A. Espeland ◽  
Carl D. Langefeld
Keyword(s):  
Single Cell ◽  
Mixed Models ◽  
Random Effect ◽  
Cell Types ◽  
Error Rates ◽  
Specific Cell ◽  
Experimental Conditions ◽  
Type 1 Error ◽  
Sample Correlation ◽  
Inflated Type

AbstractCells from the same individual share common genetic and environmental backgrounds and are not statistically independent; therefore, they are subsamples or pseudoreplicates. Thus, single-cell data have a hierarchical structure that many current single-cell methods do not address, leading to biased inference, highly inflated type 1 error rates, and reduced robustness and reproducibility. This includes methods that use a batch effect correction for individual as a means of accounting for within-sample correlation. Here, we document this dependence across a range of cell types and show that pseudo-bulk aggregation methods are conservative and underpowered relative to mixed models. To compute differential expression within a specific cell type across treatment groups, we propose applying generalized linear mixed models with a random effect for individual, to properly account for both zero inflation and the correlation structure among measures from cells within an individual. Finally, we provide power estimates across a range of experimental conditions to assist researchers in designing appropriately powered studies.

scholarly journals Thawing boar semen in the presence of seminal plasma improves motility, modifies subpopulation patterns and reduces chromatin alterations

2017 ◽  
Vol 29 (8) ◽  
pp. 1576 ◽  
Author(s):  
Rocío Fernández-Gago ◽  
Manuel Álvarez-Rodríguez ◽  
Marta E. Alonso ◽  
J. Ramiro González ◽  
Beatriz Alegre ◽  
...  
Keyword(s):  
Seminal Plasma ◽  
Chromatin Structure ◽  
Sperm Chromatin ◽  
Chromatin Compaction ◽  
Sperm Analysis ◽  
Positive Effects ◽  
Fragmentation Index ◽  
Non Linear ◽  
Boar Semen ◽  
Dna Fragmentation Index

Seminal plasma could have positive effects on boar semen after thawing. In the present study we investigated changes in the motility and chromatin structure in spermatozoa over 4 h incubation (37°C) of boar semen thawed in the presence of 0%, 10% or 50% seminal plasma from good-fertility boars. Cryopreserved doses were used from seven males, three of which were identified as susceptible to post-thawing chromatin alterations. Motility was analysed by computer-aided sperm analysis every hour, and data were used in a two-step clustering, yielding three subpopulations of spermatozoa (slow non-linear, fast non-linear, fast linear). Chromatin structure was analysed using a sperm chromatin structure assay and flow cytometry to determine the DNA fragmentation index (%DFI) as a percentage, the standard deviation of the DFI (SD-DFI) and the percentage of high DNA stainability (%HDS), indicating chromatin compaction. Thawing without seminal plasma resulted in a rapid loss of motility, whereas seminal plasma helped maintain motility throughout the incubation period and preserved the subpopulation comprising fast and linear spermatozoa. The incidence of chromatin alterations was very low in samples from non-susceptible males, whereas samples from males susceptible to post-thawing chromatin alterations exhibited marked alterations in %DFI and %HDS. Seminal plasma partly prevented these alterations in samples from susceptible males. Overall, 50% seminal plasma was the most efficient concentration to protect motility and chromatin. Some changes were concomitant with physiological events reported previously (e.g., semen thawed with 50% seminal plasma increased the production of reactive oxygen species and yielded higher fertility after AI). Thawing in the presence of seminal plasma could be particularly useful in the case of samples susceptible to post-thawing chromatin damage.

scholarly journals Fertilisation rate obtained with frozen-thawed boar semen supplemented with rosmarinic acid using a single insemination timed according to vulvar skin temperature changes

2015 ◽  
Vol 63 (1) ◽  
pp. 100-109 ◽  
Author(s):  
Victoria Luño ◽  
Lydia Gil ◽  
Maite Olaciregui ◽  
Juan Grandía ◽  
Trinidad Ansó ◽  
...  
Keyword(s):  
Skin Temperature ◽  
Artificial Insemination ◽  
Rosmarinic Acid ◽  
Intrauterine Insemination ◽  
Fertility Rates ◽  
Temperature Changes ◽  
Progressive Motility ◽  
A Value ◽  
Fertilisation Rate ◽  
Boar Semen

Artificial insemination (AI) of sows with frozen-thawed semen usually results in lower pregnancy rates and litter sizes than the use of liquid preserved semen. The present study evaluated the effectiveness of vulvar skin temperature changes as a predictor of ovulation in sows and determined the fertility rates obtained after AI with frozen-thawed semen supplemented with rosmarinic acid (RA). Semen was collected from mature boars and cryopreserved in experimental extenders supplemented with or without 105 μM of RA. Multiparous sows were inseminated with a single dose of semen when vulvar skin temperature decreased to a value below 35 °C. Intrauterine insemination was performed using 1.5 × 109 spermatozoa. The sows were slaughtered 48 h after AI and the embryos and oocytes were recovered from the oviducts. Total and progressive motility, viability and acrosome integrity were significantly (P < 0.05) higher in RA-supplemented semen samples compared with the control. Fertilisation occurred in all sows inseminated in the study, although there were no significant differences between the experimental groups. Sows inseminated with RA-supplemented semen showed a slight increase in the number of embryos recovered as compared to sows inseminated with control semen. In conclusion, insemination according to vulvar skin temperature changes resulted in successful fertilisation in all sows, although supplementation of the freezing media with RA did not improve the fertilising ability of frozen-thawed boar sperm.

Spermatozoa and seminal plasma small extracellular vesicles miRNAs as biomarkers of boar semen cryotolerance

2021 ◽  
Author(s):  
Ana Carolina Pedrosa ◽  
Mariana A. Torres ◽  
Diego V. Alkmin ◽  
Jorge E.P. Pinzon ◽  
Simone Maria M.K. Martins ◽  
...  
Keyword(s):  
Extracellular Vesicles ◽  
Seminal Plasma ◽  
Boar Semen

Seasonal changes of steroid concentrations in seminal plasma of a European wild boar

1984 ◽  
Vol 107 (3) ◽  
pp. 425-427 ◽  
Author(s):  
D. Schopper ◽  
J. Gaus ◽  
R. Claus ◽  
H. Bader
Keyword(s):  
Wild Boar ◽  
Seminal Plasma ◽  
Seasonal Pattern ◽  
Plasma Concentrations ◽  
Boar Taint ◽  
Steroid Production ◽  
European Wild Boar ◽  
Clear Seasonal Pattern ◽  
Boar Semen ◽  
Low Levels

Abstract. The influence of season on testicular steroid production as a parameter of testicular function has been studied in a wild boar. Semen was collected once weekly while it served the dummy. In seminal plasma concentrations of the following steroids were determined by radioimmunoassay: unconjugated testosterone, conjugated testosterone, unconjugated total oestrogens, conjugated total oestrogens and 5α-androst-16-en-3-one ('boar-taint steroid'). All steroids showed a clear seasonal pattern with highest concentrations in autumn and early winter and low levels from January to July. Maxima during the rutting season were 10–25 times greater than average values out of season. During a 2-month-period (mid-July until mid-September) libido was abolished and the wild boar refused to mount the dummy. These results indicate that the seasonal variation in testicular steroid production by the wild boar, regulated by photoperiod, are similar to those of the domestic boar.

scholarly journals Effect of bicarbonate on growth of the oleaginous microalga Botryococcus braunii

2019 ◽  
Vol 10 (1) ◽  
Author(s):  
Giovanna Salbitani ◽  
Carmela M.A. Barone ◽  
Simona Carfagna
Keyword(s):  
Inorganic Carbon ◽  
Algal Growth ◽  
Botryococcus Braunii ◽  
Final Concentration ◽  
Time Intervals ◽  
Experimental Conditions ◽  
Co2 Gas ◽  
Inorganic Carbon Source ◽  
Doubling Times

The effect of bicarbonate, produced by the enzymatic hydration of CO2 from postcombustion fumes, was investigated on Botryococcus braunii growth. The NaHCO3, supplied to cultures in the role of inorganic carbon source is proposed as a more eco-sustainable alternative to gaseous CO2. The salt was provided to the cultures at the final concentration of 0.5-1.5-2.5 g L- 1. The growth rate was considered for specific time intervals (T0-T5, T5-T10 and T0- T10) showing values significantly higher in the culture supplemented with 2.5 g L-1 bicarbonate. The doubling times were also considered in all experimental cultures showing a faster doubling for the period T0÷T5. The increase in pH drives the increase in growth in the experimental conditions in which the salt was added. The results suggest that bicarbonate is able to promote the algal growth, therefore it can be considered a valid alternative to CO2 gas.

scholarly journals Expression Pattern of Seminal Plasma Extracellular Vesicle Small RNAs in Boar Semen

2020 ◽  
Vol 7 ◽  
Author(s):  
Zhiqian Xu ◽  
Yanshe Xie ◽  
Chen Zhou ◽  
Qun Hu ◽  
Ting Gu ◽  
...  
Keyword(s):  
Expression Pattern ◽  
Seminal Plasma ◽  
Small Rnas ◽  
Extracellular Vesicle ◽  
Boar Semen

Pre-Adipocytes Differentiate Into Megakaryocytes (MK) by a Paracrine Thrombopoietin (TPO) Loop

Blood ◽  
2012 ◽  
Vol 120 (21) ◽  
pp. 4742-4742
Author(s):  
Yukako Ono ◽  
Yumiko Matsubara ◽  
Tatsuya Tanaka ◽  
Shinichiro Okamoto ◽  
Yasuo Ikeda ◽  
...  
Keyword(s):  
Cell Line ◽  
Board Of Directors ◽  
Terminal Differentiation ◽  
Final Concentration ◽  
Similar Degree ◽  
Potential Candidate ◽  
Advisory Committees ◽  
Experimental Conditions ◽  
Maintenance Medium ◽  
Primary Mouse

Abstract Abstract 4742 The regulation of megakaryopoiesis and thrombopoiesis are incompletely understood. Better understanding of the underlying mechanisms would be of biological import, but may also lead to novel approaches for generating of platelets for clinical application. One cell line that undergoes terminal differentiation into megakaryocytes is pre-adipocytes. These cells represent a potential candidate cell for ex vivo generation of megakaryocytes. Here we demonstrate that pre-adipocytes differentiate into MKs and platelets by using an endogenous paracrine loop involving TPO, the primary cytokine involved in megakaryopoiesis, and its receptor c-mpl. We previously reported that pre-adipocytes differentiate into MKs and form platelets when the culture medium is switched from maintenance medium to MK lineage induction (MKLI) medium. Neither media include TPO. Based on these observations, the present study tested the hypothesis that pre-adipocytes might differentiate into MKs and platelets by endogenous TPO and c-mpl expression of sufficient magnitude to drive megakaryopoiesis. We used primary mouse pre-adipocytes (CD45-, CD117-, Sca1+, CD29+, CD73+, CD90+, CD105+, CD106+) from subcutaneous adipose tissues and also the mouse pre-adipocyte line 3T3-L1 cells. The TPO levels, as assessed by ELISA, in the supernatant from 106 pre-adipocytes cultured in 2 ml MKLI medium without exogenous TPO (TPO-) were unmeasurable level on Day 0, 29±14 pg/ml on Day 7 and 8±2 pg/ml on Day 12. Similar results were obtained in the supernatant from 3T3-L1 during MK differentiation. We did not observe measureable TPO in supernatants from pre-adipocytes and 3T3-L1 cells cultured in maintenance medium on Days 0, 7 and 12. We then compared MK differentiation from pre-adipocytes in MKLI media in the absence (TPO-) or addition (final concentration, 50 ng/ml; TPO+) of TPO. The frequency of CD41-postive pre-adipocytes in culture after 6 days was 58±11% for TPO+ and 70±7% for TPO- (p>0.05), consistent with endogenous TPO being sufficient under these circumstances to stimulate MK differentiation of pre-adipocytes. DNA ploidy and c-mpl expression assessed by immunohistochemistry were also similar with or without added exogenous TPO. We next examined the number of CD41-expressing large-sized (>15 μm) cells beginning with 1.2 × 104 preadipocytes in the absence or presence of the c-mpl inhibitor rat anti-mouse c-mpl inhibitor AMM2 (final concentration, 10 μg/ml). In the absence of AMM2, 10-fold more CD41-positive, large cells were seen (6.8±1.7×103) than in its presence (0.6±0.2×103, p<0.05). These observations support that pre-adipocytes differentiate into MKs using endogenous TPO stimulation via c-mpl. We next examined the ability to release platelets from these treated pre-adipocytes in each experimental condition (pre-adipocytes and 3T3-L1 cells, each ± exogenous TPO). Infusion of 2 × 105 carboxyfluorescein diacetate succinimidyl ester (CFSE)-labeled MKs into irradiated thrombocytopenic mice (7 days post-2.0Gy exposure) led to a time-dependent appearance of CFSE+/CD41+ platelet-sized particles with a peak at 4 hours after the infusion reaching ∼2.5% of the total circulating platelet population (∼30 platelets/infused MK) under all tested experimental conditions. For platelet function, blood samples from each of these mice were perfused over a collagen-coated chip for 10 minutes, and the incorporation of CFSE+ particles into thrombi on the chip was determined (Total Thrombus-formation Analyzing System). A similar degree of platelet incorporation was observed under all experimental conditions and each was with an efficiency similar to that seen when CFSE+ control platelets were infused. These findings demonstrate that pre-adipocytes differentiate into MKs and subsequent platelets by an endogenous TPO release and a paracrine loop involving c-mpl. We propose that such pre-adipocytes could be used as a model of a continuously replicating cell line that upon switching media simultaneous expresses TPO and its receptor c-mpl to establish a paracrine loop leading to terminal differentiation into functional MKs. The basis of why this media switch induces this change needs to be clarified to further develop this pre-adipocyte strategy as a donor-independent source for platelet transfusion as well as for studying mechanism underlying megakaryopoiesis and thrombopoiesis. Disclosures: Matsubara: Medico's Hirata: Honoraria; Advisory Committees on VerifyNow: Membership on an entity's Board of Directors or advisory committees. Murata:Medico's Hirata: Honoraria; Advisory Committees on VerifyNow: Membership on an entity's Board of Directors or advisory committees.

Pengaruh Konsentrasi Sperma Terhadap Daya Tetas dan Bobot Tetas DOC Ayam KAMARAS

2017 ◽  
Vol 2 (4) ◽  
Author(s):  
Moulizar Moulizar ◽  
Aman Yaman ◽  
Muhammad Daud
Keyword(s):  
Sex Ratio ◽  
Sperm Concentration ◽  
Final Concentration ◽  
Fertility Rate ◽  
High Fertility ◽  
Female Ratio ◽  
Male And Female ◽  
Egg Fertility ◽  
Female Chicken ◽  
Local Chickens

Abstrak. Turunan hasil persilangan antara ayam lokal, ayam ras petelur dan ayam arab dihasilkan Ayam KAMARAS (Kampung - Arab – Ras).   Penelitian ini bertujuan untuk mengetahui pengaruh konsentrasi sperma terhadap fertilitas, daya tetas, bobot tetas DOC ayam KAMARAS dan Sex rasio. Konsentrasi sperma  ini berguna untuk mengetahui perbandingan jantan dan betina yang paling efisien dalam suatu pemeliharaan. Sperma ditampung dan dicampur dari 3 ekor ayam KAMARAS jantan yang berumur sekitar 7 bulan. Sperma yang telah diketahui konsentrasinya kemudian dibagi menjadi 4 bagian dan masing-masing diencerkan dengan NaCl fisiologis sehingga diperoleh konsentrasi akhir yaitu 25x106/0,5 ml (dosis 1); 50x106/0,5 ml (dosis 2); dan 75x106/0,5 ml (dosis 3) dan 100x106/0,5 ml (dosis 4). 24 ekor ayam KAMARAS betina diinseminasi untuk mengetahui pengaruh konsentrasi sperma terhadap daya tetas dan bobot tetas ayam KAMARAS. Ayam betina yang digunakan adalah ayam KAMARAS yang berumur sekitar 7 bulan.Hasil penelitian menunjukkan bahwa perlakuan konsentrasi sperma tidak berpengaruh nyata terhadap fertilitas telur namun disetiap perlakuan terdapat hasil yang berbeda-beda pada perlakuan P0: 25x106/0,5 ml tingkat fertilitas (100 %), P1: 50x106/0,5 ml (100%), P2: 75x106/0,5 ml dan pada perlakuan P3 terendah 83,33% diantara 4 perlakuan konsentrasi sperma yang terbaik untuk menghasilkan fertilitas  yang tinggi yaitu konsentrasi 50x106/0,5 ml. Pengaruh konsentrasi sperma terhadap daya tetas tidak berpengaruh nyata namun disetiap perlakuan terdapat hasil yang berbeda-beda pada perlakuan P0: 25x106/0,5 ml tingkat fertilitas (100 %), P1: 50x106/0,5 ml (100%), P2 75x106/0,5 ml (100%) dan pada perlakuan P3 terendah (83,33%) dari setiap perlakuan konsentrasi sperma yang dapat meningkatkan daya tetas telur adalah pada konsentrasi 50x106/0,5 ml dengan tingkat daya tetas (100%). Pengaruh konsentrasi sperma terhadap bobot tetas DOC KAMARAS berpengaruh nyata terhadap bobot tetas DOC pada setiap perlakuan P0 25x106/0,5 (31,1 g/butir), P1: 50x106/0,5 ml (33 g/butir), P2: 75x106/0,5 ml (32,1 g/butir) dan pada perlakuan P3 100x106/0,5 ml bobot tetas DOC sebesar (33,3 g/butir),  pengaruh konsentrasi sperma terhadap sex rasio didapat hasil pada perlakuan 50x106/0,5 ml-75x106/0,5 ml menghasilkan sex rasio DOC betina lebih dominan (66,6-75%).Effect of Sperm Concentration on Hatching Power and Hatching Weight of DAM Chicken KAMARASAbstract. Derivative results of crosses between local chickens, laying chicken and arab chicken produced Chicken KAMARAS (Kampung - Arab - Ras). This study aims to determine the effect of sperm concentration on fertility, hatchability, weight of DOC chicken KAMARAS and Sex ratio. This sperm concentration is useful for knowing the most efficient male and female ratio in a maintenance. Sperm was collected and mixed from 3 chickens KAMARAS male about 7 months old. The known sperm concentration is then divided into 4 parts and each diluted with physiological NaCl to obtain the final concentration of 25x106 / 0.5 ml (dose 1); 50x106 / 0.5 ml (dose 2); and 75x106 / 0.5 ml (dose 3) and 100x106 / 0,5 ml (dose 4). 24 chickens KAMARAS females inseminated to determine the effect of sperm concentration on hatchability and weight of hens chicken KAMARAS. The female chicken used is KAMARAS chicken which is about 7 months old.The results showed that sperm concentration treatment did not have significant effect on egg fertility but in each treatment there were different results on treatment of P0: 25x106 / 0,5 ml fertility rate (100%), P1: 50x106 / 0,5 ml (100% ), P2: 75x106 / 0.5 ml and at the lowest P3 treatment 83.33% among the 4 best sperm concentration treatment to produce high fertility ie concentration 50x106 / 0,5 ml. The effect of sperm concentration on hatchability was not significant but in each treatment there were different results on treatment of P0: 25x106 / 0,5 ml fertility rate (100%), P1: 50x106 / 0,5 ml (100%), P2 75x106 / 0.5 ml (100%) and at the lowest P3 treatment (83.33%) of each treatment of sperm concentration which can increase the hatchability of eggs at concentration 50x106 / 0,5 ml with hatchability level (100%). The influence of sperm concentration on DOC KAMARAS hatching weight significantly influenced the weight of DOC in each treatment P0 25x106 / 0,5 (31,1 g / grain), P1: 50x106 / 0,5 ml (33 g / grain), P2: 75x106 / 0,5 ml (32,1 g / grain) and at treatment of P3 100x106/ 0,5 ml of DOC hight weight (33,3 g / grain), influence of sperm concentration to sex ratio obtained result at treatment 50x106 / 0, 5 ml-75x106 / 0.5 ml result in sex ratio of female DOC is more dominant (66,6-75%).

Proteins and Heparin Binding Proteins of Spermatozoa and Seminal Plasma in Beetal Bucks: Purification, Characterization and Role in In vivo Fertility

2020 ◽  
Author(s):  
Ranjna S. Cheema ◽  
Navjot S. Dhillon ◽  
Sumit Singhal
Keyword(s):  
Seminal Plasma ◽  
Binding Proteins ◽  
Proteome Analysis ◽  
Fertility Rate ◽  
Heparin Binding ◽  
Sds Page ◽  
Sperm Extract ◽  
Sperm Membrane ◽  
Special Relevance

Background: The proteome analysis of seminal plasma and spermatozoa is of special relevance in livestock. Heparin binding proteins (HBPs) found in the seminal plasma of several mammals are shown to bind to sperm membrane and affect a series of events that contribute to normal fertility, such as sperm capacitation, formation of the oviduct reservoir and binding to the oocyte. Profiles of HBPs from seminal plasma and sperm membranes have been associated with sperm fertility. Although, HBPs present in the SP are described in several species, but little is known about HBPs in buck. Methods: Seminal plasma (SP) and sperm extracts (SE) of 13 bucks were subjected to heparin-sepharose affinity chromatography. Sperm extract, seminal plasma and purified HBPs and Non-HBPs were fractionated by SDS-PAGE. Total 78 females (6 per buck) were mated with 13 bucks. Bucks were divided into two groups, G-I (high fertile, 83.3-100% FR) and G-II (low fertile, 50-66.7% FR). Relationship between HBPs and fertility rate was observed. Result: SDS – PAGE of SP and SE resulted in resolution of 22 (10-240 kDa) and 21 (10-270 kDa) bands, respectively. Based on fertility rate 15 and 13 kDa proteins were absent in SP of higher number of GI-compared to G-II bucks. Fourteen bands ranging from 10 – 180 kDa and 10 – 150 kDa were separated from SP-NHBP and SP-HBP. SP-HBPs of 75, 35, 30, 28, 25 and 13 kDa were present in higher (28.6%, 42.5%, 26.2%, 40.5%, 14.3% and 36.2%) number of high fertile than low fertile bucks. NHBP and HBP purified from SE resolved into 11 bands ranging from 10 – 135 kDa and 10 – 120 kDa, respectively. SE-HBP of 53 kDa, 50/45 kDa and 25 kDa were present in higher percentage of high fertile than low fertile bucks.

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