111 ESTIMATION OF SPERM QUALITY IN FRESH AND FROZEN - THAWED SEMEN FROM ASTURIANA DE LOS VALLES BULLS

2006 ◽  
Vol 18 (2) ◽  
pp. 163
Author(s):  
C. Tamargo ◽  
M. Carbajo ◽  
C. Diez ◽  
D. Martin ◽  
C. O. Hidalgo
Keyword(s):  
Sperm Motility ◽  
Semen Quality ◽  
Sperm Quality ◽  
Membrane Integrity ◽  
Daily Routine ◽  
Morphological Abnormalities ◽  
Hypoosmotic Swelling Test ◽  
Motion Characteristics ◽  
The Impact ◽  
Casa System

Artificial insemination and semen cryopreservation have significantly improved the breeding potential of male animals. However, current freezing techniques commonly result in reduced semen quality (Januskauskas et al. 1999 Theriogenology 52, 641-658), and surviving cells are affected post-thaw either structurally or functionally (Nagy et al. 2004 Anim. Reprod. Sci. 80, 225-235). In this work we analyze the impact of cryopreservation on Asturiana de los Valles bull sperm. Ejaculates (n = 373) from seven adult bulls were weekly collected by means of artificial vagina. Immediately after collection, routine parameters including volume (V), mass motility (MM), and concentration (C) of sperm cells were evaluated. Then the semen was extended with a commercial extender, loaded into 0.25-mL plastic straws at a concentration of 23 � 106 per straw, frozen and stored for further analysis. Four straws per ejaculate were thawed, pooled and analyzed for motion characteristics by means of a CASA system (Sperm Class Analyzer, SCA 2002� Microptic S. L., Barcelona, Spain) added to an optical phase-contrast microscope with heatable (37�C) stage. Immediately after thawing, we analyzed the % of motile spermatozoa (MS) and the % of progressively motile spermatozoa (PMS); then samples were incubated for 3 h at 37�C and MS and PMS were measured again (MS3 and PMS3, respectively). Functional integrity of the plasmallema was evaluated by the hypoosmotic swelling test (HOST) together with the % of typical tail coiling/swelling (percentage of HOST-positive spermatozoa, HOST-PS). The % of viable spermatozoa (VS) [membrane integrity was evaluated by fluorescence microscopy with a dual staining system (propidium iodide (PI) and 6-carboxyfluorescein diacetate (CFDA)]. Sperm showing partial or complete red fluorescence (PI staining) were considered nonviable, whereas sperm showing complete green fluorescence were considered viable. Altered acrosomes (AA) and morphological abnormalities were also determined. The % of morphological abnormalities was classified according to their location in head (HA), midpiece (MA), and tail (TA). Proximal and distal cytoplasmic droplets were counted as separate abnormalities (CD). Data were analyzed by the MEANS procedure of SAS (SAS Institute, Inc., Cary, NC, USA). A significant (P < 0.05) decrease in the sperm motility was observed after freezing/thawing (MS: 80.20 � 0.75 vs. 47.36 � 1.04, and PMS: 68.73 � 0.73 vs. 42.14 � 0.96 for fresh and frozen-thawed semen, respectively). Also, the frozen-thawed sperm showed increased % of HA, MA, AA, HOST-PS, and VS (P < 0.05). These morphological abnormalities could contribute to decreasing sperm motility. The new computer and video technologies provide useful information about sperm quality and can be used in the daily routine of processing semen. This work was performed in collaboration with ASEAVA.

scholarly journals Correlations Amongst Functional and Morphological Attributes and Oxidative Markers of Fresh and Cryopreserved Semen of Gir and Murrah Bulls

2020 ◽  
Vol 15 (03) ◽  
pp. 24-29
Author(s):  
A. J. Dhami ◽  
Tapasvi M Patel ◽  
DV Chaudhari
Keyword(s):  
Plasma Membrane ◽  
Sperm Motility ◽  
Semen Quality ◽  
Sperm Quality ◽  
Membrane Integrity ◽  
Winter Season ◽  
Correlation Coefficients ◽  
Plasma Membrane Integrity ◽  
Murrah Buffalo ◽  
Oxidative Markers

This study was undertaken during the winter season on healthy mature Gir cattle and Murrah buffalo bulls (n=3 each). The semen samples (6 ejaculates/bull, total 36 ejaculates) collected in the morning using artificial vagina were evaluated for routine seminal attributes, including acrosomal and plasma membrane integrity. The samples were then diluted @ 100 million sperm/ml with tris fructose yolk glycerol extender without and with sericin @ 0.1, 0.25, 0.5 and 1.0% (w/v), filled in French mini-straws, and frozen in LN2 using biofreezer as per standard freezing protocol. Straws were thawed in water bath at 37°C for 30 sec and evaluated for post-thaw quality, viz., motility, viability, morphology, acrosome integrity and plasma membrane integrity (HOST). Lipid peroxidation (malondialdehyde - MDA production) and activities of enzymes superoxide dismutase (SOD) and glutathione peroxidase (GPx) were assessed as oxidative markers in seminal plasma of freshly diluted and frozen-thawed semen samples. Sericn at 0.5% level significantly (p less than 0.01) improved the post-thaw sperm quality with reduced oxidative stress in both the species. The breed-wise correlation coefficients (r) among sperm quality attributes and oxidative markers were studied in fresh and frozen-thawed semen of each species, and also for fresh with frozen-thawed semen. The findings revealed significant interrelationships amongst most of the attributes of fresh as well as post-thawed semen and also of fresh semen attributes with those of cryopreserved semen including oxidative markers in both the species. Sperm motility estimation in fresh, pre-freeze and post-thawed semen was a legitimately good indicator of quality of spermatozoa at various steps of semen processing/freezing, and its fertilizing potential. Thus, the sperm motility, HOS test and either MDA or SOD/GPx activity alone may be used as valuable and practical tools for routine assessment of bovine semen quality considering significant correlations found between them.

180 ASSESSMENT OF BULL SEMEN QUALITY LOADED IN NEW SensiTemp STRAWS USING SEMEN AND IN VITRO PRODUCTION TECHNOLOGIES

2016 ◽  
Vol 28 (2) ◽  
pp. 221
Author(s):  
D. Le Bourhis ◽  
S. Camugli ◽  
P. Salvetti ◽  
L. Schibler ◽  
E. Schmitt
Keyword(s):  
Sperm Motility ◽  
Semen Quality ◽  
Sperm Quality ◽  
Semen Analysis ◽  
Membrane Integrity ◽  
Computer Assisted ◽  
Cleavage Rate ◽  
Fluid Medium ◽  
And Control

SensiTemp, a new in vitro maturation (IMV) bull straw concept, presents the advantage of colour changing while the straw is thawed. The colour of frozen straws is blue and straws start to become white when the temperature reaches 33°C, with a complete change of colour at 37°C. The objective of this study is to assess sperm quality after thawing of semen frozen in SensiTemp from 2 bulls, by analysing, in experiment 1, sperm motility and membrane integrity using computer-assisted semen analysis (CASA) and flow cytometry (FC), and, in experiment 2, the in vitro embryo production (IVP) using IVP technologies [IVM, IVF, and in vitro culture (IVC)]. The ejaculates of 2 bulls, selected during preliminary experiments on high in vitro fertility, were harvested at CIA L’Aigle, France, and split ejaculates were frozen in experimental (SensiTemp) and conventional (control) straws. In experiment 1 after thawing semen from the 2 types of straws (5 pooled straws each; 2 replicates), motility was assessed using the IVOS CASA system (Hamilton Thorne Inc., Beverly, MA, USA) and membrane integrity was evaluated through FC with Cytosoft software (Millipore-Guava Technologies Inc., Hayward, CA, USA). In experiment 2, IVF was used to evaluate the non-toxicity of SensiTemp and control straws. Cumulus-oocyte complexes (COC; n = 1178; 4 replicates) collected from slaughterhouse ovaries were matured in IVM medium (TCM-199 with bicarbonate, Sigma-Aldrich, Saint Quentin Fallavier, France; 10 µg mL–1 FSH-LH, Reprobiol, Liège, Belgium; and 10% FCS, Thermo Fisher, Illkirch, France) for 22 h. After fertilization, presumptive zygotes of each group (SensiTemp and control for each bull) were cultured in synthetic oviduct fluid medium (SOF, Minitube, Tiefenbach, Germany) with 1% estrous cow serum (ECS) and 0.6% BSA (Sigma-Aldrich, France) up to 8 days. All cultures were conducted at 38.5C in 5% CO2, and 5% O2. The cleavage and blastocysts rates were evaluated on Days 3 and 7, respectively, for each group. Embryo quality was recorded on Day 7 according to the IETS evaluation. Data from each bull were analysed separately using the chi-squared test (P < 0.05). In experiment 1, neither sperm motility from bull 1 (61.2 and 60.5%) and bull 2 (66.2 and 66.5%) nor membrane integrity from bull 1 (58.6 and 52.2%) and bull 2 (61.0 and 61.9%) were different between SensiTemp and control, respectively. Results from experiment 2 showed no difference (P > 0.05) in cleavage rate between SensiTemp and control for the 2 bulls: 92.1 and 91.7% for bull 1 and 94.2 and 94.6% for bull 2 respectively. The blastocysts rate on Day 7 did not differ (P > 0.05) among groups (47.5, 47.1 and 51.3, 50.4% for SensiTemp and control bull 1 and bull 2, respectively) nor the quality of embryos retrieved in the different groups: 25.4, 23.3, and 30.8, 29.6% in grade 1 embryo for SensiTemp and control bull 1 and bull 2, respectively. Those results demonstrate, in vitro, that the new SensiTemp straws were non-toxic and did not affect the semen quality after thawing nor did the SensiTemp straws affect the ability of sperm cells to fertilize oocytes and produce 8-day-old embryos.

Semen quality and fertility of liquid stored and frozen-thawed semen in crossbred pigs of North-Eastern India

2018 ◽  
Author(s):  
G Kadirvel ◽  
M K Kalita ◽  
Raju Kr Dewry ◽  
Ashok Kumar ◽  
Nripendra Mahanta ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Sperm Motility ◽  
Semen Quality ◽  
Sperm Quality ◽  
Membrane Integrity ◽  
Eastern Region ◽  
Plasma Membrane Integrity ◽  
Frozen Semen ◽  
North Eastern ◽  
Farrowing Rate

Study was conducted to compare the semen quality and fertility of liquid stored semen for three days and frozen-thawed semen in the north-eastern region of India. For liquid semen, the semen ejaculates were extended in Beltsville Thawing Solution (BTS) extender and preserved at 17°C for three days. For cryopreservation, semen was diluted Lactose-egg yolk-glycerol extender and frozen in straw using programmable freezer with freezing rate of 40°C/min from -6 to -140°C. The preserved evaluated for sperm motility, viability, plasma membrane integrity and fertility. The results revealed that the liquid stored semen has maintained the sperm motility and viability up to day 3 without significant reduction. Similarly the plasma membrane integrity did not differ significantly up to day 2, but it was significantly (P less than 0.05) reduced on days 3 in liquid stored semen. After freezing and thawing, the mean sperm motility, viability and plasma membrane integrity were 58.25 ± 2.96%, 64.75 ± 2.47% and 47.06 ± 2.02%, respectively. These parameters were significantly (PP less than 0.01) lower as compared to the liquid stored semen from day 0 to day 3. After insemination with liquid semen, the farrowing rate was 77.7%, 80.76%, 73.07% and 69.8%, respectively from day 0, day1, day 2 and day 3. The pregnancy rate, farrowing rate and litter size did not differ significantly among different days of liquid storage. These parameters were significantly (PP less than 0.01) lower in frozen semen as compare to that of liquid stored semen. The study concluded that the liquid semen stored up to three days is more efficient than frozen-thawed semen in terms of preserving sperm quality and fertility.

scholarly journals Modified hypoosmotic swelling test for the assessment of boar and bull sperm sensitivity to cryopreservation

2014 ◽  
Vol 83 (4) ◽  
pp. 313-319 ◽  
Author(s):  
Petra Přinosilová ◽  
Věra Kopecká ◽  
Jaroslava Hlavicová ◽  
Monika Kunetková
Keyword(s):  
Semen Quality ◽  
Sperm Quality ◽  
Membrane Integrity ◽  
Membrane Resistance ◽  
Freezing Process ◽  
Sperm Analysis ◽  
Hypoosmotic Swelling Test ◽  
Sperm Plasma Membrane ◽  
Viable Sperm ◽  
Sperm Membrane

Routine methods for the evaluation of sperm quality are not sufficiently useful to determine the sensitivity of sperm cells to cold shock. The aim of our preliminary study was to determine whether the sperm plasma membrane integrity evaluated by modified hypoosmotic swelling test based on simple hypoosmotic swelling test (HOS test) and eosin-nigrosin staining could be helpful in predicting the degree of boar sperm survivability during semen cryopreservation. Ejaculates collected from 24 boars and 20 bulls were used in the experiment. Fresh ejaculates were evaluated by routine sperm analysis and a modified HOS test, and subsequently frozen. Sperm cryosurvivability was defined as the percentage of motile spermatozoa that survived the freezing process. A higher percentage of sperm was recovered after the thawing of semen with a higher percentage of HOS-positive and eosin-negative sperm (P < 0.01). Both indicators were found to be correlated (r = 0.707 and r = 0.705, respectively; P < 0.01). Moreover, the percentage of HOS-positive and eosin-negative sperm was similar to the percentage of viable sperm after thawing as determined by traditional eosin-nigrosin staining in boars (50.90 ± 9.88% and 49.31 ± 11.63%, respectively) and bulls (55.91 ± 10.34% and 55.63 ± 6.64%, respectively) and both indicators showed a positive correlation (r = 0.558 and r = 0.504, respectively; P < 0.05). In conclusion, based on the obtained results, we can assume that the modified HOS test can detect differences in sperm membrane resistance which allows assessment of semen quality from the perspective of its further use, e.g. cryopreservation.

scholarly journals Effect of Macrophages in Semen on Sperm Quality

2020 ◽  
Author(s):  
Gangxin Chen ◽  
Beihong Zheng
Keyword(s):  
Cell Membrane ◽  
Sperm Motility ◽  
Semen Quality ◽  
Sperm Quality ◽  
Sperm Concentration ◽  
Membrane Integrity ◽  
Significant Negative Correlation ◽  
High Group ◽  
Cross Sectional ◽  
Cell Membrane Integrity

Abstract Background: This was a cross-sectional study in China, we analyzed the levels of macrophages(Mφ) in semen. The study evaluated the influence of the levels of Mφ in semen on sperm quality.Methods: The subjects were 78 males between 25 and 35 years old. The samples were divided into a low group(Mφ<6×105/ml) and a high group (Mφ>6×105/ml). Evaluation included consideration of the influencing factors of male semen quality, macrophage concentration, sperm motility, morphology, membrane integrity DNA fragmentation index (DFI), anti-sperm antibodies (AsAb), IL-10, and IL-12 in semen.Results: There was no difference in the physical or chemical indices of the semen, sperm concentration, AsAb, IL-10, or IL-12 between the two groups (P>0.05). The percentage of sperm forward motility (PR%), the rate of normal sperm shape, and the integrity of cell membranes in the low group were higher than those in the high group (P<0.05), while the percentage of sperm inactivity (IM%), the rate of sperm head deformity, the rate of deformity in the neck and middle segment, the sperm malformation index (SDI), the abnormal sperm index (TZI), and the sperm DFI in the low group were lower than those in the high group (P<0.05). The concentration of Mφ in the semen was linearly correlated with sperm concentration, sperm PR%, IM%, sperm normal shape rate, head deformity rate, neck and middle deformity rate, SDI, TZI, sperm DFI, and sperm cell membrane integrity (P<0.05), but there was no linear correlation with IL-10 or IL-12 (P>0.05).Conclusions: The concentration of Mφ in semen had no significant correlation with semen volume or sperm concentration, but it did have a significant negative correlation with sperm motility, sperm morphology, cell membrane integrity, and DNA breakage rate. There was no significant correlation with the concentration of IL-10 or IL-12.

scholarly journals Influence of age on sperm characteristics evaluated by light and electron microscopies

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Giulia Collodel ◽  
Fabio Ferretti ◽  
Maurizio Masini ◽  
Giacomo Gualtieri ◽  
Elena Moretti
Keyword(s):  
Sperm Motility ◽  
Semen Quality ◽  
Sperm Quality ◽  
Semen Analysis ◽  
Sperm Concentration ◽  
Study Data ◽  
Wine Consumption ◽  
Electron Microscopies ◽  
Urogenital Infection ◽  
The Impact

AbstractThe impact of male aging on male fertility has only recently become of interest to the scientific community. This study aims to assess the relationship between age and fertility among a sample of men, considering the individual and pathological characteristics. In this retrospective study data of semen analysis and medical history of 1294 Italian male patients were considered. Semen analysis was performed by light microscopy and transmission electron microscopy mathematically elaborated. A generalized linear model was used to explore the influence of male age on semen quality, considering as confounders wine consumption, smoking habits, presence of varicocele, consanguinity and positive semen bacteriological analysis and urethral swab. The mean age of the participants was 36.41 ± 6.379. Male aging without impact of confounders was correlated with a decrease in sperm concentration and motility and an increased in sperm necrosis. Sperm concentration and progressive motility were negatively related to the presence of confounders as wine consumption (sperm motility), urogenital infection (sperm concentration and motility), varicocele (sperm concentration) and consanguinity (sperm motility). Urogenital infection, varicocele and consanguinity positively correlated with sperm necrosis. The most important finding was the observation of a negative effect of male aging on sperm parameters such as concentration, motility, and viability. It is possible to hypothesize age-dependent changes of testicular environment, probably related to reactive oxygen species production. The demonstration, in a large sample of patients, that aging influences sperm quality strongly motivates further research focused on the mechanisms involved in this phenomenon and its effects on offspring fitness.

scholarly journals Effect of macrophages in semen on sperm quality

2021 ◽  
Vol 19 (1) ◽  
Author(s):  
Gangxin Chen ◽  
Beihong Zheng
Keyword(s):  
Cell Membrane ◽  
Sperm Motility ◽  
Semen Quality ◽  
Sperm Quality ◽  
Sperm Concentration ◽  
Membrane Integrity ◽  
Cross Sectional Study ◽  
High Group ◽  
Cross Sectional ◽  
Cell Membrane Integrity

Abstract Background This was a cross-sectional study in China which analyzed the levels of macrophages (Mφ) in semen and evaluated the influence of Mφ levels in semen on sperm quality. Methods The subjects involves 78 males, 25- to 35-year old. The samples were divided into a low group (Mφ < 6 × 105/ml) and a high group (Mφ > 6 × 105/ml). Evaluation included consideration of the influencing factors of male semen quality, macrophage concentration, sperm motility, morphology, membrane integrity DNA fragmentation index (DFI), anti-sperm antibodies (AsAb), IL-10, and IL-12 in semen. Results There was no difference in the physical or chemical indices of the semen, sperm concentration, AsAb, IL-10, or IL-12 between the two groups (P > 0.05). The percentage of sperm forward motility (PR%), the rate of normal sperm shape, and the integrity of cell membranes in the low group were higher than those in the high group (P < 0.05), while the percentage of sperm inactivity (IM%), the rate of sperm head deformity, the rate of deformity in the neck and middle segment, the sperm deformity index (SDI), the teratozoospermia index (TZI), and the sperm DFI in the low group were lower than those in the high group (P < 0.05). The concentration of Mφ in the semen was linearly correlated with sperm concentration, sperm PR%, IM%, sperm normal shape rate, head deformity rate, neck and middle deformity rate, SDI, TZI, sperm DFI, and sperm cell membrane integrity (P < 0.05), but there was no linear correlation with IL-10 or IL-12 (P > 0.05). Conclusions The Mφ concentration in semen is not significantly correlated with semen volume or sperm concentration, but negatively correlated with sperm motility, morphology, cell membrane integrity, and DNA damage rate. There is no significant correlation between the macrophages and the concentration of IL-10 or IL-12.

scholarly journals Macro- and microelements in serum and seminal plasma as biomarkers for bull sperm cryotolerance

2021 ◽  
Vol 63 (1) ◽  
Author(s):  
Maja Zakošek Pipan ◽  
Petra Zrimšek ◽  
Breda Jakovac Strajn ◽  
Katarina Pavšič Vrtač ◽  
Tanja Knific ◽  
...  
Keyword(s):  
Seminal Plasma ◽  
Semen Quality ◽  
Sperm Quality ◽  
Membrane Integrity ◽  
Quality Characteristics ◽  
Freezing And Thawing ◽  
Additional Tool ◽  
Bull Semen ◽  
Bull Sperm ◽  
Fresh Semen

ABSTRACT Background Wide variation in fertility rates is observed when using frozen bull semen, even when the bulls have met quality standards for semen production. Therefore, a simple and reliable test to assess the freezing potential of bull semen based on the analysis of fresh semen or blood would be of great value. Attention is now turning to assessment of seminal plasma components such as proteins and elements. In the present study, the concentrations of macro- and microelements in fresh bull semen plasma and in serum and their correlation with quality characteristics of fresh semen and with semen quality after freezing and thawing were determined. Ejaculates were collected from 30 mature bulls, and semen volume, concentration, sperm motility, morphology, tail membrane integrity, plasma membrane permeability and DNA fragmentation were determined on the day of collection and after freezing and thawing. The concentrations of macroelements (Na, Mg, K and Ca) and microelements (Cu, Fe, Zn and Se) were determined in the seminal plasma and serum. The semen samples were classified into satisfactory and unsatisfactory groups according to the fresh semen quality. Results Zinc and Se levels measured in serum were associated with almost all fresh and frozen-thawed semen quality characteristics, while Fe levels were associated only with acrosomal defects in fresh semen. Zinc and Fe levels in fresh seminal plasma were associated with various quality characteristics of fresh and frozen-thawed semen, while Se level in fresh seminal plasma was not associated with any of the semen quality characteristics. Conclusions Microelements were shown to be useful as biomarkers involved in the analysis of bull sperm quality and could be used as an additional tool to predict bull semen quality after freezing and thawing. Our results confirm that the analysis of Zn and Se levels in serum and Zn, Cu and Fe levels in fresh seminal plasma can provide information to discriminate between bull semen samples with spermatozoa with high or low cryotolerance.

scholarly journals Spectrophotometric Analysis of the Resazurin Reduction Test as a Tool for Assessing Canine Semen Quality

2013 ◽  
Vol 57 (2) ◽  
pp. 281-285 ◽  
Author(s):  
Rafał Strzeżek ◽  
Krystyna Filipowicz ◽  
Marta Stańczak ◽  
Władysław Kordan
Keyword(s):  
Sperm Motility ◽  
Seminal Plasma ◽  
Semen Quality ◽  
Membrane Integrity ◽  
Spectrophotometric Analysis ◽  
Normal Morphology ◽  
Additional Tool ◽  
Reduction Test ◽  
Highly Correlated

Abstract The resazurin reduction test (RRT) was subjected to spectrophotometric analysis to evaluate the quality of canine semen. Twenty four samples of canine semen were analysed. The absorption peaks for resazurin and resorufin were determined at 615 and 580 nm, respectively. The RRT ratio (RRTsperm-the ratio for samples containing spermatozoa, RRTplasma-the ratio for samples containing seminal plasma) was calculated by dividing the absorbance at 580 nm by the absorbance at 615 nm. Spearman’s correlation test was used to determine the significance of correlations between the analysed sperm parameters and the results of the resazurin reduction assay. The RRT ratio was highly correlated with sperm motility (r=0.68, P<0.01), progressive sperm motility (r=0.61, P<0.01), the subpopulation of cells with rapid velocity (r=0.72, P<0.01), and the subpopulation of cells with medium velocity (r= -0.54, P<0.05). A negative correlation was observed between the reducing capacity of seminal plasma vs. sperm with plasma membrane integrity (r= -0.60, P<0.01) and sperm with normal morphology (r= -0.58, P<0.01). The RRT test can be used as an additional tool for evaluation of the quality of canine semen.

scholarly journals Effect of Supplementation of Polyvinylpyrrolidone in Extender on Buffalo Semen Parameters

2020 ◽  
Vol 53 (1) ◽  
Author(s):  
Bushra Ismail Khan ◽  
Shamim Akhter ◽  
Sanwal Aslam ◽  
Rabea Ejaz
Keyword(s):  
Sperm Motility ◽  
Semen Quality ◽  
Membrane Integrity ◽  
Bubalus Bubalis ◽  
Semen Parameters ◽  
Suction Pump ◽  
Bull Semen ◽  
Buffalo Semen ◽  
Live Sperm ◽  
And Control

The current study was planned to evaluate the supplementation of Polyvinylpyrrolidone in extender on cryopreservation of Nili-Ravi buffalo bull semen. The semen samples were collected from Nili-Ravi buffalo (Bubalus bubalis) bull kept at SPU Qadirabad, District Sahiwal, Pakistan. Qualifying semen ejaculates having motility >60%, volume >5-6ml and concentration >0.5 billion/ ml were diluted 50 × 106 motile sperm ml approximately at 37°C in Tris-citric acid extender supplemented with different concentrations of PVP (0.01, 0.05, 0.1mM). The extender without PVP was kept as control. Semen was stored at 4°C for a period of 2 h and kept at 4°C for 4h. Semen was filled in 0.5 ml French straws using suction pump at 4°C, plunged and stored in liquid nitrogen (-196°C). Semen straws were rewarmed at 37°C for 30 seconds and assessed for sperm motility, plasma membrane integrity (PMI), dead sperm percentage and the live sperm percentage. The data on the role of PVP on different parameters of semen quality were analyzed by using ANOVA and RCBD. Higher percentage (P< 0.05) of sperm motility (66.1±7.51 and 59.4±10.72) and PMI (72.9±5.39 and 75.7±6.5) was observed in extenders having 0.05 mM and 0.1mM PVP compared to extenders having 1.5mM PVP and control. The percentage acrosomal integrity was observed greater (P< 0.05) in extended semen containing 0.1mM (68.2±0.50) PVP compared to extenders having 0.01 and control.

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