Papers by Murtaza Andrabi
Animal Reproduction Science, 2016
Theriogenology, 2015
Our objectives were to study the effect of trehalose in extender on (1) antioxidant enzymes profi... more Our objectives were to study the effect of trehalose in extender on (1) antioxidant enzymes profile during cryopreservation (after dilution, before freezing, and after thawing), (2) in vitro quality (after thawing), and (3) in vivo fertility of Nili Ravi buffalo (Bubalus bubalis) bull spermatozoa. Semen samples (n = 20) from four buffalo bulls were diluted in Tris-citric acid-based extender having different concentrations of trehalose (0.0, 15, 30, 45, and 60 mM) and frozen in French straws. At post dilution, profile of sperm catalase (U/mL) was higher (P < 0.05) in extenders containing 15, 30, and 45 mM of trehalose as compared to control. Although profiles of superoxide dismutase (U/mL) and total glutathione (μM) were higher (P < 0.05) in extenders containing 15 and 30 mM of trehalose as compared to control. At prefreezing, sperm catalase, superoxide dismutase, and total glutathione profiles were higher (P < 0.05) in all the treatment groups as compared to control. At post thawing, the profiles of catalase and total glutathione were higher (P < 0.05) in extender containing 30-mM trehalose as compared to other treatment groups and control. Whereas, profile of superoxide dismutase was higher (P < 0.05) in extenders containing 30, 45, and 60 mM of trehalose as compared to control and 15mM group. Post thaw total sperm motility (%) was higher (P < 0.05) in extender containing 30-mM trehalose as compared to control and 15 and 60-mM groups. Although sperm progressive motility (%), rapid velocity (%), average path velocity (μm/s), straight line velocity (μm/s), curvilinear velocity (μm/s), plasma membrane (structural and functional, %), acrosome (%), and DNA (%) integrity were higher (P < 0.05) in extender containing 30 mM trehalose as compared to other treatment groups and control. The fertility rates (61% vs. 43%) were higher (P < 0.05) in buffaloes inseminated with semen doses cryopreserved in extender containing 30 mM of trehalose than the control. It is concluded that addition of 30-mM trehalose in extender improves the semen antioxidant enzymes activity, post thaw quality, and fertility in Nili Ravi buffaloes.
Theriogenology, 2016
The present study was carried out to assess various postthaw semen quality parameters for the pre... more The present study was carried out to assess various postthaw semen quality parameters for the prediction of fertility in buffalo bull during low-breeding season. Semen (30 ejaculates) was collected from five adult buffalo bulls with artificial vagina (42 °C). Sperm motility parameters, velocity distribution, motion kinematics, and subpopulations were analyzed by computer-aided sperm motion analyzer (CASA). Moreover, sperm visual motility, supravital plasma membrane integrity, viability/acrosome integrity, viability/mitochondrial transmembrane potential, DNA fragmentation/integrity, and morphology were analyzed by phase-contrast microscope, supravital hypoosmotic swelling test, Trypan blue/Giemsa staining, propidium iodide/"5,5',6,6'-tetrachloro-1,1',3,3'-tetraethylbenzimidazolyl carbocyanine iodide" (JC-1) fluorochromes, neutral comet assay/acridine orange assay and wet mount technique, respectively. Outcome of 528 inseminations was analyzed for in vivo fertility. Pearson's correlation coefficients revealed that sperm progressive motility (%), rapid velocity (%), average path velocity (μm/s), straight line velocity (μm/s), subpopulation one (most rapid, and progressive) of motile spermatozoa (%), supravital plasma membrane integrity (%), and viable spermatozoa with intact acrosome (%) were significantly correlated with in vivo fertility (r = 0.64, P < 0.01; r = 0.57, P < 0.01; r = 0.52, P < 0.01; r = 0.56, P < 0.01; r = 0.73, P < 0.001; r = 0.74, P < 0.001; r = 0.88, P < 0.001); whereas nonviable spermatozoa with damaged acrosome or low-mitochondrial transmembrane potential and comet length (μm) of neutral comet assay were negatively associated with in vivo fertility (r = -0.79, r = -0.75, P < 0.001, and r = -0.60, P < 0.05, respectively). Multiple regression analysis reported that combination of semen quality parameters as predictor of fertility were better (R(2) adjusted = 81.30%, P < 0.001) as compared with single parameter (R(2) adjusted = 50.20%, P < 0.007). It is concluded that assessment of CASA parameters and some other sperm structural and functional parameters, that is, integrity of plasma membrane and acrosome, and transmembrane potential of mitochondria were able to predict the in vivo fertility of water buffalo bull during low-breeding season.
Theriogenology, 2015
Our objectives were to study the effect of trehalose in extender on (1) antioxidant enzymes profi... more Our objectives were to study the effect of trehalose in extender on (1) antioxidant enzymes profile during cryopreservation (after dilution, before freezing, and after thawing), (2) in vitro quality (after thawing), and (3) in vivo fertility of Nili Ravi buffalo (Bubalus bubalis) bull spermatozoa. Semen samples (n = 20) from four buffalo bulls were diluted in Tris-citric acid-based extender having different concentrations of trehalose (0.0, 15, 30, 45, and 60 mM) and frozen in French straws. At post dilution, profile of sperm catalase (U/mL) was higher (P < 0.05) in extenders containing 15, 30, and 45 mM of trehalose as compared to control. Although profiles of superoxide dismutase (U/mL) and total glutathione (μM) were higher (P < 0.05) in extenders containing 15 and 30 mM of trehalose as compared to control. At prefreezing, sperm catalase, superoxide dismutase, and total glutathione profiles were higher (P < 0.05) in all the treatment groups as compared to control. At post thawing, the profiles of catalase and total glutathione were higher (P < 0.05) in extender containing 30-mM trehalose as compared to other treatment groups and control. Whereas, profile of superoxide dismutase was higher (P < 0.05) in extenders containing 30, 45, and 60 mM of trehalose as compared to control and 15mM group. Post thaw total sperm motility (%) was higher (P < 0.05) in extender containing 30-mM trehalose as compared to control and 15 and 60-mM groups. Although sperm progressive motility (%), rapid velocity (%), average path velocity (μm/s), straight line velocity (μm/s), curvilinear velocity (μm/s), plasma membrane (structural and functional, %), acrosome (%), and DNA (%) integrity were higher (P < 0.05) in extender containing 30 mM trehalose as compared to other treatment groups and control. The fertility rates (61% vs. 43%) were higher (P < 0.05) in buffaloes inseminated with semen doses cryopreserved in extender containing 30 mM of trehalose than the control. It is concluded that addition of 30-mM trehalose in extender improves the semen antioxidant enzymes activity, post thaw quality, and fertility in Nili Ravi buffaloes.
The objective of this study was to evaluate tris-citric egg yolk cryodiluter for cryopreservation... more The objective of this study was to evaluate tris-citric egg yolk cryodiluter for cryopreservation of Sahiwal bull epididymal spermatozoa. Sahiwal bull epididymal spermatozoa were recovered from eight slaughtered bulls. Semen retrieved from cauda of each testis pair was pooled and diluted in tris-citric egg yolk extender having 30 × 10 6 spermatozoa ml -1 . After dilution semen was filled in 0.5 ml straws, cooled to 5°C within two hours and equilibrated for four hours at 5°C. The straws were held at liquid nitrogen vapours for 10 minutes and then plunged into liquid nitrogen for storage. Sperm motility, plasma membrane and acrosomal integrity of each semen sample were assessed post thaw. Sperm motility of Sahiwal epididymal spermatozoa was 50.6±1.5, 33.8±0.9 and 18.1±1.3 percent at 0, 2 and 4 hour post thaw, respectively. Percentage of intact plasma membrane was observed 70.5±1.4, 52.9±1.0 and 40.0±1.6 at 0, 2 and 4 hour post thaw, respectively. The percentage of spermatozoa with normal acrosomes was 79.1±0.4, 75.1±0.2 and 71.5±0.2 at 0, 2 and 4 hour post thaw, respectively. In conclusion, tris-citric egg yolk cryodiluter may be used for the cryopreservation of Sahiwal bull epididymal spermatozoa.
This study was designed to identify the effect of fructose addition in skim milk extender on qual... more This study was designed to identify the effect of fructose addition in skim milk extender on quality of buffalo bull spermatozoa (motility, in vitro longevity, plasma membrane integrity, normal apical ridge and abnormalities) stored at 5°C for seven days. For this purpose, semen from five Nili-Ravi buffalo bulls was collected and diluted at 37 o C with a final concentration of 10 x 10 6 motile spermatozoa per ml in skim milk extender containing 5mM or 10mM fructose or without fructose (control). The extended semen was cooled from 37 o C to 5 o C in 2 hours and stored at 5°C for seven days. Sperm motility, in vitro longevity and plasma membrane integrity did not differ (P>0.05) in all three experimental extenders at 1 st day of storage. Sperm motility, in vitro longevity, plasma membrane integrity on 3 rd , 5 th and 7 th day of storage was significantly (P<0.05) higher in extender containing fructose 5mM and 10mM as compared to control. Sperm morphology viz; normal acrosomes; head, mid piece and tail abnormalities remained similar (P>0.05) in all three experimental extenders upto seven days of storage at 5 o C. In conclusion, addition of 5mM and 10mM fructose in skim milk extender improved the preservability of buffalo bull semen stored at 5 o C.
Veterinary and human toxicology
We determined the effect of chronic administration of furazolidone (Fz) on sexual maturation of m... more We determined the effect of chronic administration of furazolidone (Fz) on sexual maturation of male broiler breeder birds (Ross 308; Gallus domesticus). A total of 20 15-w-old birds were randomly assigned to receive 0, 150, 250 or 350 mg Fz/kg feed daily for 5-w. Blood samples were taken at weekly intervals. The birds were challenged with 500 IU human chorionic gonadotropin (hCG) i.v. at the age of 24 w before slaughtering. Concentration of testosterone in the plasma was measured by a specific radioimmunoassay. Testicular tissue was processed for morphometric studies. Testicular weights of the groups fed 250 or 350 mg Fz/kg feed/d were decreased (P &lt; 0.05). Plasma testosterone levels were affected by age (P &lt; 0.001) and dose of Fz (P &lt; 0.001). Mean plasma testosterone levels during and after drug administration were reduced (P &lt; 0.05) by all Fz doses. Human chorionic gonadotropin administration led to poor testosterone response (P &lt; 0.05) in all Fz-dosed groups, but not the control group (P &lt; 0.05). As compared to the control, Fz-dosing reduced the seminiferous tubule diameter (P &lt; 0.05) at the 350 mg/kg feed dose, seminiferous epithelial height (P &lt; 0.05) at the 250 and 350 mg/kg feed doses, and Leydig cell nuclear diameter (P &lt; 0.05) at the 350 mg/kg feed dose. This data suggest that sexual maturation in male broiler breeder birds is adversely affected by chronic Fz-administration. Actions of Fz on sexual maturation probably involve a direct effect at the testicular level.
International Journal of Agriculture and Biology
This essay illustrates the basic concepts of cryopreservation and the causes of cryoinjury to Bos... more This essay illustrates the basic concepts of cryopreservation and the causes of cryoinjury to Bos taurus and Bos indicus bull spermatozoa. The possible roles of cryoprotectants, cooling/freezing and thawing rates are discussed. Advancement in freezing technology for sexed spermatozoa of bull is also briefly described.
Turkish Journal of Veterinary and Animal Sciences
The study was conducted to evaluate the quality of cryopreserved buffalo and zebu semen thawed an... more The study was conducted to evaluate the quality of cryopreserved buffalo and zebu semen thawed and held at low temperature. Progressive motility of frozen semen thawed in a 37 °C water bath for 45 s (control) was compared with that of semen held in ice water (3-5 °C) for 180 min. Semen collected from 3 buffalo and 2 zebu bulls was used for this purpose over 3 weeks (replicates). Fertility was compared after performing 28 inseminations with buffalo semen and 100 inseminations with zebu semen either after control thawing or thawing straws in ice water for 30-60 min. Progressive motility of buffalo and zebu semen thawed and held in ice water for 30 and 90 min, respectively, was not different from that of semen thawed at 37 °C. The conception rate of buffaloes and zebu cows (69.2% and 60.0%) inseminated after control thawing was higher than that of animals inseminated with semen kept in ice water for 30-60 min (53.3% and 47.7%). However, the difference between conception rates after ins...
Pakistan journal of zoology
Vitrified-warmed oocytes are now considered to be a regular source of supply for in vitro fertili... more Vitrified-warmed oocytes are now considered to be a regular source of supply for in vitro fertilization (IVF) technique and therefore present study was undertaken to compare the best of several combinations of dimethyl sulfoxide (DMSO) and 1,2-propanediol (PROH) for the vitrification of buffalo oocytes. Dimethyl sulfoxide (DMSO) and PROH at a concentration of 6 M were used as cryoprotectant agents (CPAs) to vitrify oocytes at germinal vesicle stage (GV) or in vitro matured oocytes at metaphase II stage (M II). In vitro fertilization rate (cleavage rate and cleavage index) was compared to examine the treatment effect in experiment I. Frozen-thawed semen of four artificial insemination donor bulls was used to compare IVF rate of vitrified-warmed oocytes in experiment II. Overall oocyte surviving after vitrified-warming procedures was 89.6%. Cleavage rate and cleavage index of GV stage oocytes vitrified in PROH did not differ (P>0.05) from those of fresh oocytes (control). The GV oo...
Conditions for in vitro maturation (IVM) of primary oocytes without conventionally used fetal cal... more Conditions for in vitro maturation (IVM) of primary oocytes without conventionally used fetal calf serum and hormones were optimized in order to reduce the cost of laboratory produced bufalo embryos. Comparisons were made between oocyte recovery methods (aspiration vs. slicing) and IVM in medium 199 (static culture method vs. lux culture method) supplemented with 4-5 × 106 granulosa cells mL-1 that contained either estrus bufalo serum (EBS) or fetal calf serum (FCS). Recovery methods were compared according to yield, i.e. cumulus oocyte complexes per ovary (COCs/ovary), the expansion rate (% of COCs that expanded), and nuclear maturation rate (% of germinal vesicle breakdown [GVBD]), following IVM for 22-24 h. In vitro maturation methods (static culture with EBS or FCS and Flux culture with EBS or FCS) were compared on the basis of the expansion rate and in vitro fertilization rate (cleavage rate). COC recovery with the slicing method (2.2 COCs/ovary) was better (P < 0.05) than w...
Journal of Veterinary Science
The objective of the present study was to determine the prognostic value of sperm motility parame... more The objective of the present study was to determine the prognostic value of sperm motility parameters and motion characteristics to predict in vitro fertilization (IVF) rate in the buffalo. Sperm motility parameters (%; motile, progressive, rapid and medium) and motion characteristics (path velocity = VAP, µm/sec; progressive velocity = VSL, µm/sec; curvilinear velocity = VCL, µm/sec; amplitude of lateral head = ALH, µm and beat cross frequency = BCF, Hz) of 5 buffalo bulls (Tag no. 2, 4, 6, 11 and 12) were determined by Cell Motion Analyzer (CASA) at post dilution (PD), pre freezing (PF) and post thaw (PT) stages. IVF ability of individual bull (n = 5) was determined by cleavage rate (CR, %). Multiple regression analysis was used to determine the prognostic values of sperm motility parameters and motion characteristics for CR as a dependent variable. At PT, Bull 2 showed the highest (P<0.05) progressive and rapid motility, and VAP, VSL and VCL values as compared to other bulls. ...
The present study was designed to see whether adding glutathione to tris-citric egg yolk extender... more The present study was designed to see whether adding glutathione to tris-citric egg yolk extender improves the quality of buffalo bull spermatozoa stored at 5°C for five days. Semen collected from Nili-Ravi buffalo (Bubalus bubalis) bulls (n = 5) was diluted (37 o C; 10 x 10 6 motile spermatozoa per ml) with extender having glutathione (0.0, 0.5, 1.0, 2.0 and 3.0mM). The extended semen was cooled from 37 o C to 5 o C in 2 hours and stored at 5°C for five days. Post-extension spermatozoal quality viz; motility, viability, plasma membrane integrity and normal acrosomes were studied at 1 st , 3 rd and 5 th day of storage. There was no difference (P > 0.05) in motility (%), viability (%), plasma membrane and acrosomal integrity (%) of sperm due to glutathione at 1 st day after dilution. Highest (P < 0.05) sperm motility, viability and plasma membrane integrity was observed in extender containing glutathione 0.5 and 1.0mM at 3 rd and 5 th day of storage. Moreover, the percentage of...
The Veterinary Journal
A pilot project was initiated to introduce artificial insemination (AI) in goats at farmer level ... more A pilot project was initiated to introduce artificial insemination (AI) in goats at farmer level with chilled semen. Does (n=18) were synchronized with progesterone impregnated vaginal sponges (60 mg Medroxyprogesterone acetate; MAP) for 11 days. At 48 hrs prior to removal of the sponges, intramuscular injection of 400 IU equine chorionic gonadotropin (eCG) and cloprostenol (0.075 mg) was given. Fixed time vaginal insemination (43-45 hrs after sponge removal) was done twice (at 12 hrs interval) in 17 does with chilled Beetal buck semen (4°C) extended with Tris-citric acid (TCA) or skimmed milk (SM) based extender (75 x 106 sperm/ml). Pregnancy test was performed at 45 days post insemination through ultrasonography. An overall 94.5% (17/18) of does showed heat signs and 78% of them were detected in heat between 12 - 24 hrs after sponge removal. An overall 29.4% (5/17) pregnancy rate was recorded. Higher pregnancy rate (44.4%) was obtained in does inseminated with SM extended semen as...
Oestrus synchronization may be used to overcome poor oestrus expression and seasonality of breedi... more Oestrus synchronization may be used to overcome poor oestrus expression and seasonality of breeding in buffalo (Bubalus bubalis). The present work was conducted to compare the efficiency of an oestrus synchronization protocol “ovsynch” in Nili Ravi buffalo during peak (n=8) and low breeding seasons (n=11) by determining luteinizing hormone (LH) peak and progesterone (P4) rise in blood plasma using ELISA. Buffaloes were administered gonadotropin releasing hormone (GnRH) analogue (50 μg lecirelin; day 0) followed by prostaglandinF2α (PGF2α) analogue (150 μg cloprostenol; day 7) and again GnRH analogue at 36 hours after PGF2α. Blood sampling for LH was started 12 h after PGF2α injection and done at 3 h interval up to 108 h. An animal was considered to have responded to ovsynch protocol if it showed LH peak within 48 h after PGF2α injection (and within 3-6 h after second GnRH injection) and showed a P4 concentration of >2.0 ng/ml on day 18 after the 1st GnRH injection. 87.5% animals ...
Journal of Applied Animal Research, 2014
The effect of cysteine addition to the freezing extender on the progressive motility, viability, ... more The effect of cysteine addition to the freezing extender on the progressive motility, viability, plasma membrane and DNA integrity of buffalo bull spermatozoa was studied. Semen from three Nili-Ravi buffalo bulls of similar age group was collected with artificial vagina. Qualifying ejaculates were split in three aliquots for dilution (50 × 10 6 spermatozoa ml -1 ) with tris-citric acid extender containing either 0.0, 0.5, 1.0 mM cysteine. Extended semen samples were cooled and equilibrated before cryopreservation. Progressive motility, plasma membrane integrity, viability and DNA integrity were assessed at 0, 2 and 4 hours post-thaw incubation (37°C). Sperm progressive motility and plasma membrane integrity of buffalo bull spermatozoa were higher (P < 0.05) in extender containing 1.0 mM cysteine than those with 0.5 mM or 0.0 mM at 0, 2 and 4 hours post-thaw. Sperm viability and DNA integrity were higher (P < 0.05) in extender containing 0.5 mM and 1.0 mM cysteine than those with 0.0 mM cysteine at 0, 2 and 4 hours postthaw. The in vivo fertility rates were similar (P > 0.5) with semen cryopreserved in extender containing cysteine (1.0 mM) compared to control. It is concluded that addition of 1.0 mM cysteine to the tris-citric acid extender improved the post-thaw in vitro quality of buffalo bull spermatozoa.
Theriogenology, 1999
A pilot project was initiated to introduce artificial insemination (AI) in goats at farmer level ... more A pilot project was initiated to introduce artificial insemination (AI) in goats at farmer level with chilled semen. Does (n=18) were synchronized with progesterone impregnated vaginal sponges (60 mg Medroxyprogesterone acetate; MAP) for 11 days. At 48 hrs prior to removal of the sponges, intramuscular injection of 400 IU equine chorionic gonadotropin (eCG) and cloprostenol (0.075 mg) was given. Fixed time vaginal insemination (43-45 hrs after sponge removal) was done twice (at 12 hrs interval) in 17 does with chilled Beetal buck semen (4°C) extended with Triscitric acid (TCA) or skimmed milk (SM) based extender (75 x 10 6 sperm/ml). Pregnancy test was performed at 45 days post insemination through ultrasonography. An overall 94.5% (17/18) of does showed heat signs and 78% of them were detected in heat between 12 -24 hrs after sponge removal. An overall 29.4% (5/17) pregnancy rate was recorded. Higher pregnancy rate (44.4%) was obtained in does inseminated with SM extended semen as compared to 12.5% for TCA extended semen. Results were encouraging in the sense that to the best of our knowledge it was the first report of kidding through AI in heat induced does in Pakistan. Moreover, it indicated the feasibility of using synchronization and fixed time AI during low breeding season to enhance the reproductive efficiency in local goats.
Reproductive Biology, 2010
This experiment was designed to compare powdered egg yolk with fresh egg yolk in an extender for ... more This experiment was designed to compare powdered egg yolk with fresh egg yolk in an extender for cryopreservation of Zebu bull semen. Sperm motility, plasma membrane integrity and viability were assessed at different stages of cryopreservation (post-dilution, pre-freezing and post-thawing). Sperm plasma membrane integrity remained similar at all the stages of cryopreservation. Sperm motility and viability were significantly higher after thawing in the extender containing powdered egg yolk. In conclusion, powdered egg yolk may be used in an extender for the cryopreservation of Zebu bull spermatozoa. Reproductive Biology 2010 10 3: 235-240.
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Papers by Murtaza Andrabi