Search

Process of cryopreservation induces cell damages due to thermal and osmotic shocks. Addition of amino acids as cryoprotective agents in extender can reduce these damages. Therefore, present study was designed to evaluate the effect of glycine supplementation in extender on freezability of cryopreserved bull semen. For this purpose, three adult Sahiwal bulls were selected for semen collection by artificial vagina having 42 C (three weeks) (replicates; n=18). Collected semen was quickly shifted to laboratory for initial evaluation. Qualified semen ejaculates (>60% motility, >1mL volume, >0.5billion/mL conc.) were diluted in tris extender having 0 (control), 5, 10, 15 and 20mM of glycine. Semen was cooled to 4 C in 2 hours and kept for 4 hours for equilibration. Semen was filled in French straws at 4 C and kept in liquid nitrogen container. Highest percentages of post thaw sperm progressive motility and acrosome integrity were observed in extender containing 5mM of glycine compared to other treatments and control (P

A great diversity of termite species is found all over the world, among these 53 species are identified from Pakistan. The knowledge about morphological features in soldier cast of termites is proved to be a technical tool for taxonomy and identification. In the present studies, the morphometric variations of external morphology in soldier caste of Odontotermes obesus (Rambur) and Microtermes obesi (Holmgren) from three different areas i.e. Gujar Khan (A), Rawalpindi (B) and Islamabad (C) were studied. About 36 characters like body, thorax, abdomen, from head to mandible tip, head, pronotum, postmentum, mandible, antenna (scape, pedicle, flagellum), and legs parts were used for morphometric measurements. The data were statistically analyzed for significant differences in their mean, standard deviation, standard error, 95% confidence interval, coefficient of variability and analysis of variance. "Student t-test" was used for the comparison of mean values using Minitab version 16. The results of the present study revealed no significant differences among the population samples collected from various localities. However little variations were observed in a few characters like antennal segments (pedicle, scape) and legs (trochanter, tibia) among the soldier cast which are adaptive in nature to survive in the environment. Keywords: Morphometric analysis, soldiers cast, Odontotermes obesus, Microtermes obesi.

Aims: Bacterial contamination may occur in feces during collection and processing of semen. Bacteria not only compete for nutrients with spermatozoa but also produce toxic metabolites and endotoxins and affect sperm quality. The aim of the present study was to investigate the effect of antibiotic supplementation on the sperm quality of Indian red jungle fowl, estimation and isolation of bacterial species and their antibiotic sensitivity. Materials and Methods: Semen was collected and initially evaluated, diluted, and divided into six experimental extenders containing gentamicin (2.5 μg/mL), kanamycin (31.2 μg/mL), neomycin (62.5 mg/mL), penicillin (200 U/mL), and streptomycin (250 μg/mL), and a control having no antibiotics were cryopreserved and semen quality was evaluated at post-dilution, post-cooling, post-equilibration, and post-thawing stages (Experiment 1). A total aerobic bacterial count was carried out after culturing bacteria (Experiment 2) and subcultured for antibiotic sensitivity (Experiment 3). Results: It was shown that penicillin-containing extender improved semen quality (sperm motility, plasma membrane integrity, viability, and acrosomal integrity) compared with the control and other extenders having antibiotics. The bacteria isolated from semen were Escherichia coli, Staphylococcus spp., and Bacillus spp. Antibiotic sensitivity results revealed that E. coli shows high sensitivity toward neomycin, kanamycin, and penicillin. Staphylococcus spp. shows high sensitivity toward streptomycin, neomycin, and penicillin. Bacillus spp. shows high sensitivity toward kanamycin and penicillin. Conclusions: It was concluded that antibiotics added to semen extender did not cause any toxicity and maintained semen quality as that of untreated control samples, and penicillin was identified as most effective antibiotic. It is recommended that penicillin can be added to the semen extender for control of bacterial contamination without affecting the semen quality of Indian red jungle fowl.

The current study was aimed to evaluate the antioxidative effect of L-Carnitine at post thawing following cryopreservation of Nili-Ravi buffalo sperm. For this purpose, semen from three buffalo bulls were collected for 3 weeks using an artificial vagina (N=18; replicates). The qualified ejaculates were diluted employing tris-citric acid extender i.e., control did not receive any L-Carnitine and experimental groups having 0.5, 1.0, and 1.5 ng/mL of L-carnitine at 37 C with approximately 50 x 106 sperm/mL. The semen was cooled at 4 C and then equilibrated (4 hours), filled in straws (0.5 mL) at4 C, placed on LN2 vapours for 10 min, and kept into an LN2 container. The thawed semen was evaluated for post-thaw quality. The integrity of the sperm plasma membrane and motility (P?0.05) was highest in the extenders having 1.0 ng/mL of L-carnitine as compared to the control(received no L-Carnitine). However, sperm chromatin integrity and viability(live sperm with intact acrosome) remained similar. It was concluded that supplementing 1.0 ng/mL L-Carnitine of extender can improve the post-thaw quality of cryopreserved sperm. Based on the results of the current experiments it is recommended to include L-carnitine extender to improve post-thaw quality of buffalo sperm in terms of its motility and integrity ofits plasma membrane. Keywords: Buffalo, Sperm, Cryopreservation, Extender, L-Carnitine, Artificial insemination.

Present study aimed to evaluate field fertility rate and calf sex ratio of Nili Ravi buffalo semen sexed through modified swim-up method (Animal Reproduction Science, 182, 2017, 69). For this purpose, five mature Nili-Ravi buffalo bulls kept at semen production unit, Qadirabad, Pakistan, were selected. Two consecutive ejaculates per week were collected with artificial vaginą for 3 weeks. Qualified semen ejaculates were pooled and divided into two aliquots. The first aliquot was processed by routine procedure (control), whereas the second was processed by modified swim-up technique. After separation, semen was diluted in tris-citric acid extender and cryopreserved using standard techniques. Sexed semen was evaluated for fertility trials during peak breeding season. Artificially inseminated animals were examined for pregnancy rate through rectal palpation at least 3 months after insemination under field conditions. Calving ratio of female and male calves were recoded after Parturition. The fertility rate was higher (p .05) in X-sorted sperm (70%) as compared with control (47%). The female calf ratio was higher (p .05) in X-sorted sperm (78.58%) compared with control (53.3%). In Conclusion, conception rate and production of female calf were significantly higher with sexed semen separated through modified swim-up method compared with unsexed control.

Aim: The present study was designed to evaluate enrofloxacin (EN), a fluoroquinolone broad-spectrum antibacterial drug for use in cryopreservation of Zebu bull semen. Materials and Methods: Semen w...

Ecological modelling is very useful tool to evaluate the spatial distribution and breeding biology. In this study GIS mapping was use to study the population of Red Vented Bulbul in 8 different regions (Chikrala, Abba Khel, Chiddru, Namal Lake, Musa Khel, Sawans, Bittian, and Gulmeri) of the district Mianwali. Spatial distribution of specie, their nests structures, total eggs and hatching rate was thoroughly investigated during the survey. Results revealed that mostly this specie was found in continuous circulations around all sites except Gulmeri. Population density was recorded higher from the month of March to July and then starts deceasing. Breeding time started from late February to late July. For nest construction, 42 nests have been investigated from study sites and observed that the Red Vented Bulbul used different plant species, including Psidium guajava, Zizyphus nummularia, Dodonea viscosa, Dalbergia sissoo, Phoenix dactylifera, Berberis lyceum, Vitis vinifera, and Morus alba. The incubation time of the eggs range between 11-14 days. The incubating and fledgling success was found to be 82 and 87% respectively. The study shows that the Red Vented Bulbul is widely distributed throughout the targeted regions of the district Mianwali.

The present study was conducted to investigate the effect of omega-3 poly unsaturated fatty acid (PUFA), α-linolenic acid (ALA; 18:3 n-3) on the in vitro maturation (IVM) of buffalo oocytes and subsequent embryonic development. Buffalo cumulusoocyte complexes (COCs; n = 2282) were in vitro matured in TCM-199 (0.6% fatty acid free bovine serum albumin, 0.02 Units/ml FSH, 1 µg/ml 17-β-estradiol, 10 µg/ml epidermal growth factor, 50 µg/ml gentamicin) supplemented with 0 (control), 25, 50, 100, 150 or 300 µm ALA under an atmosphere of 5% CO2 in air at 38.5oC for 22-24 h. The matured oocytes were then fertilized in Tyrode’s Albumin Lactate Pyruvate (TALP) medium and cultured in synthetic oviductal fluid (SOF) medium. Concentrations up to 100 μm ALA improves (P ≤ 0.05) the cumulus expansion compared to control. Higher percentage of oocytes reaching MII stage was observed at 50 μm and 100 μm of ALA compared to control (P ≤ 0.05). Concentrations of 150 and 300 µm ALA were detrimental both for cumulus expansion and nuclear maturation rate of buffalo oocytes. Moreover, supplementation with 100 μm ALA improved (P ≤ 0.05) cleavage rate compared to control and treatment with 50 and 100 μm ALA yielded significantly higher morulae compared to control. The results of present study indicate that the supplementation with 100 μm ALA to the IVM medium improves nuclear maturation rate of buffalo oocytes and subsequent early embryonic development.

Overwintering larvae of the beetle Dendroides canadensis produce potent antifreeze proteins to inhibit inoculative freezing and promote supercooling. We hypothesized that addition of Dendroides canadensis recombinant antifreeze proteins (DAFPs) in the extender will improve the quality and fertility of cryopreserved Nili-Ravi buffalo (Bubalus bubalis) sperm. The study was divided into two parts: (1) Evaluation of the effect of DAFPs on the quality of frozen-thawed buffalo bull sperm and (2) Examination of the fertilizing ability of frozen-thawed buffalo bull sperm. Semen was collected from three bulls using an artificial vagina (42 °C). Qualifying ejaculates from each bull were divided into four aliquots and diluted (at 37 °C, 50 × 10(6) sperm/mL) in tris-citric acid extender containing DAFP (at 0.1, 1.0, and 10 μg/mL), and the sperm were evaluated for important characteristics relative to a control without DAFP. D canadensis recombinant antifreeze proteins at any of the three concentrations did not affect sperm progressive motility or plasma membrane integrity (PMI), either before or after the semen was cooled to 4 °C in 2 hours. However, after 24 hours of cryostorage at -196 °C, followed by thawing at 37 °C for 30 seconds, sperm progressive motility and PMI were higher (P 0.05) in extender containing DAFP at 10 μg/mL compared with control. The in vitro-fertilizing ability of cryopreserved (-196 °C) sperm supplemented with DAFP (10 μg/mL) was slightly higher (P = 0.098) compared with control, as assessed through in vitro cleavage rate of in vitro matured buffalo oocytes. Also, the in vivo fertility rate was evaluated by inseminating 100 buffaloes (50 inseminations per extender) 12 hours after standing heat. The fertility rate of cryopreserved buffalo bull sperm in terms of positive pregnancy at 90 days after insemination was clinically higher but remained statistically nonsignificant in extender containing DAFP at 10 μg/mL (52.0%) compared with control (43.8%). In conclusion, supplementation of 10 μg/mL of DAFP in the extender improved the motility and PMI of Nili-Ravi buffalo sperm after freeze-thawing, and yielded numerically higher, although statistically nonsignificant, in vitro cleavage, and in vivo fertility rate.

Two experiments were conducted to evaluate the effect of royal jelly (RJ) on post-thaw sperm quality, in vitro and in vivo fertility rate of cryopreserved buffalo bull sperm. The semen was collected from three mature regular donor buffalo bulls, ejaculates were pooled and semen evaluated initially. In Experiment 1, the ejaculates were extended in tris-citric acid diluter supplemented with different RJ concentrations (0, 0.05, 0.1, 0.2, 0.3 or 0.4%). The diluted semen was cooled to 4°C, packaged into 0.5 mL straws and frozen using standard procedure. The straws were thawed and assessed for sperm progressive motility, viability, plasma membrane, acrosome, and chromatin integrity. The results indicated that sperm progressive motility was significantly greater (P0.05) in 0.05, 0.1, 0.2 and 0.3% RJ than 0.4% RJ supplemented and control groups. The sperm viability, plasma membrane and acrosome integrity were significantly improved (P0.05) in 0.1% RJ supplemented group the compared to other treatment groups. In Experiment 2, cryopreserved sperm with 0.1% RJ supplementation and control (without RJ supplementation) were used to observe the in vitro fertilizing potential and in vivo fertility. In vitro fertilization method was applied to assess the cleavage rate; whereas, AI was performed in buffalo during in vivo fertility trial. The buffaloes were inseminated 12h after standing estrus and pregnancy diagnosis was performed through ultrasonography. The results revealed that the cleavage rate was higher (P0.05) in 0.1% RJ as compared to control group. However, the pregnancy rate was similar (P0.05) between 0.1% RJ supplemented and control groups. It is concluded that supplementation of RJ in freezing extender can improve the cryosurvival rate and in vitro fertilizing capacity of buffalo bull sperm.

Lower fertility in buffaloes with frozen-thawed semen is attributed to sperm damage that is believed to be due to formation of ice crystals during freeze/thaw process. It was hypothesized that antifreeze proteins in the extender may improve the post thaw quality of buffalo bull sperm. For this purpose, two separate experiments were conducted to evaluate antifreeze proteins III (AFP III) at 0 (control), 0.1, 1 and 10 μg/mL (Experiment I) and 0 (control), 0.01, 0.1 and 1 μg/mL (Experiment II) for its effect on post thaw quality of buffalo bull semen. Semen was collected from three Nili-Ravi buffalo (Bubalus bubalis) bulls with artificial vagina (42 °C) for three weeks (replicate) per experiment. For each experiment, qualifying ejaculates (6 ejaculates/bull) were divided into four aliquots and diluted (at 37 °C having 50 × 10(6) sperm/mL) in tris-citric acid extender containing above mentioned concentrations of AFP III. Diluted semen was cooled to 4 °C in 2 h, equilibrated for 4 h, filled in 0.5 mL straws, kept over liquid nitrogen vapors for 10 min and plunged in the liquid nitrogen. After 24 h of storage, semen straws were thawed at 37 °C for 30 s to assess sperm progressive motility (SM), plasma membrane integrity (PMI), viability (live sperm with intact acrosome) and normal epical ridge (NAR). In experiment I, improvement (P0.05) in percentage SM and sperm PMI was recorded in extender containing 0.1 μg/mL AFP III compared to control, the higher concentrations (1 μg/mL and 10 μg/mL) being inefficient. While evaluating the lower concentration (experiment II), 0.01 μg/mL of AFP III in the extender it was found to be ineffective to improve semen quality parameters, while 0.1 μg/mL AFP III in extender was found better in terms of progressive motility and plasma membrane integrity of buffalo bull semen compared to control. Sperm viability and NAR remained similar (P0.05) in extenders containing different concentrations of AFP III and control in both of experiments. In conclusion addition of AFP III in the extender at 0.1 μg/mL improved the progressive motility and plasma membrane integrity of cryopreserved buffalo bull semen.

This study was designed to compare the effect of traditional antibiotics streptomycin–penicillin (SP) with neomycin–penicillin (NP), polymyxin–penicillin (PP), and colistin–penicillin (CP) in extender against the post-thaw quality and bacterial count of buffalo (Bubalus bubalis) bull spermatozoa. Qualifying semen ejaculates (n = 18) from three Nili-Ravi buffalo bulls were splits into five aliquots and diluted (50×10−6 spermatozoa/ml; 37°C) in extender containing SP, NP, PP, CP, or without antibiotics (control). The diluted semen was cooled to 4°C in 2 hours and equilibrated for 4 hours. Cooled semen was filled in 0.5 ml French straws at 4°C, kept over liquid nitrogen vapors (5 cm) for 10 minutes and then plunged in the liquid nitrogen for storage. For the assessment of post-thaw semen quality (motility, plasma membrane integrity (PMI), sperm viability) and total aerobic bacterial count (TABC), thawing was performed at 37°C for 30 seconds. Sperm motility, PMI, sperm viability, and TABC remained similar (P>...

Background Brucella species occasionally cause spontaneous human abortion. Brucella can be transmitted commonly through the ingestion of raw milk or milk products. The objective of this study was to determine the sero-prevalence of and to identify potential risk factors for brucellosis in pregnant women from Rawalpindi, Pakistan. Methods We conducted a cross-sectional study at the Gynecology Outdoor Patient department of the Benazir Bhutto Hospital, Rawalpindi, Pakistan from March to June 2013. Data related to potential risk factors and clinical history was collected by individual interviews on the blood sampling day. The 429 serum samples collected were initially screened by Rose Bengal Plate Agglutination test for the detection of Brucella antibodies. We applied standard descriptive statistics and logistic regression analyses. Results Twenty five (5.8 %; 95 % confidence interval (CI): 3.8 % -8.5 %) serum samples were found to be seropositive. Brucellosis-related clinical symptoms were recorded in various seropositive cases. Animal contact, raw milk consumption, having an abortion history and the experience of an intrauterine fetal death were associated with seropositivity for brucellosis in univariate analyses (all p

Sperm selection techniques have been developed to get sperm suspensions enriched in motile and functional cells. Studies show that selection before cryopreservation improves post-thaw quality of cryopreserved sperm but information on buffalo bull sperm is scarce. The study was aimed to 1) perform a comparative analysis of sperm selection procedures; Swim-Up (SU), Sephadex?-G15 Filtration (S-G15) or Glass Wool Filtration (GWF) for total and motile cell recovery, 2) to assess the impact of sperm selection prior to cryopreservation on sperm quality (motility, morphology, cell membrane and normal apical ridge, viability and livability, chromatin integrity) and sperm functionality (Embryo Cleavage after IVF with selected sperm) in post-thawed suspensions of buffalo bull sperm. Semen was collected from 5 Nili Ravi buffalo bulls maintained at the Semen Production Unit Qadirabad, District Sahiwal, Pakistan. Ejaculates were divided into four aliquots for SU, S-G15 and GWF and control. After sperm selection, total and motile sperm recovery was highest in GWF samples (total sperm=84.08±8.39%; motile sperm=80.42±3.57%). An improvement (P

Experiments were conducted to evaluate the effect of Antarctic fish antifreeze glycoproteins, (AFGP) size 1-5 (34-10.5 kDa) and 7-8 (3.2 and 2.4 kDa) in extender on buffalo bull sperm at cooling (4 °C) and at post thawing. Semen was collected from three Nili-Ravi buffalo bulls with artificial vagina for 3 weeks. Qualifying ejaculates from each buffalo bull were diluted (at 37 °C having 50×10(6) sperm/mL) in tris-citric acid extender containing AFGP at 0 (control), 0.1, 1 and 10 μg/mL. An aliquot of diluted semen was evaluated for sperm progressive motility and plasma membrane integrity, while the remaining fraction was cooled to 4 °C in 2 h. Further, an aliquot of cooled semen was evaluated for the previously described variables and the remaining fraction was cryopreserved (-196 °C). After 24 h of storage, straws were thawed at 37 °C for 30 s to assess post-thaw sperm quality. Inclusion of AFGP in the extender did not affect (P0.05) sperm progressive motility and plasma membrane integrity of buffalo bull sperm at cooling stage (4 °C). However, at post thawing, improvement (P0.05) in sperm progressive motility and plasma membrane integrity was recorded in extender containing AFGP 1-5 and AFGP 7-8 at 1 μg/mL compared to the control. Percentage of live sperm with an intact acrosome remained similar (P0.05) in extenders containing different amounts of AFGP and control. In conclusion, supplementation of 1 μg/ml of AFGP in extender improved the motility and plasma membrane integrity of Nili-Ravi buffalo sperm after thawing.