• Asian-Australasian Journal of Animal Sciences
• /
• v.12 no.5
• /
• pp.702-707
• /
• 1999

A percoll density gradient technique was developed for producing high quality turkey semen and improving the fertility by removing deleterious cellular components, including spermiophages, bacteria, abnormal or dead spermatozoa, and other cellular debris. The combination of three different percoll densities, 1.05, 1.07, and 1.08 showed the best resolution and was selected to prepare a discontinuous percoll density gradient to obtain healthy spermatozoa from semen smples. Bacteria, spermiophages, and abnormal or dead spermatozoa were detected from the density range from 1.05, 1.05 to 1.07, and 1.07 to 1.08, respectively. Healthy spermatozoa were collected from the density greater than 1.08. Spermatozoa obtained from percoll density gradient centrifugation showed better sperm motility than those from unprocessed pooled semen. Bacteria including Escherichia coli, Staphylococcus aureus, and Proteus spp., were predominant contaminants in turkey semen, and the numbers of cells were approximately $5{\times}10^5$ to $1{\times}10^9cfu/ml$. The overall fertility rates in hens inseminated with processed percoll density gradient were higher than those in hens with unprocessed semen especially for unhealthy sperm. In conclusion, semen quality can be improved by percoll density gradient centrifugation, which augmented the fertility of turkey breeders.

• Korean Journal of Agricultural Science
• /
• v.50 no.4
• /
• pp.941-952
• /
• 2023

The metabolites of agrichemicals, such as organophosphorus pesticides, are known to be more hazardous than their parent pesticides. 3,5,6-trichloro-2-pyridinol (TCP) is a major degradation product of chlorpyrifos, one of the organophosphate insecticides widely used in agriculture. In vivo or in vitro exposure to chlorpyrifos has been known to interfere with male reproductive functions, leading to reduced fertility in mammals. Therefore, this study was performed to examine the changes in the fertilization competence of boar spermatozoa exposed to TCP. Sperm samples were subjected to varying concentrations of TCP (10, 50, 100, 200 µM) and different periods of incubation. Sperm motility, motion kinematics, viability, acrosome integrity, intracellular reactive oxygen species (ROS) production, and gene expression levels (ODf2, ZPBP2, AKAP3 and AKAP4) were evaluated after exposure of the sperm to TCP. A significant dose-dependent reduction in motility was observed in sperm samples incubated with TCP compared to the controls after both incubation periods. Sperm viability was significantly decreased in samples incubated with 50, 100, and 200 µM TCP in both incubation periods. A significantly lower percentage of normal acrosomes and gene expression levels were observed in sperm samples exposed to 50, 100, and 200 µM TCP after both incubation periods, compared to the controls. There was a significant increase in the ROS production in spermatozoa incubated with 100 - 200 µM TCP after both incubation periods. Consequently, the direct exposure of boar spermatozoa to TCP interferes with sperm functions and leads to decreased fertilization. In order to identify and address the various causes of reproductive decline, the impact of chemical metabolites needs to be discussed in depth.

• Animal Bioscience
• /
• v.36 no.10
• /
• pp.1530-1535
• /
• 2023

Objective: Semen cryopreservation result in decreased sperm parameters and fertilization ability. Sericin exhibits antioxidant activity by reducing lipid peroxidation resulting from free radicals, which can potentially improve cryopreservation outcomes. The present study aimed to examine the efficacy of various sericin concentrations supplemented with a rooster semen-freezing extender on post-thaw semen quality and fertilizing ability of sperm after cryopreservation. Methods: Semen samples were collected from 40 roosters (5 reps), then were pooled, and divided into four groups by the levels of sericin supplementation (0%, 0.25%, 0.50%, and 0.75%) in a freezing extender. Semen suspensions were loaded in medium straw (0.5 mL) and cryopreserved with the traditional liquid nitrogen vapor method. Post-thawed semen was evaluated for sperm motility, sperm viability, and lipid peroxidation. Also, the fertility test was determined. Results: The results showed that supplementation of the freezing extender with 0.50% to 0.75% sericin resulted in greater total motility and progressive motility and lower malondialdehyde levels than the other groups after cryopreservation (p<0.05). However, the viability of 0.75% decreased compared with the value of 0.50% sericin supplementation (p<0.05). Moreover, the fertility and hatchability of total eggs were significantly higher in the 0.50% sericin group than in the other groups (p<0.05). Conclusion: In conclusion, 0.50% sericin is recommended as an alternative component of the freezing extender to improve cryopreserved rooster semen.

• Journal of Animal Science and Technology
• /
• v.63 no.1
• /
• pp.46-57
• /
• 2021

Fruits with antioxidant enrichment can be an economically affordable supplement for mitigating oxidative damage prone spermatozoa membrane pathologies. Computer-assisted sperm analyzer and oxidative status were utilized to evaluate the impact of watermelon (Citrullus lanatus) fortification of dextrose saline as diluent for rooster semen and fertility response of hens inseminated. Watermelon juice and dextrose saline were used to formulate diluent of 7 treatments consisting of unextended semen (positive control), 10%, 20%, 30%, 40%, 50% and only dextrose saline (negative control) designated as Treatments 1-7. Pooled semen was obtained from fertile roosters and equilibrated with diluents at ratio 1:2 in the various treatments and were evaluated using computer software coupled microscope and seminal oxidative status assay. 168 laying hens randomly divided into 7 treatment of 8 replicates and 3 hen per replicate. Hen were everted, and semen (2 × 108 Spermatozoa) deposited intra-vagina and eggs collected over 8 weeks to assess fertility and hatchability of eggs laid. The result obtained revealed that watermelon-dextrose saline rooster semen diluent enhanced progressive motility, sperm kinetics and lowered non-progressive motility in T2-T6 compared to T7 over the 3 hours of evaluation. Watermelon addition to rooster semen diluent enhance the antioxidant capacity of rooster semen and lowered lipid peroxide generation. The percentage fertility was highest in T3 (81.01%) and T4 (81.24%) with lowest value obtained in T7 (73.46%). The hatchability of eggs set of hens inseminated with undiluted semen (71.46%) was lower than values for hens inseminated with watermelon inclusive extended semen (75.71%-80.39%). The optimal inclusion of 30%-40% watermelon in dextrose saline diluent enhance rooster semen kinetics, seminal oxidative stability and egg fertility.

• Journal of Animal Reproduction and Biotechnology
• /
• v.35 no.2
• /
• pp.190-197
• /
• 2020

The effects of the number of frozen-thawed ram sperm per single and double intra-cervical artificial insemination (AI) on fertility in ewes were studied. A total of 89 non-pregnant ewes were synchronized for oestrus with two doses of 100 ㎍ PGF_2α (Cloprostenol) 9 days apart. The ewes were randomly assigned to one of four groups; D200 (n = 23; double AI with 200 × 10⁶ sperm), S200 (n = 24; single AI with 200 × 10⁶ sperm), D100 (n = 24; double AI with 100 × 10⁶ sperm) and S100 (n = 18; single AI with 100 × 10⁶ sperm). Ewes were inseminated within 12 to 18 h for single AI and, within 10 to 12 h and 16 to 18 h for double AI after the onset of oestrus. The onset of oestrus ranged from 28 to 76 h (54.33 ± 1.28 h). The high percentage (29.2%) of ewes showed oestrus between 51 to 60 h. The non-return rates were highest in group D200 (56.5%) and differed significantly (p < 0.05) from group S100 (11.1%). No ewes were pregnant in group S100, and the pregnancy rates among the remaining groups did not differ. The mean gestation period was 152.8 ± 0.5 days and no difference was observed among the groups. The lambing and multiple birth rates were 100% in group D200. The single and twin lambing was highest in group D100 (33.3%) and group D200 (83.3%), respectively. Only one triplet lambing and the highest lambing size (2.2 ± 0.2) was recorded in group D200. In conclusion, double AI with 200 × 10⁶ sperm showed comparatively most practical for achieving high pregnancy rates and lambing performances in Bangladeshi ewes under field conditions.

• Toxicological Research
• /
• v.11 no.1
• /
• pp.69-75
• /
• 1995

The present study was carried out to establish several spermatotoxicity test methods. For this purpose we investigated following parameters in the fertility study of DA-125, a new anticancer agent, in rats: testicular spermatid counts, epididymal sperm counts, daily sperm production rate, sperm morphology, and serum testosterone concentration. Motility and velocity of sperms were also measured using non-treated rats. At 0.3 mg DA-125/kg, spermatids per 1g testis and daily sperm production rate per 1g testis were significantly decreased, when compared with those of control group. Several types of abnormal sperms, such as no head, pin head, double head, hook at wrong angle, no tail, and small sperm, were found in both treated and control groups at a low frequency. Serum testosterone concentration at 0.3 mg DA-125/kg was close to the control value. Sperm motility and velocity measured with non-treated rats were in a good agreement with the results of other investigators. In our study established spermatotoxicity test methods can be used as a tool not only for the close examination of the cause of drug- or chemical-induced infertility, but also for the effective evaluation of reproductive toxicity.

• Journal of Embryo Transfer
• /
• v.11 no.1
• /
• pp.81-83
• /
• 1996

This study was carried out to investigate the effect of sperm concentration of 5ml maxi-straw on farrowing rate and number of pigs born alive per litter in deep frozen boar semen. We did not find out the effect of sperm concentration on post-thaw sperm motility and NAR acrosome. However, farrowing rate and number of pigs born alive per litter of 7. 5 x 10˚ /5ml and 10.0 x 10˚ /5m1 sperm concentrations were higher than those of 5. 0 /10˚ /5ml sperm concentration.

• Proceedings of the Korean Aquaculture Society Conference
• /
• 2003.10a
• /
• pp.40-40
• /
• 2003

DNA content of the sperm of tetraploid mud loach (Misgurnus mizolepis) males and the ploidy status of progenies generated by crossing tetraploid males with diploid females are described. Reproductive performance of the induced adult tetraploid males ranged from sterility to fertility. Of 48 tetraploid males tested, 12 were sterile but the other 36 produced functional sperm. Of these 36, 26 produced haploid sperm, which on fertilizing the haploid eggs, generated diploid progenies. Seven tetraploid males were mosaics in their sperm, as indicated by the production of diploid, aneuploid and/or triploid offspring. Only 3 males produced diploid sperm rendering the production of all-triploid progenies. The DNA content of sperm of a tested tetraploid male was consistent throughout the 3 progeny tests, i.e. the haploid sperm-producing 4n males persisted to produce the haploid sperm only likewise the diploid sperm producing4n males consistently produced the diploid sperm only, when progeny testing was extended to 3 successive but alternate years. Hence, a careful and direct examination of the DNA profile of sperm from tetraploid males is a pre-requisite for reproductive containment of genetically modified fish.

• Reproductive and Developmental Biology
• /
• v.35 no.3
• /
• pp.203-207
• /
• 2011

The objective of this study was to determine the effect of semen extenders on the motility, viability and fertility in vitro of spermatozoa during storage of fresh boar semen diluted in different commercial extenders used for pig artificial insemination (AI). In this experiment, semen were diluted in Androhep plus, Beltsville Thawing Solution (BTS), Modena, Seminark and Vitasem LD. Five ejaculates were collected from three Duroc boars and sub-samples were diluted ($30{\times}10^6$ spermatozoa/ml) in different extenders. Semen was stored at $170^{\circ}C$ for 10 days. Sperm motility and viability was assessed using Computer-Assisted Semen Analysis (CASA) and flow-cytometry on 1, 3, 5 and 10 day post collection The motility of spermatozoa stored in different extenders was gradually decreased by increasing the duration of storage of semen. However, there was not significant1y different in the sperm motility and viability among other extenders. On the other hand, the in vitro-matured oocytes were fertilized and cultured in vitro to assess the fertility of boar spermatozoa stored for 3 and 10 days in different extenders. The percentage of morula and blastocyst were taken as indicators of fertility in vitro of spermatozoa. Therefore, there were no differences in the rate of embryos developed to the molular and blastocyst stage. There were no differences in the motility and fertility in vitro among 5 kinds of commercial boar semen extenders.

• Korean Journal of Animal Reproduction
• /
• v.3 no.1
• /
• pp.30-35
• /
• 1979

A, B and C dilutors were used to make Ka (A plus B (1 : 1)) and Na (B plus C(1 : 1)) dilutors in this experiment. Three aliqots of semen were respectivly diluted 1 : 1 and 1 : 2 (semen: dilutor) with Ka, Na and C dilutors and stored at 5$^{\circ}C$ for 7 days in order to study their livability during storage. Fertility was checked for the diluted semen with Ka, Na and C dilutors. Whole semen and extended semen with Na dilutos with and without DMSO were cold shocked at various temperatures for 10 min. Effects of different 1st and 2nd dilution with A, B, C and Na dilutors and of vessels on freezability of spermatozoa were investigtigated. 1. Extended semen 1 : 2 with Na and C dilutors showed highest live sperm index during storage for 7 days at 5$^{\circ}C$. 2. The components of Na dilutor per 100$m\ell$ were skim milk 2.5g, trisaminomethane 0.54g, citric acid 0.265g, glucose 2.835g, fructose 1.5g, sodium lauryl sulfate, 0.08g, penicillin 0.06g, streptomycin 0.075g, and egg yolk 10$m\ell$. 3. Fertility of diluted semen was higher than that of whole semen. Ka dilutor showed higher fertility than Na and C dilutors, and there was no difference in the fertility between Na and C dilutors. 4. Na dilutor with DMSO showed slightly higher livability than Na dilutor without DMSO during storage for 7 days at 5$^{\circ}C$. 5. Cold shock at 1$0^{\circ}C$ for 10 min. decreased greatly the sperm livalility of whole semen but not of extended semen with Na dilutor. Addition of DMSO to Na dilutor has no effect in prevention of cold shock. 6. The extended semen with C. C dilutor (1st and 2nd dilution with C and C dilutor) showed higher post-thawing sperm livability than A.A and Na. B dilutors. Na. B dilution shwed higher post-thawing sperm livability than A.A dilution. There was no difference in the post-thawing livability between semen in 1$m\ell$ straw and 10$m\ell$ aluminium package.