• Asian-Australasian Journal of Animal Sciences
• /
• v.6 no.3
• /
• pp.447-450
• /
• 1993

The effect of adding transparent fluid (TF) to fowl semen on fertilizing capacity of fowl spermatozoa and on hatchability was studied. Diluted semen and semen containing 15% TF were stored for 24 h at $3-5^{\circ}C$ and inseminated at weekly basis for 5 consecutive weeks. No significant differences were observed in fertility, hatchability and embryonic mortality among the treatments. The results suggest that TF is not necessarily detrimental to fowl spermatozoa even when mixed with semen and stored outside the body.

• Proceedings of the KSAR Conference
• /
• 2004.06a
• /
• pp.227-227
• /
• 2004

The purpose of this study was to investigate the effect of taurine on sperm characteristics and gene expressions(bax and Gpx) in fresh boar semen during in vitro storage. The motility of spermatozoa in Modena, Modana plus taurine 25 mM, Modana plus taurine 50 mM, Modana plus taurine 75 mM and Modana plus taurine 100 mM were 63.1%, 65.1%, 65.3%, 82.5% and 80.8%, respectively. (omitted)

• Journal of Embryo Transfer
• /
• v.21 no.4
• /
• pp.339-344
• /
• 2006

This study was to investigate the optimal short-term storage diluents for goat spermatozoa. Semen was collected with electro-ejaculation from two goats. The collected semen was diluted in BTS and centrifuged at 500 g for 5 minutes. The supernatant was discarded and diluted with BTS, Modena or Triladyl$^{(R)}$ and stored at 4 or $17^{\circ}C$ for 8 days. The motility of spermatozoa in BTS and Modena stored at $4^{\circ}C$ was rapidly decreased at day 1. The motility of spermatozoa in BTS at $17^{\circ}C$ was decreased to 30$\sim$45% at day 2. In Modena at $17^{\circ}C$, the inappropriate motility fur artificial insemination (AI) was reached at day 4. The spermatozoa stored in Triladyl$^{(R)}$ at 4 or $17^{\circ}C$ were more viable and higher motility up to day 4. In conclusion, liquid storage of goat spermatozoa in Triladyl$^{(R)}$ at 4 or $17^{\circ}C$ for 4 days showed permissible viability for AI.

• Asian-Australasian Journal of Animal Sciences
• /
• v.28 no.12
• /
• pp.1713-1720
• /
• 2015

The present work describes experiments undertaken to evaluate the usefulness of selected physicochemical indices of semen, cell membrane integrity and sperm chromatin structure for the assessment of boar semen sensitivity to processes connected with pre-insemination procedures. The experiments were carried out on 30 boars: including 15 regarded as providers of sensitive semen and 15 regarded as providers of semen that is little sensitive to laboratory processing. The selection of boars for both groups was based on sperm morphology analyses, assuming secondary morphological change incidence in spermatozoa as the criterion. Two ejaculates were manually collected from each boar at an interval of 3 to 4 months. The following analyses were carried out for each ejaculate: sperm motility assessment, sperm pH measurement, sperm morphology assessment, sperm chromatin structure evaluation and cell membrane integrity assessment. The analyses were performed three times. Semen storage did not cause an increase in the incidence of secondary morphological changes in the group of boars considered to provide sperm of low sensitivity. On the other hand, with continued storage there was a marked increase in the incidence of spermatozoa with secondary morphological changes in the group of boars regarded as producing more sensitive semen. Ejaculates of group I boars evaluated directly after collection had an approximately 6% smaller share of spermatozoa with undamaged cell membranes than the ejaculates of boars in group II ($p{\leq}0.05$). In the process of time the percentage of spermatozoa with undamaged cell membranes decreased. The sperm of group I boars was characterised with a lower sperm motility than the semen of group II boars. After 1 hour of storing diluted semen, the sperm motility of boars producing highly sensitive semen was already 4% lower ($p{\leq}0.05$), and after 24 hours of storage it was 6.33% lower than that of the boars that produced semen with a low sensitivity. Factors that confirm the accuracy of insemination male selection can include a low rate of sperm motility decrease during the storage of diluted semen, low and contained incidence of secondary morphological changes in spermatozoa during semen storage and a high frequency of spermatozoa with undamaged cell membranes.

• Reproductive and Developmental Biology
• /
• v.29 no.3
• /
• pp.149-154
• /
• 2005

The purpose of this study was the analysis of sperm ability in Specific Pathogen Free (SPE) miniature pig for production of bio-organ. The collected semen was diluted with extender and stored at $17^{\circ}C$t for up to 7 days. The semen samples were evaluated at 0, 1, 3, 5, and 7 days of storage for analysis of sperm ability. Sperm ability was evaluated by examining viability, progressive motility, sperm abnormality and intensity of the sperm membrane. Also, the semen was processed according to the convenient freezing method, and frozen-thawed sperm was evaluated by examining viability, capacitation and acrosome reaction using chlortetracycline (CTC) staining. Motility of spermatozoa of SPF miniature pig was significantly (P<0.05) lower on 3 days or later compared to the Duroc, Yorkshire and Landrace in domestic boar. The percentage of abnormal spermatozoa of Landrace were significantly (P<0.05) higher than in SPF miniature pig, Duroc and Yorkshire that had a similar percentage on 5 or 7 days of sperm storage. The percentage of spermatozoa with coiled tail decreased during the storage period but there were no significant difference. On the other hand, viability of frozen-thawed spermatozoa had a significantly (P<0.05) lower in SPF miniature pig than in other domestic boars. CTC patterns had no significant difference, but SPF miniature pig had higher percentage of capacitated spermatozoa and lower percentage of acrosome-reacted it than domestic boars. Therefore, this study suggest that it is necessary to develop the suitable extender and freezing methods methods for the high viable rate and fertilizing ability in vitro.

• Korean Journal of Veterinary Research
• /
• v.31 no.1
• /
• pp.11-18
• /
• 1991

The purpose of this study was designed to investigate the methods for the functional elevations of sperm-host (utero-vaginal, U-V) glands in domestic hens. The laying hens were assigned to five groups of low-, medium-, high- fecundity, gonadotrophin-, and caffeinetreated hen groups, these group hens were sacrified at interval after last artificial inseminations (AI). Number of U-V gland observed in tissue preparation of each hen U-V region were investigated, and also the appearance rates of spermatozoa-contained U-V glands were calculated. 1. In low-fecundity hen groups, the appearance rates of spermatozoa-contained U-V glands were found to be 13.5, 15.6, 11.8, 13.6, 2.3, 0, and 0% respectively at the hens of 1, 3, 7, 10, 13, 16, and 19 days after AI. 2. In medium-fecunditiy hen groups, the appearance rates of spermatozoa-contained U-V glands were found to be 21.7, 22.7, 13.4, 10.4, 10.0, 7.7 and 0% respectively at the hens of 1, 3, 7, 10, 13, 16, and 19 days after AI. 3. In high-fecundity hen groups, the appearance rates of spermatozoa-contained U-V glands were found to be 30.8, 31.8, 28.9, 13.0, 10.3, 10.8, and 0.9 respectively at the hen of 1, 3, 7, 10, 13, 16, and 19 days after AI. 4. In gonadotrophin-treated hen groups, the appearance rates of spermatozoa-contained U-V glands were found to be 31.8, 33.7, 32.3, 17.3, 12.0, 5.0, and 1.0% respectively at hens of 1, 3, 7, 10, 13, 16, and 19 days after AI. 5. In caffeine-treated hen groups, the appearance rates of spermatozoa-contained U-V glands were found to be 33.2, 29.2, 22.4, 17.8, 12.7, 0, and 1.1% respectively at hens of 1, 3, 7, 10, 13, 16, and 19 days after AI. 6. The appearance rates of completely filled U-V glands and partially filled U-V glands of spermatozoa-contained U-V glands were found to be 3.8:1. So we suggested as follows: The appearance rates of spermatozoa-contained glands tend to be high from 1 day after AI to 7 days and tend to declined rapidly from 10 days. Also higher fecundity hen groups tend to be higher in the appearance rates and longer in spermatozoa-contained duration in U-V glands than in lower fecundity hen groups. Gonadotrophin hormone tend to increase the appearance rates of spermatozoa-contained U-V glands than those in control group, whereas caffeine tend to increase those rates at 1 day and to declined more rapidly from 3 day than in control group.

• Asian-Australasian Journal of Animal Sciences
• /
• v.19 no.12
• /
• pp.1716-1721
• /
• 2006

The present study was conducted to observe the effect of osmolality of glycerolated TEST-yolk glycerol extenders on post-thawing sperm kinematics of ram spermatozoa of the native Malpura breed maintained in a semi-arid tropical environment. Good quality semen obtained from adult rams was pooled, split and diluted to 1,000 million spermatozoa per ml in complete TEST-yolk-glycerol extenders of 900, 1,200, 1,500 and 1,800 mOsm/kg osmolality. Diluted semen samples were loaded in 0.25 ml straws and cooled down to $-125^{\circ}C$ freezing temperature at the rate of $-25^{\circ}C$ per minute under controlled conditions before plunging into liquid nitrogen for storage. The thawing of straws was performed at $50^{\circ}C$ in a water bath for 10 seconds and sperm kinematics of the frozen-thawed spermatozoa were assessed by a computer-assisted sperm analysis technique. Osmolality of diluent had no significant effect on post-thawing % motility, % rapid, % medium and % slow moving frozen-thawed spermatozoa but significantly (p< 0.05) affected the % linearity and % straightness. The post-thawing % motility and % rapid motile spermatozoa were highest in samples extended in diluent of 1,500 mOsm/kg osmolality and lowest in 900 mOsm/kg. The curvilinear velocity of spermatozoa was significantly (p<0.05) higher for samples extended in 1,800 mOsm/kg, compared to those in 900 and 1,200 mOsm/kg, but the effect was not significantly different to those extended in diluent of 1,500 mOsm/kg osmolality. The study indicated that ram spermatozoa could tolerate a wide osmolality range for dilution in the complete TEST-yolk-glycerol extender for their cryosurvival. The highest recovery of motile spermatozoa following thawing was achieved in samples extended in the TEST-yolk-glycerol diluent of 1,500 mOsm/kg osmolality.

• Asian-Australasian Journal of Animal Sciences
• /
• v.33 no.7
• /
• pp.1068-1076
• /
• 2020

Objective: In the present study, we determined efficiency of incorporating caffeine, melatonin or omega-3 polyunsaturated fatty acid in the diluent on mitigating consequences of (a) liquid chilled- and (b) cryo-storage of ram spermatozoa. Methods: In the first experiment, ejaculates (n = 30) were collected from 5 adult rams and were pooled, diluted (1:10) with Tris-citric acid (base diluent) and were split into 4 aliquots assigned for: control (untreated), caffeine (0.1 mM), melatonin (0.3 mM) or omega-3 fatty acids (0.3 mM) (T₀). The diluted specimens were stored at 4℃ for 48 h, during which sperm physical and cytological properties were evaluated along with oxidative stress indices (T₂₄, T₄₈). In the second experiment, 15 ejaculates (3 per male) were pooled, diluted with glycerolized base diluent (4% glycerol, v/v) and were split corresponding to the same previous treatment groups before being processed for cryopreservation. Post-thaw physical and kinematic sperm properties were assessed by a computer-assisted sperm analysis system. Results: The results clarified superiority of both melatonin and omega-3 supplementation on maintaining (p<0.05) sperm properties, while reducing (p<0.05) lipid peroxidase reaction and enzymatic activities of alanine aminotransferase, aspartate aminotransferase, and alkaline phosphatase in preservation medium, compared to caffeine either during liquid-chilled storage or cryopreservation of spermatozoa. Conclusion: Melatonin and omega-3 are regarded efficient alternatives to caffeine when processing ram spermatozoa for application of artificial insemination or in vitro fertilization.

• Journal of Embryo Transfer
• /
• v.28 no.4
• /
• pp.349-353
• /
• 2013

The objective of this study was to compare the effect of semen extenders on the sperm motility, viability, acrosome integrity and functional integrity of plasma membrane (HOST: hypo-osmotic swelling test) during liquid preservation of Korean Native boar semen. In this experiment, semen was diluted in Androhep plus, Beltsville Thawing Solution (BTS), ModenaTM, Seminark and Vitasem LD. Sperm-rich fractions were collected from three Korean Native boars and sub-samples were diluted ($30{\times}10^6$ spermatozoa/ml) in different semen extenders. Semen samples were stored at $17^{\circ}C$ for 96 hours. On everyday (0, 24, 48, 72, 96 h) after storage, the sperm characteristics relevant for fertility, such as sperm motility, viability, acrosome integrity and HOST positive were evaluated. The motility of spermatozoa stored in different extenders was no significantly different among other extenders (P>0.05). Also, no difference was observed among samples processed with different extenders in the percentage of sperm viability, acrosome integrity and HOST positive. All extenders maintained a high percentage (70%) of sperm motility, viability and acrosome integrity through 96 h of storage. The result of this study show that there was no significant differences among extenders in their capacity to preserve motility, viability and membrane integrity of spermatozoa from normal, fertile Korean Native boars for 96 h of liquid preservation at $17^{\circ}C$.

• Proceedings of the KSAR Conference
• /
• 2004.06a
• /
• pp.277-277
• /
• 2004

The purpose of this study was to investigate the effect of different conditions (osmolity, solution, incubation times, comparison of fresh and frozen/thawed semen and storage times) on the swelling of canine spermatozoa. Employing the hypoosmotic swelling test (HOST), the membrane integrity of spermatozoa in different solutions (sucrose, fructose, latose, Na-citrate, Na-citrate plus sucrose, Na-citrate plus fructose and Na-citrate plus lactose were 61.4%, 66.2%, 62.5%, 68.1%, 62.0%, 68.5% and 60.2%, respectively. (omitted)