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Publication Open Access Supplemental effect of varying L-cysteine concentrations on the quality of cryopreserved boar semen.(2010) Kampon Kaeoket; Panida Chanapiwat; Padet Tummaruk; Mongkol Techakumphu; Mahidol University. Faculty of Veterinary Science. Semen Laboratory.Cryopreservation is associated with the production of reactive oxygen species, which leads to lipid peroxidation of the sperm membrane and consequently a reduction in sperm motility and decreased fertility potential. The aim of this studyPublication Open Access Distribution of spermatozoa in the reproductive tracts of sows after intra-uterine insemination using frozen-thawed boar semen(2012) Panida Chanapiwat; Kampon Kaeoket; Padet Tummaruk; Mahidol University. Faculty of Veterinary ScienceThe aims of the present study were to determine the number of spermatozoa in the reproductive tract of sows after intra-uterine insemination (IUI) with frozen-thawed (FT) boar semen and to investigate the influence of adding seminal plasma in the thawing medium on the sperm transport in the female reproductive tract. Fourteen multiparous sows were divided into 3 groups: group I (n= 5), insemination with 80 ml of extended fresh semen, group II (n= 4), insemination with 40 ml of FT semen diluted with ModenaTM extender and group III (n= 5), insemination with 40 ml of FT semen diluted with combination of ModenaTM and seminal plasma. All the sows were inseminated once with 2x109 motile spermatozoa at 36.1±2.8 hours after human Chorionic Gonadotropin (hCG) administration. The reproductive tract was collected from the sows at 12.4±2.2 hours after insemination and was divided into 5 parts, i.e. ampulla, cranial isthmus, caudal isthmus, utero-tubal junction (UTJ) and cranial uterine horn. All parts of the reproductive tract were flushed by phosphate buffer solution and the number of spermatozoa was determined using a Neubauer hemocytometer. Spermatozoa were found in all parts of the sows’ reproductive tracts at 12.4 hr after IUI using either fresh (group I) or FT boar semen (group II and III). Most of the spermatozoa were found in the UTJ (47.8%, 47.8% and 38.1% in group I, II and III, respectively, p> 0.05) and the caudal isthmus (27.2%, 26.4% and 28.1% in group I, II and III, respectively, p> 0.05). The total number of recovered spermatozoa in group III (409,420 sperm) tended to be higher than group I (286,750 sperm, p= 0.109) and II (287,000 sperm, p= 0.139). Number of spermatozoa in the cranial isthmus in group III (76,400 sperm) tended to be higher than that in group I (35,750 sperm) and II (33,333 sperm) (p> 0.05). It can be concluded that at 12.4 hr after IUI using FT semen, spermatozoa were found in all parts of the reproductive tracts of the sows similar to that using extended fresh semen. Supplementation of seminal plasma in the thawing medium of FT boar semen tended to increase the transportation of spermatozoa toward the cranial isthmus. This might be due to the suppression of PMN cells in the female reproductive tract and the reduction in cryoinjury of the FT boar sperm caused by seminal plasma.Publication Open Access The sperm DNA damage after cryopreservation of boar semen in relation to post-thawed semen qualities, antioxidant supplementation and boars effects.(2010) Panida Chanapiwat; Kampon Kaeoket; Padet Tummaruk; Mahidol University. Faculty of Veterinary ScienceThe objectives of the present study were to evaluate the damage of DNA of the frozen-thawed (FT) boar spermatozoa and to investigate the effect of various concentrations of L-cysteine supplementation on the sperm DNA damage. A total of 104 cryopreserved semen samples from twenty-six ejaculates of 16 proven boars were analyzed. Of these samples, each semen sample contained a different concentration of L-cysteine i.e., 0 (n=41), 5 (n=41), 10 (n=11) and 15 (n=11) mM. All of the semen samples were cryopreserved by controlled-rate freezer. The semen was thawed at 50oC for 12 sec and the damage to the sperm DNA was determined using acridine orange (AO) staining. The results revealed that, on average, the DNA damage was observed in 0.5% of the FT boar spermatozoa. DNA damage varied among the boars from 0.0% to 4.0%. The levels of DNA damage were 0.6%, 0.4%, 0.5% and 0.9% in the extenders supplemented with 0, 5, 10 and 15 mM of L-cysteine, respectively (p>0.05). In conclusion, the DNA damage of the FT boar spermatozoa was relatively low. No adverse effect of L-cysteine supplementation up to 10 mM on the damage of the sperm DNA was found. Boar characteristic is the most important factor affecting the damage of the sperm DNA.Publication Open Access Effect of L-cysteine on Chilled carp (Cyprinus carpio) semen qualities.(2013) Kan Kledmanee; Somrat Taweedet; Prawporn Thaijongruk; Panida Chanapiwat; Kampon Kaeoket; Mahidol University. Faculty of Veterinary Science. Department of Clinical Sciences and Public Health. Semen LaboratoryThe aim of the present study was to study the effects of L-cysteine on chilled carp (Cyprinus carpio) semen qualities. Pooled semen samples were prepared from eight fish, and divided into five groups according to the concentrations of L-cysteine as follows: 0 (T1), 0.5 (T2), 1.0 (T3), 1.5 (T4) and 2 (T5) mM. The sperm motility, duration of sperm motility and sperm viability were evaluated at 0, 12, 24, 48, 72 hours after chilled storage. Comparing between treatment and control groups, the percentage of sperm motility, duration of sperm motility and the percentage of sperm viability in the treatment groups were significantly higher than the control group (p < 0.05). Considering over a period of time after chilled storage, modified Kurokura’s extender plus L-cysteine groups were able to maintain carp semen qualities (motility, duration of motility and viability) up to 24 hour. Comparing all concentrations of L-cysteine, at 24 hour after chilled storage, the optimal concentration of L-cysteine was found at 1 mM. In conclusion, supplementation of L-cysteine at 1 mMol in modified Kurokura’s extender can be recommended for carp semen chilled storage.Publication Open Access Effects of adding melatonin on the quality of frozen-thawed boar semen(2017) Natcha Thongrueang; Nutchanat Chaibangyang; Panida Chanapiwat; Kampon Kaeoket; Mahidol University. Faculty of Veterinary Science. Department of Clinical Science and Public HealthThe aim of this study was to study the effects of adding melatonin at different concentrations on the quality of cryopreserved boar semen. Semen samples (n = 6) were collected by hand-glove technique. Semen samples were diluted with Modena? and divided into 6 groups. According to the concentrations of melatonin at 0 (control, group A), 0.1 (group B), 0.5 (group C), 1.0 (group D), 1.5 (group E) and 2.0 mM (group F) to the lactose-egg yolk extenders used to freeze boar semen with traditional method. Progressive motility, viability and acrosome integrity were evaluated both before and after cryopreservation. The results showed that here was no significant difference in percentage of progressive motility among groups. However, a higher percentage of progressive motility was found in group D (39.17%). A higher percentage of sperm viability and acrosome integrity were found in group F (28.33%) and group F (34.50%), respectively. In conclusion, the present results suggest that adding melatonin between 0.1 and 1 mM during freezing yield a superior post-thawed semen qualities than freezing without melatonin.Publication Open Access Comparative study on six different long term commercial extenders for fresh boar semen(2010) Kampon Kaeoket; Titisa Srisowanna; Ubonrat Wichaidit; Panida Chanapiwat; Sukanya Manee-in; Mahidol University. Faculty of Veterinary Science. Semen LaboratoryIn Thailand, there are numerous commercial extenders for preservation of boar semen; however, no study has been carried out to compare these extenders in view of maintaining of sperm motility, plasma membrane integrity and acrosome integrity during cold storage. The aim of this study was to assess the percentage of sperm motility, HOST positive, viability and acrosome integrity of boar spermatozoa extended in Merck-III, Androstar®Plus, ModenaTM, NUTRIXcell®, VITASEM LD, Duragen and Dofu goldTM. Ejaculated from boars (n=6, one ejaculate from each boar) were collected and sub-samples (splited samples) were diluted (3x109 spermatozoa/100 ml) in the different extenders and stored for 10 days at 18ºC. On every second day (days 0, 2, 4, 6, 8, 10) after storage, the sperm parameters such as sperm motility, HOST positive, viability and acrosome integrity were evaluated. There were significant differences in characteristic of sperm motility and acrosome integrity between extenders. For instance, on day 4, Merck III (short term extender) and other long term extenders are able to maintain the semen qualities as claimed by the manufacturing. On day 8, the percentages of sperm motility, viability and acrosome integrity were highest (p< 0.01) in Androstar®Plus (72%) and ModenaTM (75%), Androstar®Plus (67%) and ModenaTM(65.6%), and Androstar®Plus (74.6%), Duragen (72.4%) and VITASEM LD (71.6%), respectively. In conclusion, changes in motility, viability and acrosome integrity during storage were affected by the extender utilized, however long term extenders maintained a high percentage (70%) of sperm motility (i.e. Androstar®Plus, ModenaTM and Duragen) and acrosome integrity (i.e. Androstar®Plus, VITASEM LD and Duragen) through 8 days of storage.Publication Open Access DHA analysis in different types of egg yolks: Its possibility of being a DHA source for boar semen cryopreservation.(2013) Kampon Kaeoket; Panida Chanapiwat; Mahidol University. Faculty of Veterinary Science. Department of Clinical Sciences and Public Health. Semen LaboratoryThe aim of this study was to investigate the concentrations of Docosahexaenoic acid (DHA) in DHA enriched hen egg yolk (H-DHA) in comparison with egg yolk from chicken, duck, quail, ostrich and crocodile in order to find a source of DHA for boar semen cryopreservation. A pool (10 eggs in each) of DHA enriched hen egg yolk and egg yolks from chicken, duck, quail, ostrich and crocodile were analyzed for fatty acids profile at the Nutrition Laboratory, Mahidol University. Comparing all egg yolks, the highest DHA concentration in egg yolk was found in H-DHA egg yolk (3.7% of total fatty acid) and the lowest was found in ostrich egg yolk (0.4% of total fatty acid). Comparing DHA concentration in all egg yolks (not including H-DHA egg yolk), the highest DHA concentrations were found in duck egg yolk (1.8% of total fatty acid) and quail egg yolk (1.5% of total fatty acid),respectively. In conclusion, H-DHA, duck and quail egg yolks can be an abundant source of DHA for boar semen cryopreservation.