The effect of Thai ginger (Kaempferia parviflora) extract orally administration on sperm production, semen preservation, and fertility in Thai native chickens under heat stress.

Thai indigenous roosters are exposed to unsuitable temperatures and humidity, resulting in a lower reproductive potential. Kaempferia parviflora (KP) extract containing methoxyflavones was fed to roosters to improve their reproductive performance. Thirty-two Thai native roosters were orally administered KP extract at 300, 450, and 600 mg, calculated according to their average body weight, for at least 14 d before semen collection and continued supplementation until the end of the experiment. The nonsupplemented group served as the control. Fresh semen in terms of semen volume, sperm concentration, mass movement score, and sperm viability were evaluated. Semen preservation at 5°C and fertility test were examined for total motility (MOT), progressive motility (PMOT), sperm viability, and lipid peroxidation up to 48 h of storage. Testosterone concentrations and testicular function were also determined. The results showed that the highest sperm concentration and sperm motility of fresh semen were observed in KP extract at 600 mg (P < 0.001). KP extract at 600 mg resulted in higher sperm viability than the control and KP extract at 300 mg (P < 0.05), but was not different from KP at 450 mg (P > 0.05). The highest MOT, PMOT, and viability were found in the roosters that received 600 mg oral KP extract (P < 0.05), while those of the roosters that received oral KP extract 300 mg and the control were the lowest (P < 0.05) at all storage times. Lipid peroxidation was significantly lower in the KP extract up to 24 h (P < 0.05). The fertility and hatchability of the KP extract at 600 mg at T48 showed a minor decrease compared to the control at T0. These results might be inferred as a result of good spermatogenesis, as revealed by the results of histological examination and testosterone activity. In summary, oral administration of 600 mg KP extract improved sperm production and successfully preserved rooster semen for a long duration of up to 48 h of storage.

Supplementation of Freezing Medium with Ginseng Improves Rooster Sperm Quality and Fertility Relative to Free Radicals and Antioxidant Enzymes.

To the best of our knowledge, this study is the first to determine the effect of ginseng as an antioxidant supplement in freezing extenders on the quality of cryopreserved rooster semen. Semen samples were collected from 40 Thai native roosters (Pradu Hang Dum) using the dorso-abdominal massage method and then pooled and divided into five groups according to the concentrations of ginseng supplementation (0, 0.25, 0.50, 0.75, and 1 mg/mL) in a freezing extender. The semen suspensions were loaded into a medium straw and cryopreserved using the liquid nitrogen vapor method. The post-thaw semen was evaluated for sperm quality (sperm motility and membrane integrity), seminal plasma characteristics (lipid peroxidation, superoxide dismutase [SOD], catalase [CAT], and glutathione peroxidase [GPx]), and fertility. The results showed that ginseng extract supplementation at 0.25 mg/mL yielded the highest total motility, progressive motility, and membrane integrity (59.47%, 30.82%, and 48.30%, respectively; p < 0.05) in cryopreserved rooster semen. Higher malondialdehyde concentrations were observed in the control group than in the other groups (p < 0.05). SOD, CAT, and GPx increased compared with those in the control group (p < 0.05). The results showed that the fertility rate with 0.25 mg/mL of ginseng was higher than that of the control group (62.80% vs. 46.28%: p < 0.05). In conclusion, supplementation with 0.25 mg/mL of ginseng is recommended as an alternative component to the freezing extender to improve rooster semen cryopreservation.

Increasing sperm production and improving cryosurvival of semen in aged Thai native roosters as affected by selenium supplementation.

OBJECTIVE: Aging roosters typically exhibit subfertility with decreasing semen quality, furthermore Thai native roosters reared in rural areas are raised for a longer duration than their usual lifespan. The present study therefore aimed to assess the effect of selenium supplementation as an antioxidative substance in diets to improve the semen cryopreservation of aged roosters. METHODS: Semen samples were collected from young (n = 20) and aged (n = 20) Thai native roosters (Pradu Hang Dum) at 36 and 105 weeks of age when starting the experiment, respectively. They were fed diets either non-supplemented or supplemented with selenium (0.75 ppm). Fresh semen quality and lipid peroxidation of fresh semen was evaluated before cryopreservation using the traditional liquid nitrogen vapor method. Post-thaw sperm quality and fertility potential were determined. RESULTS: Advancing age is unrelated to decreasing fresh semen quality (p>0.05). However, lipid peroxidation in rooster semen depended on age, and the malondialdehyde (MDA) concentration increased in aged roosters (p<0.05). Selenium supplementation in diets significantly decreased the MDA concentration and increased the sperm concentration (p<0.05). In contrast, cryopreserved semen was affected by advancing rooster age, and selenium influenced sperm quality (p<0.05). Younger roosters had higher post-thaw sperm quality and fertility potential than aged roosters (p<0.05). Likewise, diet selenium supplements improved post-thaw sperm quality and fertility compared with the non-supplement group. CONCLUSION: Rooster's age does not influence the rooster sperm quality of fresh semen, while sperm cryotolerance and fertility were greater in young roosters than in aged roosters. However, sperm of aged roosters could be improved by dietary selenium supplementation.

Improvement of rooster semen freezability and fertility rate after sericin supplementation in freezing semen extender.

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.

Fertilizing ability and survivability of rooster sperm diluted with a novel semen extender supplemented with serine for practical use on smallholder farms.

Semen extenders are essential for maintaining the quality of sperm during storage and assuring the success of fertility after insemination. The objective of this study was to examine the effect of a new rooster semen extender (hereinafter, NCAB) supplemented with serine on the survivability and fertilizing ability of sperm following storage. NaCl solution, the NCAB extender, and IGGKPh extender were used as treatments. In Experiment 1, different storage temperatures (5°C and 25°C) and durations of storage (0, 12, and 24 h) were used to compare the semen quality and determine the suitable storage temperature. The fertility test was performed in in-station tests and on-farm experimental trials in Experiments 2 and 3, respectively. The results indicated that the interaction effect of duration, treatment, and storage temperature on all sperm parameters was highly significant. The NCAB extender significantly improved rooster semen quality during storage for 24 h at 5°C (P < 0.01). The sperm diluted in saline solution could not survive 24 h of storage at 25°C. The fertility and hatchability rates obtained for sperm diluted with the NCAB extender were higher than those diluted with other extenders. In addition, the fertilizing capacity of the NCAB extender-diluted sperm under field conditions was significantly higher than that of the saline solution-treated sperm. In conclusion, the NCAB extender supplemented with serine and stored at a low temperature (5°C) positively affects sperm quality and fertilization.

The Melatonin Treatment Improves the Ovarian Responses After Superstimulation in Thai-Holstein Crossbreeds Under Heat Stress Conditions.

The effect of heat stress with melatonin treatment on the superovulatory responses and embryo characteristics in Thai-Holstein crossbreeds under heat stress conditions was examined. Six non-lactating cows (replication = 4; n = 24) were assigned to one of 2 treatments in double cross-over design. All cows were superstimulated with traditional treatment. Melatonin group (n = 12): cows received intramuscularly injection 18 mg/50 kg. simultaneously with GnRH injection, while those in the control group (n = 12) received none. Bloods samples were taken to determine lipid peroxidation (MDA) and the activity of the antioxidant enzymes (superoxide dismutase; SOD). The experiment was conducted from April to September, which determined severe heat stress (the mean temperature-humidity index above 77). The results revealed that numbers of large follicles and corpora lutea were higher in the melatonin group than in the control group (p < 0.01). Numbers of recovered ova/embryos, fertilized ova, and transferable embryos were higher in the melatonin group (p < 0.01); meanwhile, more degenerated embryos were found in the control group (p < 0.01). Increased activity of the antioxidant enzymes SOD after melatonin administration decreased MDA levels (p < 0.05). In summary, melatonin administration benefited the ovarian response and embryo quality in superstimulated Thai-Holstein crossbreed affected by heat stress.

Ovarian responses and FSH profiles at superovulation with a single epidural administration of gonadotropin in the Thai-Holstein crossbreed.

The conventional method of ovarian superstimulation requires multiple injections of gonadotropins which is time-consuming and may be stressful for the cows. This study was designed to determine whether a single epidural injection of FSH (EI group) would induce the superovulatory response in the Thai-Holstein crossbreed and evaluate FSH plasma hormone concentrations. Eight cows (replication = 3; n=24) were assigned to one of 2 treatments in switch back design. Control group (n=12): cows were received 400 mg FSH twice daily by intramuscularly for 4 days (80, 80, 60, 60, 40, 40, 20 and 20 mg), EI group (n=12): cows were received 400 mg FSH by single epidural injection. Data were collected in term of ovarian follicle responses, superovulatory responses, ova/embryo collection. FSH concentrations were examined using ELISA. The total follicular responses during oestrus were not different between treatments; however, the large follicles were less frequent (P < 0.01) while the medium follicle sizes were higher (P < 0.05) in the EI group. The plasma concentration of FSH in EI was dramatically increased within 2 hours before decreasing sharply thereafter (P < 0.01) and did not remain above baseline after 10 hours of administration. The embryo quality was better in the control than the EI groups (P < 0.05). Interestingly, the number of ovulation cysts in the EI group was 50%. The ovarian responses and embryo quality in the cows with cysts were worse compared with the non-cyst groups (P < 0.05). In conclusion, alternative protocols decreased the superovulatory response and increased poor embryo quality in Thai-Holstein crossbred. Also, the incidence of ovarian follicular cysts is higher in the EI group.

Relationship of the Temperature-Humidity Index (THI) with Ovarian Responses and Embryo Production in Superovulated Thai-Holstein Crossbreds under Tropical Climate Conditions.

Heat stress strongly negatively affects reproductive traits in dairy cattle. The purpose of the present study was to investigate the effect of heat stress in superstimulated Thai-Holstein crossbreds under tropical climate conditions. Data included 75 records from 12 superovulated Thai-Holstein crossbreds between 2018 and 2020. Cows were superstimulated with conventional treatment. The mean temperature-humidity index (THI) was evaluated for three data collection periods: during the 9, 21 and 42-day periods before the insemination day to determine the period in which THI mostly affected superstimulation responses. The THI levels/thresholds were determined and interpreted together with the superovulatory response. A significantly negative correlation was obtained for the THI during the period 9 days before insemination. Negative effects on the number of large follicles and corpora lutea began at a THI of 72 and were apparently severe after a THI of 77, similar to the ovulation rate, fertilized ova and transferable embryos (p < 0.05). Meanwhile, more degenerated embryos were found with THI values (p < 0.05). The superovulatory response in Thai-Holstein crossbreds under tropical climate conditions is highly affected by heat stress starting at a THI of 72 and becomes more severe at a THI higher than 77.

A novel route of follicle-stimulating hormone administration with a split-single ischiorectal fossa in Thai-Holstein crossbred superovulation programs under heat stress conditions.

The objectives of this study were to compare the efficiency of a split single injection of follicle-stimulating hormone (FSH) given by either intramuscular (split-single IM) or ischiorectal fossa (split-single IRF) injection to the traditional treatment and to determine the concentrations of FSH. The temperature and humidity index (THI) values were interpreted together with the ovarian responses and embryo characteristics. The ovarian responses in the split-single IRF group were similar to those of the control group (p > .05) but higher compared with the split-single IM group (p < .05). Higher peak levels of plasma FSH in the split-single IRF group did not differ compared with the control group (p > .05) but were lower in split-single IM administration (p < .05). The results showed a significant decrease in the numbers of large follicles and corpora lutea (CLs) in the moderate THI compared with low and high THI (p < .05). The high THI affected ovulation rate as well as the numbers of transferable embryos and degenerated embryos (p < .05). In conclusion, the split-single IRF administration had a comparable superovulatory response to the traditional twice-daily protocol. Moreover, the ovulation rate, ovarian follicle responses, and embryo quality were affected by heat stress.

Lipid profile of sperm cells in Thai native and commercial roosters and its impact on cryopreserved semen quality.

Plasma membrane lipids are the key factor in the ability of chicken sperm to be frozen. They ensure fluidity and flexibility of the sperm membrane for effective viability and motility during in vitro storage. The objective of this study was to compare the lipid profiles of different native and commercial chicken breeds: native Thai (Pradu Hang Dam) roosters and commercial (Rhode Island Red) roosters, with respect to their frozen sperm quality. In addition, lipid peroxidation and antioxidant enzymes superoxidase dismutase (SOD) and catalase (CAT) were also examined. Semen was collected from 12 roosters of each breed. For fresh semen, parameters assessed include semen volume, pH, sperm concentration, sperm motility, and viability, while for frozen semen, the parameters assessed were sperm motility and viability. Moreover, other parameters assessed included malondialdehyde (MDA) concentration, activities of SOD and CAT, and fatty acid profile. We found that sperm viability and motility of frozen semen were higher in the commercial breed than in the native breed (P < 0.05). The commercial chicken breed had higher MDA concentration than the native breed (P < 0.05), but antioxidant enzymes remained unchanged in both. Levels of arachidonic acid (AA; C20:4n-6) and docosahexaenoic acid (DHA; C22:6n-3) were significantly higher (P < 0.05) in the commercial than in the native breed; however, n-6 to n-3 ratios were not different. In conclusion, our study found that lipid profiles have an influence on frozen sperm viability and motility between the breeds. Polyunsaturated fatty acids, particularly AA and DHA, are beneficial to sperm quality.

Evaluation of cryopreserved boar semen after supplementation sericin form silkworm (Bombyx mori) in semen extender.

Boar cryopreserved semen is scarcely used for artificial insemination due to its quality which is largely reduced by membrane lipid peroxidation. This present study was designed to improve the post-thawed boar semen quality by determining the optimal level of sericin supplementation (antioxidants) in semen extender. Five levels of sericin supplementation between 0% and 1% (w/v) were examined. Semen was frozen by the liquid nitrogen vapor method, thawed slowly at 5°C for 5 min, and used for the evaluation of sperm quality. The results indicated 0.5%-1% sericin supplementation was more effective on maintenance of sperm viability, acrosome integrity, and mitochondrial functions during freezing-thawing. Moreover, 0.75% sericin supplementation was most protective toward total sperm motility and sperm progressive motility. Additionally, 0.25%-0.75% sericin supplementation significantly suppressed increases in the index of lipid peroxidation. In conclusion, 0.75% sericin is recommended as an alternative component of the freezing extender to improve cryopreserved boar semen. However, further research using AI will be necessary to demonstrate that this indication can be applied to the production of offspring in the farms.