Fertility and Insemination Characteristics of Sperm Storage Tubules in Old Thai-Native Hens.

We aimed to evaluate the effects of sperm concentration (150-250 × 106 spz/dose) and insemination frequency (once, twice, and thrice weekly) on fertility and sperm storage tubule (SST) characteristics. The SSTs were classified into five categories: namely, SSTs having an unscorable (SST1), empty (SST2), low (SST3), medium (SST4), and high (SST5) sperm count after insemination. The results showed that only insemination frequency affected the fertility rate (p < 0.05). The highest fertility was found in the thrice-weekly insemination group; however, this rate was not significantly different from that for the twice-weekly insemination group, except on day 7, while the once-weekly insemination group showed the lowest fertility rate (p < 0.05) from day four onward. On day 1, the SST characteristics showed no differences among the various insemination frequencies. On day 4, the SST2 and SST3 categories increased in the once-weekly insemination group (p < 0.05), while the SST4 and SST5 categories decreased compared to the twice- and thrice-weekly insemination groups (p < 0.05). On day 7, only the thrice-weekly insemination group maintained a level of SST5 category tubules like that measured on day 1 (p > 0.05). In summary, the insemination dose of 150 × 106 sperm was enough for fertilization, and thrice-weekly insemination was the appropriate frequency in old Thai native hens for maintaining a high sperm density in the SSTs throughout the week.

Solid storage supplemented with serine of rooster semen enhances higher sperm quality and fertility potential during storage at 5°C for up to 120 h.

Information on prolonging the storage duration of cold semen with acceptable fertility in roosters is limited. This study aimed to determine the efficiency of solid storage with the addition of various concentrations of serine to the Thai native rooster (Pradu Hang Dum) semen extender on semen quality and fertility potential during storage at 5°C for up to 120 h. Pooled semen was diluted with a base extender and a gelatin extender containing 0, 2, 4, and 6 mM serine, then stored at 5°C for 120 h. In Experiment 1, the semen quality and malondialdehyde (MDA) concentrations were assessed at 0, 24, 72, and 120 h after storage. In Experiment 2, fertility potential in terms of fertility and hatchability rates was determined using the most effective solid-storage semen from Experiment 1. Sperm quality decreased with increasing storage time (P < 0.05). The lowest semen quality was observed in the control group since T24 of storage compared with the other groups (P < 0.05). Progressive motility, viability, and mitochondrial function were higher (P < 0.05) in the extender supplemented with gelatin and serine groups than those in the gelatin alone group at T72 and T120. In the extender supplemented with gelatin and serine groups, the highest semen quality was observed in the gelatin with 4 mM serine groups. The differences among extenders supplemented with serine were insignificant (P > 0.05), and the lowest MDA was observed in the gelatin with 4 mM serine groups. The fertility and hatchability rates in gelatin with 4 mM serine at T24 were comparable to those in fresh semen (83.87 and 86.12% vs. 86.66 and 88.3%; P > 0.05). Those of T72 were significantly better than those of the control at the same hour of storage (64.08 and 71.61% vs. 52.38 and 64.48%), while those of T120 were not different among groups. In summary, a semen extender as a solid medium supplemented with 4 mM serine successfully preserved the rooster semen for a long duration up to 72 h of storage time.

Antifreeze protein type III addition to freezing extender comprehensively improves post-thaw sperm properties in Okinawan native Agu pig.

Sperm cryopreservation often leads to physical cell damage through ice crystal formation. This study evaluates the improvements to freezing extender cryoprotective activity due to antifreeze protein (AFP) addition, which primarily acts on ice crystal formation, through investigating the post-thaw sperm properties of Okinawan native Agu pig. Six individual boar sperm samples were diluted with the freezing extender supplemented with 1 µg/mL of AFP I or AFP III and then subjected to cryopreservation. Treatment with AFP I during the freezing procedure had no improvement for any characteristics after thawing compared to untreated sperm. In contrast, the addition of AFP III to the freezing extender strongly increased sperm motility, mitochondria and cell membrane integrity, and the acrosomal proteolytic activity of frozen-thawed sperm in 5 of 6 individuals (P < 0.05). Furthermore, cryoinjury prevention by AFP III significantly enhanced sperm viability (by ATP content), and maintained DNA quality and in vitro sperm penetrability compared with AFP I treatment (P < 0.05). These findings demonstrate that AFP III addition to the freezing extender of boar sperm is more effective in maintaining sperm characteristics than the extender without AFP III or AFP I, despite individual differences in response.

Influence of Semen Collection Frequency and Seasonal Variations on Fresh and Frozen Semen Quality in Thai Native Roosters.

This study aimed to determine the effects of the frequency of semen collection (once, twice, and thrice weekly) and seasonal variations on the fresh and frozen semen quality of Thai native roosters throughout the year. Data on temperature and humidity were collected and used to calculate the temperature-humidity index (THI). The average temperature and THI were lower in the winter than in the rainy season and the summer (p < 0.05). In contrast, the average relative humidity was not different among the seasons but was higher in the rainy season (p > 0.05). None of the fresh or frozen semen quality parameters were influenced by the frequency of semen collection, but the season did have an effect. The highest sperm concentration was obtained in the winter (p < 0.05). In contrast, the lowest sperm concentration was found during the rainy season, which presented the highest humidity. Regarding the frozen semen quality, the highest malondialdehyde concentration and the lowest motility were found in the summer (p < 0.05). In conclusion, semen collection can be conducted thrice per week for a consecutive year without affecting semen quality while maximizing sperm production. However, the highest sperm production was obtained in the winter, which is also a suitable season for producing semen for cryopreservation.

Characterization of Oviduct Lining, with Emphasis on the Sperm Storage Tubule Region (Uterovaginal Junction), Correlated with Fertility in Mature and Old Thai Native Hens.

The effect of age on fertility was investigated in Thai native chickens. The objective of this study was to determine the effects of age (mature and old) on the morphological characteristics of the reproductive organs and the histological characteristics of the uterovaginal junction (UVJ) tissues, resident sperm in the UVJ, and fertility duration in Thai native hens. We found no differences in the morphological characteristics of the reproductive organs, except for the number of follicles and the sizes of the fifth large yellow follicle in mature hens, which were greater than those in old hens (p < 0.05). The diameter of the sperm storage tubules (SSTs) epithelium was larger in old hens than in mature hens (p < 0.05), whereas the epithelium height was lower in old hens (p < 0.05). The number of sperm in the SSTs was greater in mature hens compared with old hens (p < 0.05). Mature hens showed a higher fertility rate than old hens. Our results suggest that, in old hens, the function of the SSTs is impaired, and sperm cannot be retained. Such a deterioration of the SSTs may be one of the factors involved in the decline in fertility.

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.