Storage Of Semen Research Articles

Полисахариды базидиальных грибов повышают качество репродуктивных клеток быков при замораживании

The global decline in the animal and plant species diversity is among the key environmental problems of the modern world. One possible way to solve this problem is to develop advanced reproductive technologies using gametes to restore genetic diversity. Cryobanks play an important role in this. The traditional method of long-term storage of reproductive cells is their cryopreservation at –196оC using cryoprotective media based on synthesized substances (glycerin, dimethyl sulfoxide, etc.). The reason for searching new effective substitutes is cytotoxicity, impaired fertility and other disadvantages when using traditional cryoprotectors. In addition, it is relevant to expand the temperature range of biomaterial storage, in particular using the conditions of electric freezers. The wide range of biological effects of polysaccharides of basidiomycetes allowed us to make an assumption about the effectiveness of their use in the composition of the medium for cell cryopreservation. This paper presents the results of a comparative study of the effectiveness of polysaccharides of xylotrophic basidiomycetes in the composition of the medium for storage of the Holstein bull semen. The semen was frozen to –5оC and storaged at the specified temperature for 7 days in AndroMed® (Minitübe, Germany) – certified extender for freezing the semen of bulls and other ruminants. Additional introduction into its composition of polysaccharides from the basidiomycetes Hericium erinaceus BP16 / Ganoderma applanatum GA significantly improves the quality of reproductive cells in the used cryopreservation technology.

Efek Penyimpanan Semen Beku Terhadap Kualitas Spermatozoa Sapi Simental Pada Suhu 5°C

This study aimed to determine the effect of frozen semen storage on sperm quality. quality of Simmental cattle spermatozoa at 5° C. This research was conducted laboratory at the UPTD Laboratory of the Livestock Breeding Center of the Jambi Provincial Food Crops Horticulture and Livestock of Jambi Province, from July 10 to August 22, 2023. August 22, 2023. Completely Randomized Design (CRD) was used in the study. with 5 treatments and 5 replications so there were 25 experimental units. Treatment PO (control) = direct thawing, P1 = storage for 15 minutes, P2 = storage for 30 minutes. storage for 30 minutes. P3 = storage for 45 minutes, P4 = storage for 60 minutes. storage for 60 minutes. The variables observed were spermatozoa motility, spermatozoa viability, spermatozoa abnormality. The results of analysis of variance showed that the storage of frozen semen of Simmental cattle at 5 ° C with different times had a significant effect (P <0.05) on the motility of spermatozoa and spermatozoa viability and not significantly different (P>0.05) on spermatozoa abnormality. Based on the research results obtained It is concluded that storage of frozen semen inside at 5 ° C can maintain the quality of frozen semen of Simental cattle with a storage time of 45 minutes.

Quality of Sperm Simmental Bulls and Success of Artificial Insemination with the Addition of Nanocalcium Phosphate in Tris Aminomethane Egg Yolk Diluent Using Semen Storage Ampoules from Nanocalcium Silicophosphate Biomaterials

Quality of Sperm Simmental Bulls and Success of Artificial Insemination with the Addition of Nanocalcium Phosphate in Tris Aminomethane Egg Yolk Diluent Using Semen Storage Ampoules from Nanocalcium Silicophosphate Biomaterials

Effect of cooling rate, extender type, and storage temperature on chilled sperm quality and pregnancy rate in Kail sheep

This study aimed to evaluate how cooling rates, extender type, and storage temperatures affect sperm quality and fertility in Kail sheep. In the first experiment, pooled semen collected from 5 adult Kail rams was extended with a Tris-based extender and divided into three aliquots. The temperature was reduced from 37 °C to 5 °C in 30 min (rapid cooling), 60 min (moderate cooling), and 90 min (slow cooling). In the second experiment, pooled semen was extended with three extenders: Tris, skim milk, and sodium citrate-based, and stored at 5 °C and 12 °C for three days. In the third experiment, ewes (n = 63) were synchronized for estrus using a progestin-based protocol, and fixed-time cervical insemination (FTCI) was performed at 48 h (n = 25) and 60 h (n = 38) after the sponge removal using semen extended in Tris-based extender. Results revealed that slow cooling rates had less (P < 0.05) detrimental impact on sperm motility and kinematics when compared to both moderate and rapid cooling rates. Moreover, semen extended with Tris-based extenders had a higher (P < 0.05) percentage of motile, progressive motile, rapid progressive, and medium progressive motile sperm compared to extenders containing skim milk and sodium citrate. Storage of semen at 5 °C better preserved sperm quality compared to 12 °C. Storage duration exerted a negative influence (P < 0.05) on sperm quality across all three extender types. The overall 55.5% pregnancy rate was achieved. The season of insemination had a significant impact while the timing of insemination did not influence the pregnancy rate. In conclusion, sperm quality is better preserved when semen is stored at 5 °C and subjected to a slow cooling rate using a Tris-based extender, and the success of FTCI is contingent upon the season of insemination in Kail sheep.

Turkey hen sperm storage tubule transcriptome response to artificial insemination and the presence of semen.

Introduction: Sperm storage within the uterovaginal junction (UVJ) of avian species occurs in specialized structures termed sperm storage tubules (SSTs) and allows for prolonged storage of semen, though the molecular mechanisms involved in semen preservation are not well understood. Little work has been done examining how function of the SSTs is impacted by insemination and by semen present in the SSTs. Methods: Transcriptome analysis was performed on isolated SSTs from turkey hens receiving no insemination (control), sham-insemination, or semen-insemination at three timepoints (D1, D30, and D90 post-insemination). Bioinformatic and functional annotation analyses were performed using CLC Genomics Workbench, Database for Annotation, Visualization, and Integrated Discovery (DAVID), and Ingenuity Pathway Analysis (IPA). Pairwise comparisons and k-medoids cluster analysis were utilized to decipher differential expression profiles in the treatment groups. Results: The SST transcriptome of the semen inseminated group exhibited the greatest differences within the group, with differences detectable for up to 90 days post insemination, while control and sham-inseminated groups were more similar. In the semen-inseminated samples, upregulation of pathways relating to classical and non-classical reproductive signaling, cytoskeletal remodeling, physiological parameters of the local UVJ environment, and cellular metabolism was observed. In the sham-inseminated samples, upregulation of immune pathways and non-reproductive endocrine hormones was observed. Discussion: This work provides insights into the molecular level changes of the SST in response to insemination as well as to the presence of semen. Results from this study may have direct implications on fertility rates as well as potential strategies for avian semen cryopreservation protocols.

Short- and long-term effects of essential oils on swine spermatozoa during liquid phase refrigeration.

The application of essential oils as potential alternatives to antibiotics in swine semen storage is promising, due to their antioxidant and antibacterial properties. However, detrimental effects on spermatozoa should be clarified first. The aim of this study was to evaluate 9 essential oils (EOs; Satureja montana, Pelargonium graveolens, Cymbopogon nardus, Melaleuca leucadendron, Eucaliptus globulus, Citrus limon, Lavandula angustifolia, Lavandula hybrida, Mentha piperita) and a blend (GL mix) on key morpho-functional parameters of swine spermatozoa. Test compounds were firstly chemo-characterized and experimental doses were prepared by suspending a fixed number of spermatozoa with 3 different concentrations (0.1, 0.5, 1mg/mL) of EOs. Experimental doses were stored at 16°C and sampled after 3 and 120h for analysis. Overall, S. montana, P. graveolens and L. angustifolia EOs induced the strongest alterations, with C. nardus and E. globulus EOs being the best tolerated. Swine spermatozoa represent a good preliminary testing platform to screen toxicity and its different patterns. The comprehensive overview on the potential mechanisms of action of some of the most common EOs, despite of the direct aim of the study being swine reproduction, may be exploited in other fields of research within both veterinary and human medicine.

Effects of liquid storage of buck semen at refrigeration temperatures on sperm viability and fertility to develop ready to use goat semen diluent

It is generally acknowledged that lower pH throughout liquid storage can decrease sperm motility and energy production, which may be advantageous for maintaining sperm viability during cryopreservation. Hence, the purpose of this study was to see how pH affected sperm viability, motility, membrane integrity, and oxidative changes after 5 days of storage in chemically specified extenders at refrigerator temperatures to formulate a ready-to-use goat semen diluent. Ejaculates were extended with tris-citric acid-fructose diluent with 20% or without egg yolk and stored at 5°C for 5 days. Sperm motility, live count, acrosome integrity, hypo-osmotic swelling positive spermatozoa, pH, and malondialdehyde all substantially differed significantly (P &lt; 0.05) during different days of liquid storage. The pH of diluted semen decreased considerably (P &lt; 0.05) from its original pH of 6.46 on 0 day to pH 6.06 on the 5th day of liquid storage at refrigeration temperature. Diluted semen at refrigeration temperature having 20% egg yolk had significantly higher (P &lt; 0.05) seminal characteristics compared to liquid semen without egg yolk. The liquid storage of semen quality became worse day by day due to a drop in pH and egg yolks acted as a cryoprotectant against the harmful effects of chilling. Hence, 20% egg yolk may be used to enhance the keeping quality of diluted buck semen at refrigeration temperature for up to 5 days. Based on this result, we formulated a ready to use goat semen diluent for liquid semen storage at room temperature and refrigeration temperature.

Androloji laboratuvarlarında semen analizi standardizasyonun Dünya Sağlık Örgütü (DSÖ) kriterlerine göre değerlendirilmesi

OBJECTIVE: Due to its complexity and subjectivity, semen analysis is difficult to standardize across laboratories. Our aim in this study is to collect basic information about semen analysis of andrology laboratories in Türkiye and to determine the level of standardization within and between laboratories for semen analysis. MATERIAL and METHODS: A survey consisting of 41 questions covering all aspects of semen analysis was sent to 145 andrology laboratories that perform semen analysis in Türkiye. Of all, 106 laboratories filled out and returned the survey. The content of the survey was prepared according to the procedures specified in the 6th edition of the 2021 World Health Organization (WHO) laboratory handbook for the examination and processing of human semen. The survey was designed to obtain information about the reported parameters, semen collection and storage, macroscopic-microscopic evaluation, advanced tests and internal-external quality control of the laboratory. RESULTS: In relation to the laboratories, 84.1% of the parameters reported in semen analysis, 75.8% of the semen collection phase, 76.5% of the storage of semen in the laboratory and macroscopic analysis, 78.07% of the microscopic analysis of semen, 49.8% of advanced tests and 73% of the parameters related to quality criteria were compatible with WHO 2021 criteria. CONCLUSION: There appear to be differences between laboratories in Türkiye in the reporting and standardization of semen analysis according to the WHO 2021 criteria. Many different standards and criteria are used for semen analysis in andrology laboratories. This makes it difficult to compare results from one laboratory to another. Therefore, it should be recommended that semen analysis be evaluated according to the criteria set by international standards and guidelines. Keywords: semen analysis, andrology laboratory, microscopic analysis

Current Semen Extenders for Bulls

Artificial insemination is the most widely used biotechnological application for animal breeding in cattle breeding. It is crucial to properly store the sperm obtained from the breeding bulls while maintaining their spermatological characteristic using suitable methods. During both long or short-term storage of spermatozoa, an ideal storage medium must be employed. For this purpose, diluents have been developed to meet the needs of spermatozoa. An ideal semen diluent contains ingredients that spermatozoa need, such as energy substances, protective agents against cold shock, buffering solutions that protect against pH changes, cryoprotectants to reduce damage to spermatozoa during freezing and antibiotics against microbial contamination. Semen dilution also allows for increasing the available semen volume to obtain more straws. Maintaining spermatological parameters at the best possible level during semen storage has important economic implications in this industry. For this reason, scientists continue to develop new diluents to achieve the optimum benefits from semen diluents. This review is aims to provide information about semen diluents used in bulls.

Protective effects of l-cysteine and N-acetyl-l-cysteine on boar sperm quality during hypothermic liquid storage with bovine serum albumin as a protectant

Hypothermic liquid storage at 4–5 °C has emerged as a novel approach for preserving boar semen, offering innovative possibilities for semen preservation. However, this method also presents challenges, including cold shock and excessive reactive oxygen species (ROS) production. Therefore, reducing oxidative damage induced by low temperatures becomes essential while supplementing appropriate protectants. In this study, we investigated the efficacy of Bovine Serum Albumin (BSA) compared to Polyvinylpyrrolidone (PVP) and Skim Milk Powder (SMP) in maintaining boar sperm motility and progressive motility using computer-assisted sperm analysis (CASA). Among the tested concentrations, 4 g/L of BSA exhibited the best protective effect. Subsequently, we supplemented different concentrations of l-cysteine (LC) and N-acetyl-l-cysteine (NAC) as additives in the presence of BSA as a protectant. Our results demonstrated that 1 mmol/L of LC and 0.5 mmol/L of NAC exhibited superior protection of sperm quality compared to other concentrations. Furthermore, the 1 mmol/L LC and 0.5 mmol/L NAC groups showed significantly improved plasma membrane integrity and acrosome integrity compared to the control group. These groups also exhibited enhanced antioxidant capacity, evidenced by increased mitochondrial membrane potential (MMP), ATP production, total superoxide dismutase (T-SOD) activity, total antioxidant capacity (T-AOC), glutathione (GSH), glutathione peroxidase (GSH-PX), and GPX-4 levels. Additionally, they demonstrated decreased reactive oxygen species (ROS) and malondialdehyde (MDA) levels, as well as reduced oxidized glutathione (GSSG) and glutathione reductase (GR) levels. Furthermore, LC and NAC treatment enhanced AMP-activated protein kinase (AMPK) phosphorylation. However, inhibiting AMPK using compound C did not inhibit the protective effects of LC and NAC on low-temperature preserved boar sperm. These findings suggest that 4 g/L BSA can serve as an effective protectant for hypothermic liquid storage of boar semen. Additionally, LC and NAC supplementation reduces oxidative damage by enhancing antioxidant capacity rather than through AMPK-mediated ATP supplementation. These results contribute to advancing the application of LC and NAC in hypothermic liquid storage of boar semen.

Post-Thaw Storage Temperature Influenced Boar Sperm Quality and Lifespan through Apoptosis and Lipid Peroxidation.

Cryopreservation deteriorates boar sperm quality and lifespan, which restricts the use of artificial insemination with frozen-thawed boar semen in field conditions. The objective of this study was to test the effects of post-thaw storage time and temperature on boar sperm survival. Semen ejaculates from five Landrace boars (one ejaculate per boar) were collected and frozen following a 0.5 mL-straw protocol. Straws from the five boars were thawed and diluted 1:1 (v:v) in BTS. The frozen-thawed semen samples were aliquoted into three parts and respectively stored at 5 °C, 17 °C, and 37 °C for up to 6 h. At 0.5, 2, and 6 h of storage, sperm motility, viability, mitochondrial membrane potential, and intracellular reactive oxygen species (ROS) levels and apoptotic changes were measured. Antioxidant and oxidant levels were tested in boar sperm (SPZ) and their surrounding environment (SN) at each timepoint. The results showed significant effects of post-thaw storage time and temperature and an impact on boar sperm quality (total and progressive motility, VCL, viability, acrosome integrity), early and late sperm apoptotic changes, and changes in MDA levels in SPZ and SN. Compared to storage at 5 °C and 37 °C, frozen-thawed semen samples stored at 17 °C displayed better sperm quality, less apoptotic levels, and lower levels of SPZ MDA and SN MDA. Notably, post-thaw storage at 17 °C extended boar sperm lifespan up to 6 h without obvious reduction in sperm quality. In conclusion, storage of frozen-thawed boar semen at 17 °C preserves sperm quality for up to 6 h, which facilitates the use of cryopreserved boar semen for field artificial insemination.

Liquid semen storage-induced alteration in the protein composition of turkey (Meleagris gallopavo) spermatozoa

Liquid storage of turkey semen without the loss of fertilizing ability is of practical interest to the poultry industry. However, fertility rates from liquid-stored turkey semen decline within a few hours. A clear cause of the decline in spermatozoa quality remains unidentified. Therefore, the purpose of the present study was to monitor the dynamics of proteomic changes in spermatozoa during 48 h of liquid storage by 2-dimensional difference in-gel electrophoresis coupled with matrix-assisted laser desorption/ionization mass spectrometry. A total of 57 protein spots were differentially expressed between fresh and stored spermatozoa; 42 spots were more and 15 were less abundant after 48 h of semen storage. Raw proteomic data are available via ProteomeXchange with identifier PXD043050. The selected differentially expressed proteins (DEPs) were validated by western blotting and localized in specific spermatozoa structures by immunofluorescence, such as the head (acrosin and tubulin α), midpiece (acrosin, aconitate hydratase 2, and glycerol-3-phosphate dehydrogenase) and tail (tubulin α). Most of the DEPs that changed in response to liquid storage were related to flagellum-dependent cell motility, energy derivation through oxidation of organic compounds and induction of fertilization, suggesting the complexity of the processes leading to the decrease in stored semen quality. The damaging effect of liquid storage on spermatozoa flagellum manifested as more microtubule proteins, such as tubulins and tektins, most likely formed by posttranslational modifications, tubulin α relocation from the tail to the sperm head, which appeared after 48 h of semen storage, and decreases in fibrous shelf proteins at the same time. Motility could be affected by dysregulation of Ca2+-binding proteins and disturbances in energy metabolism in spermatozoa flagellum. Regarding sperm mitochondria, DEPs involved in energy derivation through the oxidation of organic compounds indicated disturbances in fatty acid beta oxidation and the tricarboxylic acid cycle as possible reasons for energy deficiency during liquid storage. Disturbances in acrosin and 1-phosphatidylinositol 4,5-bisphosphate phosphodiesterase zeta may be involved in rapid declines in the fertility potential of stored turkey spermatozoa. These results showed the complexity of the processes leading to a decrease in stored semen quality and broadened knowledge of the detrimental effects of liquid storage on turkey spermatozoa physiology.

Artificial Insemination: A Review

After the issue of practical Human insemination and its application to humans was resolved, remarks and worries surfaced that didn't seem appropriate given the significance or frequency of the process. Endoscopic artificial insemination is a sophisticated reproductive method that was first used on small ruminants, in which semen is injected into the uterus using a specialized device. By using endoscopic artificial insemination techniques in place of natural mating methods and Trans cervical artificial insemination techniques for locally produced progeny, this research seeks to boost the mass population of Indonesian indigenous. The conditions of artificial insemination have completely changed as a result of the development of semen storage, particularly with regard to application flexibility and safety. Even if fresh semen is still used, it is now evident that semen preservation will play a significant role in artificial insemination going forward. Increasing the gamete density at the fertilization location is there asoning behind artificial insemination. Alot of procedures might cause a miss formed pregnancy, which could damage the physical entity of the pregnancy from the moment it formed—in the womb or in a test tube—until it was delivered, so one of a pregnancy's rights is tube preserved and protected until the baby is born, and that conception can happen even in the absence of natural spouse contact using techniques like artificial insemination which will accelerate the rate of genetic improvement. &amp; to conceive a woman using a method other than sexual contact by Artificial Insemination.

Seasonal stress on semen quality profiles, seminal biochemical and oxidative stress attributes in endangered Teressa goat of Andaman and Nicobar Islands

Objective: To measure seasonal effects on semen quality profiles, seminal biochemical and oxidative stress attributes in fresh and liquid stored semen in monsoon and dry seasons. Methods: A total of 10 Teressa bucks (3-4 years) were selected from breeding farm, ICAR-Central Island Agricultural Research Institute, Port Blair, Andaman and Nicobar Islands, India. Semen samples (n=25 per season) were collected through artificial vagina method and preserved at refrigerated temperature (5°C) for 48 h using Tris citrate glucose based extender. We detected semen quality parameters [volume, mass activity, pH, sperm concentration, total motility, viability, total sperm abnormality, and plasma membrane, acrosomal and nuclear integrities], biochemical profiles [aspartate amino transferase (AST), alanine amino transferase (ALT) and total cholesterol], and oxidative stress markers [total antioxidant capacity (TAC) and malondialdehyde (MDA)] during monsoon and dry seasons. Results: Semen quality parameters significantly differed between seasons (P&lt;0.05) and among storage periods (P&lt;0.05). Volume, pH, mass activity, motility, viability, acrosomal, plasma membrane and nuclear integrities, and TAC were significantly higher (P&lt;0.05). Sperm concentration, sperm abnormalities, MDA, AST, ALT and total cholesterol were significantly lower in fresh semen of monsoon than dry season (P&lt;0.05). Motility, viability, acrosomal, plasma membrane and nuclear integrities, and TAC were significantly decreased (P&lt;0.05) while sperm abnormality, AST, ALT, total cholesterol and MDA were significantly increased as liquid semen storage period advanced (P&lt;0.05). Conclusions: Monsoon season has higher beneficial effects on semen quality profiles and liquid stored semen remained usable for upto 48 h. Good quality ejaculates with higher TAC and lower MDA can be cryopreserved and will be used for artificial insemination.

Storage of boar semen at 17°C without conventional antibiotics in an extender containing an organic bactericidal substance

IntroductionFacing the global threat of antimicrobial resistance, the reduction of antibiotic use in semen extenders is a main goal in artificial insemination (AI) of pigs. The aim of this study was to investigate the potential of a commercial extender containing an organic bactericidal supplement in the absence of conventional antibiotics to control bacterial growth and to maintain the quality of boar spermatozoa during long-term semen storage for up to 144 h at 17°C.MethodsSemen from 233 boars housed at 16 European AI centers was split and diluted in the long-term extender “Androstar Plus without antibiotics + organic bactericidal supplement” (APlus) and in the control extender Beltsville Thawing Solution (BTS) with gentamicin, which is routinely used in many AI centers. Sperm motility was assessed with computer-assisted semen analysis (CASA) and membrane integrity was evaluated with flow cytometry. The number of bacteria was determined by counting colonies on agar plates.ResultsAt the end of storage, bacterial counts were ≥ 106 CFU/mL in 10.7% of the APlus and in 0.4% of the BTS samples. At the same time, bacterial counts were only weakly correlated with sperm motility (r = −0.23, p &amp;lt; 0.05), and there was no correlation with sperm membrane integrity (p &amp;gt; 0.05). Among the 12 identified bacterial species in APlus samples, loss of sperm quality was exclusively observed in the presence of &amp;gt;106 CFU/mL Serratia marcescens and Klebsiella oxytoca. Both these bacterial species, despite their known multi-drug resistance and the continuous use of gentamicin in Europe, proved sensitive to this antibiotic, thus indicating an efficient quality assurance program and responsible antibiotic use.ConclusionLong-term storage of boar semen at 17°C without conventional antibiotics in an extender containing an organic bactericidal supplement is an option if semen samples are regularly tested for the presence of S. marcescens and K. oxytoca, and the source of contamination is eliminated.

Spermatozoa quality of Landrace Boar in Beltsville thawing solution diluent with various levels of moringa leaf extract

This study aims to find the best level of sun-dried Moringa leaf extract (SDMLE) in Beltsville Thawing Solution (BTS) diluent to preserve the spermatozoa of Landrace boars. Semen from a two-year-old Landrace boar was collected using the massage method. Semen with spermatozoa motility ≥70% and spermatozoa abnormalities &lt;20% was diluted with BTS (P0, control), BTS + 3% SDMLE (P1), BTS + 6% SDMLE (P2), BTS + 9% SDMLE (P3), and BTS + 12% SDMLE (P4). Storage of liquid semen using a cool box with a temperature of 15-20oC, while evaluating the quality of semen every 8 hours for spermatozoa motility, viability, abnormality, and longevity. The results showed that spermatozoa stored for 40 hours in BTS + 3% SDMLE (P1) diluent had higher quality (P&lt;0.05) than the other four diluents, namely motility (54.00%), viability (61.16%), abnormality (3.62%), and longevity (45.77%). The results of this study concluded that 3% SDMLE added to BTS diluent was the best level for the preservation of spermatozoa in landrace boars.

Effect of semen extenders, dilution rates and storage periods on spermatozoa quality of Horasi chicken ecotype

Numerous semen extenders have been recommended for storing poultry semen. This study was conducted to assess a suitable extender, storage period and dilution rate for semen collected from the Horasi chicken ecotype. To accomplish the objectives, a 4 x 4 x 4 factorial experiment in a completely randomized design employing four extenders (BPSE, LAKE, CARI and garlic extract mixture), four dilution rates (1:2, 1:3, 1:4 and 1:5) and four storage periods (0, 12, 24 and 48 hours) was used. Semen samples were collected from ten Horasi cockerels using the abdominal massage technique, kept at 4°C and subsequently examined for semen motility, viability and normalcy after extension with specific diluent, each dilution rate and storage period. Sperm motility and viability were significantly (p&lt;0.05) reduced across all extenders and dilution rates examined, while defective spermatozoa increased with storage period. The Beltsville Poultry Semen Extender (BPSE) maintained high spermatozoa motility, viability, and normalcy at all dilution rates and storage periods tested, followed by the CARI and LAKE extenders, whereas the Garlic extender performed worse at all dilution rates and storage periods than the other extenders, especially at a 1:5 dilution rate and 48 hours of storage. Based on our findings, a high percentage of sperm motility, viability, and normalcy for Horasi chicken ecotype semen were observed while using BPSE at a dilution rate of 1:2, and the quality was maintained throughout the storage period tested. Thus, it is concluded that BPSE extender at a 1:2 dilution rate is a suitable extender for Horasi chicken ecotype semen.

Effect of Ferulic Acid on Semen Quality of Goat Bucks during Liquid Storage at 17 °C.

This study investigated the effect of different concentrations of ferulic acid (FA) on the quality of goat semen preserved at 17 °C. First, semen was collected from three black-headed goat bucks using an artificial vagina. Then, the mixed semen was diluted with basal dilutions containing different concentrations of FA (0, 25, 50, 100, and 200 μmol/L) and stored at 17 °C. Sperm total motility, plasma membrane integrity, acrosome integrity, reactive oxygen species (ROS) levels, malondialdehyde (MDA) content, and total antioxidant capacity (T-AOC) were measured during semen storage. The results showed that sperm total motility, plasma membrane integrity, and acrosome integrity were significantly improved in the 50 μmol/L FA group compared with the control group (0 μmol/L) on days 1-5, and the level of T-AOC significantly increased, while the contents of ROS and MDA significantly reduced. Meanwhile, the goats' conception rate showed that supplementing semen with 50 μmol/L FA preserved at 17 °C for 3 days had no significant effect on fertility. Taken together, our findings suggest that adding 50 μmol/L FA in dilution at 17 °C can improve goat bucks' semen quality.

Analysis of changes in the morphological structures of sperm during preservation of liquid boar semen in two different seasons of the year

Animal fertility is the result of a combination of genetic, physiological and environmental factors. Assessment of semen quality plays a key role in determining the reproductive performance of boars. The aim of the study was to determine the effect of two seasons of the year on the morphology, morphometry, cell membrane integrity and mitochondrial activity of sperm during storage of liquid boar semen at 17 °C. The study was carried out using 20 boars. Four ejaculates were collected from each boar in each of two seasons of the year. All tests were carried out five times: at 1, 24, 48, 96 and 168 h during storage. The results showed that sperm in ejaculates collected in summer are more sensitive to storage conditions than those from ejaculates obtained in winter. The percentage of sperm with morphological defects was shown to increase with the storage time of the diluted ejaculates, particularly between 96 and 168 h of preservation. In summer, the percentage of sperm with an intact cell membrane and the percentage with high mitochondrial membrane potential are much lower than in winter, at every hour of semen preservation. In the case of boars used for artificial insemination, it is worth taking into account the season when the semen is collected, especially if it is to be stored in liquid form. Assessment of sperm cell structures during storage of liquid semen should be implemented at insemination stations and should be carried out more often in summer.

Trehalose as a Stabilizer of the Lipid Composition of Membranes and the Composition of the Cytosol of Frozen/Thawed Rooster Spermatozoa

Low-temperature semen storage technologies are already being used in poultry conservation programs, but the quality of reproductive material stored in cryobanks varies greatly and cannot always be successfully used for practical purposes. Therefore, it is necessary to improve the compositions of cryoprotective media to improve their quality. This study aimed to investigate the composition of membrane lipids and carbohydrates in the cytosol of rooster spermatozoa, to explain the dose-dependent effect of a combination of trehalose and fructose in cryoprotective media on the preservation of their morphological and kinetic parameters during freezing/thawing, and to determine the most effective diluent composition. Ejaculates were collected from Rhode Island Red roosters (n = 10). The effectiveness of three diluents containing trehalose was evaluated: LCM-control (0 mM), Treh20 (9.5 mM), and Treh30 (13.4 mM). Chromatographic analysis of membrane lipids, carbohydrates, and polyols of the spermatozoa cytosol was performed. A decrease in the content of glycolipids in the plasma membranes of spermatozoa from 2.0% in native spermatozoa to 1.1–1.4% (frozen/thawed) and phospholipids from 71.2% (native) to 70.5% (frozen/thawed) reduced the progressive sperm motility from 65.7% in native spermatozoa to 12.6–27.6% (frozen/thawed). The same dynamics were observed for the viability parameter of 90.4% (native) and 27.0–41.2% (frozen/thawed). The Treh20 diluent, using a combination of fructose (36 mM) and trehalose (9.5 mM) saccharides, maximally preserved the lipid profile of plasma membranes and the composition of the cytosol of frozen/thawed rooster spermatozoa, which positively affected the indicators of general and progressive mobility and viability.