Storage Of Semen Research Articles

Semen cryopreservation and storage in liquid nitrogen: Impact on chromatin compaction.

Sperm cryopreservation is a consolidate option for long-term male fertility preservation. The freezing/thawing procedure causes detrimental effects to spermatozoa, including damage to viability, motility, membrane composition, and DNA, whereas the effect on sperm chromatin compaction is less studied. The primary aim of this study was to investigate the impact of cryopreservation on sperm chromatin compaction. Furthermore, the effect of cryopreservation on sperm parameters (motility, viability, chromatin compaction, and DNA fragmentation) was also assessed in relation to the storage time in liquid nitrogen. Semen samples, collected from 126 (92 normozoospermic and 34 oligozoospermic) patients undergoing routine semen analysis in the Andrology Laboratory of Careggi University Hospital of Florence, were frozen by conventional fast vapor freezing method. Sperm motility, viability, kinematic parameters (by computer-aided sperm analysis [CASA]), chromatin compaction (by staining with both aniline blue [AB] and Chromomycin A3 [CMA3]), and sperm DNA fragmentation (sDF, by TUNEL/Propidium Iodide [PI]) were evaluated before freezing and after thawing at different timepoints. After 7 days of storage, a significant decline in sperm motility, viability, and kinematics parameters, as well as a significant increase in the percentage of sperm positivity to CMA3, AB, and sDF, were observed. It is noteworthy that while motility and viability decreased in almost all subjects, the increase in CMA3 and AB positivity was observed in 68.0% and 79.2% of samples, respectively. A progressive deterioration of sperm motility and viability, less evident for chromatin structure, was observed at longer times of storage (28 and 180 days). Our results indicate that freezing/thawing procedures can alter chromatin structure. A reduction in protamine content and/or a modification in chromatin assembly can be hypothesized. Furthermore, the length of storage in liquid nitrogen appears to progressively affect sperm parameters, although it should be confirmed in larger cohort of subjects. Current sperm cryopreservation protocols need to be improved with new strategies and personalized procedures aimed to minimize the damage.

Evaluating the effect of semen storage and dilution rate on boar sperm quality

ABSTRACT This study analyzed the effect of boar semen storage and dilution rate on sperm motility, swimming patterns and kinematics. It used 35 sexually mature, healthy boars, with an average age of 19.0 ± 10.34 months and known fertility. Samples were separated into four treatments according to dilution ratio (1:1, 1:4, 1:8 and 1:16; v:v) with Androstar Plus® and refrigerated at 17°C in the dark for 96 h. Subsamples were analyzed at 12, 24, 28, 72 and 96 h using the ISAS®v1 Computer-Assisted Semen Analysis System (CASA). Total and progressive motility decreased (P < 0.05) as semen storage time increased. The percentage of morphological abnormalities increased after 48 h of semen storage. Samples diluted 1:1 (v:v) had higher values of total motility and faster spermatozoa compared to other dilution rates. It was concluded that sperm motility, swimming patterns and plasma membrane integrity showed a significant downward pattern as semen storage time and dilution rate increased.

Supplementation of Epimedium polysaccharide (EPS) improves goat semen characteristics following cryopreservation

Cryopreservation facilitates long-term semen storage and enables the exchange of genetic material among elite livestock over extensive distances. A decrease in sperm quality is an unavoidable outcome of the cryopreservation process. Prior research has established that incorporating cryoprotectants into the diluent can mitigate freeze-induced damage and enhance sperm quality. This study aims to assess the impact of Epimedium polysaccharide (EPS) on the cryopreservation of goat semen. Samples were obtained from six healthy goats following an initial examination. One portion of the semen was diluted with a base solution containing EPS for treatment purposes, whereas another was diluted without EPS, serving as the control. Results indicated that varying concentrations (1, 2, 3, 4, 5mg/mL) of EPS in the diluent enhanced both physiological characteristics and antioxidant enzyme activities in cryopreserved goat sperm. Further analysis showed that the 3mg/mL EPS concentration significantly improved sperm motility (52.10%), plasma membrane integrity (57.01%), tail plasma membrane integrity (52.37%), acrosome integrity rate (52.45%), and antioxidant enzyme activities relative to other groups (P<0.05). Additionally, the inclusion of 3mg/mL EPS substantially improved the sperm's fertilization capability. In conclusion, our experiments confirm that EPS supplementation significantly enhances sperm quality post-freezing, with 3mg/mL identified as the optimal concentration.

Kombinasi Sari Buah Semangka dalam Pengencer Kuning Telur dan Larutan Tris Terhadap Kualitas Spermatozoa Babi Landrace

in egg yolk diluent (EY) on the quality of spermatozoa in landrace pigs. In this study, the semen source was obtained from 1 male landrace pig aged 2 years in healthy condition, and had been trained in semen storage which was then diluted with treatment diluent, namely: T0: WMJ 80%+EY 20%, T1: WMJ 60% +EY 20%+TS 20%, T2: WMJ 40%+EY 20%+TS 40%, T3: WMJ 20%+EY 20%+TS 60%, T4: EY 20%+TS 80%. Diluted semen is stored at 15-20 ̊C and evaluated every 8 hours for motility, viability, abnormalities and survival. The results showed that the diluent WMJ 20%+EY 20%+TS 60% had higher quality (P&lt;0.05) compared to the other four treatments, namely with motility (62.00±2.74%), viability (70.20±2.77%), abnormality (3.30±0.27%) and survival (32.00±0.00) hours. It was concluded that the diluent WMJ 20%+EY 20%+TS 60% could have a good influence in maintaining the quality of spermatozoa in landrace pigs.

Evaluation of the use of L-carnosine in short-term storage of ram semen with spermatological and flow-cytometric analyzes

This study was conducted to investigate the effect of adding different doses of L-carnosine to semen in vitro on the short-term storage of semen. Semen taken from 6 Akkaraman rams was used in the research. Semen taken from each animal was pooled. It was diluted with tris+egg yolk extender containing L-carnosine at different doses (1, 5, 10, 20, 50, 100 and 200 mM) and stored at +4°C for 96 hours. It was determined that 1, 5, 10, 20 mM L-carnosine doses increased total, progressive and rapid motility, kinematic parameters VCL, VSL, VAP values and high mitochondrial membrane potential level during short-term storage. It was also determined that these doses protected membrane integrity, reduced the rate of abnormal spermatozoon, acrosomal damage and low mitochondrial membrane potential level. It was determined that a dose of 200 mM L-carnosine had a toxic effect and negatively affected the parameters. As a result, when ram semen is desired to be stored for a short time (for 96 hours), it may be recommended to add 1, 5, 10 and 20 mM L-carnosine amino acid to the semen extenders in order to preserve spermatological parameters.

Morphological characterization of spermatozoa in Apis mellifera and the effect of processing and semen storage

The aim of this study was to establish a standardized approach for evaluating sperm morphology in the honey bee and to investigate the impact of processing techniques and semen storage on the incidence of sperm morphoanomalies. The first experiment was designed to characterize the sperm morphoanomalies in unaltered honey bee semen, examining if they are compatible with sperm motility and viability and determining their occurrence in the ejaculates of mature and healthy honey bee drones. The different forms of sperm morphoanomalies were described in detail. In the morphological analysis of fresh drone ejaculates, 7.77% of spermatozoa showed abnormal morphology on average, with 2.20%, 4.75% and 0.81% of head, tail and multiple defects, respectively. The method also allowed us to determine the status of acrosome integrity. The second experiment was designed to assess the impact of smearing and air-drying on the occurrence of sperm morphoanomalies, showing a significant increase in the incidence of both head and tail sperm defects when comparing to wet samples. In the third and fourth experiments, the effect of semen storage at room temperature and semen cryopreservation, respectively, on the occurrence of sperm morphoanomalies were investigated. The preservation of semen at 22 °C led to a significant increase in spermatozoa with coiled tails after day 1, whereas the remaining anomalies did not exhibit significant variations over time. The freezing-thawing process showed a more pronounced effect on the incidence of morphological defects, with an increase in the percentage of spermatozoa with deformed heads and coiled tails.

Effect of Three Commercially Available Extenders Containing Phospholipids of Different Sources on Skopelos Buck Liquid-Stored Sperm Quality.

The effect of four extenders on buck semen quality parameters was examined during a 48 h liquid storage. Semen was collected from six Skopelos bucks and diluted in the following extenders, containing: soy lecithin (SL, OviXcell®), plant phospholipids (PP, AndroMed®), egg yolk lecithin (EY, Steridyl®), or no phospholipids (basic extender). Samples were stored at 5 °C for 48 h and assessed at 0, 24 and 48 h for viability (eosin-nigrosin), acrosome integrity (SpermBlue®), membrane functional integrity (HOST), mitochondrial function (Rhodamine 123/SYBR-14/PI) and motility parameters (CASA). No significant reduction in total or progressive spermatozoa motility and mitochondrial function was observed at 24 h, whereas they all dropped significantly at 48 h, in all extenders. Spermatozoa viability, cell membrane functionality and acrosome integrity dropped progressively (0 h > 24 h > 48 h) in all groups. No significant difference among extenders was observed concerning spermatozoa mitochondrial function. Overall, spermatozoa viability, cell membrane functionality and acrosome integrity were higher in the three commercial extenders, compared to the basic extender. SL and EY extenders (OviXcell® and Steridyl®, respectively) preserved viability more effectively than the PP extender (AndroMed®). Total motility was higher in the PP extender, compared to the SL extender. Spermatozoa acrosome integrity tended to be higher in the EY extender compared to all the other extenders. Further investigation of the protective potential of different types of cryoprotectants on liquid buck semen storage is important.

Investigation of optimal sperm storage conditions for short-term storage

Objective The quality of sperm cells is important role in the success rates of assisted reproduction technology (ART) treatments. The quality of the sperm cells shows variations depending on the temperature of short-term semen storage as well as the methods of semen preparation. Thus, this study aimed to investigate the sperm viability, motility and DNA fragmentation following different sperm preparation methods and short-term storage conditions, respectively. Materials and Methods A total of 25 semen samples were evaluated. In the first part of this study, different incubation temperatures were investigated in two groups, in such the first group involved the semen samples and the second group involved the sperm cells separated by density gradient centrifugation method, respectively. The samples in each group were incubated at 4°C, room temperature (21°C) and 37°C for 24 hours, respectively. The sperm cell qualities were evaluated by mobility analysis, DNA fragmentation by acridine orange staining and sperm cell viability by propidium iodide staining. Results and Discussion The analysis outcome demonstrated that the mobility, DNA fragmentation and viability of the sperm cells were statistically different when incubated at RT (21°C) in both groups. Furthermore, samples prepared by the density gradient centrifugation method were shown to have better quality. The optimum short-term storage temperature was detected to be the room temperature. Conclusion The conclusion of this investigation is crucial to assess storage conditions in ART clinics. This study provides essential data for short-term sperm storage and preparation methods to improve the success rates of ART clinics. Bangladesh Journal of Medical Science Vol. 23 No. 04 October’24 Page : 1137-1141

Exploratory Metabolomics and Lipidomics Profiling Contributes to Understanding How Curcumin Improves Quality of Goat Semen Stored at 16 °C in Tropical Areas.

Reactive oxygen species (ROS) exert a vital role in sperm quality during semen preservation, where excessive ROS leads to oxidative damage and undermines sperm integrity. Curcumin, a botanical extract, is capable of neutralizing ROS and enhancing the activity of antioxidant enzymes. This study was aimed at evaluating the effects of curcumin on sperm viability, acrosome integrity, and antioxidant levels, as well as metabolomic and lipidomic profiles. The results demonstrated that curcumin at 25 µmol/L significantly enhanced sperm motility, plasma membrane, and acrosome integrity, elevated the levels of antioxidant enzymes (T-AOC, CAT, SOD), and decreased ROS production (p < 0.05). Metabolomic analysis identified 93 distinct metabolites that showed significant differences between the control and curcumin-treated groups. KEGG pathways emphasized the participation of these metabolites in key metabolic processes such as the citric acid cycle, cholesterol metabolism, and fatty acid metabolism. Curcumin treatment brought about notable variations in lipid profiles, including increased levels of phosphatidylcholine, acylcarnitine, and triglyceride over the storage time, suggesting enhanced lipid anabolic activity. Overall, the supplementation of curcumin at 25 µmol/L effectively mitigates oxidative stress and prolongs the viability of semen storage at 16 °C by modulating specific metabolic and lipid profiles.

452 Data-driven strategic sustainability initiatives of beef and dairy genetics consortiums: A comprehensive landscape analysis for USA, Brazil, and Europe

Abstract The past two decades have witnessed beef and dairy genetic consortiums implementing data-driven germplasm propagation programs in global cattle systems. These programs optimize for multi-dimensional economic, environmental, and animal welfare related sustainability goals and are driven by consumer demands and shifts in supply chains. To summarize these programs and gain insights into the sustainability journey of industry, we did a comprehensive landscape analysis of the strategic initiatives of key global genetics companies, their germplasm offerings, and data platforms offered in three major cattle producing geographies, namely the USA, Brazil, and Europe. Within these locales, they followed selection index-based genomic and reproductive management strategies for both beef, dairy, and beef-on-dairy production, tailored to relevant market and climatic conditions (Figure 1). While individual traits in selection indices ranged between 40 to 90 traits, all traits could be broadly grouped into three: namely production, longevity, health and reproduction. In the USA, companies like ABS Global, Select Sires, Genex Cooperative, STGenetics, Zoetis, Vytelle offer genomic selection programs encompassing germplasm products, and genetic and production data tracking platforms across the beef, dairy, and beef-on-dairy value chains. These companies have implemented reproductive management programs such as in-vitro fertilized embryo transfer, sexed semen, along with genomic selection with a focus on future replacements based on multi-trait sustainability indices. These initiatives are customized for each of the cow-calf, backgrounding, and feedlot sector of beef production. In dairy, these companies have diversified data-driven production programs customizable to conventional and organic production systems. In Europe, companies like CRV, Viking Genetics, and Alta Genetics prioritize traits related to resource efficiency, animal welfare and environmental impact. Semen companies also offer data management tools tailored to European market requirements, alongside services for on-farm semen production and storage to reduce transportation-related emissions. In Brazil, companies such as ABS Pecplan, CRV Brazil, and Semex Brazil, developed herd management solutions focusing on traits like heat tolerance, tropical environmental adaptability, and disease resistance. The landscape analysis revealed a dynamic industry, evolving to meet sustainability goals and market demands. These enterprises collaborate with stakeholders across the value chain, including producers, policymakers, and industry associations, to scale data-driven production systems that balance profitability and environmental footprint of cattle production. As such, they have positioned themselves to capitalize on the growing interest in sustainable livestock production practices worldwide by aligning scientific and technical strategies with evolving regulatory requirements and consumer demands. The global cattle industry is positioned to venture forward by leveraging niche carbon markets and new marketing channels propelled by emerging principles of environmental, social, and corporate governance. Strategic germplasm and data-tracking platform initiatives by these groups thus become an integral part of the livestock sustainability solution palette and will contribute its share to a climate-neutral Earth by 2050.

How do taurine and ergothioneine additives improve the parameters of high- and low-quality turkey semen during liquid preservation?

Abstract The study aimed to examine the parameters of turkey ejaculates (n=40) of high and low quality (HQ and LQ, respectively), preserved with the addition of taurine (TAU; 5 mM, 10 mM) and ergothioneine (EGT; 5 mM, 10 mM) for 48 h at 5°C in a liquid state. The motility, plasma membrane integrity (PMI), mitochondrial function, apoptotic and necrotic spermatozoa percentage, and sperm cells that generated NO were determined after 2, 24, and 48 h of storage. The preserved semen was also analysed for the activity of superoxide dismutase (SOD), glutathione peroxidase (GPx), catalase (CAT), glutathione (GSH) content, and malondialdehyde (MDA) production. Taurine, in both doses, may improve the antioxidant status of stored turkey semen as well as sperm motility, viability and functionality regardless of ejaculate quality, as manifested by increased SOD and CAT activities, reduced MDA levels, and enhanced sperm characteristics i.e. plasma membrane integrity (PMI), mitochondrial membrane potential (MMP), total motility (TMOT), and progressive motility (PMOT). In turn, the addition of EGT increased GSH content in the external environment and suppressed lipid peroxidation in turkey spermatozoa, in particular those from low-quality (LQ) ejaculates. However, it appears that EGT could deliver more satisfactory results when added at lower concentrations.

Supplementation of sperm cooling medium with Eurycoma longifolia extract enhances native Thai chicken sperm quality and fertility potential.

Cooled semen storage methods result in oxidative stress generated by an imbalance between oxidation rates, specifically reactive oxygen species production, and sperm cell antioxidants, leading to degradation of semen quality. We aimed to investigate the impact of adding Eurycoma longifolia (EL) extract as an antioxidant supplement in semen storage medium (IGGKPh semen extender) on semen quality and fertility potential. EL extract at concentrations of 5, 10, 15, and 20 mg/mL was assessed for its antioxidant capacity in IGGKPh semen extender. Our findings revealed that the total phenolic content in the EL extract did not vary significantly across the various concentrations and temperatures tested. However, incubation at 5°C was found to be the most effective temperature for increasing the EL extract antioxidant capacity as assessed via the 2,2-diphenyl-1-picrylhydrazyl inhibition assay in a dose-dependent manner. Supplementation of the IGGKPh semen extender with 15 mg/mL EL extract was found to enhance semen quality during cold storage for up to 48 h (p < 0.05), as evidenced by decreased malondialdehyde levels in cooled semen (p < 0.05). However, antioxidant enzyme activities showed no significant differences among the various experimental groups (p > 0.05). The fertility test showed that the 15 mg/mL EL extract group stored for 24 h had a higher percentage than the control group (p < 0.05). However, there was no significant difference in percentage between the two groups at 48 h of storage (p > 0.05). The hatchability showed no significant difference in both 24 and 48-h storage periods (p > 0.05). Our results indicated that supplementing the IGGKPh semen extender with 15 mg/mL EL extract may positively influence semen quality during storage, suggesting potential applications for enhancing semen quality.

Carvacrol and Thymol Enhance the Quality of Beni Arouss Buck Semen Stored at 4 °C Thanks to Their Antimicrobial Properties.

This study aims to investigate the impact of carvacrol and thymol on the quality of Beni Arouss buck semen stored in skim milk at 4 °C. Ejaculates were collected from eight Beni Arouss bucks weekly for 11 weeks, pooled, and then divided into three equal parts. Samples were diluted to 400 × 106 sperm/mL in skim milk (control) and skim milk supplemented with a single dose of 200 µM carvacrol and thymol each. Evaluations of sperm motility, viability, abnormalities, membrane integrity, lipid peroxidation, and bacterial growth were conducted at 0, 6, 24, and 48 h of liquid storage at 4 °C. After 48 h of storage, the results indicate that the addition of carvacrol positively influences total and progressive motility and viability. However, it also leads to a decrease in lipid peroxidation and bacterial growth compared to the control group (p < 0.05). Thymol showed similar results to carvacrol, except for progressive motility (p > 0.05). Bacterial growth was negatively correlated with total and progressive motility and viability (p < 0.05), while no correlation between lipid peroxidation and these parameters was observed (p > 0.05). In conclusion, the addition of carvacrol and thymol to skim milk extender moderately improves the quality of Beni Arouss buck semen after 48 h storage at 4 °C due to its antimicrobial activity.

Long-term cryopreservation of bull semen as a main strategy for the ex situ conservation of endangered Polish Red cattle

Abstract Regular verification of the quality of cryopreserved semen derived from native cattle is one of the tasks performed at the bank as recommended by the Food and Agriculture Organization. The purpose of the present study was to evaluate the quality of semen from PR bulls stored for 40, 50 and 60 years in the BMB using standard evaluation parameters such as sperm motility as well as structural-functional parameters such as plasma membrane integrity, transmembrane mitochondrial potential and sperm chromatin damage. Semen pellets from 27 PR bulls (3 ejaculates/bull) were tested. The data were analysed by one-way and two-way ANOVA, and the significance of the difference (P≤ 0.01) between the means was determined using Duncan’s test. Our study results revealed that the long-term storage of semen had no effect on sperm characteristics after thawing. However, statistically significant differences (P≤0.01) in sperm plasma membrane integrity and transmembrane mitochondrial potential, following storage in liquid nitrogen were noted between bulls at all time points. However, there were no significant differences (P&gt;0.01) in sperm chromatin damage between breeds or between different storage times, and the degree of DNA fragmentation ranged from 0.4 to 0.8%.

Fertility after photodynamic inactivation of bacteria in extended boar semen.

Antimicrobial resistance is an increasing challenge in semen preservation of breeding animals, especially in the porcine species. Bacteria are a natural component of semen, and their growth should be inhibited to protect sperm fertilizing capacity and the female's health. In pig breeding, where semen is routinely stored at 17°C in the liquid state, alternatives to conventional antibiotics are urgently needed. Photodynamic inactivation (PDI) of bacteria is a well-established tool in medicine and the food industry but this technology has not been widely adopted in semen preservation. The specific challenge in this setting is to selectively inactivate bacteria while maintaining sperm integrity and functionality. The aim of this study was to test the principle of PDI in liquid stored boar semen using the photosensitizer 5,10,15,20-tetrakis(N-methyl-4-pyridyl)-21H,23H-porphine (TMPyP) and a white light LED-setup. In the first step, photophysical experiments comprising singlet oxygen phosphorescence kinetics of TMPyP and determination of the photosensitizer triplet time revealed a sufficiently high production of reactive singlet oxygen in the Androstar Premium semen extender, whereas seminal plasma acted as strong quencher. In vitro experiments with extended boar semen showed that the established PDI protocol preserves sperm motility, membrane integrity, DNA integrity, and mitochondrial activity while efficiently reducing the bacteria below the detection limit. A proof-of-concept insemination study confirmed the in vivo fertility of semen after photodynamic treatment. In conclusion, using the PDI approach, an innovative tool was established that efficiently controls bacteria growth in extended boar and maintains sperm fertility. This could be a promising contribution to the One Health concept with the potential to reduce antimicrobial resistance in animal husbandry.

Unveiling the Potential of Aloe vera Gel Supplementation in a Cooling Extender: A Breakthrough in Enhancing Rooster Sperm Quality and Fertility Ability.

The cooling of semen storage at 5 °C from a Thai native rooster (Pradu Hang Dum), supplemented with herbs possessing antioxidant properties, provided limited research. This study was conducted to evaluate the efficiency of Aloe vera (AV) gel supplementation at various levels on the quality of cooled semen and subsequent fertility after artificial insemination. Sixty-four chickens had semen pooled, diluted, and supplemented with different levels of AV gel (0% as control, 0.25%, 0.50%, 1.0%, 2.5%, 5.0%, 10%, and 20%), and then stored for 72 h. In Experiment 1, semen quality, malondialdehyde (MDA) levels, and pH values were assessed at 0, 24, 48, and 72 h after storage. Experiment 2 assessed fertility potential using the most effective cooled storage semen from Experiment 1. Results showed a decrease in semen quality with prolonged storage time (p < 0.001). The highest semen quality was observed in the group supplemented with 1.0% AV gel (p < 0.001), whereas the lowest was noted in the 20% AV gel group (p < 0.001). Furthermore, the 1.0% AV gel group exhibited the highest semen quality at 24, 48, and 72 h of storage. The evaluation of fertility and hatchability rates revealed a statistically significant improvement in fertility potential (p < 0.05) in the group supplemented with 1.0% AV gel. In summary, this study represents the first investigation of stored Thai native rooster semen using a semen extender supplemented with Aloe vera gel at 5 °C, demonstrating its efficacy for storage up to 72 h. The addition of 1% AV gel was recommended as an antioxidant supplementation during the semen storage process at 5 °C to enhance semen quality and fertility rates.

Update of the cooling protocol for antibiotic-free storage of boar semen at 5°C improves sperm quality and maintains low bacterial counts.

Preserving boar semen at 5°C instead of the conventional storage temperature of 17°C would enable a reduction of antibiotic use in pig insemination. To protect the chilling-sensitive boar spermatozoa, holding the extended semen at a higher temperature before cooling could be beneficial and facilitate the implementation of the innovative preservation concept in practice, provided that bacterial growth is kept at a low level. The aim of this study was to introduce a holding time (HT) at 17°C before cooling and to examine the effect on sperm quality and bacterial growth compared to the original cooling protocol for antibiotic-free 5°C semen storage. A series of experiments with semen doses from eight boars extended in Androstar® Premium without conventional antibiotics revealed that sperm kinematics and the integrity of sperm plasma membranes and acrosomes were improved with HT between 16 and 24 h followed by delayed cooling with 0.04°C/min when compared to the original protocol for semen preservation at 5°C (p < 0.05). Both a shorter HT of 6 h and a faster cooling rate of 0.07°C/min reduced sperm quality (p < 0.05). The HT for 24 h did not compromise the inhibitory effect on bacterial growth during long-term semen storage at 5°C, not even in semen doses spiked with Serratia marcescens. In conclusion, semen storage at 5°C with the modified cooling protocol improved sperm quality and is antimicrobially efficient. It thus presents a ready-to-use tool for a reduction or replacement of antibiotics in pig insemination.

Antimicrobial peptides and proteins as alternative antibiotics for porcine semen preservation

BackgroundAntimicrobial resistance (AMR) is nowadays a major emerging challenge for public health worldwide. The over- and misuse of antibiotics, including those for cell culture, are promoting AMR while also encouraging the research and employment of alternative drugs. The addition of antibiotics to the cell media is strongly recommended in sperm preservation, being gentamicin the most used for boar semen. Because of its continued use, several bacterial strains present in boar semen have developed resistance to this antibiotic. Antimicrobial peptides and proteins (AMPPs) are promising candidates as alternative antibiotics because their mechanism of action is less likely to promote AMR. In the present study, we tested two AMPPs (lysozyme and nisin; 50 and 500 µg/mL) as possible substitutes of gentamicin for boar semen preservation up to 48 h of storage.ResultsWe found that both AMPPs improved sperm plasma membrane and acrosome integrity during semen storage. The highest concentration tested for lysozyme also kept the remaining sperm parameters unaltered, at 48 h of semen storage, and reduced the bacterial load at comparable levels of the samples supplemented with gentamicin (p > 0.05). On the other hand, while nisin (500 µg/mL) reduced the total Enterobacteriaceae counts, it also decreased the rapid and progressive sperm population and the seminal oxidation-reduction potential (p < 0.05).ConclusionsThe protective effect of lysozyme on sperm function together with its antimicrobial activity and inborn presence in body fluids, including semen and cervical mucus, makes this enzyme a promising antimicrobial agent for boar semen preservation.

Potential use of ashwagandha extract in storage of liquid boar semen

Potential use of ashwagandha extract in storage of liquid boar semen

Evaluation of Dry Ice for Short-Term Storage and Transportation of Frozen Boar Semen.

To address the safety problems posed by the transportation of boar semen using LN, this study was conducted on the short-term storage of frozen boar semen in dry ice (-79 °C). Boar semen frozen in LN was transferred to dry ice, kept for 1 day, 3 days, 5 days, 7 days, or 8 days, and then moved back to LN. The quality of frozen semen stored in LN or dry ice was determined to evaluate the feasibility of short-distance transportation with dry ice. The results showed that 60 °C for 8 s was the best condition for thawing frozen semen stored in dry ice. No significant differences in spermatozoa motility, plasma membrane integrity, or acrosome integrity were observed in semen after short-term storage in dry ice compared to LN (p > 0.05). There were no significant changes in antioxidant properties between storage groups either (p > 0.05). In conclusion, dry ice could be used as a cold source for the short-term transportation of frozen boar semen for at least 7 days, without affecting sperm motility, morphological integrity, or antioxidant indices.