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In the most species studied sperm abnormalities have long been associated with male infertility and sterility. This study evaluated the sperm morphology (normality and abnormality) of beef cattle at several Artificial Insemination centers in Indonesia. Total of 142 bulls were used in this study; an ejaculate from each bull was examined. A drop of semen was placed on 3-4 glass slides, and smears were prepared and air-dried. The smears were stained with carbolfluchsin-eosin (Williams stain). Types of morphological abnormalities were recorded from 500 cells on each sample. It was recorded that 77.46% samples had low primary sperm abnormalities (10%) was found in 5.63% samples. Pear shaped was the most frequently type of abnormality found on examined samples (2.24 ± 2.94%); while double head was the lowest (0.01 ± 0.04%). (Animal Production 12(1): 44-49 (2010) Key words : sperm abnormality, beef catlle, artificial insemination center

The objective of the research was to evaluate the speed and duration of centrifugation and seminal plasma levels on the viability of stallion spermatozoa. Semen was collected from three stallions twice a week. The collected semen then evaluated macro- and microscopically. Exp I, the semen was diluted 1:1 with skim milk extender and then centrifuged at 2000 and 3000 rpm for 15 and 20 minutes each. The seminal plasma was removed, and the sperm pellet was re-diluted with skim milk extender. The extended semen then divided in two tubes, stored in 5oC and in a room temperature. The semen then was examined every 3 hours for room temperature and 12 hours for 5oC. Exp II, the semen diluted 1:1 with skim milk extender and then centrifuged at 3000 rpm for 15 minutes. The seminal plasma was removed and the pellet was re-diluted with skim milk extender consisted of 0%, 25%, 50% and 75% of the seminal plasma. The extended semen then was stored in 5oC and examined daily. Results showed that speed and duration of centrifugation had no effect on the quality of stallion semen. The liquid semen without seminal plasma showed 47.50% motile sperms and 61.00% life sperms, wich were significantly higher (P

Generally, the problem of semen freezing process is cold shock effect and intracelullar change due to water release which is related to ice crystal formation. One factor to solve the problem is finding out optimal equilibration time and thawing method so there is only a little damage spermatozoa during freezing process. The research has been done to explore the influence of equilibration time and thawing methode on frozen semen quality of garut rams. The results shown that the mean of percentages of progressive motile sperm, percentages of viable sperm, percentages of plasma membrane and acrosomal intact on four hours equilibration (52.50% ; 62.33%; 57.17% and 56.42%) were not significantly different (P>0.05) from five hours equilibration (52.27%; 63.67%; 56.92% and 57.58 %) and six hours equilibration (54.17%; 61.00%; 59.42% and 58.58%) respectively. The percentages of progressive motile sperm, percentages of viable sperm, percentages of plasma membrane and acrosomal intact on the thawing method on the temperature of 370C for 30 seconds (53.33% ; 62.39%; 57.94% dan 58.61) were not significantly different (P>0.05) from the thawing methode on the temperature of 250C for 45 seconds (52.22% ; 62.28%; 57.72% dan 56.44) respectively. The conclusion shown that the treatment of four hours equilibration, five hours equilibration, six hours equilibration and also the treatment of thawing method on the temperature of 370C for 30 seconds and the thawing methode on the temperature of 250C for 45 seconds do not have effect on garut ram freezing semen quality. (Animal Production 7(2): 74-80 (2005) Key Words : Equilibration, Thawing, Semen, Garut ram

In estrous synchronization, the hormone progesterone is an important element. Various hormone preparations currently available have limitations, especially those related to environmental impact issues. Various alternatives are being studied, and using biodegradable polymeric materials (chitosan-polyethylene glycol combination) to develop new devices is considered one of the solutions. This contribution aims to design and evaluate intravaginal implants that can release progesterone and be degraded in the body and the environment. Implants are made by melting and molding techniques. In vitro drug release studies using dyes as drug models. Implant degradation studies tested in compost. Changes in the shape of the implant, while it is in the vagina, are observed by ultrasound. Blood collection was performed three days before and during implantation to obtain a blood progesterone profile. In vitro drug release studies using dye as a drug model showed a chitosan-PEG profile that released the drug faster at first, then slowed down. Implant degradation studies in compost and vagina demonstrated a gradual degradation process. The blood progesterone profile increased during implantation, as high as 15 ng/mL on the third day. In conclusion, the chitosan-PEG intravaginal implant formulation designed using the melting and molding technique proved to be degraded in the compost environment. It released the hormone progesterone for four days according to the degradation period of the implant in the vagina.

Bali cattle are the indigenous cattle well known attributed to their high fertility. Various proteomes in sperm are substances that can determine sperm quality and fertility. One of which is alpha enolase (ENO1). This study aims to assess the presence of ENO1 in the plasma membrane of sperm and its relationship with the semen quality of Bali cattle. The study used 30 ejaculates from 5 bulls for sperm quality assessment and detection of ENO1. Sperm quality is indicated by total motility, progressive motility, kinematics motility, viability, plasma membrane integrity, and ENO1 quantity. Sperm motility and kinematics motility were measured using CASA, while viability was assessed by eosin-nigrosine differential staining. The HOS test was used to determine plasma membrane integrity, and ENO1 quantity was measured by the ELISA method. Data were analyzed using ANOVA (randomized group design), linear regression, Pearson’s correction, and the t-Student test. The results showed that ENO1 was detected in the plasma membrane at 1.27 ng/106 sperms. The amount of alpha-enolase in the plasma membrane of Bali cattle sperm is affected by sperm concentration (p

This study aims to determine the effect of the age difference of Simmental bulls on motion characteristics, capacitation status, and DNA fragmentation of post-thawing sperms. The frozen semen used was collected from twelve bulls, which were divided into four groups of age, which include a group of two, four, ≥ 10 years old with high semen rejection (≥ 10 HR), and ≥ 10 years old with low semen rejection (≥ 10 LR). Computer Assisted Sperm Analysis (CASA) was used to determine sperm motion characteristics, capacitation status by chlortetracycline (CTC) staining, plasma membrane integrity, and viability using eosin-nigrosine staining. In contrast, the DNA fragmentation index was determined using the Sperm-Bos-halomax kit. The results showed that the four year old group had a higher total and progressive motility percentage than the others (p

Diluents are substances added to the semen to increase the semen volume, reduce sperm density, and maintain sperm viability. This study aimed to compare the effects of various semen diluents on the quality of frozen semen of Limousin bull. Three mature Limousin bulls were used as semen sources. The semen was collected using an artificial vagina and then evaluated for its quality. Sperms with motility less than 70% and abnormality greater than 20% were excluded from this study. After semen evaluation, each ejaculate was individually divided into four equal tubes and diluted with skimmed milk-egg yolk (SMEY), Tris–egg yolk (TEY), Tris–egg yolk–skimmed milk (TEYSM), or Andromed®. Diluted semen was loaded into a mini straw and then equilibrated at 5 °C for 4 h. Following equilibration, the straws were frozen. The percentages of sperm motility, viability, intact membrane, and sperm DNA integrity were evaluated after freezing. Furthermore, malondialdehyde (MDA) and aspartate aminotransferase (AST) enzyme concentrations were assessed after thawing. The sperm motility with TEY, TEYSM, and Andromed® did not differ significantly but was higher than with SMEY. Sperm viability, membrane intactness, and sperm DNA integrity did not differ among treatments. By contrast, the MDA levels and concentration of AST in the Andromed® diluent semen showed the lowest values compared with SMEY semen (p

The aim of the study was to produce polyclonal antibody (rabbit anti-ovPAG) which could detectPAG in the urine of pregnant ewes. Twelve rabbits were immunized against ovPG DEAE-TrisHCl (DT),DEAE-NaCl 20mM (DN2), DEAE-NaCl 40mM (DN4), DEAE-NaCl 80mM (DN8), DEAE-NaCl160mM (DN16), DEAE-NaCl 320mM (DN32) and DEAE-NaCl 1M (DN1) and NaCl 0.9 % as aplacebo. The 0.5 ml of isolate (purified from ovine cotyledon) was emulsified in equal volume withcomplete and incomplete Freud’s adjuvant. The mixture of each isolate and adjuvant was injected atmutiple sites along the dorsal area of rabbits by subcutaneous route. Blood were collected from marginalear vein, starting before first injection (baseline) and every 14 days. Rabbit anti-ovPAG were measuredusing Modified ELISA Technique. By using Western Blot Technique, DN32 showed the best immuneresponse among others and also could differenciate ovPAG in the urine of pregnant ewes It could beconcluded that ovPAG DN32 is a specific source of rabbit anti-ovPAG production. Protein of ovPAG atmolecular weight 31 kDa is a pregnancy protein marker of garut sheep and could be developed as amajor protein for producing antibodi.

The aims of this study was to compare Tris egg yolk and Citrate egg yolk extender andsupplementation of fructose on citrate egg yolk on the quality of Friesian Holstein (FH) bull semenstored at 5 oC. Semen was collected from 5 FH bulls using an artificial vagina. The semen wereevaluated macroscopic and microscopically. The semen divided into three tubes and extended with Trisegg yolk (TEY), Citrate egg yolk (CEY) or Citrate fructose egg yolk (CFEY). Extended semen wasstored at 5 oC and evaluate daily for sperm motility and viability. There was no significant differences(P>0.05) on the sperm viability among three extender, for every time observation during 144 hours ofstorage. This similar finding found on the sperm motility in all extender for 48 hours of storage. Thesperm motility in TEY demonstrated significantly greater (P

This study was conducted to evaluate the effects of sugars supplementation in skim milk based (SM) and dimitropoulos (DV) extenders on the sperm motility and viability in stallion semen storage at 5 oC and ambient temperature (24-29 oC). Semen samples were collected from 3 stallions; evaluate individually and each of them divided into 8 aliquots. Four out of eight aliquots were diluted 1:1 with SM, while the remaining four were diluted 1:1 with DV; all were then centrifuged at 1006 g (3000 RPM) for 15 min. The supernatants were discarded, and each pellet was re-diluted with SM (control), SM trehalose (SMT), SM-raffinose (SMR), SM-fructose (SMF), DV (control), DV-trehalose (DVT), DV-raffinose (DVR), and DV-fructose (DVF). The diluted semen were divided into 2 aliquots and stored at 5 oC or ambient temperature. The sperm motility and viability were evaluated every 3 h on chilled semen stored at ambient temperature, and every 12 h on those stored at 5 oC. Results of the experiments demonstrated that sperm motility and viability in DV extender significantly higher (P

The aims of study was to compare the glycerol concentration in Tris glucose egg yolk (TGEY) diluents on the quality of deer frozen semen. Semen was collected from 5 Timor deer using electroejaculator. Immediately after collection the semen was evaluated macroscopic and microscopically. After initial evaluation, the semen was divided into three tubes and extended with Tris egg yolk with three different glycerol concentrations, which were 10% (TGEY10); 12% (TGEY12) and 14% (TGEY14). The sperm motility, viability, acrosome intact and membrane intact were evaluated in raw semen, after equilibration and after thawing. The results showed that there were no differences (p>0.05) on the sperm motility, viability as well as sperm acrosome intact. Sperm membrane intact in TGEY10 (52.50±5.89%) and TGEY14 (51.50±4.12 %) were higher (p

Anoa is Indonesia endemic fauna that included into Appendix I by CITES. Anoa breeding efforts have not succeed, due to their agressivity, soliter, wildness and monogamous. The aim of this experiment was to observe morphology and biometry of anoa’s sperm stained using William’s (W) and eosin-nigrosin (EN). Semen was collected from two male anoas by electroejaculator. Slides was made for morphology and biometry observation of ejaculate. The results showed that abnormal sperm morphology of the mature anoa (10 years) was 32.27%-35.00%, meanwhile the young one (3 years) was 24.03%–31.89%. In mature anoa, measurement for head width, and midpiece and tail length of sperm were not signficantly different between W and EN staining, but significantly different (P

Semen used for artificial insemination (AI) can be prepared in different ways, fresh extended fresh or chilled, or frozen-thawed (FT). The aim of the experiment was to evaluate the use of preserved semen to inseminate the oestrus mares. Nineteen mares were used in this research. The mares were synchronized with double injection of PGF2α 14 days apart. The follicle size was monitored using ultrasound scanner during the third day of oestrus, and 2500 IU hCG was administered at the same time. The AI was conducted 35 hours after hCG injection with total motile sperm 200x106 for chilled semen and 250-300x106 for frozen semen. The result demonstrated that the response of the oestrus with double injection of PGF2α was 73.7%. The conception rate (CR) was 14.3% (1/7) with frozen semen and 42.9% (3/7) with chilled semen. It is concluded that AI with chilled semen resulted higher conception rate than frozen semen.

The objective of this experiment was to evaluate the effects of isoflavone-riched soybean flour, zinc (Zn) and vitamin E on quality of rats spermatozoa as animal model. Diet was given as isonitrogen and isocaloric with 15% of dietary protein. Thirty male Sprague Dawley post weaning rats (21 days old) were divided into six groups and treated with isoflavone-riched soybean flour, Zn and vitamin E in different combination. Isoflavone-riched soybean flour (3 mg/day) was given by oral administration, whereas Zn and vitamin E were mixed with the basic diet. The treatments were given for 2 months. Thirty female Sprague Dawley post weaning rats (21 days old) were used to observe male fertility. Results indicated that synergic interaction between isoflavone-riched soybean flour, Zn and vitamin E increased motility and concentration of rats spermatozoa. The treatment was not significant on relative weight of testis, conception rate and foetus size. The best results was shown by the group which was given isoflavone-riched soybean flour with diet containing both Zn and vitamin E, i.e. motility rate of 79.50±1.12%, sperm concentration of 1636.90±87.95 million/ml, and abnormality rate of 9.00±0.70%. Conception rate and the number of foetus on female rats were 100% and 11±0.71, respectively. It is concluded that combination treatment of isoflavone-riched soybean flour, Zn and vitamin E on male rats showed the best fertility in comparison with single treatment and the other combination.

The study was conducted to evaluate the effect of intervention of skim milk based functional drink supplemented with 100 mg of soy isoflavone and 8 mg of Zn-sulfate (susumeno) on serum estradiol and thymulin hormones levels of permenopausal women. After 2 month of intervention, the estradiol levels did not change, and the levels were in the range of 39,16 – 41,99 pg/ml. However the thymulin hormone levels increased significantly from 2,37 µg/ml to 3,31 µg/ml. Increase in thymulin levels shows that the disfuction of immune system of premenopausal could be improved by the functional drink.

Ongole Grade cattle is one of local cattle in Indonesia.This study was aimed to analyze the effect of bull age and season on semen quality of Ongole Grade cattles macroscopically (sperm motility, and sperm concentration) and microscopically (semen ejaculate volume) to support the policy and management on improvement of semen production. Data were collected from 533 ejaculates of 10 bulls aged 3, 4, and 5 yr at Singosari National Artificial Insemination Center(SNAIC). The 4-year-old bulls hadhighestsperm motiliy and semen ejaculate volume. The lowest ejaculate volume was reported in Ongole Gradecattlesaged 3 yr. Sperm motility and concentrationweresignificantly higher (P

Aim: This study aimed to analyze the individual factors influencing the sperm quality of Bali bulls at Baturiti Artificial Insemination (AI) center. Materials and Methods: Semen that was ejaculated from nine Bali bulls was collected using artificial vaginas (n=5/bull). Semen ejaculates were evaluated immediately after collection to measure the quality of the fresh semen, including semen volume, sperm concentration, progressive motility, membrane integrity (MI), and abnormal morphology. Frozen semen was evaluated for progressive sperm motility, concentration, viability, MI, abnormal morphology, and deoxyribonucleic acid (DNA) fragmentation. Other secondary data, focusing on semen quantity (semen volume and sperm concentration), were also collected from frozen the semen production data of the Baturiti AI center from 2017 to 2019. Data were analyzed statistically using a completely randomized design, and one-way analysis of variance was applied to find differences among individual bulls. Results: Significant differences (p

The objective of this study was to determine histomorphological change of seminiferous tubules, sperm quality and testosterone level after administration and cessation of pegagan extract. The research used 4x2 factorial complete randomized design, the first factor was dose-administration duration and the second factor was cessation length of pegagan extract. Parameters consisted of degree of spermatogenesis, sperm quality (motility, concentration and abnormality), and testosterone level. The data were processed using analysis of variance and Duncan’s test. Results showed that there was no interaction between dose-administration duration and cessation length. There was no significant difference of dose-administration duration of pegagan on degrees of spermatogenesis, sperm motility and testosterone level. Cessation length did not significantly affect degrees of spermatogenesis, sperm motility, and testosterone level although these results indicated that cessation of pegagan extract for 12 days increased degrees of spermatogenesis and sperm motility, indicated the antifertility potency of pegagan extract. In conclusion, Pegagan extract has potency as anti-fertility agent although the degree of spermatogenesis, sperm quality and testosterone levels were still in the normal range for the reproductive process and not irreversible, and pegagan could be explored as an reversible anti-fertility agent.

The aims of study was to compare the glycerol concentration in Tris glucose egg yolk (TGEY)diluents on the quality of deer frozen semen. Semen was collected from 5 Timor deer usingelectroejaculator. Immediately after collection the semen was evaluated macroscopic andmicroscopically. After initial evaluation, the semen was divided into three tubes and extended with Trisegg yolk with three different glycerol concentrations, which were 10% (TGEY10); 12% (TGEY12) and14% (TGEY14). The sperm motility, viability, acrosome intact and membrane intact were evaluated inraw semen, after equilibration and after thawing. The results showed that there were no differences(p>0.05) on the sperm motility, viability as well as sperm acrosome intact. Sperm membrane intact inTGEY10 (52.50±5.89%) and TGEY14 (51.50±4.12 %) were higher (p

s to assess different CPAs on stallion semen cryopreservation. Skim milk (SM) and Dimitropoulos(DV) were the extenders used in this study; each was added by glycerol (Gly), combination of ethyleneglycol-glycerol (EG+Gly) or dimethilformamide (DMF). Each semen sample was evaluated and dividedequally into six tubes; semen in the three tubes was diluted 1:1 with (SM), while in the remaining tubesthe semen was diluted 1:1 by DV. After being diluted, all tubes were centrifuged at 1006xg for 10minutes. The supernatan discarded, the pellet was rediluted by SM trehalosa or DV trehalose, and addedby G, EG+Gly, or DMF to reach the final sperm concentration of 200x106/ml. The extended semen wasindividually packed in 0.3 ml minitube, equilibrated at 4oC for 2 hours, frozen in liquid nitrogen vaporfor 10 minutes, and then was stored in liquid nitrogen container at -196 oC. After 24 hours, the semenwas thawed at 37 oC for 30 second. There were no significantly different (p>0.05) on the percentages ofmotile and viable sperm in SMT (21.7% and 43.4%, respectively) compared with those extended withDV T extender (26.9% and 50.8%, respectively). DMF demonstrated better results as CPA compared tothe others; and DVTDMF combination had the best protection during cryopreservation in this study.

Morphologically abnormal sperm in semen has been associated with the sub fertility and sterilityfor many years. This study assessed the sperm morphology of Friesian Holstein bull semen which wascollected from three Artificial Insemination centers in Indonesia. Total of 22 bulls were used in thisstudy; an ejaculate from each bull was examined. Three to four glass slides were prepared for each bullsample; a drop of semen was placed on each glass slide, smeared, and air-dried. The smeared sampleswere stained with carbolfuchsin-eosin (Williams stain). Morphological abnormality types were recordedfrom total of 500 spermatozoa. Results demonstrated that all 22 bulls had low sperm abnormality(

Anoa, which is the endemic animal in Indonesia, its population is unevitably decreasing, therefore anoa included in the list of endangered species by International Union for Conservation of Nature (IUCN). The experiment was aimed to apply the artificial insemination (AI) technique for anoa and to examine their parturition behavior. The experiment involved 2 males and 5 females anoa at Taman Safari Indonesia Bogor. Semen was collected by electroejaculator, then evaluated and diluted in Tris-egg yolk extender. The anesthetized estrous females were inseminated intracervically with extended semen (100x106 sperm/1.0 mL). Females which did not show estrous signs during 2-3 cycles after AI were predicted pregnant, and therefore confirmed by ultrasonography. As a comparison, another predicted-pregnant female after natural mating was scanned. The results showed that intracervical AI resulted pregnancy in one female, and the gestation period of anoa was ranged from 313 d (AI) to 324 d (natural mating). There were three stages of parturition process was observed: Stage 1 was characterized by the abdominal contraction and amniotic membrane rupture, Stage 2 was characterized by the fetal expulsions, and Stage 3 was characterized by the placental expulsions and ingestion of placenta by the dam. The duration of each stage of parturition was 6-8 h (stage 1), 30-60 min (stage 2), and 15-180 min (stage 3). It is concluded that AI technique was applicable to anoa in captivity, and the parturation behavior of anoa was comparable to buffalo and cattle. However, the fetal delivery occurred when the dam was in standing position and the dam ingested the placenta.

The aim of the study was to produce polyclonal antibody (rabbit anti-ovPAG) which could detect PAG in the urine of pregnant ewes. Twelve rabbits were immunized against ovPG DEAE-TrisHCl (DT), DEAE-NaCl 20mM (DN2), DEAE-NaCl 40mM (DN4), DEAE-NaCl 80mM (DN8), DEAE-NaCl 160mM (DN16), DEAE-NaCl 320mM (DN32) and DEAE-NaCl 1M (DN1) and NaCl 0.9 % as a placebo. The 0.5 ml of isolate (purified from ovine cotyledon) was emulsified in equal volume with complete and incomplete Freud’s adjuvant. The mixture of each isolate and adjuvant was injected at mutiple sites along the dorsal area of rabbits by subcutaneous route. Blood were collected from marginal ear vein, starting before first injection (baseline) and every 14 days. Rabbit anti-ovPAG were measured using Modified ELISA Technique. By using Western Blot Technique, DN32 showed the best immune response among others and also could differenciate ovPAG in the urine of pregnant ewes It could be concluded that ovPAG DN32 is a specific source of rabbit anti-ovPAG production. Protein of ovPAG at molecular weight 31 kDa is a pregnancy protein marker of garut sheep and could be developed as a major protein for producing antibodi.

In order to improve Timor deer (Cervus timorensis) frozen semen quality, the influence of sugar and glycerol concentration on semen characteristics of sperm was investigated. The semen was collected from five sexually mature Timor deer using an electroejaculator. The semen was evaluated and divided into six equal tubes and diluted with Tris sucrose glycerol 10% (TSG10); Tris sucrose glycerol 12% (TSG12); Tris sucrose glycerol 14% (TSG14); Tris glucose glycerol 10% (TGG10); Tris glucose glycerol 12% (TGG12); and Tris glucose glycerol 14% (TGG14). The diluted semen was packed in 0.3 ml minitub straw, equilibrated at 5 oC for 4 hours and frozen on liquid nitrogen vapor for 10 minutes. The total of forward motility, viability, acrosome integrity and membrane integrity were assessed in fresh, after equilibration and after thawing. Results demonstrated that the percentage of sperm motility in TSG10 was higher (P

Molecular-based biomarkers are believed to be more accurate in determining bulls' fertility and sperm's various fertility characteristics. The sperm acrosome-associated 1 (SPACA1) molecule, located in the anterior acrosomal and equatorial segments of the sperm head, is thought to be related to the function of binding and fusion between sperm and oocyte. This study aims to analyze the association of SPACA1 mRNA and protein with the fertility rate and semen quality of Bali bulls (Bos sondaicus) and assess its potential as a potential molecular biomarker determining bull fertility and sperm quality. Frozen semen from 20 Bali bulls was used in the research, which was then divided into two groups: high (HF) and low fertility (LF). SPACA1 mRNA abundance was analyzed using qRT-PCR, and SPACA1 protein abundance was analyzed using EIA. The semen quality parameters analyzed were motility (CASA), plasma membrane integrity (HOS test), sperm head morphology abnormalities (William staining), sperm viability (SYBR 14-PI), sperm capacitation (CTC assay), and sperm acrosome integrity (FITC-PNA). The results showed that there was a significant difference (P < 0.05) between high fertility (HF) and low fertility (LF) bulls, both in the abundance of SPACA1 at the mRNA, protein, and semen quality levels. Overall, the correlation test results showed a close relationship (P < 0.01) between the abundance of SPACA1 mRNA and protein with field fertility and various semen quality parameters tested in the study. A low level of SPACA1 molecules indicates low fertility and semen quality in Bali bulls. The SPACA1 molecule has the potential to be developed through further research to become a biomarker for determining fertility and semen quality in bulls., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Increasing the age of bulls results in a decrease in reproductive function, including a reduction in sperm quality, which plays a vital role in determining the fertility of bulls. Through a proteomic approach, this research aims to analyze the influence of age factors on various proteomes contained in bull sperm. Frozen semen samples from Simmental Bulls were categorized into three age groups: two, four, and ≥10 years old. Subsequently, the post-thaw sperm cells obtained were separated based on molecular weight using 1D-SDS-PAGE. Peptides extracted from the bands produced in each age group were subjected to LC-MS/MS analysis. A total of 72 protein types were identified, with 45 being detected in the 4-year-old group and 41 expressed in both the 2 and ≥10-year-old groups. The results provided insights into proteins' role in sperm metabolism across all age groups. Specifically, the 2-year-old group exhibited the expression of proteins associated with acrosome assembly and spermatid development (SPACA1). In contrast, those in the 4-year-old group were linked to motility (PEBP4) and sperm decapacitation factor (PEBP1). Proteins expressed in the 2 and -year-old groups were discovered to be involved in fertilization processes (TEX101). In contrast, the ≥10-year-old age group was associated with hyperactive movement related to capacitation (Tubulin). In conclusion, age influenced the differences observed in the proteomic profile of post-thaw Simmental bull sperm using the 1D-SDS-PAGE tandem LC-MS/MS approach., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Satrio, Karja, Setiadi, Kaiin, Pardede and Purwantara.)