Research Article

Impact of Urea Moringa Molasses Multi-Nutrient Block Supplementation on Liquid Semen Quality and Fertility in Etawah Crossbreed Goats

Nursyam Andi Syarifuddin1*, Muhammad Rizal1, Muhammad Riyadhi1, Anis Wahdi1, Habibah Habibah1, Athhar Manabi Diansyah2, A. Ni’mahtul Churriyah1, Rahmat Rahmat1

1Department of Animal Science, Faculty of Agriculture, University of Lambung Mangkurat, Jl. Jend. A. Yani, Banjarbaru, 70714, Indonesia; 2Department of Animal Production, Faculty of Animal Science, Hasanuddin University, Jl. Perintis Kemerdekaan, Makassar, 90245, Indonesia.

Received | September 12, 2025; Accepted | December 26, 2025; Published | February 07, 2026

*Correspondence | Nursyam Andi Syarifuddin, Department of Animal Science, Faculty of Agriculture, University of Lambung Mangkurat, Jl. Jend. A. Yani, Banjarbaru, 70714, Indonesia; Email: [email protected]

Citation | Syarifuddin NA, Rizal M, Riyadhi M, Wahdi A, Habibah H, Diansyah AM, Churriyah AN, Rahmat R (2026). Impact of urea moringa molasses multi-nutrient block supplementation on liquid semen quality and fertility in Etawah crossbreed goats. J. Anim. Health Prod. 14(1): 289-296.

DOI | https://dx.doi.org/10.17582/journal.jahp/2026/14.1.289.296

ISSN (Online) | 2308-2801

Copyright: 2026 by the authors. Licensee ResearchersLinks Ltd, England, UK.

This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).

INTRODUCTION

The Etawah Cross-breed (EC) goat is produced by crossing the Etawah (Jamnapari) goat with the Kacang goat. EC goats serve dual purposes, providing both meat and milk, which makes them popular among Indonesian farmers (Handayani et al., 2024; Hariyono and Endrawati, 2023). These goats are capable of giving birth three times within two years, with each lactation period lasting 4-6 months and a dry period of 2-3 months, achieved by implementing a breeding program every eight months. Optimal reproductive performance, such as this, can be supported by effective reproductive management practices, including the use of artificial insemination technology.

Providing high-quality semen is essential for the success of artificial insemination programs. Liquid semen technology serves as an alternative method for semen supply and storage, especially to address the shortage of frozen semen in rural regions. Semen can be refrigerated at temperatures between 3-5°C, maintaining sperm viability for 2-4 days (Yang et al., 2018), making it a more practical, faster, and cost-effective option compared to frozen semen, which requires the use of liquid nitrogen. A major challenge in semen preservation is the reduction in sperm quality caused by free radicals (Rizal et al., 2025) and Reactive Oxygen Species (ROS) (Liang et al., 2023), which can damage sperm membranes, proteins, and nucleic acids, ultimately leading to sperm death.

The quality of semen during storage can be preserved by providing nutritious feed ingredients, such as moringa leaves. These leaves are rich in a wide range of essential nutrients, including protein, fat, calcium, magnesium, potassium, selenium, vitamin C, vitamin E, and polyphenolic compounds (Gangwar et al., 2024), all of which play a vital role in maintaining sperm viability. Additionally, moringa leaves contain phenolic compounds like flavonoids (Syarifuddin et al., 2025), which help reduce sperm damage during semen processing and enhance sperm fertility (Shah et al., 2022).

The use of moringa leaves as a feed supplement in the form of Moringa Multinutrient Block (MMNB) was developed by Alfian et al. (2025) and is well-suited for implementation by farmers in rural communities. Research by Syarifuddin et al. (2022) demonstrated that supplementing with Urea Moringa Molasses Multi-nutrient Block (UMMMB) enhances libido (including mounting and ejaculation), total sperm motility, and viability in male Etawah Crossbreed (EC) goats. Building on these findings, this study aims to assess the quality and fertility of liquid semen from EC goats fed with UMMMB supplements. The expected outcome is to offer goat farmers in rural areas an alternative source of high-quality liquid semen.

MATERIALS AND METHODS

Experimental design and ethical approval

This research involved eight Etawah Cross (EC) bucks with an average age of 36.72 ± 1.06 months and an average body weight of 54 ± 2.97 kg, sourced from the Pelaihari Breeding Centre. In addition, 64 adult female goats (does) belonging to the Rukun Jaya Makmur Farmer Group in Landasan Ulin District, Banjarbaru City, South Kalimantan Province, were included. The selected does were clinically healthy, aged 18–24 months, exhibited regular estrous cycles, and had previously given birth at least once.

The independent variable was the type of dietary supplementation (UMMB vs. UMMMB), while the dependent variables were fresh semen characteristics, liquid semen longevity, and conception rate. The number of experimental animals was determined based on previous studies with similar designs (Syarifuddin et al., 2025), ensuring sufficient statistical power to detect treatment differences.

All experimental procedures complied with ethical standards and were approved by the Animal Ethics Committee of the Faculty of Agriculture, University of Lambung Mangkurat (Approval No.:017/UN8.1.23.5/PG/2025) and adhered to Indonesian animal welfare guidelines.

Animals, feeding, and management

The bucks were housed individually in stage cages (1.25 m × 1.00 m, elevated 1 m above the ground) for eight weeks. Diets were formulated to meet the standard nutritional requirements for male goats, with fresh native grass provided as the basal diet and unlimited access to drinking water.

Four bucks were randomly assigned to receive native grass plus a 200 g block of UMMB (control), while the other four bucks received native grass plus a 200 g block of UMMMB (treatment). The feed block was offered first and forage was provided only after complete consumption of the block. The formulation of UMMB and UMMMB followed Syarifuddin et al. (2025), and their chemical composition, together with that of the native grass, is presented in Tables 1 and 2.

 

Table 1: Compositions of the multinutrient block used as supplements.

Feed ingredients (%)	UMMB (Control)	UMMMB (Treatments)
Moringa leaf powder	-	30
Palm kernel meal	25	-
Concentrate Laying Ducks (CP144®)	5	-
Fine rice bran	33	33
Molasses	20	20
Limestone	5	5
Common Salt	5	5
Urea (46% N)	5	5
Mineral Mix	2	2
Total	100	100

 

UMMB: Urea molasses multi-nutrient block; UMMMB: Urea moringa molasses multi-nutrient block.

 

Semen collection and evaluation

Semen was collected weekly during weeks 4–8 using an artificial vagina (AV) between 07:00 and 10:00 AM prior to feeding. Ejaculates were first evaluated macroscopically (volume, color, consistency), and subsequently examined microscopically for sperm concentration, motility, viability, and morphology. Concentration was determined using a hemocytometer.

 

Table 2: Nutrient Content of Native Grass, UMMB, and UMMMB (as % DM*).

Nutrient composition (%)	Native grass	UMMB	UMMMB
Dry matter	30.77	60.11	59.62
Ash	11.92	12.18	11.40
Crude Protein	10.79	19.75	21.41
Crude Fiber	31.52	3.11	2.59
Ether Extract	0.62	2.82	2.51
Calcium (Ca)	0.78	3.61	3.78
Phosphorus (P)	0.44	2.05	2.81
Nitrogen Free Extract	45.15	64.34	62.09
TDN (calculated)	55.38	83.44	83.14

 

Note: * Values were obtained from analysis conducted by Laboratory of Nutrition and Animal Feedstuff, Animal Science Department, Faculty of Agriculture, University of Lambung Mangkurat. UMMB: Urea molasses multi-nutrient block; UMMMB: Urea moringa molasses multi-nutrient block.

 

Motility was assessed subjectively under a phase-contrast microscope at 400× magnification, with ≥200 sperm observed per sample. Viability was evaluated using eosin–nigrosin staining. Abnormalities were determined from 200 spermatozoa per smear (Diansyah et al., 2025a).

Processing of liquid semen and quality evaluation

In week 8, ejaculates meeting the quality threshold (≥70% motility, ≥2 ml volume) were diluted with Andromed® extender. The diluent was prepared by mixing 5 ml of Andromed® with 20 ml of distilled water (1:4 ratio), homogenized, and equilibrated in a water bath at 38°C prior to use (Diansyah et al., 2025b). Semen was diluted to a final concentration of 100 × 10⁶ sperm/ml and stored at 3–5 °C.

Sperm quality parameters (motility, viability, and plasma membrane integrity by hypo-osmotic swelling test) were assessed at 30 minutes post-dilution (day 0) and at 24-hour intervals for five consecutive days (Nirmala et al., 2025).

Artificial insemination (AI) and conception rate

Estrus synchronization was performed twice in all does using Lutalyse® (11-day interval). Does showing estrus 72 hours after the second injection were inseminated intracervically with 0.25 ml mini-straws containing liquid semen collected on day 0 (week 8) from UMMB- or UMMMB-supplemented bucks.

In total, 21 does were inseminated with semen from UMMB-fed bucks and 43 does with semen from UMMMB-fed bucks. Pregnancy was determined by the absence of estrus over two consecutive cycles and subsequently confirmed by birth data, following standard field practice. All 64 does underwent the same synchronization protocol without any superovulation treatment; therefore, the difference in the number of inseminated does (21 vs. 43) resulted solely from variation in estrus response after synchronization rather than from experimental manipulation. The conception rate was analyzed using a chi-square test, which appropriately accommodates unequal group sizes, and therefore the comparison remained statistically valid despite the unequal number of does.

Statistical analysis

All quantitative data (fresh and liquid semen traits) were expressed as mean ± standard error (SE). A repeated-measures ANOVA was applied to evaluate semen longevity across storage days, with treatment (UMMB vs. UMMMB) as a fixed factor and time as a repeated measure. Independent-sample t-tests were used for fresh semen characteristics. Conception rate (%) was analyzed by chi-square test. All analyses were conducted using SPSS® version 26 (IBM Corp., Armonk, NY, USA). A p-value < 0.05 was considered statistically significant.

Although mixed-effects models could provide a more detailed representation of hierarchical data structures (e.g., repeated ejaculates within bucks and conception outcomes nested by sire), the limited number of experimental animals restricted the use of such complex models. Therefore, the present analysis focused on fixed treatment effects using repeated-measures ANOVA, which remains appropriate for the dataset and study objectives.

RESULT AND DISCUSSION

Fresh semen quality

Bucks supplemented with UMMMB exhibited significantly higher sperm motility and viability compared with UMMB-fed bucks (p < 0.05; Table 3, Figure 1). All semen quality traits from both groups remained within the acceptable limits defined by the Indonesian National Standard (SNI 4869-3:2023), confirming their suitability for further processing into liquid semen. However, semen characteristics can vary among individual bucks, and the small sample size (n = 4 per group) may contribute to part of the observed variation thus, random allocation was used to minimize potential bias arising from pre-existing individual variation. The observed improvement in motility and

 

Table 3: The fresh semen quality traits of Etawah Crossbreed (EC) bucks.

Semen quality traits	UMMB (Control)	UMMMB (Treatment)	p-Value	References#
	(Mean ± SEM)
Volume (ml)	0.94 ± 0.07	1.01 ± 0.07	0.45	1.0 (0.5 – 1.2)[1]
Consistency	Medium to Thick	Thick	-	Medium to Thick[2]
pH	6.6 ± 0.07	6.7 ± 0.12	0.43	5.9 – 7.3[3]
Mass Movement	++ (30%), +++ (70%)	+++ (100%)	-	++ to +++[2]
Sperm Motility (%)	73.75 ± 1.25a	79.29 ± 1.25b	0.04	60 – 80[3]
Concentration (×106/ml)	3.070 ± 35.38	3.470 ± 39.41	0.52	250 – 500[4]
Viability (%)	81.48 ± 1.02a	85.92 ± 0.77b	0.00	60 – 80[4]
Abnormality (%)	5.61 ± 0.56	5.41 ± 0.36	0.71	5 – 20[3]
Intact Plasma Membrane (%)	87.00±1.11	88.94±0.63	0.14	≥60[5]

 

Note: ab Mean in the same row without common letters are different at p<0.05. #Reported range of values for semen of EC bucks: 1Syarifuddin et al., 2025, 2Susilawati, 2011, 3Garner and Hafez, 2000, 4Ax et al., 2000, 5Saratsi et al., 2023. UMMB: Urea molasses multi-nutrient block; UMMMB: Urea moringa molasses multi-nutrient block.

 

viability among UMMMB-fed bucks may reflect enhanced nutrient intake and antioxidant activity, which supports spermatogenesis and protects spermatozoa against oxidative stress. Similar findings have been reported in small ruminants supplemented with phytogenic or plant-based feed additives that improved semen traits through antioxidant and metabolic pathways (Yusuf et al., 2026; Nurlatifah et al., 2025a).

 

Liquid semen quality

Sperm motility and viability declined progressively during cold storage (p < 0.05; Table 4). According to the Indonesian National Standard (SNI 4869-3:2023), semen should maintain a minimum motility of 40% during storage. Semen from UMMMB-fed bucks retained motility above the minimum threshold of 40% until day 4, whereas semen from UMMB-fed bucks fell below this limit earlier. Viability followed a similar pattern, with UMMMB maintaining values >50% up to day 4, which is considered the minimum requirement for AI (Kumar et al., 2024). Plasma membrane integrity also remained higher in UMMMB semen (61.06% on day 4) compared with UMMB (59.67%), indicating better tolerance to cold storage. According to Rahayu et al. (2025), intact membrane integrity above 60% is essential for AI success, suggesting that UMMMB-fed bucks provided semen of superior preservability.

The superior performance of the UMMMB group can be attributed to the inclusion of Moringa oleifera leaf meal, which is rich in bioactive antioxidants such as flavonoids, polyphenols, vitamin E, and selenium (Alfian et al., 2025). These compounds neutralize reactive oxygen species (ROS) generated during semen storage, thus protecting sperm membranes from lipid peroxidation and maintaining mitochondrial energy production. The block matrix facilitates a slow and steady nutrient release, improving nutrient utilization and antioxidant defense over time. This physiological mechanism explains why UMMMB-fed bucks produced semen with higher motility, viability, and membrane integrity during cold preservation (Syarifuddin et al., 2025).

The progressive decline in motility and viability is consistent with previous reports attributing storage-related deterioration to lactic acid accumulation, pH reduction, and membrane destabilization under cold temperatures (Ibáñez-Arancibia et al., 2021; Mohajer et al., 2024). Membrane lipid phase transitions impair permeability and ATP synthesis, leading to loss of viability (Iqbal et al., 2022).

 

Table 4: The sperm motility, viability, and intact plasma membrane of liquid semen stored for 5 days at 3-5 °C.

Semen quality	Day 1	Day 2	Day 3	Day 4	Day 5	References#
	Means ± SEM
Sperm motility (%)
UMMB	72.86±1.07a	67.81±0.64	59.67±1.65	41.33±0.91a	23.33±1.98	>40[1]
UMMMB)	78.75±1.11b	69.06±0.50	62.50±0.79	45.94±1.84b	29.17±2.53
p-Value	0.03	0.14	0.13	0.04	0.08
Viability (%)
UMMB	81.03±0.85a	78.44±0.56a	62.50±0.79	55.33±1.33	39.67±1.69	>50[2]
UMMMB)	85.48±1.19b	80.38±0.54b	69.67±1.65	59.63±1.76	45.58±2.40
p-Value	0.03	0.02	0.12	0.06	0.06
Intact plasma membrane (%)
UMMB	83.25±0.85	79.88±1.34	68.80±1.22	59.67±1.50	42.25±1.05a	≥ 60%[3]
UMMMB)	86.00±0.91	80.44±0.97	71.19±0.98	61.06±1.65	50.50±1.57b
p-Value	0.14	0.74	0.13	0.54	0.00

 

Note: ab Mean in the same column without common letters are different at p<0.05. # Reported range of values for semen of EC bucks: 1Syarifuddin et al., 2025, 2Ax et al., 2000, 3Revell and Mrode, 1994. UMMB: Urea molasses multi-nutrient block; UMMMB: Urea moringa molasses multi-nutrient block.

 

Table 5: The conception rate of the does inseminated with liquid semen from bucks supplemented with UMMB and UMMMB.

Treatment	Number of acceptors (head)	Pregnant (head)	Not pregnant (head)	Conception rate (%)	References#
UMMB	21	6	15	28.57a	50 – 80%[1]
UMMMB	43	28	15	65.12b

 

Note: ab Mean in the same column without common letters are different at p<0.05. # Reported range of values for semen of EC bucks: 1Nurlatifah et al., 2025b. UMMB: Urea molasses multi-nutrient block; UMMMB: Urea moringa molasses multi-nutrient block.

 

However, the higher baseline semen quality in UMMMB-fed bucks appears to confer greater resilience against cold stress. Recent studies confirm that dietary antioxidants and phytogenic compounds can stabilize sperm membranes during preservation, thereby extending functional lifespan of spermatozoa (Song et al., 2025; Nurlatifah et al., 2025a).

Fertility of liquid semen

Artificial insemination with liquid semen collected from UMMMB-fed bucks yielded significantly higher conception rates compared with UMMB-fed bucks (65.12% vs. 28.57%, p < 0.05; Table 5). The conception rate achieved with UMMMB supplementation falls within the standard range for goat AI (50–80%) (Suyadi et al., 2021), whereas UMMB semen produced substandard fertility outcomes. A total of 21 does were inseminated with semen from bucks fed UMMB and 23 does with semen from bucks fed UMMMB, resulting in an unbalanced distribution between groups due to the field availability of synchronized females rather than selection bias. Conception rates were based on semen collected from four bucks per group, and although random allocation of does was applied to minimize bias, the potential influence of individual “buck effect” could not be entirely excluded. While the chi-square test used accommodates unequal sample sizes, slight variations in statistical precision may occur; however, the marked difference in conception rates still indicates a clear biological advantage of UMMMB supplementation.This improvement in conception is likely attributable to the higher motility, viability, and membrane integrity observed in UMMMB semen, all of which are critical determinants of fertilizing capacity (Bianchi and Wright, 2016; Shan et al., 2021). Sperm motility, in particular, is strongly correlated with fertilization success (Tanga et al., 2021; Pardede et al., 2020; Diansyah et al., 2025c), while plasma membrane integrity ensures proper ionic regulation, capacitation, and oocyte penetration. The superior fertility outcomes in the UMMMB group are therefore consistent with the enhanced semen preservation quality observed in this study.

Overall interpretation

The present findings demonstrate that UMMMB supplementation enhances both fresh and liquid semen quality in Etawah Cross bucks, which in turn translates into higher conception rates under field conditions. These improvements are consistent with reports highlighting the role of dietary bioactive compounds in improving male fertility parameters in small ruminants (Al-Bulushi et al., 2019; Nurlatifah et al., 2025b). The novelty of this study lies in demonstrating that a modified UMMB formulation can sustain semen quality during storage and improve AI outcomes, offering a practical strategy for goat reproduction programs in tropical environments.

Unlike conventional UMMB formulations that primarily provide energy and nitrogen sources, the modified UMMMB introduces functional phytochemicals from Moringa oleifera that improve reproductive performance through antioxidant and metabolic pathways. This methodological improvement demonstrates a practical innovation for on-farm semen quality enhancement in tropical smallholder systems.

CONCLUSION

This study demonstrated that dietary supplementation with UMMMB containing moringa leaf meal significantly improved semen quality and fertility outcomes in Etawah Cross bucks. Bucks receiving UMMMB exhibited higher sperm motility, viability, plasma membrane integrity, and conception rates compared with those fed standard UMMB. These findings highlight the potential of UMMMB as a practical nutritional strategy to enhance semen preservability and artificial insemination success in goats under tropical field conditions. Nevertheless, the limited number of experimental animals and potential variability among individual bucks warrant cautious interpretation of the findings. Further research involving larger populations, repeated experimental cycles, and biochemical profiling of antioxidant and metabolic markers is recommended to validate these results and elucidate the underlying physiological mechanisms.

ACKNOWLEDGEMENTS

The authors gratefully acknowledge the Ministry of Research, Technology, and Higher Education of Indonesia for financial support through the Applied Research Program. Appreciation is also extended to the Rukun Jaya Makmur Farmers Group, Banjarbaru City, South Kalimantan, for providing facilities and assistance in animal management during this study.

Novelty Statement

This study demonstrates that dietary supplementation of Etawah Cross bucks with a urea moringa molasses multi-nutrient block (UMMMB) improves fresh semen quality while significantly enhancing the preservability and fertility of liquid semen during cold storage under field conditions. In contrast to conventional UMMB formulations, the inclusion of Moringa oleifera leaf meal supplies functional antioxidant compounds that help maintain sperm motility, viability, and plasma membrane integrity for up to four days at 3–5 °C. These findings provide a practical and scalable nutritional approach to improve artificial insemination success in smallholder goat production systems in tropical regions.

AUTHOR’S CONTRIBUTION

NAS, MR, MR, AW, and HH, conceived and designed the experiment. NAS, MR, MR, and AW performed the experimental procedures. NAS, and MR, supervised and coordinated the research and provided clinical data. Statistical analysis was conducted by AMD, ANC and RR. The initial draft of the manuscript was prepared by NAS, AMD, ANC and RR. All authors critically reviewed and approved the final version of the manuscript.

Generative AI and AI-assisted technology statement

The authors declare that no Generative AI was used in the creation of this manuscript.

Conflict of interest

The authors have declared no conflict of interest.

REFERENCES

Al-Bulushi S, Manjunatha BM, Bathgate R, Rickard JP, Graaf SP (2019). Artificial insemination with fresh, liquid stored and frozen thawed semen in dromedary camels. PLoS One, 14: e0224992. https://doi.org/10.1371/journal.pone.0224992

Alfian A, Yusuf M, Ako A, Toleng A, Diansyah A, Amrullah M, Rahmat R, Hasrin H (2025). The effects of mineral mix supplementation on reproductive performance of Bali bulls. Egypt J. Vet. Sci., pp. 1–8. https://doi.org/10.21608/ejvs.2025.335655.2502

Ax RL, Dally M, Didion BA, Lenz, RW, Love CC, Varner DD, Hafez B, Bellin ME (2000). Semen evaluation. In: Hafez, E.S.E and Hafez, B. (Eds). Reproduction in Farm Animals. Seventh edition. Lippincott Williams and Wilkins. USA. pp. 365-375. https://doi.org/10.1002/9781119265306.ch25

Bianchi E, Wright GJ (2016). Sperm meets egg: The genetics of mammalian fertilization. Ann. Rev. Genet., 50: 93–111. https://doi.org/10.1146/annurev-genet-121415-121834

Diansyah AM, Yusuf M, Rahmat R, Alfian AM (2025a). Enhanced sperm sexing efficiency and quality preservation in Bali bulls using freeze-dried albumin separation media. Open Vet. J., 15: 2416–2426. https://doi.org/10.5455/OVJ.2025.v15.i6.14

Diansyah AM, Santoso S, Herdis H, Yusuf M, Priyatno TP, Maulana T, Toleng A. L, Dagong MIA, Said S, Iskandar H, Nurlatifah A, Lestari P, Affandy L, Baharun A (2025b). Identification of reproductive performance in Bali-polled bulls using computer-assisted semen analysis and plasma seminal proteomics. Vet. World, 18: 102–109. https://doi.org/10.14202/vetworld.2025.102-109

Diansyah AM, Nurlatifah A, Damaris C, Rahmat R, Saili T, Amrullah MF, Alfian AM, Adam AAS, Kaiin EM, Damaynti E (2025c). Evaluation of sperm sexing technique using BSA column medium in Garut rams: Enhancing sperm quality, kinematics, and fertility performance for improved breeding efficiency. Open Vet. J., 15: 3727–3736. https://doi.org/10.5455/OVJ.2025.v15.i8.37

Gangwar C, Kumar A, Gururaj K, Mishra AK, Ranjan R, Kumar M, Rai A. Mittal N (2024). Impact of varying doses of moringa leaf extract supplementation in the cryopreservation media on sperm quality, antioxidant capacity and antimicrobial activity of frozen-thawed buck spermatozoa. Indian J. Anim. Sci., 94: 362-368. https://doi.org/10.56093/ijans.v94i4.146821

Garner H, Hafez ESE (2000). Spermatozoa and seminal plasma. In: (eds. E.S.E. Hafez and B. Hafez). Reproduction in farm animals. Seventh edition. Lippincott Williams & Wilkins. USA. pp. 96-109. https://doi.org/10.1002/9781119265306.ch7

Handayani M, Darwanto DH, Jamhari J (2024). Etawah crossbreed goat farming for income enhancement of households in Purworejo District, Indonesia. Adv. Anim. Vet. Sci., 12: 1273–1280. https://doi.org/10.17582/journal.aavs/2024/12.7.1273.1280

Hariyono DH, Endrawati E (2023). Indigenous goat genetic resources in Indonesia: Current status and future improvement. J. Adv. Vet. Res., 13: 141-149. https://advetresearch.com/index.php/AVR/article/view/1111.

Ibáñez-Arancibia E, Farías JG, Valdebenito I (2021). Use of antioxidants and time of cold storage: Effects over viability parameters and enzymatic levels in semen of rainbow trout (Oncorhynchus mykiss, Walbaum, 1792). Braz. J. Biol., 83: e245329. https://doi.org/10.1590/1519-6984.245329

Iqbal S, Naz S, Bhutta MF, Sufyan A, Awan MA (2022). Antioxidant effect of Moringa oleifera leaves extract in extender improves post-thaw quality, kinematics, lipid peroxidation, total antioxidant capacity and fertility of water buffalo bull semen. Andrologia, 54: e14300. https://doi.org/10.1111/and.14300

Kumar M, Ranjan R, Bhardwaj A (2024). Effects of liquid storage of buck semen at refrigeration temperatures on sperm viability and fertility to develop ready to use goat semen diluent. J. App. Biol. Biotech., 12: 114-118. https://doi.org/10.7324/JABB.2024.171758

Liang J, Gao Y, Feng Z, Zhang B, Na Z, Li D (2023). Reactive oxygen species and ovarian diseases: Antioxidant strategies. Redox. Biol., 62: 102659. https://doi.org/10.1016/j.redox.2023.102659

Mohajer M, Asadzadeh N, Javaheri BH, Dadashpour DN, Masoudi R (2024). Preservation of buck semen quality during chilling storage using coenzyme Q10. Iran. J. Vet. Med., 18: 429-434.

National Research Council (2007). Nutrient requirement of small ruminant (sheep, goat, cervids, and new world camelids). The National Academies Press, Washington, D.C. pp. 271-293.

Nirmala A, Toleng AL, Yusuf M, Herdis H, Diansyah AM, Amrullah MF, Rahmat R, Rajamuddin R, Alfian AM, Hasrin H (2025). Effect of adding kasumba turate (Carthamus tinctorius L.) extract in tris-egg yolk diluent on sperm quality in Bali bulls. J. Anim. Health Prod., 13: 154-159. https://doi.org/10.17582/journal.jahp/2025/13.1.154.159

Nurlatifah A, Prihantoko KD, Menassol JB, Herdis H, Diansyah AM, Khan FA, Maidin MS (2025). Water hyacinth: A promising functional feed ingredient to optimize reproductive performance in Garut rams. Open Vet. J., 15: 4681–4690. https://doi.org/10.5455/OVJ.2025.v15.i9.72

Nurlatifah A, Diansyah AM, Churriyah AN, Yusuf M, Sahiruddin S, Masturi M, Damayanti E, Irawan F, Azis IU, Alfian AM, Adam AAS, Rahmat R, Amrullah MF (2025a). Dietary supplementation of phytogenic leaf powder enhances reproductive parameters in Bali bulls. J. Anim. Health Prod, 13: 1252-1260. https://doi.org/10.17582/journal.jahp/2025/13.4.1252.1260

Pardede BP, Agil M, Yudi Y, Supriatna I (2020). Relationship of frozen-thawed semen quality with the fertility rate after being distributed in the Brahman cross breeding program. Vet. World, 13: 2649–2657. https://doi.org/10.14202/vetworld.2020.2649-2657

Pereira GR, Becker EG, Siqueira LC, Ferreira R, Severo CK, Truzzi VS, Oliveira JFC, Gonçalves PBD (2010). Assessment of bovine spermatozoa viability using different cooling protocols prior to cryopreservation. Ital. J. Anim. Sci., 9: 465-470. https://doi.org/10.4081/ijas.2010.e88

Rahayu J, Purwantara B, Said S, Arifiantini R (2025). Comparison of two methods for sperm plasma membrane integrity assessment in frozen Murrah buffalo semen. J. Ilmu Ternak Vet., 30: 19-27. https://doi.org/10.14334/jitv.v30i1.3488

Revell SG, Mrode RA (1994). An osmotic resistance test for bovine semen. Anim. Reprod. Sci., 36: 77-86. https://doi.org/10.1016/0378-4320(94)90055-8

Rizal M, Syarifuddin NA, Sumantri I, Wahdi A, Riyadhi M, Rizqiana S, Rifani MA (2025). Cryopreservation of boer crossbreed goat semen with andromed extender and moringa leaf extract. Acta Vet. Indones., 13(2).

Saratsi A, Samartzi F, Panagiotidis I, Basioura A, Tsiokos D, Ligda C, Rekkas CA (2023). Post-thaw parameters of buck semen quality after soy lecithin extender supplementation with fumaric acid. Vet. Sci., 10: 569. https://doi.org/10.3390/vetsci10090569

Shah SAH, Haider MS, Ahmed H, Fayyaz MH, Andrabi SMH (2022). Validation of double freezing protocol for Beetal buck (Capra hircus) spermatozoa. Reprod. Domest. Anim., 57: 1615–1622. https://doi.org/10.1111/rda.14238

Shan S, Xu F, Hirschfeld M, Brenig B (2021). Sperm lipid markers of male fertility in mammals. Int. J. Mol. Sci., 22: 8767. https://doi.org/10.3390/ijms22168767

SNI 4869.3-2023 Semen beku – Bagian 3: Kambing dan Domba. Badan Standardisasi Nasional (BSN), Jakarta.

Song M, Jia B, Dai D, Xu X, Cao J, Guo J, Wang L, Zhong T, Zhan S, Li L, Zhang H (2025). Effect of chitosan on buck semen quality and semen plasma metabolites during low-temperature storage. Front. Vet. Sci., 13:1544234.

Sukmawati E, Arifiantini RI, Purwantara B (2014). Freezing capacity of sperm on various type of superior bulls. J. Ilmu Ternak Vet., 19: 168-175. https://doi.org/10.14334/jitv.v19i3.1079

Susilawati T (2011). Spermatologi (In Indonesian). Universitas Brawijaya Press (UB Press), Malang. pp.91-140.

Suyadi S, Wahjuningsih S, Septian WA, Furqon A, Putri RF, Nugraha CD (2021). Reproductive performance and fertility index of Etawah-crossbred goats based on several parities at goat breeding station-Singosari, Malang, Indonesia. IOP Conf. Ser. Earth Environ. Sci., 788: 012136. https://doi.org/10.1088/1755-1315/788/1/012136

Syarifuddin NA, Rizal M, Riyadhi M, Wahdi A (2022). Libido and sperm quality of the etawah cross-breed fed urea moringa molasses multi-nutrient block supplement. J. Hunan Univ. Nat. Sci., 49: 131-140. https://doi.org/10.55463/issn.1674-2974.49.3.14

Syarifuddin NA, Rizal M, Riyadhi M, Wahdi A, Yusuf M, Syamsu JA, Diansyah AM, Rahmat R (2025). Moringa oleifera enriched UMMB improves semen quality and fertility in Etawah cross bucks. Adv. Anim. Vet. Sci., 13: 2225-2233. https://doi.org/10.17582/journal.aavs/2025/13.10.2225.2233

Tanga BM, Qamar AY, Raza S, Bang S, Fang X, Yoon K, Cho J (2021). Semen evaluation: methodological advancements in sperm quality-specific fertility assessment: A review. Anim. Biosci., 34: 1253–1270. https://doi.org/10.5713/ab.21.0072

Yang DH, Standley NT, Xu ZZ (2018). Application of liquid semen technology under the seasonal dairy production system in New Zealand. Anim. Reprod. Sci., 194: 2–10. https://doi.org/10.1016/j.anireprosci.2018.01.004

Yusuf M, Diansyah AM, Sahiruddin S, Masturi M, Maulana T, Said S, Rahmat R, Alfian AM, Adam AA, Yusri AN, Nurlatifah A (2026). Integrated Metabolomic and Functional Assessment of Sexed Frozen Semen in Holstein Friesian Bulls. Trop. Anim. Sci. J., 49: 19-29. https://doi.org/10.5398/tasj.2026.49.1.19