Solo Black Garlic fermented whey kefir improves sperm quality of male mice (Mus musculus) induced by lead acetate
Abstract
Globally, men experience a decline in total fertility, characterized by a significant decline in sperm quality in terms of concentration, motility, and morphology. Environmental factors can modulate this through exposure to toxic elements, such as the heavy metal lead, in the workplace. Lead can accumulate in nature and contaminate drinking water supply systems. A series of negative responses can occur and disrupt spermatozoa production and quality. Natural ingredients, such as solo black garlic (SBG), have the potential to maintain conditions in reproductive function against the adverse effects of lead. This study utilized SBG fermented with whey kefir to investigate its effect and optimal dosage on improving spermatozoa quality in male mice induced by lead acetate. The test animals were 25 Swiss Webster male mice aged 12-14 weeks, divided into five treatment groups (n = 5 per group), namely negative control (KN), distilled water; positive control (KP), lead acetate 75 mg/kg BW; and three SBG treatment groups given lead acetate (SBG-60, SBG-120, and SBG-180). SBG was given together with lead acetate at doses of 60, 120, and 180 mg/kg BW for 30 days. The results showed SBG-120 restored concentration (31.00 × 106/ml) and motility (80.83%) toward negative-control levels; morphology abnormalities decreased by 30.50%, which were not significantly different (p > 0.05) from the negative control. Therefore, SBG with a dose of 120 mg/kg BW is optimum for maintaining spermatozoa quality in male mice exposed to lead acetate.
Keywords
Full Text:
PDFReferences
Abilash, D., & Sridharan, T. (2024). Impact of air pollution and heavy metal exposure on sperm quality: A clinical prospective research study. Toxicology Reports, 13. https://doi.org/10.1016/J.TOXREP.2024.101708
Afiati, F., Setiyoningrum, F., Priadi, G., & Handoyo, C. H. (2020). Characteristics of Solo Black Garlic Fermented in Kombucha Black Tea. IOP Conference Series: Earth and Environmental Science, 439(1), 012053. https://doi.org/10.1088/1755-1315/439/1/012053
Afzaal, M., Saeed, F., Rasheed, R., Hussain, M., Aamir, M., Hussain, S., Mohamed, A. A., Alamri, M. S., & Anjum, F. M. (2021). Nutritional, biological, and therapeutic properties of black garlic: a critical review. International Journal of Food Properties, 24(1), 1387–1402. https://doi.org/10.1080/10942912.2021.1967386
Agarwal, A., Sharma, R., Gupta, S., Finelli, R., Parekh, N., Selvam, M. K. P., Henkel, R., Durairajanayagam, D., Pompeu, C., Madani, S., Belo, A., Singh, N., Covarrubias, S., Darbandi, S., Sadeghi, R., Darbandi, M., Vogiatzi, P., Boitrelle, F., Simopoulou, M., Shah, R. (2021). Sperm Morphology Assessment in the Era of Intracytoplasmic Sperm Injection: Reliable Results Require Focus on Standardization, Quality Control, and Training. The World Journal of Men’s Health, 40(3), 347. https://doi.org/10.5534/WJMH.210054
Agarwal, A., Sharma, R., Gupta, S., Finelli, R., Parekh, N., Selvam, M. K. P., Pompeu, C. P., Madani, S., Belo, A., Darbandi, M., Singh, N., Darbandi, S., Covarrubias, S., Sadeghi, R., Arafa, M., Majzoub, A., Caraballo, M., Giroski, A., McNulty, K., Henkel, R. (2021). Standardized Laboratory Procedures, Quality Control and Quality Assurance Are Key Requirements for Accurate Semen Analysis in the Evaluation of Infertile Male. The World Journal of Men’s Health, 40(1), 52. https://doi.org/10.5534/WJMH.210022
Ai, T. T., & Huong, N. T. (2018). Research on the Production of Black Garlic Juice. International Journal of Pharmaceutical Science Invention, 7(5), 07–14. www.ijpsi.org
Amida, N., Lisdiana, L., Christijanti, W., & Iswari, R. S. (2021). Efek ekstrak black garlic terhadap kualitas spermatozoa tikus setelah dipapar asap rokok. Prosiding Seminar Nasional Biologi, 298–303.
Amor, S., González-Hedström, D., Martín-Carro, B., Inarejos-García, A. M., Almodóvar, P., Prodanov, M., García-Villalón, A. L., & García, M. G. (2019). Beneficial Effects of an Aged Black Garlic Extract in the Metabolic and Vascular Alterations Induced by a High Fat/Sucrose Diet in Male Rats. Nutrients, 11(1), 153. https://doi.org/10.3390/NU11010153
Andriani, D., & Murtisiwi, L. (2018). Penetapan Kadar Fenolik Total Ekstrak Etanol Bunga Telang (Clitoria ternatea L.) dengan Spektrofotometri Uv Vis. Cendekia Journal of Pharmacy, 2(1), 32–38. https://cjp.jurnal.stikescendekiautamakudus.ac.id/index.php/cjp/article/view/15
Aprianti, H. (2023). Perbandingan senyawa fitokimia, kadar flavonoid total dan aktivitas antibakteri bawang putih (Allium sativum L.) dan bawang hitam terhadap Streptococcus sanguinis. UIN Sunan Gunung Djati Bandung.
Asadpour, R., Azari, M., Hejazi, M., Tayefi, H., & Zaboli, N. (2013). Protective effects of garlic aquous extract (Allium sativum), vitamin E, and N-acetylcysteine on reproductive quality of male rats exposed to lead. Veterinary Research Forum : An International Quarterly Journal, 4(4), 251.
Bani, I. I., Zulkarnain, Z., Gholib, G., Syahrizal, D., Nugraha, T. P., Ramadhan, A., Akmal, M., & Maulina, N. (2024). Effect of black garlic (Allium sativum) on gonadosomatic index, follicle-stimulating hormone level and spermatozoa quality: A study in monosodium glutamate-exposed rat model. Narra J, 4(2), e617–e617. https://doi.org/10.52225/NARRA.V4I2.617
Becerril-Chávez, H., Colín-González, A. L., Villeda-Hernández, J., Galván-Arzate, S., Chavarría, A., Eduarda de Lima, M., Túnez, I., & Santamaría, A. (2017). Protective effects of S-allyl cysteine on behavioral, morphological and biochemical alterations in rats subjected to chronic restraint stress: Antioxidant and anxiolytic effects. Journal of Functional Foods, 35, 105–114. https://doi.org/10.1016/J.JFF.2017.05.034
Behnejad, G., Mohammadi, T., & Soleimanzadeh, A. (2025). Allicin and hesperidin protect sperm production from environmental toxins in mice. Scientific Reports, 15(1), 1–14. https://doi.org/10.1038/S41598-025-97730-9
Bhattacharjee, N. V., Schumacher, A. E., Aali, A., Abate, Y. H., Abbasgholizadeh, R., Abbasian, M., Abbasi-Kangevari, M., Abbastabar, H., ElHafeez, S. A., Abd-Elsalam, S., Abdollahi, M., Abdollahifar, M. A., Abdoun, M., Abdullahi, A., Abebe, M., Abebe, S. S., Abiodun, O., Abolhassani, H., Abolmaali, M., Vollset, S. E. (2024). Global fertility in 204 countries and territories, 1950–2021, with forecasts to 2100: a comprehensive demographic analysis for the Global Burden of Disease Study 2021. The Lancet, 403(10440), 2057–2099. https://doi.org/10.1016/S0140-6736(24)00550-6
Bocu, K., Boeri, L., Mahmutoglu, A. M., & Vogiatzi, P. (2024). Can lifestyle changes significantly improve male fertility: A narrative review? Arab Journal of Urology, 1–11. https://doi.org/10.1080/20905998.2024.2421626
Chao, H. H., Zhang, Y., Dong, P. Y., Gurunathan, S., & Zhang, X. F. (2023). Comprehensive review on the positive and negative effects of various important regulators on male spermatogenesis and fertility. Frontiers in Nutrition, 9, 1063510. https://doi.org/10.3389/FNUT.2022.1063510/BIBTEX
Choi, I., Cha, H., & Lee, Y. (2014). Physicochemical and Antioxidant Properties of Black Garlic. Molecules, 19(10), 16811–16823. https://doi.org/10.3390/molecules191016811
Fauziyah, A., & Dwijananti, P. (2013). Pengaruh Radiasi Sinar X terhadap Motilitas Sperma Pada Tikus Mencit (Mus musculus). Jurnal Pendidikan Fisika Indonesia, 9(1), 1–5. https://doi.org/10.15294/jpfi.v9i1.2585
Ghafil, J. S., & Al-Taee, N. S. N. (2020). Effect of Garlic and Celery Extracts on Lead Toxicity in Male Mice. Medico-Legal Update, 20(2). https://doi.org/10.37506/mlu.v20i2.1178
Ghalehkandi, J. G. (2014). Garlic (Allium sativum) juice protects from semen oxidative stress in male rats exposed to chromium chloride. Anim. Reprod, 11(4), 526–532.
Giulioni, C., Maurizi, V., De Stefano, V., Polisini, G., Teoh, J. Y. C., Milanese, G., Galosi, A. B., & Castellani, D. (2023). The influence of lead exposure on male semen parameters: A systematic review and meta-analysis. Reproductive Toxicology, 118. https://doi.org/10.1016/j.reprotox.2023.108387
Haryanto, B. (2016). Lead exposure from battery recycling in Indonesia. Reviews on Environmental Health, 31(1), 13–16. https://doi.org/10.1515/reveh-2015-0036
Huang, B., Wang, Z., Kong, Y., Jin, M., & Ma, L. (2023). Global, regional and national burden of male infertility in 204 countries and territories between 1990 and 2019: an analysis of global burden of disease study. BMC Public Health, 23(1). https://doi.org/10.1186/s12889-023-16793-3
Hur, S. J., Lee, S. Y., Kim, Y. C., Choi, I., & Kim, G. B. (2014). Effect of fermentation on the antioxidant activity in plant-based foods. Food Chemistry, 160, 346–356. https://doi.org/10.1016/J.FOODCHEM.2014.03.112
Ihsani, N., Hernahadini, N., Pertiwi, L., N.F., M. K., & Fadhillah, S. N. (2019). Pengaruh Pemberian Air Jeruk Nipis (Citrus aurantifolia) Terhadap Konsentrasi dan Motilitas Spermatozoa Mencit (Mus musculus) Setelah Terpapar Asap Rokok. Jurnal Kedokteran YARSI, 27(1), 035–042. https://doi.org/10.33476/JKY.V27I1.779
Irvanto, R., Paramita, W., Susilowati, S., Damayanti, T. L., & Safitri, E. (2021). Kualitas Motilitas dan Viabilitas Spermatozoa Dari Semen Afkir Sapi Limousin Pada Pengencer Susu Skim Kuning Telur Sitrat dengan Penambahan Berbagai Kadar Glukosa. Ovozoa: Journal of Animal Reproduction, 7(2), 96–101. https://doi.org/https://doi.org/10.20473/ovz.v7i2.2018.96-101
Kowalczyk, A. (2021). The Role of the Natural Antioxidant Mechanism in Sperm Cells. Reproductive Sciences, 29(5), 1387. https://doi.org/10.1007/S43032-021-00795-W
Kumar, N., & Singh, A. (2015). Trends of male factor infertility, an important cause of infertility: A review of literature. Journal of Human Reproductive Sciences, 8(4), 191. https://doi.org/10.4103/0974-1208.170370
Leary, S. L., Underwood, W., Anthony, R., Cartner, S., Grandin, T., Greenacre. Cheryl, Brant-Gwaltney, S., McCrackin, M. A., Meyer, R., MIller, D., Shearer, J., Turner, T., & Yanong, R. (2020). AVMA guidelines for the euthanasia of animals : 2020 edition (2020 Edition). American Veterinary Medical Association.
Lestari, V. A. (2017). Pengaruh Konsentrasi Fenol dan Sumber Sinar terhadap Degradasi Fenol Menggunakan Fotokatalis Lapis Tipis TiO2/N-Kitosan. Universitas Brawijaya.
Libero, M. L., Montero-Hidalgo, A. J., Recinella, L., Luque, R. M., Generali, D., Acquaviva, A., Orlando, G., Ferrante, C., Menghini, L., Di Simone, S. C., Nilofar, N., Chiavaroli, A., Brunetti, L., & Leone, S. (2024). The Protective Effects of an Aged Black Garlic Water Extract on the Prostate. Nutrients , 16(17), 3025. https://doi.org/10.3390/NU16173025/S1
Mai, H., Ke, J., Li, M., He, M., Qu, Y., Jiang, F., Cai, S., Xu, Y., Fu, L., Pi, L., Zhou, H., Yu, H., Che, D., Gu, X., Zhang, J., & Zuo, L. (2023). Association of living environmental and occupational factors with semen quality in chinese men: a cross-sectional study. Scientific Reports, 13, 15671. https://doi.org/10.1038/S41598-023-42927-Z
Martinez, G. (2022). First-line Evaluation of Sperm Parameters in Mice (Mus musculus). Bio-Protocol, 12(20). https://doi.org/10.21769/BIOPROTOC.4529
Massányi, P., Massányi, M., Madeddu, R., Stawarz, R., & Lukáč, N. (2020). Effects of Cadmium, Lead, and Mercury on the Structure and Function of Reproductive Organs. Toxics, 8(4), 94. https://doi.org/10.3390/TOXICS8040094
Mathur, P., Rani, K., Bhatnagar, P., & Flora, S. J. S. (2023). Incidence of Morphological Defects in Sperm of Mice Exposed to Hospital Effluent. Toxics, 11(5), 418. https://doi.org/10.3390/TOXICS11050418
Montoto, L. G., Magaña, C., Tourmente, M., Martín-Coello, J., Crespo, C., Luque-Larena, J. J., Gomendio, M., & Roldan, E. R. S. (2011). Sperm Competition, Sperm Numbers and Sperm Quality in Muroid Rodents. PLoS ONE, 6(3), e18173. https://doi.org/10.1371/JOURNAL.PONE.0018173
Mustafa, H. N. (2023). Ameliorative potential of the quercetin on lead-induced testicular damage: morphohistometric and biochemical analysis. African Journal of Urology, 29(1), 1–8. https://doi.org/10.1186/S12301-023-00369-Z
Nasr, N. E., Elmadawy, M. A., Almadaly, E. A., Abdo, W., & Zamel, M. M. (2017). Garlic Powder Attenuates Apoptosis Associated with Lead Acetate-Induced Testicular Damage in Adult Male Rats. Alexandria Journal of Veterinary Sciences, 54(1), 70–70. https://doi.org/10.5455/AJVS.268149
Nguyen, T. T. A., Le, C. T., Nankervis, L., Van, C. H., Vu, S. Van, Dao, M. H., Thi Pham, K., & Van Phan, U. (2025). Black garlic (Allium sativum) powder, a promising feed supplementation improves hematological and immunological indices, body composition, and growth performance of golden pompano (Trachinotus blochii). Regional Studies in Marine Science, 82, 104034. https://doi.org/10.1016/J.RSMA.2025.104034
Omotoso, G. O., Oyewopo, A. O., Kadir, R. E., Olawuyi, S. T., & Jimoh, A. A. G. (2009). Effects Of Aqueous Extract Of Allium Sativum (Garlic) On Semen Parameters In Wistar Rats. The Internet Journal of Urology, 7(2). https://doi.org/10.5580/A5412
Panner Selvam, M. K., Agarwal, A., Henkel, R., Finelli, R., Robert, K. A., Iovine, C., & Baskaran, S. (2020). The effect of oxidative and reductive stress on semen parameters and functions of physiologically normal human spermatozoa. Free Radical Biology and Medicine, 152, 375–385. https://doi.org/10.1016/J.FREERADBIOMED.2020.03.008
Raj, K., & Das, A. P. (2023). Lead pollution: Impact on environment and human health and approach for a sustainable solution. Environmental Chemistry and Ecotoxicology, 5, 79–85. https://doi.org/10.1016/J.ENCECO.2023.02.001
Ramos-Treviño, J., Bassol-Mayagoitia, S., Hernández-Ibarra, J. A., Ruiz-Flores, P., & Nava-Hernández, M. P. (2018). Toxic Effect of Cadmium, Lead, and Arsenic on the Sertoli Cell: Mechanisms of Damage Involved. DNA and Cell Biology, 37(7), 600–608. https://doi.org/10.1089/DNA.2017.4081
Rana, M. M., Shiozawa, K., Mukai, K., Takayanagi, K., Eguchi, K., Sultana, H., Ohsaki, Y., Komai, M., & Shirakawa, H. (2021). S-allyl Cysteine Enhances Testosterone Production in Mice and Mouse Testis-Derived I-10 Cells. Molecules, 26(6), 1697. https://doi.org/10.3390/MOLECULES26061697
Rombe, I. S., Purwaningsih, & Tethool, A. N. (2023). Kualitas Spermatozoa Cauda Epididimis Sapi Bali Pada Kelompok Umur Berbeda. Jambura Journal of Animal Science, 5(2), 23–31. https://doi.org/10.35900/JJAS.V5I2.19236
Saeidpour, Z., Bouloorchi Tabalvandani, M., Javadizadeh, S., & Badieirostami, M. (2024). Enhanced sperm isolation via bulk acoustic waves for high-throughput motility screening. Scientific Reports, 14(1), 1–11. https://doi.org/10.1038/s41598-024-78536-7
Saputri, M. (2019). Perbandingan Kadar Flavonoid Total Ekstrak Daun Sirsak (Annona muricata L.) Berdasarkan Kekeringan Bahan. Repository Poltekkes Kemenkes Palembang. https://repository.poltekkespalembang.ac.id/items/show/1069
Senapati, S. K., Dey, S., Dwivedi, S. K., & Swarup, D. (2001). Effect of garlic (Allium sativum L.) extract on tissue lead level in rats. Journal of Ethnopharmacology, 76(3). https://doi.org/10.1016/S0378-8741(01)00237-9
Setiyoningrum, F., Priadi, G., & Afiati, F. (2022). Chemical properties of solo black garlic fermented by Saccharomyces cerevisiae. IOP Conference Series: Earth and Environmental Science, 976(1), 012044. https://doi.org/10.1088/1755-1315/976/1/012044
Sharma, A., Sharma, V., & Kansal, L. (2010). Amelioration of lead-induced hepatotoxicity by Allium sativum extracts in Swiss albino mice. Libyan Journal of Medicine, 5(1). https://doi.org/10.4176/091107
Sitepu, S. A., Marisa, J., & Farid, M. M. (2023). Abnormalitas Spermatozoa Semen Segar Kambing dengan Pemberian Pakan Suplemen Ekstrak Kunyit. Scenario (Seminar of Social Sciences Engineering and Humaniora), 260–266. https://jurnal.pancabudi.ac.id/index.php/scenario/article/view/4524
Solichah, A., & Herdyastuti, N. (2021). Pengaruh Lama Pemanasan Proses Fermentasi terhadap Kadar Fenolik Total dan Aktivitas Antioksidan Bawang Hitam. Unesa Journal of Chemistry, 10(3), 280–287. https://doi.org/10.26740/UJC.V10N3.P280-287
Suru, S. M. (2008). Onion and garlic extracts lessen cadmium-induced nephrotoxicity in rats. BioMetals, 21(6), 623–633. https://doi.org/10.1007/s10534-008-9148-5
Susila Ningsih, I., Chatri, M., & Advinda, L. (2023). Flavonoid Active Compounds Found In Plants Senyawa Aktif Flavonoid yang Terdapat Pada Tumbuhan. Serambi Biologi, 8(2), 126–132.
Takemura, S., Ichikawa, H., Naito, Y., Takagi, T., Yoshikawa, T., & Minamiyama, Y. (2014). S-allyl cysteine ameliorates the quality of sperm and provides protection from age-related sperm dysfunction and oxidative stress in rats. Journal of Clinical Biochemistry and Nutrition, 55(3), 155–161. https://doi.org/10.3164/JCBN.14-39
Thalia, C. U., Chrisnasari, R., & Dewi, A. D. R. (2020). Pengaruh Pengolahan Terhadap Nilai Fungsional Bawang Putih (Allium sativum). Keluwih: Jurnal Sains Dan Teknologi, 1(1), 1–14. https://doi.org/10.24123/SAINTEK.V1I1.2782
Turner, K. A., Rambhatla, A., Schon, S., Agarwal, A., Krawetz, S. A., Dupree, J. M., & Avidor-Reiss, T. (2020). Male Infertility is a Women’s Health Issue—Research and Clinical Evaluation of Male Infertility Is Needed. Cells, 9(4), 990. https://doi.org/10.3390/CELLS9040990
Xie, J., Yu, J., Fan, Y., Zhao, X., Su, J., Meng, Y., ... & Wang, Z. (2020). Low dose lead exposure at the onset of puberty disrupts spermatogenesis-related gene expression and causes abnormal spermatogenesis in mouse. Toxicology and Applied Pharmacology, 393, 114942. https://doi.org/10.1016/j.taap.2020.114942
Yahya, Y. (2015). Perbedaan Tingkat Laju Osmosis Antara Umbi Solonum tuberosum dan Doucus carota. Jurnal Biology Education, 4(1). https://doi.org/10.32672/JBE.V4I1.145
Yeni, Y., Cicek, B., Yildirim, S., Bolat, İ., & Hacimuftuoglu, A. (2024). Ameliorating effect of S-Allyl cysteine (Black Garlic) on 6-OHDA mediated neurotoxicity in SH-SY5Y cell line. Toxicology Reports, 13, 101762. https://doi.org/10.1016/J.TOXREP.2024.101762
Zeng, X., Wang, Y., Yang, S., Liu, Y., Li, X., & Liu, D. (2023). The functionalities and applications of whey/whey protein in fermented foods: a review. Food Science and Biotechnology, 33(4), 769. https://doi.org/10.1007/S10068-023-01460-5
Zi, T., Liu, Y. N., Zhang, Y. S., Wang, Z. L., Wang, Z. X., Zhan, S., Peng, Z., Li, N., Liu, X. X., & Liu, F. J. (2022). Protective effect of melatonin on alleviating early oxidative stress induced by DOX in mice spermatogenesis and sperm quality maintaining. Reproductive Biology and Endocrinology, 20(1), 1–11. https://doi.org/10.1186/S12958-022-00977-4
DOI: http://dx.doi.org/10.30821/biolokus.v8i2.4708
Refbacks
- There are currently no refbacks.
Copyright (c) 2025 Jurnal Biolokus : Jurnal Penelitian Pendidikan Biologi dan Biologi
indexed by :










