The Drosera Extract as an Alternative In Vitro Supplement to Animal Semen: Effects on Bovine Spermatozoa Activity and Oxidative Balance

Eva Tvrdá


In vitro storage and processing of animal semen is considered to be a risk factor to spermatozoa activity, possibly leading to reduced fertility and litter sizes following artificial insemination (AI). A variety of substances isolated from natural resources have the potential to exhibit protective or antioxidant properties on the spermatozoon, thus they may extend the lifespan of stored semen. Drosera (Drosera rotundifolia L.) has been shown to possess antimicrobial, anti-inflammatory and antioxidant properties, making the plant extract a potential candidate for preserving liquid animal semen during in vitro storage. This study compared the ability of different concentrations of Drosera extract on the motility, viability and superoxide production of bovine spermatozoa during different time periods (0, 2, 6, 12 and 24h) of in vitro culture. Spermatozoa motility was assessed using the SpermVisionTM CASA (Computer aided sperm analysis) system. Cell viability was examined using the metabolic activity MTT assay and the nitroblue-tetrazolium (NBT) test was applied to quantify the intracellular superoxide formation. The CASA analysis revealed that Drosera extract supplementation was able to prevent a rapid decline of spermatozoa motility, especially in the case of concentrations ranging between 1 and 5 mg/mL (P<0.001 with respect to Times 6h, 12h and 24h). At the same time, concentrations ranging between 1 and 10 mg/mL of the extract led to a significant preservation of the cell viability throughout short-term (P<0.05 in case of Time 6h) as well as long-term periods of the experiment (P<0.01 with respect to Time 12h, and P<0.001 in case of Time 24h). 10 and 5 mg/mL of the extract exhibited antioxidant characteristics, translated into a significant reduction of the intracellular superoxide production, particularly notable at Times 12h (P<0.01) and 24h (P<0.001). The results indicate that the Drosera extract is capable of delaying the damage inflicted to the spermatozoon by the in vitro environment.


Drosera, spermatozoa, motility, viability, superoxide production

Full Text:



Donnelly, E.T., McClure, N., Lewis, S.E.M., The effect of ascorbate and α-tocopherol supplementation in vitro on DNA integrity and hydrogen peroxide-induced DNA damage in human spermatozoa. Mutagenesis, 1999, 14(5), 505-512.

Agarwal, A., Sekhon, L., The role of antioxidant therapy in the treatment of male infertility. Human Fertility, 2010, 13(4), 217–225.

Saleh, R.A., Agarwal, A., Oxidative stress and male infertility: from research bench to clinical practice. Journal of Andrology, 2002, 23(6), 737-752.

Alarcón de la Rastra, C., Curcumin: a promising spice for therapeutics. Molecular Nutrition & Food Research, 2008, 52, 985.

Asirvatham, R., Christina, A.J.M., Anticancer activity of Drosera indica L., on Dalton’s Lymphoma Ascites (DLA) bearing mice. Journal of Intercultural Ethnopharmacology, 2012, 1(3), 9-14.

Asirvatham, R., Arockiasam, J.M.C., Murali, A., In vitro antioxidant and anticancer activity studies on Drosera indica L. (Droseraceae). Advanced Pharmaceutical Bulletin, 2013, 3(1), 115-120.

Fukushima, K., Nagai, K., Hoshi, Y., Masumoto, S., Mikami, J., Takahashi, Y., Oike, H., Kobori, M., Drosera rotundifolia and Drosera tokaiensis suppress the activation of HMC-1 human mast cells. Journal of Ethnopharmacology, 2009, 125, 90-96.

Kačániová, M., Ďurechová, D., Vuković, N., Kántor, A., Petrová, J., Hleba, L., Vatľák, A., Antimicrobial activity of Drosera rotundifolia L. Scientific Papers: Animal Science and Biotechnologies, 2014, 47(2), 366-369.

Michalko, J., Matušíková, I., The role of glucanases in digestion of carnivorous plant Drosera rotundifolia L. Journal of Microbiology, Biotechnology and Food Sciences, 2012, 1, 671-678.

Massanyi, P., Chrenek, P., Lukáč, N., Makarevich, A.V., Ostro, A., Živčák, J., Bulla, J., Comparison of different evaluation chambers for analysis of rabbit spermatozoa motility using CASA system. Slovak Journal of Animal Science, 2008, 41, 60-66.

Mosmann, T., Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. Journal of Immunological Methods, 1983, 65, 55–63.

Knazicka, Z., Tvrda, E., Bardos, L., Lukac, N., Dose- and time-dependent effect of copper ions on the viability of bull spermatozoa in different media. Journal of Environmental Science and Health Part A, 2012, 47, 1294-1300.

Esfandiari, N., Sharma, R.K., Saleh, R.S., Thomas, Jr., A.J., Agarwal, A., Utility of the nitroblue tetrazolium reduction test for assessment of reactive oxygen species production by seminal leukocytes and spermatozoa. Journal of Andrology, 2003, 24(6), 862-870.

Tvrdá, E., Lukáč, N., Lukáčová, J., Kňažická, Z., Massányi, P., Stimulating and protective effects of vitamin E on bovine spermatozoa. Journal of Microbiology, Biotechnology and Food Sciences, 2013, 2(1), 1386-1395.

Asirvatham, R., Arockiasam, J.M.C., Murali, A., Antitumor activity of ethanol and aqueous extracts of Drosera burmannii vahl. in EAC bearing mice. Spatula, 2012, 2(2), 83-88.

Krychowiak, M., Grinholc, M., Banasiuk, R., Krauze-Baranowska, M., Głód, D., Kawiak, A., Królicka, A., Combination of silver nanoparticles and Drosera binata extract as a possible alternative for antibiotic treatment of burn wound infections caused by resistant Staphylococcus aureus. PLoS One, 2014, 9(12), e115727.

Tvrdá, E., Lukáč, N., Lukáčová, J., Jambor, T., Hashim, F., Massányi, P., Dose- and time-dependent in vitro effects of quercetin on bovine spermatozoa activity and superoxide production. Folia Veterinaria, 2014, 58(4), 224-231.

Aravindakshan, M., Chauhan, P. S., Sundaram, K., Studies on germinal effects of quercetin, a naturally occurring flavonoid. Mutation Research, 1985, 144, 99-106.

Taepongsorat, L., Tangpraprutgul, P., Kitana, N., Malaivijitnond, S., Stimulating effects of quercetin on sperm quality and reproductive organs in adult male rats. Asian Journal of Andrology, 2008, 10, 249-258.

Córdoba, M., Pintos, L., Beconi, M. T., Differential activities of malate and isocitrate NAD(P)-dependent dehydrogenases are involved in the induction of capacitation and acrosome reaction in cryopreserved bovine spermatozoa. Andrologia, 2005, 37, 40-46.

Li, M. W., Yudin, A. I., VandeVoort, C. A., Sabeur, K., Primakoff, P., Overstreet, J. W., Inhibition of monkey sperm hyaluronidase activity and heterologous cumulus penetration by flavonoids. Biology of Reproduction, 1997, 56, 1383-1389.

Çeribaş, A.O., Sakin, F., Türk, G., Sönmez, M., Ateşşahin, A., Impact of ellagic acid on adriamycin-induced testicular histopathological lesions, apoptosis, lipid peroxidation and sperm damages. Experimental and Toxicologic Pathology, 2012, 64(7-8), 717-724.

Ateşşahin, A., Türk, G., Yilmaz, S., Sönmez, M., Sakin, F., Ceribasi, A.O., Modulatory effects of lycopene and ellagic acid on reproductive dysfunction induced by polychlorinated biphenyl (Aroclor 1254) in male rats. Basic & Clinical Pharmacology & Toxicology, 2010, 106(6), 479-489.

Sukardi, S., Yaakub, H., Ganabadi, S., Ahmad, Z., Abdul Hamid, R., Impaired sperm parameters of Balb/c mice fed plumbagin. Planta Medica, 2009, 75, PJI123.

Madhavan, V., Kumar, B.H.P., Murali, A., Yoganarasimhan, S.N., Antifertility activity of Drosera burmannii. Pharmaceutical Biology, 2009, 47(2), 128-131.

Anandakumar, A.M., Paulsamy, S., Sathishkumar, P., Senthilkumar, P., Preliminary phytochemical studies for the quantification of secondary metabolites of medicinal importance in the plant Acalypha fruticosa Forssk. Journal of Applied and Natural Science, 2009, 1(1), 41-43.

Klein, S.M., Cohen, G., Cederaum, A.I. Production of formaldehyde during metabolism of dimethyl sulphoxide by hydroxyl radical generating system. Biochemistry, 1991, 20, 6006-6012.

Thambiraj, J., Paulsamy, S., Sevukaperumal, R., Evaluation of in vitro antioxidant activity in the traditional medicinal shrub of western districts of Tamilnadu, India, Acalypha fruticosa Forssk. (Euphorbiaceae). Asian Pacific Journal of Tropical Biomedicine, 2012, 2(1), S127-130.

Gulcin, I., Beydemir, S., Alici, H.A., Elmastas, M., Buyukokuroglu, M.E., In vitro antioxidant properties of morphine. Pharmacological Research, 2004, 49(1), 59-66.


  • There are currently no refbacks.

Copyright (c)

Scientific Papers Animal Science and Biotechnologies publish open access articles under the terms of the Creative Commons Attribution 4.0 Licence (read full legal code).


ISSN print 1841 - 9364
ISSN online 2344 - 4576
ISSN-L  1841 - 9364
(former ISSN 1221-5287, E-ISSN 1841-9364)

PAPER ACCESS: Full text articles available for free
FREQUENCY: Semiannual

Banat´s University of Agricultural Sciences and Veterinary Medicine "King Michael I of Romania" from Timisoara
Bioengineering Faculty of Animal Resources
300645, Timişoara, Calea Aradului 119, Romania

E-mail: spasb [at]
Phone: +40-256-277160, Fax.: +40-256-277110