Department of Biological Sciences, Faculty of Science, King Abdulaziz University, Jeddah, Saudi Arabia

Corresponding author email: iullah@kau.edu.sa

Article Publishing History

INTRODUCTION

Nigera sativa (Black seed) is a herbaceous plant, grows annually; however, is also cultivated in different parts of the world, including Asia, Africa, and America. Black seeds are derived from the common fennel flower plant (Nigella sativa) of the Kennebug family (Ranuncula family) (Ahmad et al., 2021; Soltane et al., 2021). Black seeds have long been known and used for medicines ( Ali and Meitei, 2011). The black seeds have been  also  used and  recommended for the treatment of minor disorders such as skin conditions and allergies (Prashar et al., 2004). Prophet Muhammad (PBUH) illumined in the medicinal importance of the black seed. A large number of studies  have been  reported in a  detailed  review  on the  medicinal use of  super foods of  plant origin, based on the recommendations of  Prophet Muhammad, (PBUH)  by ). In recent years, research in the field of agriculture has focused on the economy of plants influenced by selective minor elements. Secondary metabolites from plant extract acts as reducing and stabilizing agents to produce new metallic nanoparticles through bio-reduction reaction (Jang et al., 2020). Comparatively, using a biological method for synthesizing nanoparticles is superior to a non-biological (chemical, physical) one, the former is inexpensive easy, and eco-friendly while the latter is highly toxic and hazardous for living organisms, ( Bharali et al., 2013 Ullah et al  2020).

The mosquitoes are the carrier of several diseases known as vector-borne diseases. The diseases are including yellow fever, malaria, dengue fever, and many others. Black seeds have reported being toxic against the larvae as well as adults of the T. castaneum even at extremely lower concentrations (Dagli et al., 2015). Moreover, plants source are a potent source for treating several diseases such as Cancer, Malaria, Human immunodeficiency virus (HIV), Hepatitis, and many other serious ailments (Dris et al., 2017). it has been mentioned in one study about the punctuality and application of plant extract in different fields including medicine, nutrition, and energy (Enan, 2005).

On the other side, the application for curing different bacterial and viral diseases in pharmacological industries is a remarkable advantage (Sabbah et al., 2020; Helaly et al., 2020).  Several studies have demonstrated the antibacterial activity of black seed oil against a variety of microorganisms (Ait Said et al., 2015). This effect is shown against Salmonella spp, E. coli, Shigellas pp. The biological method is cost-effective than others due to the abundant availability of biological entities easy procedure, hence highly explored for syntheses of nanoparticles (Zuzarte et al., 2012). The present study was designed to determine the larvicidal and antimicrobial activities of the extract of black seeds.

MATERIAL AND METHODS

Methanolic extraction: The black seeds were rinsed in distilled water for removing surface dust particles. After washing the seeds were kept under sunlight for drying. The seeds were crushed with the help of liquid nitrogen and mortar. The powered seeds were transferred to the flask containing 70% methanol. After homogenization, the mixture was incubated on a magnetic stirrer at room temperature for two days. The mixture was filtered by using Whatman filter papers no.1. Furthermore, the filtrate was dried and was dissolved in deionized distilled water.

Antimicrobial activity of the extract: The fungicidal activity of the biogenic silver nanoparticles was assessed against the infectious fungi Candida albicans. This activity was determined using the colony-forming method. The C. albicans was cultured on PDA (potato dextrose agar) at 37°C, for 48 hours. Potato dextrose agar media were prepared and autoclaved. The autoclave media was transfer to the Petri plates and wait until solidification. The extract was applied on the plates followed by the transfer of C. albicans culture. The plates were incubated for 48 hours at 37°C.

Larvicidal Bioassay of the extract:The larvae of the mosquito were obtained from the Mosquito Research Laboratory, King Abdulaziz University Saudi Arabia. The larvae were reared in containers (25 × 35 × 6 cm) filled with up to the marked water supplemented with artificial food. The colony was maintained under the following conditions: 27^°C ± 2^°C, 70% ± 5% relative humidity. Different dilutions of an extract of black seed i.e., 20, 40, 60, 80, 100% (w/v) were used to assess the larvicidal activities.  The larvae were transferred to each well of the six-well culture plates. The larvae were given standard larval food and the temperature was kept 28 ± 2^°C. After 24 h of exposure, percentage mortality was noted. The experiment was repeated three times.

RESULT AND DISCUSSION

Antimicrobial activity of the extract:The spread plate method was used to assess the antimicrobial activity of the extract against C. albicans. The spread plate method is widely employed to check the susceptibility of chemical components of extracts against the microbes. The results revealed that the extract consisted of metabolites, responsible for antimicrobial activity against C. albicans (Fig. 1A and 1B). The surface of the plates i.e., control and treated showed the efficacy of the extract. No colony was observed on the plate treated with the extract.  Hazardous effects associated with the usage of synthetic chemicals, it is important to introduce natural products, of plant origin. The plant origin compounds will reduce the harmful effects in the environment. The results of our study revealed that the extract showed 100% fungicidal activity against C. albicans. It has been reported that the aqueous extract of N. sativa exhibits an inhibitory effect against C. albicans (Ait Said et al., 2015). In the other study, Al-Jabra et al. reported that the ether extract of N. sativa seed showed inhibitory effects against T. interdigitale, T. rebrand (Zuzarte et al., 2012).

Figure 1: Antimicrobial activity of the extract of black seed against Candida albicans grown on potato dextrose Agar. (A) Candida albicans grown without application of extract represents control. (B)  Candida albicans has grown with the application of extract.

Larvicidal activity of the extract:The extract showed great larvicidal activity against the larvae of the mosquitoes. The rate of mortality shown in results (Fig. 2) represented the range of mortality in the mosquito larvae and revealed the insecticidal activity of ethanol extracts. The per cent mortality of the extract was directly related to the concentration of the extract as well as the time of exposure. The results exhibited that the maximum larvicidal activity was recorded by 60, 80, and 100% dilutions after 72 h of exposure. However, more than 50% mortality was recorded in all dilutions even after 24 h. In addition, the extract showed great larvicidal activity against the larvae of the mosquitoes in a dose-dependent manner. The percent mortality of the extract was directly related to the concentration of the extract as well as the time of exposure (Abutaha et al., 2018). It has been reported that extracts of plants such as Ageratum conyzoides and Cassia sophera induced mortality in mosquitoes (Ghosh et al., 2012). Moreover, a Mentha piperita extract resulted in maximum insecticidal activity against B. brassicas with increased concentration and time exposure (Mersha et al., 2014). Similarly, essential oil from Cinnamomum zeylannicum was found to be effective against B. brassicae. The results showed that the extract was extremely effective against mosquitos and also showed antimicrobial activities.

Figure 2: Larvicidal activities of different dilutions of the extract of black seeds.

CONCLUSION

The results showed that the extract was extremely effective against mosquitos and also showed antimicrobial activities. The methanolic extract was dried and dissolved in different concentrations in water e.g., 20, 40, 60, 80, and 100%. All the dilutions were effective and killed the mosquito at their larval stages even within 24 h. similarly, the extract showed great antimicrobial activities against albicans.

ACKNOWLEDGE

REFERENCES

Abutaha, N., Al-Mekhlafi, F.A., Al-Keridis, L.A., Farooq, M., Nasr, F.A., Al-Wadaan, M., (2018) Larvicidal potency of selected xerophytic plant extracts on Culex pipiens (Diptera: Culicidae). Entomological Research, 48, 362-371.

Ahmad, M.F., Ahmad, F.A., Ashraf, S.A., Saad, H.H., Wahab, S., Khan, M.I., Ali, M., Mohan, S., Hakeem, K.R., Athar, M.T., (2021). An updated knowledge of Black seed (Nigella sativa Linn.): Review of phytochemical constituents and pharmacological properties. Journal of Herbal Medicine, 25, pp 100-404.

Ait Said, L., Zahlane, K., Ghalbane, I., El Messoussi, S., Romane, A., Cavaleiro, C., Salgueiro, L., (2015) Chemical composition and antibacterial activity of Lavandula coronopifolia essential oil against antibiotic-resistant bacteria. Natural Product Research, 29, 582-585.

Ali, S.A., Meitei, K.V., (2011). Nigella sativa seed extract and its bioactive compound thymoquinone: the new melanogens causing hyperpigmentation in the wall lizard melanophores. J Pharm Pharmacol, 63, 741-6. https://www.ncbi.nlm.nih.gov/pubmed/21492177

Ali, S.A., Parveen, N., Ali, A.S., (2018). Links between the Prophet Muhammad (PBUH) recommended foods and disease management: A review in the light of modern superfoods. International Journal of Health Sciences, 12, 61-69. . https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5870322/

Bharali, P., Saikia, J.P., Ray, A., Konwar, B.K., (2013) Rhamnolipid (RL) from Pseudomonas aeruginosa OBP1: a novel chemotaxis and antibacterial agent. Colloids and surfaces. B, Biointerfaces 103, 502-509.

Dagli, N., Dagli, R., Mahmoud, R.S., Baroudi, K., (2015) Essential oils, their therapeutic properties, and implication in dentistry: A review. J Int Soc Prev Community Dent, 5, 335-340.

Dris, D., Tine-Djebbar, F., Bouabida, H., Soltani, N., (2017) Chemical composition and activity of an Ocimum basilicum essential oil on Culex pipiens larvae: Toxicological, biometrical and biochemical aspects. South African Journal of Botany, 113, 362-369.

Enan, E.E., (2005) Molecular and pharmacological analysis of an octopamine receptor from American cockroach and fruit fly in response to plant essential oils. Archives of insect biochemistry and physiology, 59, 161-171.

Ghosh, A., Chowdhury, N., Chandra, G., (2012) Plant extracts as potential mosquito larvicides. Indian J Med Res 135, 581-598.

Helaly, W.M.M., Elyamani, E.M., Saad, M.S.I., (2020). Influence of Some Plant Extracts as Antiseptics to Control Bacterial and Fungal Diseases of Silkworms, Bombyx mori L. Zagazig Journal of Agricultural Research, 47, 707-717.

Jang, E.K., Kwon Jung, B., Park, G.S., Rahim Khan, A., Hong, S.J., Park, Y.J., Kim, W.C., Shin, J.H., Al-Ghamdi, M.S.K., Oudh Al-Johny, B., Anwar, Y., Faisal Siddiqui, M., Ullah, I. (2020). Cloning and expression of the insecticidal toxin gene “tccB” from Photorhabdus temperata M1021 in Escherichia coli expression system. Journal of Asia-Pacific Entomology, 23, 172-176.

Mersha, W., Ayele, N., Fentahun, G., Getinet, M., Kassu, K., Nagappan, R., (2014). Repellent and insecticidal activity of Mentha piperita (L.) plant extracts against cabbage aphid [Brevicoryne brassicae Linn. (Homoptera: Aphididae)]. American-Eurasian Journal of Scientific Research 9, 150-156.

Prashar, A., Locke, I.C., Evans, C.S., (2004) Cytotoxicity of lavender oil and its major components to human skin cells. Cell Proliferation 37, 221-229.

Sabbah, M., Altamimi, M., Di Pierro, P., Schiraldi, C., Cammarota, M., Porta, R. (2020). Black Edible Films from Protein-Containing Defatted Cake of Nigella sativa Seeds. Int. J. Mol. Sci. 21, 832.

Soltane, R., Mtat, D., Chrouda, A., Alzahrani, N., Al-Ghamdi, Y.O., El-Desouky, H., Elbanna, K. 2021. Nigella sativa Seed Extract in Green Synthesis and Nanocomposite. In: Fawzy Ramadan, M. (ed.) Black cumin (Nigella sativa) seeds: Chemistry, Technology, Functionality, and Applications. Cham: Springer International Publishing.

Ullah, I., Al-Ghamdi, K.M.S., Anwar, Y., Mahyoub, A.J. (2020). Exploring the Insect Control and Plant Growth Promotion Potentials of Endophytes Isolated From Calotropis procera Present in Jeddah KSA. Natural Product Communications, 15, 19-24.

Zuzarte, M., Gonçalves, M.J., Cruz, M.T., Cavaleiro, C., Canhoto, J., Vaz, S., Pinto, E., Salgueiro, L., (2012) Lavandula luisieri essential oil as a source of antifungal drugs. Food Chemistry 135, 1505-1510.MENTS

The authors are extremely thankful to all those who supported the study during experimentation and drafting.