DOI: http://dx.doi.org/10.18203/2319-2003.ijbcp20173732

Synergistic antimicrobial activity of crude ethanolic extracts of garlic and neem leaves against bovine mastitis pathogens: an in vitro assay

Makhulu Edmond Edward, Nyaga Nderitu Samuel, Wambugu Stanley, Areba Gerald

Abstract


Background: Treatment of bovine mastitis (BM) is highly expensive necessitating better and affordable control strategies. Synergy of plant extracts against microbes associated with BM stand unexploited. Therefore, this study sought to evaluate the sensitivity of major bovine mastitis bacteria- Staphylococcus aureus and Escherichia coli to combination of two traditional herbal plants, garlic and neem, widely used in East Africa.

Methods: Maceration and soxhlet processes were used to obtain crude ethanolic extracts of neem extracts (NE) and garlic (GE) respectively. The extracts were used to determine Minimum Inhibition Concentration (MIC) against test pathogens. Synergy was investigated by combining the two extracts at different concentrations and determining colony forming units per liquid medium (CFU/ml) using turbidity as a measure of inoculum growth.

Results: NE was effective against S. aureus at 200µg/ml and 100µg/ml (average zone of inhibitions were 11±0.5774 mm and 10±0.5774 mm respectively). E. coli was resistant to NE. GE was effective against S. aureus at 200µl/ml and 100µl/ml (average zones of inhibition were 14±0.8819 mm and 12±0.8819 mm respectively). GE was effective against E. coli only at 200µl/ml with an average zone of inhibition of 11±0.5774 mm.

Conclusions: There was synergy when garlic MIC was combined with NE MIC. Increase in concentration of GE above its MIC increased the rate of decrease of the CFU. No significant change was shown when NE concentration was increased above its MIC. The combination of NE and GE can have additive effect in treatment S. aureus mastitis while GE can supplement NE resistance.


Keywords


Colony forming units, Escherichia coli, Minimum inhibition concentration, Staphylococcus aureus, Turbidity

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References


Ministry of Livestock Development (MLD)-Kenya. Development of a workable disease control strategy in cattle. Smallholder Dairy Commercialization Programme: 2010.

Somba CN. Marketing Strategies To Be Adopted By The Dairy Industry To Improve Company Competitiveness: A Case Study Of Kinangop Dairy Ltd (Doctoral dissertation, University of Nairobi): 2016.

Gitau GK, Bundi RM, Vanleeuwen J, Mulei CM. Mastitogenic bacteria isolated from dairy cows in Kenya and their antimicrobial sensitivity. Jour Sou Afri Vet Ass. 2014;85(1):752-7.

Alekish MO, Al-qudah KM, Al-saleh A. Prevalence of antimicrobial resistance among bacterial pathogens isolated from bovine mastitis in northern Jordan. Vet Med. 2013;164(6):319-26.

Olajuyigbe OO, Afolayan AJ. Synergistic interactions of methanolic extract of Acacia mearnsii de wild. with antibiotics against bacteria of clinical relevance. Int Jour Mol Sci. 2012;13:8915-32.

Thenmalar M. Molecular studies on biofilm associated Staphylococcus aureus infections in mastitis in cattle From Tamilnadu. Thesis Submitted to Periyar University. 2014. Available at: http://hdl.handle.net/10603/17446.

Tiwari JG, Babra C, Tiwari HK, Williams V, Wet SD, Gibson J, et al. Trends in therapeutic and prevention strategies for management of bovine mastitis: An overview. Jour of VACC. 2013;4(2):2157-7560.

Sivananthan M. Antibacterial activity of 50 medicinal plants used in folk medicine. Int Jour Biosci. 2013;3(4):104-21.

Uwimbabazi F, Uwimana J, Rutanga JP. Assessment of antibacterial activity of Neem plant (Azadirachta indica) on Staphylococcus aureus and Escherichia coli. Jour of Med Plants Stud. 2015;3(4):85-91.

Ncube NS, Afolayan AJ, Okoh AI. Assessment techniques of antimicrobial properties of natural compounds of plant origin: current methods and future trends. Afr Jour Biotech. 2008;7(12):1797-806.

Ogbuewu IP, Odoemenam VU, Obikaonu HO, Opara MN, Emenalom OO, Uchegbu MC, et al. The growing importance of neem (Azadirachta indica A. Juss) in agriculture, industry, medicine and environment: A review. Res Jour Med Plant. 2011;5(3):230-45.

Goncagul G, Ayaz E. Antimicrobial effect of garlic (Allium sativum) and traditional medicine. Jour Ani Vet Advs. 2010;9(1):1-4.

Dzulkarnain SH, Rahim IA. Antimicrobial activity of methanolic Neem extract on wound infection bacteria. Int Conf Bio Chem Enviro Sci. 2014;4(1).

Gafar MK, Itodo AU, Warra AA, Abdullahi L. Extraction and physicochemical determination of garlic (allium sativum L) oil. Int Jour Food Sci Nutr 2012;1(2):1-7.

Sadek PC. Miscibility and Solubility. In P. C. Sadek, The HPLC solvent guide. 2nd Ed. New York, Wiley-Interscience; 2002:22-23.

Maragathavalli S, Brindha S, Kaviyarasi NS, Annadurai B, Gangwar SK. Antibacterial activity in leaf extract of Neem (Azadirachta indica Linn.) Int Jour Sci Nat. 2012;3(1):110-3.

Panda SK. Screening methods in the study of antimicrobial properties of medicinal plants. Int Jour Biotech Res. 2012;2(1):1-35.

Okemo PO, Mwatha WE, Chhabrab SC, Fabry W. The kill kinetics of Azadirachta indica a. Juss. (meliaceae) extracts on Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa and Candida albicans. Afr Jour Sci Tech. 2001;2(2):113-8.

Ankri S, Mirelman, D. Antibacterial properties of allicin from garlic. Micro Inf. 1999;1(2):125-9.

Miron T, Rabinkov A, Mirelman D, Wilchek MQ, Weiner L. The mode of action of allicin: its ready permeability through phospholipid membranes may contribute to its biological activity. Biochimica et Biophysica Acta (BBA) - Biomem. 2000;1463(1):20-30.