Isolation, morphological identification and in vitro antibacterial activity of endophytic bacteria isolated from Azadirachta indica (neem) leaves

Vet World Vol.10 May-2017 Article-7

Research Article

Veterinary World, 10(5): 510-516

https://doi.org/10.14202/vetworld.2017.510-516

Ankit Kumar Singh¹, Rajesh Kumar Sharma¹, Varsha Sharma², Tanmay Singh¹, Rajesh Kumar³, and Dimple Kumari⁴

1. Department of Veterinary Pharmacology and Toxicology, College of Veterinary Science and Animal Husbandry, Nanaji Deshmukh Veterinary Science University, Jabalpur - 482 001, Madhya Pradesh, India.
2. Department of Veterinary Microbiology, College of Veterinary Science and Animal Husbandry, Nanaji Deshmukh Veterinary Science University, Jabalpur - 482 001, Madhya Pradesh, India.
3. Department of Animal Nutrition, College of Veterinary Science and Animal Husbandry, Nanaji Deshmukh Veterinary Science University, Jabalpur - 482 001, Madhya Pradesh, India.
4. Department of Veterinary Biotechnology, College of Veterinary Science and Animal Husbandry, Nanaji Deshmukh Veterinary Science University, Jabalpur - 482 001, Madhya Pradesh, India.

Background and Aim: The objective of this study was to isolate endophytic bacteria from Azadirachta indica (neem) leaves, their identification and investigate their antibacterial activity against three Gram-positive bacteria, Staphylococcus aureus, Streptococcus pyogenes and Bacillus cereus and Gram-negative bacteria Escherichia coli, Salmonella Typhimurium and Klebsiella pneumoniae.

Materials and Methods: Fresh leaves of A. indica (neem) was procured from the Department of Botany, JNKVV, Jabalpur. Five samples were taken, and each sample was divided into five subsamples and separated for further isolation of endophytic bacteria. For sterilization leaves were treated with double distilled water, 0.1% sodium hypochlorite, 0.01% bavistin, 0.05% and 70% ethanol. Sterilized leaves of the plants were embedded in Kings B (KB) petri plates and incubated at 37°C for 24 h. Characterization of the bacteria was done according to its morphology and by Gram-staining. After that, a single colony was transferred into brain heart infusion (BHI) broth and incubated at 37°C for 24 h. The antibacterial effect was studied by the disk diffusion method with known antibiotic ciprofloxacin (Ci) as standard.

Results: A total of 25 bacterial isolates from A. indica (neem) were obtained and identified morphologically. Most of the samples on KB media depicted irregular shape, flat elevation, undulated, rough, opaque, and white in color. Most of the samples on blood agar showed irregular, raise elevation, undulated, smooth, opaque and all the isolates were nonhemolytic and nonchromogenic. The growth of endophytic bacteria in BHI broth were all isolates showed turbidity. The microscopic examination revealed that maximum isolates were Gram-positive and rod shaped. Good antibacterial activity was observed against S. aureus, Streptococcus pyogenes, E. coli, Salmonella Typhimurium, and K. pneumoniae.

Conclusion: Endophytic bacteria are present in leaves of A. indica (neem) and it possesses antibacterial activity against few Gram-positive and Gram-negative bacteria. Keywords: antibacterial activity, Azadirachta indica (neem), ciprofloxacin, endophytic bacteria, leaves.

Keywords: antibacterial activity, Azadirachta indica (neem), ciprofloxacin, endophytic bacteria, leaves.

How to cite this article: Singh AK, Sharma RK, Sharma V, Singh T, Kumar R, Kumari D (2017) Isolation, morphological identification and in vitro antibacterial activity of endophytic bacteria isolated from Azadirachta indica (neem) leaves, Veterinary World, 10(5): 510-516.

Received: 13-06-2016 Accepted: 23-03-2017 Published online: 11-05-2017

Corresponding author: Rajesh Kumar Sharma E-mail: drrksvet2012@gmail.com

DOI: 10.14202/vetworld.2017.510-516

Copyright: Singh, et al. This article is an open access article distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http:// creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.