IN VITRO ANTIBACTERIAL ACTIVITY OF NEEM LEAVES ON EXTENDED-SPECTRUM BETA-LACTAMASE AND AMBLER CLASS C COPRODUCERS FROM SKIN AND SOFT-TISSUE INFECTIONS.
Â Objective: Clinical isolates coproducing extended-spectrum beta-lactamase (ESBL) and Ambler Class C (AmpC) beta-lactamase are a therapeutic challenge. The aim of the present study was to evaluate the antibacterial activity of Azadirachta indica methanolic leaf extract against lactose fermenting coproducers of ESBL and AmpC isolated from skin and soft-tissue infections (SSTIs).
Methods: A total of 88 non-duplicate lactose fermenting strains isolated from SSTIs specimens were tested for their antibiotic susceptibility pattern by Kirby-Bauer disk diffusion method. Ceftazidime and cefoxitin-resistant strains were screened for coproduction of ESBL and AmpC beta-lactamase by phenotypic confirmatory disc diffusion test and E-test. Antibacterial activity of methanolic neem leaf (NLM) extract was examined by the disc diffusion method and the minimum inhibitory concentration (MIC) of NLM was determined by the agar dilution technique.
Results: Thirty-seven lactose fermenting strains were confirmed to be coproducers of ESBL and AmpC, out of which 27 were Escherichia coli and 10 were Klebsiella spp. The nimbin content in the neem leaf extract was found to be 0.007% (w/w) by high-performance liquid chromatography (HPLC) analysis. The NLM extract showed inhibitory activity against all the 37 lactose fermenting isolates from SSTIs, coproducing ESBL and AmpC. The MIC of NLM extract was determined to be in the range of 4â€“8% (40â€“80 mg/ml) against the selected isolates.
Conclusion: It can be stated that NLM extract might have therapeutic significance against coproducers of ESBL and AmpC beta-lactamase isolated from SSTIs.
2. Sasirekha B, Shivakumar S. Occurrence of plasmid-mediated AmpC Î²-lactamases among Escherichia coli and Klebsiella pneumoniae clinical isolates in a tertiary care hospital in Bangalore. Indian J Microbiol 2012;52:174-9.
3. Sundin DR. Hidden beta-lactamases in the Enterobacteriaceae-dropping the extra disks for detection, Part II. Clin Microbiol Newslett 2009;31:47-52.
4. Mohanty S, Kapil A, Das BK, Dhawan B. Antimicrobial resistance profile of nosocomial uropathogens in a tertiary care hospital. Indian J Med Sci 2003;57:148-54.
5. Yuvaneswaran K, Wong NK. Cytotoxicity and antimicrobial properties of neem (Azadirachta indica) leaf extracts cytotoxicity and antimicrobial properties of neem (Azadirachta indica) leaf extracts. Int J Pharm Pharm Sci 2015;7:179-82.
6. Koona S, Budida S. Antibacterial potential of the extracts of the leaves of Azadirachta indica Linn. Notulae Sci Biol 2011;3:65-9.
7. Colle JG, Miles RS, Watt B. Test for the identification of bacteria. In: Collee JG, Faser AG, Marmion BP, Simmons A, editors. Mackie and McCartney Practical Medical Microbiology. 14th ed. London: Churchill Livingstone; 1996. p. 131-45.
8. Clinical and Laboratory Standards Institute. Performance Standards for Antimicrobial Disc Susceptibility Tests: Approved Standard, CLSI Document M02-A11. 11th ed. Wayne PA: CLSI; 2012.
9. Shah PJ, Williamson MT. Antibacterial and synergistic activity of Calendula officinalis methanolic petal extract on Klebsiella pneumoniae co-producing ESBL and AmpC Beta-lactamase. Int J Curr Microbiol Appl Sci 2015;4:107-17.
10. Rathod SJ, Williamson MT. Antibacterial activity of green tea extract in combination with cefotaxime on diarhea causing ESBL producing E.coli. Int J Pharm Pharm Sci 2015;7:258-62.
11. Shah PJ, Williamson MT. Antibacterial activity of honey against ESBL producing Klebsiella pneumoniae from burn wound infections. Int J Curr Pharm Res 2015;7:32-6.
12. Bimakr M, Rahman RA, Taip FS, Ganjloo A, Salleh LM, Selamat J, et alComparison of different extraction methods for the extraction of major bioactive flavonoid compounds from spearmint (Mentha spicata L.) leaves. Food Bioprod Process 2011;89:67-72.
13. Shah PJ, Williamson MT, Rathod SJ. Antibacterial effect of methanolic neem leaf extract on ESBL, AmpC and MBL producers from skin and soft tissue infections. Int J Pharm Res Biosci 2016;5:117-28.
14. Ward PB, Carson M, Dodd JS, Pavillard ER. Prediction of sulfamethoxazole-trimethoprim synergistic action against members of the family Enterobacteriaceae with a two-plate agar dilution breakpoint MIC system. J Clin Microbiol 1984;19:899-901.
15. Afroz Z, Metri BC, Jyothi P. Bacteriological profile and antimicrobial susceptibility pattern of skin and soft tissue infections among gram negative Bacilli in a tertiary care hospital of south India. J Pharm Sci Res 2015;7:397-400.
16. Shinde S, Nataraj G, Mehta P. Multiple Beta-lactamase resistance mechanism in Escherichia coli and Klebsiella pneumoniae. Bombay Hosp J 2013;55:32-9.
17. Mugnier P, Poirel L, Pitout M, Nordmann P. Carbapenem-resistant and OXA-23-producing Acinetobacter baumannii isolates in the United Arab Emirates. Clin Microbiol Infect 2008;14:879-82.
18. Pai H, Kang C, Byeon J, Lee K, Park W, Kim H, et al. Epidemiology and clinical features of bloodstream infections caused by AmpC Type Î²-lactamase producing Klebsiella pneumoniae. Antimicrob Agents Chemother 2004;48:3720-8.
19. Gomase PV, Rangari VD, Verma PR. Phytochemical evaluation and hepatoprotective activity of fresh juice of young stem (tender) bark of Azadirachta indica. Int J Pharm Pharm Sci 2011;3 Suppl 2:55-9.
20. Chaturvedi P, Bag A, Rawat V, Jyala N, Satyavali V, Jha PK. Antibacterial effects of Azadirachta indica leaf and bark extracts in clinical isolates of diabetic patients. Natl J Integr Res Med 2011;2:5-11.
21. Dahiya P, Purkayastha S. Phytochemical screening and antimicrobial activity of some medicinal plants against multi drug resistant bacteria from clinical isolates. Indian J Pharm Sci 2012;74:443-50.
22. Emmanuel N, Ifeanyichukwu I, Chika E, Emmanuel E, Nwankwo C. Inhibitory effects of neem (Azadirachta indica Linn.) and bitter Kola (Garcinia kola Heckel) leaves on selected pathogenic bacteria. Afr J Pharm Pharmacol 2013;7:2763-7.
The publication is licensed under CC By and is open access. Copyright is with author and allowed to retain publishing rights without restrictions.