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Published in: BMC Complementary Medicine and Therapies 1/2017

Open Access 01-12-2017 | Research article

Inhibition of biofilm formation in Mycobacterium smegmatis by Parinari curatellifolia leaf extracts

Authors: Benjamin Bhunu, Ruvimbo Mautsa, Stanley Mukanganyama

Published in: BMC Complementary Medicine and Therapies | Issue 1/2017

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Abstract

Background

Tuberculosis (TB) is a serious public health problem worldwide. Mycobacterium tuberculosis (M. tuberculosis) grows as drug tolerant pellicles. Agents that inhibit biofilm formation in M. tuberculosis have the potential to reduce the disease treatment period and improve the quality of tuberculosis chemotherapy. Parinari curatellifolia (P. curatellifolia) leaf extracts are claimed to treat symptoms similar to tuberculosis in ethnomedicinal practices. Mycobacterium smegmatis (M. smegmatis) is a surrogate organism used in antimycobacterial drug discovery assays. In this study, the effect of the leaf extracts of P. curatellifolia on M. smegmatis growth and biofilm formation was investigated in order to determine the basis of its use in traditional medicinal use.

Methods

Phytochemicals from P. curatellifolia leaves were prepared using a mixture of 50% dichloromethane (DCM): 50% methanol and by serial exhaustive extraction using different solvents of decreasing polarity. The solvents were used in the following order, hexane > dichloromethane > ethyl acetate > acetone >ethanol > methanol > water. The micro-broth dilution method was used as an antimycobacterial susceptibility test to screen for the extract that effectively inhibited M. smegmatis growth and biofilm formation. Biofilm quantification was performed by staining the biofilms with crystal violet and determining the amount of the stain using a spectrophotometer. In addition, the effects of combining the most active extract with kanamycin were also investigated.

Results

The minimum inhibitory concentrations (MIC) of the extracts were found to be 6.2 μg/ml for the acetone extract, 12.5 μg/ml for both the ethanol and the total extract and 50 μg/ml for both the methanol and ethyl acetate extracts. The ethanol extract, dichloromethane extract and water extract were the only extracts that effectively inhibited biofilm formation in M. smegmatis. Combining the ethanol extract with kanamycin enhanced the effect of the ethanol extract in terms of inhibition of biofilm formation.

Conclusions

P. curatellifolia leaves contain phytochemicals that have the potential to be used both as antimycobacterial and anti-biofilm formation compounds.
Literature
3.
go back to reference Maher D, Raviglione M. Global epidemiology of tuberculosis. Clin Chest Med. 2005:183–95. Maher D, Raviglione M. Global epidemiology of tuberculosis. Clin Chest Med. 2005:183–95.
4.
go back to reference Lienhardt C, Raviglione M, Hafner R, Jaramillo E, Hoelcher M, Zumla A, et al. New drugs for the treatment of tuberculosis: need, challenges, promise and prospects for future. J Infect Dis. 2012;205:S241–9. doi:10.1093/infdis/jis034.CrossRefPubMed Lienhardt C, Raviglione M, Hafner R, Jaramillo E, Hoelcher M, Zumla A, et al. New drugs for the treatment of tuberculosis: need, challenges, promise and prospects for future. J Infect Dis. 2012;205:S241–9. doi:10.​1093/​infdis/​jis034.CrossRefPubMed
5.
go back to reference Kwan CK, Ernst JD. HIV and tuberculosis: a deadly human Syndemic. Clin Microbiol Rev. 2011:351–76. Kwan CK, Ernst JD. HIV and tuberculosis: a deadly human Syndemic. Clin Microbiol Rev. 2011:351–76.
7.
go back to reference Veleyati AA, Masjedi RM, Forma P, Tabarsa P, Ghavani J, Ziazariffi A, et al. Emergency of new forms of totally drug resistant tuberculosis bacilli: super extensively drug resistant tuberculosis or totally drug resistant strains in Iran. Chest. 2009;136(2):420–5. doi:10.1378/chest.08-2427.CrossRef Veleyati AA, Masjedi RM, Forma P, Tabarsa P, Ghavani J, Ziazariffi A, et al. Emergency of new forms of totally drug resistant tuberculosis bacilli: super extensively drug resistant tuberculosis or totally drug resistant strains in Iran. Chest. 2009;136(2):420–5. doi:10.​1378/​chest.​08-2427.CrossRef
8.
go back to reference Blanco D, Perez-Herran E, Cacho M, Ballell L, Castro J, González Del Río R, et al. Mycobacterium tuberculosis gyrase inhibitors as a new class of antitubercular drugs. Antimicrob Agents Chemother. 2014;59:1868–75. doi:10.1128/AAC.03913-14.CrossRef Blanco D, Perez-Herran E, Cacho M, Ballell L, Castro J, González Del Río R, et al. Mycobacterium tuberculosis gyrase inhibitors as a new class of antitubercular drugs. Antimicrob Agents Chemother. 2014;59:1868–75. doi:10.​1128/​AAC.​03913-14.CrossRef
9.
go back to reference Ackart D, Hascal-Dove L, Ceres SM, Kirk N, Podell B, Maled C, et al. Expression of antimicrobial tolerance by attached communities of Mycobacterium tuberculosis. Pathog Dis. 2013;70:359–69.CrossRef Ackart D, Hascal-Dove L, Ceres SM, Kirk N, Podell B, Maled C, et al. Expression of antimicrobial tolerance by attached communities of Mycobacterium tuberculosis. Pathog Dis. 2013;70:359–69.CrossRef
10.
go back to reference Lee W, Vanderven BC, Fahey RS, Russel DG. Intracellular Mycobacterium tuberculosis exploits host derived fully acids to limit metabolic stress. J Biol Chem. 2013;288:6788–800.CrossRefPubMedPubMedCentral Lee W, Vanderven BC, Fahey RS, Russel DG. Intracellular Mycobacterium tuberculosis exploits host derived fully acids to limit metabolic stress. J Biol Chem. 2013;288:6788–800.CrossRefPubMedPubMedCentral
11.
go back to reference Hett EC, Rubin EJ. Bacterial growth and cell division: a mycobacterial perspective. Microbiol Mol Biol Rev. 2008; 72: 126–156. doi: 10.1128/MMBR.00028-07 PMCID: PMC2268284. Hett EC, Rubin EJ. Bacterial growth and cell division: a mycobacterial perspective. Microbiol Mol Biol Rev. 2008; 72: 126–156. doi: 10.​1128/​MMBR.​00028-07 PMCID: PMC2268284.
12.
go back to reference Brosch R, Gondon SV, Brodin PA. A new evolutionary scenario for the Mycobacterium tuberculosis complex. Proc Natl Acad Sci U S A. 2002;99:3680–9.CrossRef Brosch R, Gondon SV, Brodin PA. A new evolutionary scenario for the Mycobacterium tuberculosis complex. Proc Natl Acad Sci U S A. 2002;99:3680–9.CrossRef
13.
go back to reference Anderson GG, Parkermo JJ, Schilling JD, Roth R, Henster J. Intracellular bacterial biofilm-like pods in urinary tract infections. Science. 2003;301:105–7.CrossRefPubMed Anderson GG, Parkermo JJ, Schilling JD, Roth R, Henster J. Intracellular bacterial biofilm-like pods in urinary tract infections. Science. 2003;301:105–7.CrossRefPubMed
14.
go back to reference Ojha A, Anand M, Jacobs WR, Bhatt A, Kreme L, Hatfull GF. GroEli: a dedicated chaperone involved in mycolic acid biosynthesis during mycobacterial growth in biofilms. Cell. 2015;123:861–73.CrossRef Ojha A, Anand M, Jacobs WR, Bhatt A, Kreme L, Hatfull GF. GroEli: a dedicated chaperone involved in mycolic acid biosynthesis during mycobacterial growth in biofilms. Cell. 2015;123:861–73.CrossRef
15.
go back to reference Mu H, Guo F, Niu N, Wang S, Duan V. Chitosan improves anti-biofilm efficacy of gentamicin through facilitating antibiotic penetration. Int J Mol Sci. 2007;15:22296–308. doi:10.3390/ijms151222296.CrossRef Mu H, Guo F, Niu N, Wang S, Duan V. Chitosan improves anti-biofilm efficacy of gentamicin through facilitating antibiotic penetration. Int J Mol Sci. 2007;15:22296–308. doi:10.​3390/​ijms151222296.CrossRef
16.
go back to reference Pandey A, Tripathi S. Concept of standardization, extraction and pre-phytochemical screening strategies for herbal drug. J Pharmacogn Phytochem. 2004;2:115–9. Pandey A, Tripathi S. Concept of standardization, extraction and pre-phytochemical screening strategies for herbal drug. J Pharmacogn Phytochem. 2004;2:115–9.
17.
go back to reference Srivastav DNS, Suresh T. Antimicrobial activity of certain medical plants using in folkloric medicine. J Ethnopharmacol. 2001;74:217–22.CrossRef Srivastav DNS, Suresh T. Antimicrobial activity of certain medical plants using in folkloric medicine. J Ethnopharmacol. 2001;74:217–22.CrossRef
18.
go back to reference Chigora P, Masocha R, Mutenheri F. The role of indigenous medicinal knowledge (IMK) in the treatment of ailments in rural Zimbabwe: the case of Mutirikwi communal lands. J Sust Dev Afr. 2007;9:1509–20. Chigora P, Masocha R, Mutenheri F. The role of indigenous medicinal knowledge (IMK) in the treatment of ailments in rural Zimbabwe: the case of Mutirikwi communal lands. J Sust Dev Afr. 2007;9:1509–20.
19.
go back to reference Mukhopadhyay MK, Banerjee P, Nath D. Phytochemicals – biomolecules for prevention and treatment of human diseases-a review. IJSER. 2012;3(7):1–32. Mukhopadhyay MK, Banerjee P, Nath D. Phytochemicals – biomolecules for prevention and treatment of human diseases-a review. IJSER. 2012;3(7):1–32.
20.
go back to reference Tochukwu HE, Usman WA. Curative effect of Parinari curatellifolia leaf extract on epiglottitis. J Pharm Biol Sci. 2013;5(6):46–51. Tochukwu HE, Usman WA. Curative effect of Parinari curatellifolia leaf extract on epiglottitis. J Pharm Biol Sci. 2013;5(6):46–51.
21.
go back to reference Halilu ME, Yebpella GG, Hassan LG, Achor M. Preliminary phytochemical screening, antibacterial activity and elemental analysis of the leaves and the root bark of Parinari curatellifolia planch ex. Benth (Chrysobalanaceae). C J Pharm Sci. 2010;4:1–5. Halilu ME, Yebpella GG, Hassan LG, Achor M. Preliminary phytochemical screening, antibacterial activity and elemental analysis of the leaves and the root bark of Parinari curatellifolia planch ex. Benth (Chrysobalanaceae). C J Pharm Sci. 2010;4:1–5.
22.
go back to reference Sousa S, Bandeira M, Carvalho PA, Duarte A, Jordao L. Nontuberculous mycobacteria pathogenesis and biofilm assembly. Int J Mycobacteriol. 2015;4(1):36–43.CrossRefPubMed Sousa S, Bandeira M, Carvalho PA, Duarte A, Jordao L. Nontuberculous mycobacteria pathogenesis and biofilm assembly. Int J Mycobacteriol. 2015;4(1):36–43.CrossRefPubMed
23.
go back to reference Barua CC, Singh A, Sen S, Barua AG, Barua IC. In vitro antioxidant and antimycobacterial activity of seeds of Piper longum Linn: A comparative study. SAJ Pharmacy and Pharmacology, 2014. 1: 101. doi: 10. 18875/2375–2262.1.101. Barua CC, Singh A, Sen S, Barua AG, Barua IC. In vitro antioxidant and antimycobacterial activity of seeds of Piper longum Linn: A comparative study. SAJ Pharmacy and Pharmacology, 2014. 1: 101. doi: 10. 18875/2375–2262.1.101.
25.
go back to reference Kisangau DP, Hosea KM, Joseph CC, Lyaruu HVM. In vitro antimicrobial assay of plants used in traditional medicine in Bukoba rural district, Tanzania. Afr J Tradit Complement Altern Med. 2007;4:510–23.CrossRefPubMedPubMedCentral Kisangau DP, Hosea KM, Joseph CC, Lyaruu HVM. In vitro antimicrobial assay of plants used in traditional medicine in Bukoba rural district, Tanzania. Afr J Tradit Complement Altern Med. 2007;4:510–23.CrossRefPubMedPubMedCentral
27.
go back to reference Vipra A, Desai SN, Junjappa RP, Roy P, Poonacha N, Ravinder P, et al. Determining the minimum inhibitory concentration of bacteriophages: potential advantages. Adv Microbiol. 2013;3:181–90.CrossRef Vipra A, Desai SN, Junjappa RP, Roy P, Poonacha N, Ravinder P, et al. Determining the minimum inhibitory concentration of bacteriophages: potential advantages. Adv Microbiol. 2013;3:181–90.CrossRef
28.
go back to reference Hawser PS, Douglas JL. Biofilm formation by Candida species on the surface of catheter material in vitro. Infect Immun. 1994;62:915–21.PubMedPubMedCentral Hawser PS, Douglas JL. Biofilm formation by Candida species on the surface of catheter material in vitro. Infect Immun. 1994;62:915–21.PubMedPubMedCentral
29.
go back to reference Nguta JM, Regina AO, Nyarko AK, Manu DY, Addo PGA. Current perspectives in drug discovery against tuberculosis from natural products. Int J Mycobacteriol. 2015;4:165–83.CrossRefPubMed Nguta JM, Regina AO, Nyarko AK, Manu DY, Addo PGA. Current perspectives in drug discovery against tuberculosis from natural products. Int J Mycobacteriol. 2015;4:165–83.CrossRefPubMed
30.
go back to reference Basaraba RJ. Experimental tuberculosis: the role of comparative pathology in the discovery of improved tuberculosis treatment strategies. Tuberculosis (Edinburg). 2008:S135–47. Basaraba RJ. Experimental tuberculosis: the role of comparative pathology in the discovery of improved tuberculosis treatment strategies. Tuberculosis (Edinburg). 2008:S135–47.
32.
go back to reference Casterton JW, Greenberg EP, Sterwart PS. Bacterial biofilms: a common cause of persistent infections. Science. 1991;284:1318–22.CrossRef Casterton JW, Greenberg EP, Sterwart PS. Bacterial biofilms: a common cause of persistent infections. Science. 1991;284:1318–22.CrossRef
33.
go back to reference Wu H, Moser C, Wang H-Z, Høiby N, Song Z-J. Strategies for combating bacterial biofilm infections. Int J Oral Sci. 2014;7:1–7.CrossRefPubMedCentral Wu H, Moser C, Wang H-Z, Høiby N, Song Z-J. Strategies for combating bacterial biofilm infections. Int J Oral Sci. 2014;7:1–7.CrossRefPubMedCentral
34.
go back to reference Matongo K. Conservation and use-values of medicinal plants in rural Eastern Zimbabwe: a study of selected medicinal plants. University of the Western Cape, Magister Artium Thesis. University of the Western Cape, South Africa. 2012. Matongo K. Conservation and use-values of medicinal plants in rural Eastern Zimbabwe: a study of selected medicinal plants. University of the Western Cape, Magister Artium Thesis. University of the Western Cape, South Africa. 2012.
35.
go back to reference Chimponda T, Mukanganyama S. Antimycobacterial activities of selected medicinal plants from Zimbabwe against Mycobacterium aurum and Corynebacterium glutamicum. Trop Biomed. 2010;27:595–610.PubMed Chimponda T, Mukanganyama S. Antimycobacterial activities of selected medicinal plants from Zimbabwe against Mycobacterium aurum and Corynebacterium glutamicum. Trop Biomed. 2010;27:595–610.PubMed
36.
go back to reference Chukwudi U, Ayodeji F, Amos J, Olawunmi O. Antibacterial activity and phytochemical evaluation of the leaf root and stem bark extracts of Parinari curatellifolia (planch. Ex benth). Int. J Adv Chem. 2014;2:178–81.CrossRef Chukwudi U, Ayodeji F, Amos J, Olawunmi O. Antibacterial activity and phytochemical evaluation of the leaf root and stem bark extracts of Parinari curatellifolia (planch. Ex benth). Int. J Adv Chem. 2014;2:178–81.CrossRef
38.
go back to reference Barchan A, Bakkali M, Arakrak A, Pagán R, Laglaoui A. The effects of solvents polarity on the phenolic contents and antioxidant activity of three Mentha species extracts. Int J Curr Microbiol App Sci. 2014;3:399–412. Barchan A, Bakkali M, Arakrak A, Pagán R, Laglaoui A. The effects of solvents polarity on the phenolic contents and antioxidant activity of three Mentha species extracts. Int J Curr Microbiol App Sci. 2014;3:399–412.
39.
go back to reference Lekganyane MA, Matsebatlela TM, Howard RL, Shai LJ, Masoko P. The phytochemical, antibacterial and antioxidant activity of five medicinal plants against the wound infecting bacteria. Afr J Biotechnol. 2012;11:13210–9. Lekganyane MA, Matsebatlela TM, Howard RL, Shai LJ, Masoko P. The phytochemical, antibacterial and antioxidant activity of five medicinal plants against the wound infecting bacteria. Afr J Biotechnol. 2012;11:13210–9.
40.
go back to reference Zumla A, Nahid P, Cole ST. Classification of drugs used to treat drug-susceptible and drug-resistant tuberculosis from advances in the development of new tuberculosis drugs and treatment regimens. Nat Rev Drug Discov. 2013;12:388–404. doi:10.1038/nrd4001.CrossRefPubMed Zumla A, Nahid P, Cole ST. Classification of drugs used to treat drug-susceptible and drug-resistant tuberculosis from advances in the development of new tuberculosis drugs and treatment regimens. Nat Rev Drug Discov. 2013;12:388–404. doi:10.​1038/​nrd4001.CrossRefPubMed
41.
go back to reference Taniguchi H, Chang B, Abe C, Nikaido Y, Mizuguchi Y, Yoshida SI. Molecular analysis of kanamycin and viomycin resistance in Mycobacterium smegmatis by use of the conjugation system. J Bacteriol. 1997;179:4795–801.CrossRefPubMedPubMedCentral Taniguchi H, Chang B, Abe C, Nikaido Y, Mizuguchi Y, Yoshida SI. Molecular analysis of kanamycin and viomycin resistance in Mycobacterium smegmatis by use of the conjugation system. J Bacteriol. 1997;179:4795–801.CrossRefPubMedPubMedCentral
42.
go back to reference Danilchanka O, Pavlenock M, Niederwis M. Role of porins for uptake of antibiotics by mycobacterium smegmatis. Antimicrob Agents Chemother. 2008;52:3127–34.CrossRefPubMedPubMedCentral Danilchanka O, Pavlenock M, Niederwis M. Role of porins for uptake of antibiotics by mycobacterium smegmatis. Antimicrob Agents Chemother. 2008;52:3127–34.CrossRefPubMedPubMedCentral
43.
go back to reference Sant'Anna da Silva AP, Nascimento da Silva LC, Martins da Fonseca CS, de Araújo JM, dos Santos Correia MT, da Silva Cavalcanti M, and Menezes Lima VL. Antimicrobial Activity and Phytochemical Analysis of Organic Extracts from Cleome spinosa Jaqc. Front Microbiol. 2016; 7: 963. Sant'Anna da Silva AP, Nascimento da Silva LC, Martins da Fonseca CS, de Araújo JM, dos Santos Correia MT, da Silva Cavalcanti M, and Menezes Lima VL. Antimicrobial Activity and Phytochemical Analysis of Organic Extracts from Cleome spinosa Jaqc. Front Microbiol. 2016; 7: 963.
44.
go back to reference Sasidharan S, Chen Y, Saravanan D, Sundram KM, Yoga LL. Extraction, isolation and characterization of bioactive compounds from Plants' extracts. Afr J Tradit Complement Altern Med. 2011;8(1):1–10.PubMed Sasidharan S, Chen Y, Saravanan D, Sundram KM, Yoga LL. Extraction, isolation and characterization of bioactive compounds from Plants' extracts. Afr J Tradit Complement Altern Med. 2011;8(1):1–10.PubMed
45.
go back to reference Tsao R, Romanchuk FE, Peterson CJ, Coats JR. Plant growth regulatory effect and insecticidal activity of the extracts of the tree of heaven (Ailanthus altissima L.). BMC Ecol. 2002;2:1.CrossRefPubMedPubMedCentral Tsao R, Romanchuk FE, Peterson CJ, Coats JR. Plant growth regulatory effect and insecticidal activity of the extracts of the tree of heaven (Ailanthus altissima L.). BMC Ecol. 2002;2:1.CrossRefPubMedPubMedCentral
46.
go back to reference Upadhyay A, Upadhyaya I, Kollanoor-Johny A, Venkitanarayanan K. Combating pathogenic microorganisms using plant-derived antimicrobials: a Minireview of the mechanistic basis. Biomed Res Int. 2014;2014:761741.PubMedPubMedCentral Upadhyay A, Upadhyaya I, Kollanoor-Johny A, Venkitanarayanan K. Combating pathogenic microorganisms using plant-derived antimicrobials: a Minireview of the mechanistic basis. Biomed Res Int. 2014;2014:761741.PubMedPubMedCentral
48.
go back to reference Samy RP, Gopalakrishnakone P. Therapeutic potential of plants as anti-microbials for drug discovery. Evid Based Complement Alternat Med. 2010;7:283–94.CrossRef Samy RP, Gopalakrishnakone P. Therapeutic potential of plants as anti-microbials for drug discovery. Evid Based Complement Alternat Med. 2010;7:283–94.CrossRef
49.
go back to reference Rabin N. Zheng1 Y, Opoku-Temeng C, du Y, Bonsu E, Sintim HO. Agents that inhibit bacterial biofilm formation. Future med. Chem. 2015;7:647–71. Rabin N. Zheng1 Y, Opoku-Temeng C, du Y, Bonsu E, Sintim HO. Agents that inhibit bacterial biofilm formation. Future med. Chem. 2015;7:647–71.
50.
go back to reference Vassea M, Robert J. Noble RJ, Akhmetzhanov AR, Gurney CT-BJ, Benateau S, Gougat-Barbera C, Kaltz O, Hochberg ME. Antibiotic stress selects against cooperation in the pathogenic bacterium Pseudomonas aeruginosa. 546–551 PNAS. 2017; 114: 546–551. Vassea M, Robert J. Noble RJ, Akhmetzhanov AR, Gurney CT-BJ, Benateau S, Gougat-Barbera C, Kaltz O, Hochberg ME. Antibiotic stress selects against cooperation in the pathogenic bacterium Pseudomonas aeruginosa. 546–551 PNAS. 2017; 114: 546–551.
Metadata
Title
Inhibition of biofilm formation in Mycobacterium smegmatis by Parinari curatellifolia leaf extracts
Authors
Benjamin Bhunu
Ruvimbo Mautsa
Stanley Mukanganyama
Publication date
01-12-2017
Publisher
BioMed Central
Published in
BMC Complementary Medicine and Therapies / Issue 1/2017
Electronic ISSN: 2662-7671
DOI
https://doi.org/10.1186/s12906-017-1801-5

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