1887

A Laboratory Activity Demonstrating the Antibacterial Effects of Extracts from Two Plant Species, and (Garlic)

    Authors: Grace J. Miller1, Anna M. G. Cunningham1, Yui Iwase1, Nicole L. Lautensack1, W. Matthew Sattley1,*
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    Affiliations: 1: Indiana Wesleyan University, Division of Natural Sciences, Marion, IN 46953-4974
    AUTHOR AND ARTICLE INFORMATION AUTHOR AND ARTICLE INFORMATION
    Source: J. Microbiol. Biol. Educ. October 2017 vol. 18 no. 3 doi:10.1128/jmbe.v18i3.1306
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    Abstract:

    A variety of plants synthesize natural products that either kill or inhibit the growth of various microorganisms. These plant products may serve as useful natural alternatives to synthetic antimicrobial pharmaceuticals and can be especially important in regions where commercial drugs are often not available. Despite this, the role of plants as producers of natural antimicrobial agents is often understated or even ignored in undergraduate biology curricula. In this laboratory exercise, students extract water-soluble constituents from two plants, (moringa) and (garlic), and determine their activity against both a gram-positive ( strain 971) and a gram-negative ( strain K12) bacterium using a disk diffusion assay on Mueller-Hinton agar. Disks infused with commercially available antibiotics (e.g., penicillin and tetracycline) serve as controls. Following an incubation period of 24 hours, students obtain quantitative data by measuring zones of growth inhibition that develop as a result of strain sensitivity. To determine the effectiveness of the learning objectives, an unannounced quiz was administered both before and after the activity, and the students showed significant gains in their understanding of key concepts. Because this activity combines aspects of two major branches of biology—plant biology and microbiology—it is suitable for use as a laboratory exercise in courses related to either discipline, or it may be used as a laboratory component of a general biology course.

Key Concept Ranking

Cell Wall Biosynthesis
0.48821712
Antibacterial Agents
0.46327084
0.48821712

References & Citations

1. Abdull Razis AF, Ibrahim MD, Kntayya SB 2014 Health benefits of Moringa oleifera Asian Pac J Cancer Prev 15 20 8571 8576 10.7314/APJCP.2014.15.20.8571 25374169 http://dx.doi.org/10.7314/APJCP.2014.15.20.8571
2. Fahey J 2005 Moringa oleifera: a review of the medical evidence for its nutritional, therapeutic, and prophylactic properties. Part 1 Trees Life J 1 5
3. Jonkers D, van den Broek E, van Dooren I, Thijs C, Dorant E, Hageman G, Stobberingh E 1999 Antibacterial effect of garlic and omeprazole on Helicobacter pylori J Antimicrob Chemother 43 6 837 839 10.1093/jac/43.6.837 10404325 http://dx.doi.org/10.1093/jac/43.6.837
4. Peixoto JR, Silva GC, Costa RA, de Sousa Fontenelle JR, Vieira GH, Filho AA, dos Fernandes Vieira RH 2011 In vitro antibacterial effect of aqueous and ethanolic Moringa leaf extracts Asian Pac J Trop Med 4 3 201 204 10.1016/S1995-7645(11)60069-2 21771453 http://dx.doi.org/10.1016/S1995-7645(11)60069-2
5. Posmontier B 2011 The medicinal qualities of Moringa oleifera Holist Nurs Pract 25 2 80 87 10.1097/HNP.0b013e31820dbb27 21325908 http://dx.doi.org/10.1097/HNP.0b013e31820dbb27
6. Rahman MM, Sheikh MMI, Sharmin SA, Islam MS, Rahman MA, Rahman MM, Alam MF 2009 Antibacterial activity of leaf juice extracts of Moringa oleifera Lam. against some human pathogenic bacteria CMU J Nat Sci 8 219 227
7. Vieira G, Mourao J, Angleo M, Costa R, Vieira R 2010 Antibacterial effect (in vitro) of Moringa oleifera and Annona muricata against gram-positive and gram-negative bacteria Rev Inst Med Trop Sao Paulo 52 3 129 132 10.1590/S0036-46652010000300003 20602021 http://dx.doi.org/10.1590/S0036-46652010000300003
8. Vinoth B, Manisvasagaperumal R, Balamurugan S 2012 Phytochemical analysis and antibacterial activity of Moringa oleifera Lam Int J Res Biol Sci 2 3 98 102
9. Cai Y, Wang R, Pei F, Liang BB 2007 Antibacterial activity of allicin alone and in combination with [beta]-lactams against Staphylococcus spp. and Pseudomonas aeruginosa J Antibiot 60 5 335 338 10.1038/ja.2007.45 17551215 http://dx.doi.org/10.1038/ja.2007.45
10. Harris JC, Cottrell S, Plummer S, Lloyd D 2001 Antimicrobial properties of Allium sativum (garlic) Appl Microbiol Biotechnol 57 282 286 10.1007/s002530100722 http://dx.doi.org/10.1007/s002530100722
11. Bauer AW, Kirby WM, Sherris JC, Turck M 1966 Antibiotic susceptibility testing by a standardized disk method Amer J Clin Path 45 4 493 496
12. Emmert EAB the ASM Task Committee on Laboratory Biosafety 2013 Biosafety guidelines for handling microorganisms in the teaching laboratory: development and rationale J Microbiol Biol Educ 14 78 83 10.1128/jmbe.v14i1.531 23858356 3706168 http://dx.doi.org/10.1128/jmbe.v14i1.531

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2017-10-30
2019-03-21

Abstract:

A variety of plants synthesize natural products that either kill or inhibit the growth of various microorganisms. These plant products may serve as useful natural alternatives to synthetic antimicrobial pharmaceuticals and can be especially important in regions where commercial drugs are often not available. Despite this, the role of plants as producers of natural antimicrobial agents is often understated or even ignored in undergraduate biology curricula. In this laboratory exercise, students extract water-soluble constituents from two plants, (moringa) and (garlic), and determine their activity against both a gram-positive ( strain 971) and a gram-negative ( strain K12) bacterium using a disk diffusion assay on Mueller-Hinton agar. Disks infused with commercially available antibiotics (e.g., penicillin and tetracycline) serve as controls. Following an incubation period of 24 hours, students obtain quantitative data by measuring zones of growth inhibition that develop as a result of strain sensitivity. To determine the effectiveness of the learning objectives, an unannounced quiz was administered both before and after the activity, and the students showed significant gains in their understanding of key concepts. Because this activity combines aspects of two major branches of biology—plant biology and microbiology—it is suitable for use as a laboratory exercise in courses related to either discipline, or it may be used as a laboratory component of a general biology course.

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Figures

Image of FIGURE 1

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FIGURE 1

Images of laboratory procedures. (A) Materials needed for the activity; (B) Moringa seed with hull removed; (C) Mortar and pestle grinding of moringa seed; (D) Ground moringa seed slurry being poured into a 15-mL centrifuge tube; (E) Slurry supernatant sterilized through a membrane filter (0.2–0.45 μm); (F) Sterile disks soaked in sterile extracts; (G) Plating extract-soaked disks onto inoculated plates; (H) Disk placement on plates of either (left) or (right); (I) Zones of inhibition apparent on plates following 24-hour incubation (37°C).

Source: J. Microbiol. Biol. Educ. October 2017 vol. 18 no. 3 doi:10.1128/jmbe.v18i3.1306
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Image of FIGURE 2

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FIGURE 2

Measuring zones of inhibition after 24-hour incubation. Inhibition of bacterial growth was evaluated by measuring the diameter (mm) of each zone of inhibition. Comparisons of individual antibacterial agents can be made between the gram-negative (A) and the gram-positive (B). Disks clockwise from top left: P, penicillin; T, tetracycline; G, garlic; M, moringa.

Source: J. Microbiol. Biol. Educ. October 2017 vol. 18 no. 3 doi:10.1128/jmbe.v18i3.1306
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Image of FIGURE 3

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FIGURE 3

Average improvement of student performance on the pre- and post-lab activity quiz. Significant gains in student understanding were evident for questions 3, 4, and 5. The questions presented to the students are given in Table 1 and in Appendices 1 and 2 . Data were compiled from the responses of 29 students. Statistical differences were determined by converting the pre- and post-lab quiz scores to percentages, which were then normalized for the statistical analysis. In this form, the chi-squared test could be applied using the CHITEST function of Excel. The resulting value calculated for these 29 sets of pre- and post-lab scores was 4 × 10 (<0.05).

Source: J. Microbiol. Biol. Educ. October 2017 vol. 18 no. 3 doi:10.1128/jmbe.v18i3.1306
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Image of FIGURE 4

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FIGURE 4

Student evaluation of the laboratory activity. The survey results showed that almost 90% of the students strongly agreed or agreed that they enjoyed the activity (question 1), the instructions were easy to follow (question 2), and they were interested in the results (question 3). Nearly 94% of the students strongly agreed or agreed that the techniques were relevant and useful (question 4). Perhaps most importantly, about 85% of the students agreed (nearly one-half strongly agreeing) that the exercise highlights the value of medicinal plants as potential producers of antibacterial agents (question 5), which was identified as a specific goal of the activity (Objective 6). The questions as presented to the students are given in Appendix 5 .

Source: J. Microbiol. Biol. Educ. October 2017 vol. 18 no. 3 doi:10.1128/jmbe.v18i3.1306
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