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 SB2014Health benefits of Moringa oleiferaAsian Pac J Cancer Prev15208571857610.7314/APJCP.2014.15.20.857125374169 http://dx.doi.org/10.7314/APJCP.2014.15.20.8571
2. Fahey J2005Moringa oleifera: a review of the medical evidence for its nutritional, therapeutic, and prophylactic properties. Part 1Trees Life J15
3. Jonkers D, van den Broek E, van Dooren I, Thijs C, Dorant E, Hageman G, Stobberingh E1999Antibacterial effect of garlic and omeprazole on Helicobacter pyloriJ Antimicrob Chemother43683783910.1093/jac/43.6.83710404325 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 RH2011In vitro antibacterial effect of aqueous and ethanolic Moringa leaf extractsAsian Pac J Trop Med4320120410.1016/S1995-7645(11)60069-221771453 http://dx.doi.org/10.1016/S1995-7645(11)60069-2
5. Posmontier B2011The medicinal qualities of Moringa oleiferaHolist Nurs Pract252808710.1097/HNP.0b013e31820dbb2721325908 http://dx.doi.org/10.1097/HNP.0b013e31820dbb27
6. Rahman MM, Sheikh MMI, Sharmin SA, Islam MS, Rahman MA, Rahman MM, Alam MF2009Antibacterial activity of leaf juice extracts of Moringa oleifera Lam. against some human pathogenic bacteriaCMU J Nat Sci8219227
7. Vieira G, Mourao J, Angleo M, Costa R, Vieira R2010Antibacterial effect (in vitro) of Moringa oleifera and Annona muricata against gram-positive and gram-negative bacteriaRev Inst Med Trop Sao Paulo52312913210.1590/S0036-4665201000030000320602021 http://dx.doi.org/10.1590/S0036-46652010000300003
8. Vinoth B, Manisvasagaperumal R, Balamurugan S2012Phytochemical analysis and antibacterial activity of Moringa oleifera LamInt J Res Biol Sci2398102
9. Cai Y, Wang R, Pei F, Liang BB2007Antibacterial activity of allicin alone and in combination with [beta]-lactams against Staphylococcus spp. and Pseudomonas aeruginosaJ Antibiot60533533810.1038/ja.2007.4517551215 http://dx.doi.org/10.1038/ja.2007.45
10. Harris JC, Cottrell S, Plummer S, Lloyd D2001Antimicrobial properties of Allium sativum (garlic)Appl Microbiol Biotechnol5728228610.1007/s002530100722 http://dx.doi.org/10.1007/s002530100722
11. Bauer AW, Kirby WM, Sherris JC, Turck M1966Antibiotic susceptibility testing by a standardized disk methodAmer J Clin Path454493496
12. Emmert EABthe ASM Task Committee on Laboratory Biosafety2013Biosafety guidelines for handling microorganisms in the teaching laboratory: development and rationaleJ Microbiol Biol Educ14788310.1128/jmbe.v14i1.531238583563706168 http://dx.doi.org/10.1128/jmbe.v14i1.531
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2017-10-30
2017-11-19

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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