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HomeMy WebLinkAbout02. Presentation (Handout)Handout Exploring the Effect of Curcumin on E. coli Growth as a Tool to Battle Diarrheal Disease in Developing Countries anakumar How does curcumin, organically found in turmeric, affect E. coli growth as the environmental determinant of E. coli-related-diarrheal-diseases becomes a growing health issue in developing countries? E. coli bacterial colony growth will decrease in turmeric -infused agar when compared to the control agar culture. O • Escherichia coli, more often referred to as E. coli is a gram -negative anaerobic bacteria that is commonly found in the lower intestine of healthy humans and animals. • While most types of E. coli are harmless, a few strains such as E. coli O157:H7 can cause severe symptoms of bloody diarrhea, vomiting, and potential kidney failure. • There are six strains of E. coli that can cause diarrhea. • In countries such as Africa where treated water is inaccessible, E. coli-caused death is an issue. For example, in 2017, 606,024 deaths were caused by diarrheal diseases in Sub -Saharan Africa. • These diarrheal diseases are majorly correlated with the presence of E. coli. • Curcuma, more commonly known as turmeric is a biologically active polyphenolic compound in turmeric, and plays a significant role in the antioxidative property of turmeric. • Curcumin has antioxidant, anti-inflammatory, and antimicrobial properties. • 99% ethanol • Distilled water • Agar Agar Type 1 • Mueller Hinton agar • Turmeric root • 2 Petri dishes • E. coli culture "° • Inoculation tubes • Bunsen burner • Incubator • Autoclave • Mortar and pestle • Micropipette • Sanitize all working surfaces with 99% ethanol • Medium preparation: 0 0.65g of nutrient broth 0 50 mL of distilled water • Sterilization to mix the medium 0 Autoclave medium at 121 degrees C for 20 mins • Curcumin extract o Grind raw turmeric with mortar and pestle o Add 20 mL ethanol to the 2g raw turmeric mixture o Filter the mixture to extract the curcumin • Media prep 0 Mix 1 g Agar Agar Type 1 and 3.8g Mueller Hinton agar with 100 mL distilled water • Sterilization of media 0 Autoclave media at 121 degrees C for 20 mins • Pour 20 mL of the media into 2 sterile Petri plates • After solidification of the media, swab 20-25µl suspension of bacterial inoculums uniformly. • Make wells on the agar medium using sterile forceps. • Pour turmeric extract into the wells according to the 2%, 5%, 10%, and 20% concentrations using a pipette. • Pour 10-40 µl of the turmeric extract into the wells and Chloramphenicol antibiotic was used as standard. • Incubate Petri plates at 37-C for 24 hours. • Measure the zone of inhibition from the edge of the well to the zone in mm. Zones of Inhibition According to Concentration of Turmeric Extract Turmeric 2 pL 5 pL 10 pL 20 pL Plate 1 Extract in Wells (IuL) Zone of 2mm 3mm 5mm 8mm Inhibition Turmeric 2 pL 5 pL 10 pL 20 pL Plate 2 Extract in Wells (pQ Zone of 1 mm 1 mm 3 mm 5 mm Inhibition Result Analysis 0 The curcumin concentration in turmeric averages at 3%. • This low concentration of curcumin is likely responsible for the low measurements for zones of inhibition. • If saturated curcumin was used in the same concentrations, there would be a greater inhibition zone. Zones of Inhibition According to Concentration of Turmeric Extract ® Plate 1 0 Plate 2 8 6 E 0 4 s 6 2 N 0 2 5 10 20 Amount of Turmeric Extract in Wells (NL) • The data supports my hypothesis. • However, the low concentration of curcumin found in turmeric is responsible for the lower -than -standard inhibition zones. • If higher concentrations of turmeric were used with concentrated amounts of curcumin, my hypothesis would be further supported. • As seen in the data, as the amount of turmeric extract increases, the zones of inhibitions increase, proving my claim that higher concentrations of curcumin would work better. • Using high concentrations of curcumin can be used to inhibit E. coli growth. • The curcumin's antibacterial activity hasn't been evaluated in clinical trials with an aim of using it as a future antibiotic. Designing medical drugs designed to treat E. coli growth using curcumin extracted from turmeric as an alternative to antibiotics can be looked into. • Additionally, exploring the inhibition of turmeric in antibiotic -resistant E. coli would further prove curcumin to be a strong alternative. Thank you FOR LISTENING