ArticlePDF Available

The Antiseptic Effect of Turmeric (Curcuma longa) Extraction on the Bacterial Growth of Escherichia Coli (K-12) and Salmonella Typhi which Cause Food Poisoning Council for Innovative Research

Authors:

Abstract and Figures

Previous research has confirmed that turmeric compounds have antiseptic properties. In this study, the effect of freshly turmeric powder on Escherichia coli strain K-12 and Salmonella typhi was examined. Fresh turmeric powder were mixed with ethanol and centrifuged to produce a supersaturated turmeric solution. The supersaturated turmeric solution with several concentrations was added to macconky agar, which was used to culture Escherichia coli strain K-12, and deoxycholate citrate agar (DC agar)which was used to culture Salmonella typhi. The growth of the bacteria was studied by microscopic inspection of bacterial colonies. It was found that turmeric powder is an effective antiseptic agent against Escherichia coli K-12 and Salmonella typhi. It was determined that even at low concentrations of 50 micro liters of turmeric solution there is a noticeable antiseptic effect. It was also determined that higher concentrations of turmeric had a greater antiseptic property. There were no Escherichia coli K-12 and Salmonella typhi growth beyond a turmeric concentration of 800 micro liters 400 micro liters respectively.
No caption available
… 
No caption available
… 
No caption available
… 
Content may be subject to copyright.
ISSN 2347-6893
546 | Page June 24, 2 0 1 4
The Antiseptic Effect of Turmeric (Curcuma longa) Extraction on the
Bacterial Growth of Escherichia Coli (K-12) and Salmonella Typhi which
Cause Food Poisoning
Zeayd Fadhil Saeed
Foundation of Technical Education, Technical Institute of Samawa, Nursing department
zeaydfadhil@gmail.com
key word: freshly turmeric powder; Escherichia coli strain K-12; Salmonella typhi ;
deoxycholate citrate agar; macconky agar.
ABSTRACT
Previous research has confirmed that turmeric compounds have antiseptic properties. In this study, the effect of freshly
turmeric powder on Escherichia coli strain K-12 and Salmonella typhi was examined. Fresh turmeric powder were mixed
with ethanol and centrifuged to produce a supersaturated turmeric solution. The supersaturated turmeric solution with
several concentrations was added to macconky agar, which was used to culture Escherichia coli strain K-12, and
deoxycholate citrate agar (DC agar)which was used to culture Salmonella typhi. The growth of the bacteria was studied by
microscopic inspection of bacterial colonies. It was found that turmeric powder is an effective antiseptic agent against
Escherichia coli K-12 and Salmonella typhi. It was determined that even at low concentrations of 50 micro liters of turmeric
solution there is a noticeable antiseptic effect. It was also determined that higher concentrations of turmeric had a greater
antiseptic property. There were no Escherichia coli K-12 and Salmonella typhi growth beyond a turmeric concentration of
800 micro liters 400 micro liters respectively.
Council for Innovative Research
Peer Review Research Publishing System
Journal: JOURNAL OF ADVANCES IN BIOLOGY
Vol 4, No.3
editor@cirjab.org
www.cirjab.com , editorsjab@gmail.com
ISSN 2347-6893
547 | Page June 24, 2 0 1 4
INTRODUCTION
Turmeric Curcuma longa, a perennial herb and member of the Zingiberaceae (ginger) family, grows to a height of three to
five feet and is cultivated extensively in Asia, India, China, and other countries with a tropical climate. It has oblong,
pointed leaves and funnel-shaped yellow flowers[1]. The rhizome, the portion of the plant used medicinally, is usually
boiled, cleaned, and dried, yielding a yellow powder. Dried Curcuma longa is the source of the spice turmeric, the
ingredient that gives curry powder its characteristic yellow color. Turmericis used extensively in foods for its flavor and
color, as well as having a long tradition of use in the Chinese and Ayurveda systems of medicine, particularly as an anti-
inflammatory and for the treatment of flatulence, jaundice, menstrual difficulties, hematuria, hemorrhage, and colic.
Turmeric can also be applied topically in poultices to relieve pain and inflammation[2]. Current research has focused on
turmeric’s antioxidant, hepatoprotective, anti-inflammatory, anticarcinogenic, and antimicrobial properties, in addition to its
use in cardiovascular disease and gastrointestinal disorders.
The active constituents of turmeric are the flavonoid curcumin (diferuloylmethane) and various volatile oils, including
tumerone, atlantone, and zingiberone. Other constituents include sugars, proteins, and resins. The best researched active
constituent is curcumin, which comprises 0.3-5.4 percent of raw turmeric [2].
Curcumin is a potent anti-inflammatory with specific lipoxygenase- and COX-2- inhibiting properties. Animal, in vitro, and
in vivo studies demonstrate turmeric’s effectiveness at decreasing both acute and chronic inflammation[3-6].Turmeric
extract and the essential oil of Curcuma longa inhibit the growth of a variety of bacteria, parasites, and pathogenic fungi. A
study of chicks infected with the caecal parasite Eimera maxima demonstrated that diets supplemented with 1% turmeric
resulted in a reduction in small intestinal lesion scores and improved weight gain [7]. Turmeric’s protective effects on the
cardiovascular system include lowering cholesterol and triglyceride levels, decreasing susceptibility of low density
lipoprotein (LDL) to lipid peroxidation [8] and inhibiting platelet aggregation[9]. These effects have been noted even with low
doses of turmeric.
The goal of this project is to study the antiseptic effect of different concentrations of turmeric on a particular strain of
Escherichia coli (K-12) and Salmonella typhi. Alcohols, like ethanol was found to be suitable solvent for turmeric as
turmeric solution in alcohol is soluble in water. For this project, 200 proof ethanol was used as a solvent for turmeric.
MATERIAL AND METHOD:
The materials that were used to conduct this experiment are:
1- Centrifuge (Beckman Brand, CPKR type).
2- Ethanol (Decond Labs Brand, standard 200 proof DSP-MD.43).
3- Laboratory test tubes (standard).
4- Autoclave (Fisher Scientific Brand).
5- Laboratory fume hood (Fisher Scientific Brand, EF-5-8).
6- Beaker (500mL capacity, standard).
7- Macconky agar, which was used to culture Escherichia coli strain K-12, and deoxycholate citrate agar (DC agar)which
was used to culture Salmonella typhi.
8- Nutrient broth tubes.
9- Sterile loops (Fisher Scientific Brand).
10- Incubator (standard).
A- Curcuma extraction preparation:-
Different amounts of the supersaturated turmeric solution, prepared by centrifuging a mixture of 200 proof ethanol and 100
grams of finely turmeric powder (which collected from local markets) was added to an agar solution to study the effect of
turmeric on Escherichia coli strain K-12 and Salmonella typhi.
200 proof ethanol was added to the beaker containing the turmeric powder (100 grams) for a total volume of 180 milliliters,
and mixed the mixture to create a supersaturated solution the beaker was placed on a lab bench. Then the ethanol and
turmeric solution in the beaker was then distributed evenly and poured into four laboratory test tubes. Each test tube had
about 45 milliliter of the supersaturated solution and whirled counterclockwise by hand a few times.
The test tubes were then placed into the centrifuge so that both sides of the centrifuge were balanced. The centrifuge was
set to the specifications: temperature of twenty-two degrees centigrade, speed of 3870 rotations per minute, timed for
seven minutes.
After seven minutes in the centrifuge, the clear turmeric solution was visible at the top and debris settled at the bottom in
each of the four test tubes, and the clear solution at the top was carefully poured in a set of new test tubes so that all
debris remained in the original test tubes. The clear solution of turmeric in ethanol served as a stock of turmeric for
experimentation. This solution was put in a cold room at four degrees centigrade to avoid evaporation of the ethanol.
ISSN 2347-6893
548 | Page June 24, 2 0 1 4
B- Culture media preparation and bacterial incubation:-
Prepare 800 ml of the macconky agar according to the industrial company in beaker to use it to culture the Escherichia
coli strain K-12 after sterile it by autoclave, the agar solution was cooled until the temperature of the agar solution was
measured to be 40 degrees. And then 100 ml of macconky agar solution was poured into another beaker and then poured
into four Petri dishes, these four Petri dishes were marked as control. The process of pouring 100 ml of macconky agar
into a second beaker was repeated; each time a different amount of turmeric stock solution was added to the macconky
agar and poured into four already marked Petri dishes. The amount of turmeric solution added were 50 micro liters, 100
micro liters, 200 micro liters, 400 micro liters, 800 micro liters, 1000 micro liters, and 2000 micro liters respectively, yielding
32 dishes including the control, each with a different concentration amount of turmeric extraction.
So prepare 800 ml of the deoxycholate citrate agar (DC agar) according to the industrial company in beaker to use it to
culture the Salmonella typhi after sterile it by autoclave and use the same method to prepare 32 Petri dishes of
deoxycholate citrate agar (DC agar) with a different concentration amount of turmeric extraction.
The stock for the Escherichia coli K-12 bacteria was prepared by sterilized swab was used to scrape the bacteria from
the agar plate from the bacterial bank of sciences collage / Biology department of Al-Qadisiya University and The bacteria
was Injection in nutrient broth tubes with nutrients and incubated for forty hours at 37 degrees Celsius, and then make
decimal dilution by taking 1 ml of bacterial broth and add to tub which contain 9 ml of distal water and then use 1 ml of
second bacterial diluted tub (10-2) by Pipette to culture it by spread on each Petri dishes of macconky agar which
prepared (the control Petri dish and the dishes with different concentrations of turmeric extraction) and incubate it in
incubator at 37degrees Celsius for 24 hour and then notice the bacterial growth on macconky agar with different
concentrations of turmeric extraction.
So The stock for the Salmonella typhi bacteria was prepared by sterilized swab was used to scrape the bacteria from the
agar plate from the bacterial bank of sciences collage / Biology department of Al-Qadisiya University and The bacteria was
Injection in nutrient broth tubes with nutrients and incubated for forty hours at 37 degrees Celsius, and then make decimal
dilution by taking 1 ml of bacterial broth and add to tub which contain 9 ml of distal water and then use 1 ml of second
bacterial diluted tub (10-2) by Pipette to culture it by spread on each Petri dishes of Deoxycholate Citrate Agar (DC agar)
which prepared (the control Petri dish and the dishes with different concentrations of turmeric extraction) and incubate it in
incubator at 37 degrees Celsius for 24 hour and then notice the bacterial growth on Deoxycholate Citrate Agar with
different concentrations of turmeric extraction.
RESULT AND DISCUSSION:
After incubation the Petri dishes in incubator at 37 degrees Celsius for 24 hour and then notice the bacterial growth of
Salmonella typhi on Deoxycholate Citrate Agar and the growth of Escherichia coli strain K-12and enumerate the total
bacterial account on each Petri dish which contain different concentrations of turmeric extraction.
The salmonella appear like yellow or colorless colonies on Deoxycholate Citrate Agar, and the Escherichia coli strain K-12
appear like deep red colonies on macconky agar.
Table (1): Explain the total bacterial account of Escherichia coli strain K-12 and Salmonella typhi and its
relationship with the concentrations of turmeric extraction.
concentrations of
turmeric extraction
The average of Escherichia coli strain
K-12 growth on macconky agar
The average of Salmonella typhi
growth on DC agar
Turmeric 1
Control
12 x 10-2
20 x 10-2
Turmeric 2
50 micro liters
10 x 10-2
16 x 10-2
Turmeric 3
100 micro liters
9 x 10-2
5 x 10-2
Turmeric 4
200 micro liters
7 x 10-2
1 x 10-2
Turmeric 5
400 micro liters
2x 10-2
Nil
Turmeric 6
800 micro liters
Nil
Nil
Turmeric 7
1000 micro liters
Nil
Nil
ISSN 2347-6893
549 | Page June 24, 2 0 1 4
Turmeric 8
2000 micro liters
Nil
Nil
The results of this study reveal that the antiseptic property and inhibition effect of turmeric is fairly consistent on different
batches of the turmeric laced agar plates. So we notice from the table above that the both species of bacteria grow
normally on the control dishes, but we see the Escherichia coli strain K-12 can't grow with Turmeric 6 (800 micro liters),
and Salmonella typhi can't grow with Turmeric 5 (400 micro liters) which is determine it as the maximum concentration of
turmeric extraction which inhibit the Escherichia coli strain K-12 growth and Salmonella typhi respectively.
Based on the data presented in table, it can be concluded that turmeric has antiseptic property even at low concentrations
of50 micro liters.
RECOMINDATIONS:
1- In future extension of the experiment, the four major molecules of turmeric (curcumin, demethooxycurcumin,
bisdemethoxoycurcumin, and 2, 5-xylenol) will be isolated and the same experiment as described above will be repeated
for each of the molecules to establish the most effective molecule and its concentration against Escherichia coli and
Salmonella typhi.
2- The turmeric can be used as a food and meat preservative because it has the antiseptic effect, anti-inflammation effect,
antibacterial effect, antiviral effect and antifungal effect.
3- The turmeric based pharmaceutical drugs can be prescribed to patients where all other antibiotics have failed.
REFERENCES:
1- Dobelis IN, ed. 1986. Magic and Medicine of Plants. Pleasantville, NY: Reader’s Digest Association, Inc. 1986.
2- Leung A. 1980. Encyclopedia of Common Natural Ingredients Used in Food, Drugs, and Cosmetics. New York, NY: John
Wiley; 1980:313-314.
3- Chandra D, Gupta S. 1972. Anti-inflammatory and anti-arthritic activity of volatile oil of Curcuma longa (Haldi) Indian J Med
Res1972;60:138-142.
4- Arora R, Basu N, Kapoor V, et al. 1971. Anti-inflammatory studies on Curcuma longa(turmeric). Indian J Med
Res1971;59:1289-1295.
5- Mukhopadhyay A, Basu N, Ghatak N, et al. 1982. Anti-inflammatory and irritant activities of curcuminana-logues in rats.
Agents Actions1982;12:508-515.
6- Srivastava R. 1989. Inhibition of neutrophil response by curcumin. Agents Actions1989;28:298-303.
7- Allen PC, Danforth HD, Augustine PC. 1998. Dietary modulation of avian coccidiosis. Int J Parasitol1998;28:1131-1140.
8- Ramirez-Tortosa MC, Mesa MD, Aguilera MC, et al. 1999. Oral administration of a turmeric extract inhibits LDL oxidation
and has hypocholesterolemic effects in rabbits with experimental atherosclerosis. Athero-sclerosis 1999;147:371-378.
9- Srivastava R, Puri V, Srimal RC, Dhawan BN. 1986. Effect of curcumin on platelet aggregation and vascular prostacyclin
synthesis. Arzneimittelforschung1986;36:715-717.
Article
Blood neutrophils, when exposed to appropriate stimuli, aggregate, degranulate and generate superoxide anion. Curcumin, a non-steroidal antiinflammatory agent, modulated these functions, depending upon the kind of stimulus used. It inhibited monkey neutrophil aggregation induced by chemotactic peptide fmlp and zymosan activated plasma (ZAP) but did not affect that induced by serum treated zymosan (STZ) and arachidonic acid (AA). Generation of O 2− radical was inhibited by curcumin, when cells were stimulated by AA, STZ and fmlp. Curcumin inhibited the release of myeloperoxidase, an azurophilic granule marker enzyme. Release of lysozyme was less susceptible to inhibition by curcumin. The results suggest that curcumin interferes with neutrophil responses to various physiological stimuli and a part of its antiinflammatory action is mediated via inhibition of neutrophil function. Inhibition of neutrophil function by curcumin appears to be mediated via calcium dependent mechanisms.
Article
In vitro and ex vivo effects of 1,7-bis(4-hydroxy-3-methoxyphenyl)-1,6-heptadiene-3,5-dione (diferuloylmethane, curcumin) and acetylsalicylic acid (ASA) on the synthesis of prostacyclin (PGI2) and on platelet aggregation has been studied in rat. Both drugs inhibited adenosine diphosphate (ADP)-, epinephrine (adrenaline)- and collagen-induced platelet aggregation in monkey plasma. Pretreatment with ASA (25-100 mg/kg), but not curcumin (100-300 mg/kg), inhibited PGI2 synthesis in rat aorta. In the in vitro system, too, curcumin caused a slight increase in the synthesis of PGI2, while ASA inhibited it. Curcumin may, therefore, be preferable in patients prone to vascular thrombosis and requiring antiarthritic therapy.
Article
The anti-inflammatory activity of curcumin (C), sodium curcuminate (NaC), diacetyl curcumin (DAC), triethyl curcumin (TEC), tetrahydro curcumin (THC) and ferulic acid (FA) was compared with that of phenylbutazone (PB) using the carrageenin-induced rat paw edema and cotton pellet granuloma tests. The rank order of potencies of curcumin analogues and PB in carrageenin-induced inflammation were NaC greater than THC greater than C greater than PB greater than TEC. The curcumin analogues de decreased carrageenin-induced paw edema at low doses, however, at higher doses this effect was partially reversed. FA and DAC were devoid of anti-inflammatory activity. Curcumin analogues were less effective in inhibiting the granulomatous tissue formation. Maximum activity was observed with TEC whereas C, NaC and PB were almost half as effective as TEC. C and NaC possess both anti-inflammatory and irritant properties as was evident from experiments in which drugs were incorporated into carrageenin and the cotton pellets for inducing inflammation. The anti-inflammatory action of NaC is not mediated through release of steroids from the adrenal cortex or inhibition of the biosynthesis of prostaglandins from arachidonic acid. The results of the present study support the rationale for the use of powdered rhizome of tumeric (contains 0.6% of curcumin) for conditions of sprain and inflammation.
Article
During the past several years, our laboratory has been investigating the anticoccidial activities of various natural products that have potential use as dietary supplements for coccidiosis control. Sources of fats containing high concentrations of n-3 fatty acids such as menhaden oil and flaxseed oil and flaxseed, when added to starter rations and fed to chicks from one day of age, effectively reduce lesions caused by the caecal parasite Eimeria tenella, but not lesions caused by Eimeria maxima. Our results are consistent with reports of effects of diets high in n-3 fatty acids on other protozoan parasites which suggest that the state of oxidative stress induced by these diets in the cells of both host and parasites is responsible for their parasitic actions. Artemisinin, a naturally occurring (Artemisia annua) endoperoxide and effective antimalarial significantly lowers lesions from E. tenella when given at low levels as a feed additive. The mechanism of its action is also considered to involve induction of oxidative stress. Diets supplemented with 8 p.p.m. gamma-tocopherol (abundant in flaxseeds) or with 1% of the spice tumeric, reduce mid-small intestinal lesion scores and improve weight gains during E. maxima infections. These compounds may exert their anticoccidial activity because they are effective antioxidants. Betaine, a choline analogue found in high concentrations in sugar beets, improves nutrient utilisation by animals under stress. When provided as a dietary supplement at a level of 0.15% it has enhanced the anticoccidial activity of the ionophore, salinomycin. Betaine may act as an osmoprotectant whereby it improves the integrity and function of the infected intestinal mucosa. In in vivo studies, betaine plus salinomycin significantly inhibit invasion of both E. tenella and E. acervulina. However, subsequent development of E. acervulina is inhibited more effectively with this combination treatment than development of E. tenella.
Article
The oxidation of low-density lipoproteins (LDL) plays an important role in the development of atherosclerosis. Curcumin is a yellow pigment obtained from rhizomes of Curcuma longa and is commonly used as a spice and food colouring. Curcumin and turmeric extracts have several pharmacological effects including antitumour, anti-inflammatory, antioxidant and antiinfectious activities although the precise mechanisms involved remain to be elicited. We evaluated the effect of an ethanol-aqueous extract obtained from rhizomes of C. longa on LDL oxidation susceptibility and plasma lipids in atherosclerotic rabbits. A total of 18 rabbits were fed for 7 weeks on a diet containing 95.7% standard chow, 3% lard and 1. 3% cholesterol, to induce atherosclerosis. The rabbits were divided into groups, two of which were also orally treated with turmeric extract at doses of 1.66 (group A) and 3.2 (group B) mg/kg body weight, respectively. A third group (group C) acted as a control. Plasma and LDL lipid composition, plasma alpha-tocopherol, plasma retinol, LDL TBARS, LDL lipid hydroperoxides and analysis of aortic atherosclerotic lesions were assayed. The low but not the high dosage decreased the susceptibility of LDL to lipid peroxidation. Both doses had lower levels of total plasma cholesterol than the control group. Moreover, the lower dosage had lower levels of cholesterol, phospholipids and triglycerides in LDL than the 3.2-mg dosage. In conclusion, the use of this extract could be useful in the management of cardiovascular disease in which atherosclerosis is important.
Magic and Medicine of Plants. Pleasantville, NY: Reader's Digest Association, Inc. 1986. 2-Leung A. 1980. Encyclopedia of Common Natural Ingredients Used in Food, Drugs, and Cosmetics
  • I N Dobelis
Dobelis IN, ed. 1986. Magic and Medicine of Plants. Pleasantville, NY: Reader's Digest Association, Inc. 1986. 2-Leung A. 1980. Encyclopedia of Common Natural Ingredients Used in Food, Drugs, and Cosmetics. New York, NY: John Wiley; 1980:313-314.