Publication Ref No.

: IJPRD/2010/PUB/ARTI/VOV-2/ISSUE-9/NOV/031

ISSN 0974 9446

ANTI TYPHOID ACTIVITY OF AQUEOUS EXTRACT OF FRUIT PEEL CITRUS SINENSIS (L.) Vivek Kumar R1*, Nandini1, Shashidhara S1, Anitha S1 Dept of pharmacognosy, Government College of pharmacy, Bangalore,Karnataka,India. Email:vivek.jsspharma@gmail.com
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Vivek Kumar

ABSTRACT Typhoid fever (TF) is an acute systemic infection caused by Salmonella Typhi, Salmonella Para Typhi A and Salmonella Para Typhi B. It is transmitted by the fecal oral route mainly via contaminated food and water. The developing countries have high rate of morbidity and mortality due to Typhoid fever, epidemics take place in developed world also. There are increased incidences of multi drug resistant in S. typhi strains that has further complicated its management and only a few antibiotics are now effective in treatment of typhoid. We report that the aqueous extracts of fruit peel Citrus sinensis (L.) confer anti typhoid activity against Salmonella Typhi, Salmonella Para Typhi A and Salmonella Para Typhi B respectively on comparison with ciprofloxacin. Key Words: Typhoid fever, Salmonella Typhi, Salmonella Para Typhi A , Salmonella Para Typhi B, anti typhoid activity, Citrus sinensis.. INTRODUCTION Typhoid fever (TF) caused by Salmonella enterica serotype Typhi (S. typhi) is a major public health problem, particularly in developing countries. TF caused 21,650,974 illnesses and 216, 510 deaths annually (Crump, 2004). In Western countries, the disease has been brought very close to eradication levels. Multidrug resistant (MDR) S. typhi strains have been reported from different parts of India (Lakshmi, 2006; Achla, 2005; Dutta, 2005; Walia, 2005;Madhulika, 2004; Safdar, 2004; Chande, 2002) that have created a significant therapeutic problem. In spite of the increasing resistant to the antibiotics and vaccines, it is pertinent to evaluate the efficacy of the natural plant products in the treatment of typhoid by evaluating anti typhoid activity against Salmonella Typhi, Salmonella Para Typhi A and Salmonella Para Typhi B . The Botanical name of fruit is Citrus sinensis (L.) belong to the Family RUTACEAE.

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Vernacular name is sweet orange, blood orange(english), musambi, malta, mosambi(hindi), nagaranga( Sanskrit). An evergreen shrub or tree, up to 12 m high; twigs angled when young, often with thick spines. Leaves are oval, 5-15 cm long and 2-8 cm wide, shiny dark green above; stalk narrowly winged. Flowers very fragrant, one or many in a leaf axil, 2-3 cm across, five white petals, stamens in groups, style with round stigma. Fruit rather variable in colour and shape, rounded green-yellow-orange, 4-12 cm in diameter, relatively thin skin, hard to remove. Bekele-Tesemma, West Indies and West Africa; widely cultivated in the sub-tropics. Dried peel admixed with cowpea and dried fish chips produced an LD50 of 4 percent (w/w) for C. maculatus and 14 percent (w/w) for D. maculatus. In repellency trials, 10 percent (w/w) admixed with cowpeas or fish chips repelled both insect species. At 18 percent (w/w) on fish chips, larval emergence of D. maculatus was reduced by 60 percent; only 37 percent of the larvae developed into adults compared with 88 percent in the untreated controls Dried leaves Three percent (w/w) admixed with wheat reduced oviposition (3.7 eggs), adult emergence (9.7 insects) and prolonged development of R. dominica (52.2 days) with regards to the control (17.1 eggs, 77.3 insects and 40 days respectively). Constituents Include flavonoids like citacridone, citbrasine and noradrenaline. The present investigation review the aqueous extracts of fruit peel Citrus sinensis (L.) against Salmonella Typhi, Salmonella Para Typhi A and Salmonella Para Typhi B respectively on comparison with ciprofloxacin. MATERIAL AND METHODS Fruit peel: Fruit peels of sweet orange were procured from local juice stall of Bangalore.

Prior to use, it was insured that the fruit peel were free from contamination, and had no microbial growth. PREPARATION OF EXTRACT: Fruit peels of sweet orange were collected from local juice stall and dried in hot air oven at temperature not more than 55C. Powdered to mesh 60.About 100g of powdered fruit peels were taken in maceration flask and kept for maceration for a period of 6 days with 100 ml of water. Then collected the extract filtered and concentrated on a hot plate. This brownish crystalline powder was used to screen anti typhoid activity. BACTERIA CULTURES: The standard strains of S.typhi, S.Para typhi A, and S.Para Typhi B were procured from the microbiology laboratory of government college of GCP, Bangalore. The bacterial strains were subculured on nutrient agar followed by incubation at 371C for 24 hours and then used for study. PHYTOCHEMICAL ANALYSIS The freshly prepared extracts were subjected to standard phytochemical analyses to test for the presence of the phytoconstituents flavonoids, saponins, and alkaloids. DETERMINATION ACTIVITY: OF ANTIMICROBIAL

Antimicrobial activity of the aqueous extracts of fruit peel Citrus sinensis (L.) was evaluated by the agar diffusion method. Aqueous extract of fruit peel Citrus sinensis (L.) were dissolved in dimethyl sulphoxide (DMSO) to get the concontration of 100mg/ml.Ciprofloxacin was used as postive control and DMSO was used as blank. Agar 218

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plates containing typhoid causing organisms were bored with the use of steril borer of 6mm radius. 2omg/ml,40mg/ml,60mg/ml,80mg/ml,100mg/ml of Aqueous extract and 100g/ml of Ciprofloxacin and blank(DMSO) was added to weii and incubated for 24hours at 371c. The diameter of the zones of inhibition around each of the well was taken as measure of the antibacterial activity. Each experiment was carried out in triplicate and mean diameter of the inhibition zone was recorded. (Refer Table No. 01 and Figure No.01)

DISCUSSION Phytochemical constituents such as flavonoids, saponins, alkaloids are secondary metabolites of plants that may serve as defense mechanisms against S.typhi, S.paratyphiA, and S.paratyphiB .This may therefore explain the demonstration of anti typhoid activity by the aqueous extracts of fruit peel Citrus sinensis (L.) OSBECK. The demonstration of anti typhoid activity S.typhi, S.paratyphiA, and S.paratyphiB may be indicative of the presence of broad spectrum antibiotic compounds. This will be of immense advantage in fighting the menace of antibiotic refractive pathogens that are so prevalent in recent times. CONCLUSION The demonstration of broad spectrum of anti typhoid activity of aqueous extracts of fruit peel Citrus sinensis (L.) OSBECK may help to discover new chemical classes of antibiotic substances that could serve as selective agents for infectious disease chemotherapy and control. This investigation has opened up the possibility of the use of this plant in drug development for human consumption possibly for the treatment of typhoid fever. The effect of fruit peel extracts on more pathogenic organisms and toxicological investigations and further purification however, needs to be carried out. REFERENCES: 1. Bibitha B, Jisha VK, Salitha CV, Mohan S, Valsa AK.Antibacterial activity of different plant extracts.Short Communiaction. Indian J Microbiol 2002; 42:361-363. 2. Marjorie MC. Plant products as antimicrobial agents. Clin Microbiol Rev1999., 12(4): 564-582. 3. Srinivasan D, Perumalsamy LP, Nathan S, Sures T.Antimicrobial activity of certain

Graphical representation of various concentrations of aqueous extracts of fruit peel Citrus sinensis (L.) on S. Typhi, S. Paratyphi A, S. Paratyphi B RESULTS Phytochemical constituents present in the fruit peel extract included flavonoids, saponins, alkaloids Results of the antimicrobial activity of the fruit extracts are shown in Table 1. The highest activity (diameter of zone of inhibition 180 mm) was demonstrated by aqueous extracts of fruit peel Citrus sinensis (L.) OSBECK against Salmonella Para TyphiA while the lowest activity(diameter of zone of inhibition 10 mm) was demonstrated against Salmonella Typhi. Gradual increase in activity with concentration and results shown in figure 2A, 2B, and 2C.

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Indian medicinal plants used in folkloric medicine. J Ethnopharm 2001; 94: 217-222. 4. Molan, PC. The antibacterial activity of honey 1. The nature of antibacterial activity. Bee World 1992;473: 59 76. 5. JH Doughari. Antimicrobial Activity of Tamarindus indica Linn. Tropical Journal of Pharmaceutical Research, December 2006; 5 (2): 597-603. 6. A. Madani and S. K. Jain. Protection against experimental salmonellosis by Terminalia belerica and Punica granatum extracts: immunological evaluation. Indian Journal of Science and Technology 2008;( 3). 7. Achla P, Grover SS, Bhatia R and Khare S Sensitivity index of antimicrobialagents as a simple solution for multidrug resistance in TABLES AND FIGURES:

Salmonella typhi. Ind. J. Med.Res.2005;121; 185-93. 8. Trivedi VB and Kazmi SM Kachnar and anar as antibacterial drugs. Indian Drugs.1979;16;295. 9. Ahmad K Experts call for surveillance of drug-resistant typhoid at a global level. Lancet.2002;16;592. 10. Ahmad I and Beg AZ Antimicrobialand phytochemical studies on 45 Indian medicinal plants against multi-drug resistant human pathogens. JEthnopharmacol. 2001;74;113-23. 11. Singh amandeep, ahmed r. bilal,anil devgun. In vitro antibiotic activity of isolated volatile oil of citrus sinensis.ijprd.2001;07.

Table 1: Zone of inhibition of various concentrations of aqueous extracts of fruit peel Citrus sinensis (L.) on typhoid organism.

Sample S.TYPHI 36 6 10 16 20 24

Zone of inhibition in mm S.PARA TYPHI A 38 8 12 18 24 30 S.PARA TYPHI B 40 7 14 20 26 31

Ciprofloxacin Aqueous exract 20mg/ml 40mg/ml 60mg/ml 80mg/ml 100mg/ml

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Fig 2- Effect of various concentrations of aqueous extracts (1- 20mg/ml, 2- 40mg/ml,3- 60mg/ml, 4- 80mg/ml, 5- 100mg/ml, B- blank, Cip ciprofloxacin.) on A S. Typhi, B S. Paratyphi A, C- S. Paratyphi B

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