Inhibitory effect of some traditional hair washing substances on hair borne bacteria

Transcription

1 Available online at Scholars Research Library Der Pharmacia Lettre, 2012, 4 (1): ( ISSN USA CODEN: DPLEB4 Inhibitory effect of some traditional hair washing substances on hair borne bacteria S. Tharmila*, E. Christy Jeyaseelan and A.C. Thavaranjit Department of Botany, Faculty of Science, University of Jaffna, Sri Lanka _ ABSTRACT Traditionally people use several plant materials and their extracts as hair washing substances. Plant based natural substances are not harmful as well as they are found to be more effective compared to synthetic one. Therefore, the present study was carried out to evaluate the inhibitory effect of some traditionally using herbal hair washing substances on different hair borne bacterial isolates in vitro. The test bacteria were isolated from the hair of randomly selected fifty students attached to the Faculty of science, University of Jaffna, Sri Lanka, and the bacteria were identified based on morphological, physiological, and biochemical characters. Aqueous extracts of leaves of Clitoria ternatea and Ocimum gratissimum, flower of Hibiscus rosa-sinensis and an herbal hair washing powder (mixture of Trigonella corniculata, Phaseolus mungo, and Trigonella foenum-graecum) were tested against isolated bacteria using agar well diffusion method. A commercial hair washing shampoo was used as standard and sterile distilled water was as control. All the test extracts except O. gratissimum were able to inhibit the growth of at least two of the test bacteria, and their antibacterial activity differed significantly (P < 0.05). The herbal hair washing powder inhibited all the test bacteria except Escherichia coli and Bacillus sphaericus and it produced significantly (P < 0.05) higher inhibition compared to other test extracts. The inhibition produced by all the test extracts except O. gratissimum against Erwinia herbicola and herbal hair washing powder against Aerococcus sp. were relatively higher than that produced by the standard on respective bacteria. Present study has experimentally proved the effectiveness of above plant based hair washing substances and enriched the value of traditional knowledge. Key words: Medicinal plants, hair washing substance, hair borne bacteria, antibacterial activity. _ INTRODUCTION Naturally human hair harbours many pathogenic bacteria, and also it acts as a potential source of cross infections. Bacteria like Staphylococcus aureus, Escherichia coli, Streptococcus viridians, β haemolytic streptococci, the Proteus group, Pseudomonas pyocyanea and Streptococcus faecalis are normally present in the hair [1]. It has been also demonstrated that in hospital 199

2 operating rooms bioaerosol contaminants were mainly associated with emissions from human hair [2]. A recent study showed that about 25% of the cutaneous bacterial population was localized within the hair follicles [3]. Nowadays there are number of modern synthetic hair washing substances are available in market. Although they show greater antibacterial activity for short term, they are extremely harmful to hair in long run. On the other hand, several people still rely on plant materials as natural hair washing substances. There is a wide spread belief that green plants sources are healthier and more harmless or safer than synthetic ones [4]. Flower of Hibiscus rosa-sinensis and leaves of Clitoria ternatea and Ocimum gratissimum are well known hair washing substances in Jaffna, Sri Lanka. In addition to that, home made herbal hair powder also popular, it is made by combination of three different plant powders. Hibiscus rosa-sinensis (Malvaceae) is a large spreading shrub with a brownish bark, young stems reddish brown. Leaves are simple, alternate, ovate, irregularly and coarsely serrate along the upper 2/3 of the blade. Flowers are regular, bisexual, solitary, axillary, bright red. Clitoria ternatea (Leguminosae) is a perennial twining herb with long, cylindrical stems covered with scantily adpressed hair. Leaves alternate, stipulate, imparipinnate compound rachis. Flowers are irregular, blue or white, solitary on a drooping legume. Ocimum gratissimum (Labiatae) is a semi woody, perennial herb, stem m tall, often woody below, much branched. Leaves are simple and opposite. Flowers are irregular, bisexual and pale greenish yellow. Herbal hair powder mixture containing fruit of Trigonella corniculata, seeds of Phaseolus mungo and Trigonella foenum-graecum. All are belongs to the family Leguminosae [5]. The present study was carried out to experimentally prove the antibacterial activity of commonly using hair washing substances, H. rosa-sinensis, C. ternatea, O. gratissimum and herbal powder (mixture of fruit of T. corniculata, seeds of P. mungo and T. foenum-graecum) against hair borne bacterial isolates. MATERIALS AND METHODS Collection of plant materials The plant parts used in this study, Flower of Hibiscus rosa-sinensis, leaves of Clitoria ternatea, and Ocimum gratissimum were collected from the Botanical garden, Department of Botany, University of Jaffna, Sri Lanka. The fresh plant materials were washed under running tap water. Herbal hair powder was prepared by grinding the dry fruits of Trigonella corniculata, seeds of Phaseolus mungo and Trigonella foenum-graecum in a ratio of 4: 2: 1 / kg. Preparation of aqueous extracts 10 g of each of the fresh plant parts of H. rosa-sinensis, C. ternatea and O. gratissimum was crushed with 20 ml distilled water by using motor and pestle separately. The aqueous extracts were then filtered through four layered muslin cloths and the filtrates were collected into sterile Mac-Cartney bottles. The extracts were immediately used for the assay. Simultaneously, aqueous extract of herbal hair powder was prepared by adding 20 ml of boiled distilled water to 10 g of fine powdered mixture in a beaker and stirred well for 10 minutes. Then it was filtered through four layered muslin cloth, and used immediately for the test. Test bacteria Bacterial species used in the current study were isolated by cotton swap method under aseptic condition from hairs of randomly selected fifty pupil of the Faculty of Science, University of 200

3 Jaffna. These isolated bacteria were collected and incubated in sterile saline water at 37 0 C for 15 minutes and then streaked separately on nutrient agar media. Plates were allowed to incubate at 37 0 C for 24 hours. Then the bacteria were identified and characterized based on morphological, physiological and biochemical tests. The test organisms were stored on nutrient agar slants at 4 C and these were sub cultured before 24 hours of the experiment. Determination of antibacterial activity Antibacterial activity of the extracts was tested by agar well diffusion method according to Jeyaseelan et al. [6], briefly, 1 ml of the test culture [10 6 CFU/ml] was mixed with 20 ml sterile nutrient agar. It was then allowed to solidify. 8 mm diameter wells were made on the surface of agar plates using a sterile cork borer, and were filled with 100 µl of each extracts. 100 µl of Meera herbal shampoo (50% v/v) and 100 µl of sterile distilled water were used as standard and control respectively. The plates were incubated at 37 C for 24 hours. After incubation, the diameter of the inhibition zones was measured. The experiment was repeated thrice. Statistical analysis The data were analyzed based on one-way analysis of variance (P < 0.05). Then mean values were compared using least significant difference test at α=0.05 by using software the SPSS system for windows (version 13.0). RESULTS AND DISCUSSION The morphological and biochemical tests for the identification of bacteria revealed the presence of eight different bacteria (Bacillus coagulans, Yersinia sp., Erwinia herbicola, Aerococcus sp., Bacillus. brevis, Escherichia coli, Bacillus sphaericus and Alcaligenes sp.) in the samples collected from randomly selected pupils (Table 1&2). Among these B. coagulans showed higher frequency, and it was followed by E. coli and B. brevis. E. herbicola was present only in 4% of the samples (Table 3). Table 1: Morphological, biochemical and physiological characters of bacterial isolates Sample Shape Simple staining Arrangement Grams staining capsule catalase Endospore Facultative anaerobc Strickly aerobic Oxidase Motility Starch hydrolysis E. coli Small rod Strepto (-) +3 X - (-) (-) Yersinia sp Small rod Strepto (-) X +3 X - (-) (-) Alcaligenes sp Very tiny rod Strepto (-) +4 X - Aerococcus sp Spherical Tetra coccus +4 X - (-) X (-) B. coagulans Small rod Strepto +4 central - (-) E. herbicola Small rod Diplo (-) + X - (-) B. sphaericus Small rod Strepto + B. brevis Rod Strepto + terminal central - (-) - present, X absent, /+ - positive/yes, (-)/- - negative/no Strong (-) (-) 201

4 Table 2: Biochemical characters of bacterial isolates Sample Casein Lecithin Gelatin Hydrolysis Indole test VP Test MR Test Citrate Urea Hydrolysis Nitrate reduction Oxidative Test Fermentation Test Sucrose Fermentation Glucose Fermentation H 2S Formation Alkali slant Acid butt H2S (g) E. coli (-) (-) (-) (-) strong No (2+) (-) (-) X Yersinia sp (-) (-) (-) (-) (-) (-) strong (3+) (-) (2+) (-) (-) (-) X Alcaligenes sp (-) (-) (-) (-) (-) slight (-) (2+) (-) (-) (-) X Aerococcus sp (-) (-) (-) (-) (-) (-) (2+) (2+) (-) (-) (-) (-) X B. coagulans (-) (-) (-) (-) (-) (-) very slight less (-) No No (-) (-) X E. herbicola (-) (-) (-) (-) (-) (-) B. sphaericus (-) (-) (-) (-) (-) (-) B. brevis (-) (-) (-) slight Slight very less (-) No (-) X (-) (-) (-) X strong (2+) (-) No (-) (-) X present, X absent, /+ - positive/yes, (-)/- - negative/no, VP test- Vo Proskauer test, MR test- methyl red test Table 3: Frequency of bacterial isolates in randomly selected hair samples Bacteria Frequency (%) Bacillus coagulans 36 Yersinia sp. 6 Bacillus brevis 16 Escherichia coli 30 Bacillus sphaericus 10 Alcaligenes sp. 6 Erwinia herbicola 4 Aerococcus sp. 6 The antibacterial study of hair washing substances revealed that all the test extracts except O. gratissimum had inhibitory effect on test bacteria and the diameter of inhibition zone differed significantly (P < 0.05). The herbal hair powder expressed better inhibition among the test extracts, which inhibited the growth of six test bacteria out of tested eight bacteria. It revealed higher activity on B. coagulans, E. herbicola and Aerococcus sp. with 21.8 mm, 20.9 mm and 18 mm mean diameter of clear zones respectively. However, other three bacteria, Yersinia sp., B. brevis and Alcaligenes sp. showed between 10 to 11 mm mean diameter of clear zones. C. ternatea also had inhibitory effect on four tested bacteria and the highest inhibition was produced against E. herbicola and it was followed by B. coagulans, Aerococcus sp. and Yersinia sp. H. rosa-sinensis was only able to inhibit the growth of E. herbicola and B. coagulans, where E. herbicola was highly inhibited (20 mm) than B. coagulans (12 mm). O. gratissimum failed to inhibit any of the test bacteria (Table 4). 202

5 Table 4: Antibacterial activity of different herbal extracts on hair borne bacteria Zone of inhibition (mm) Aqueous extracts B. coagulans Yersinia sp E. herbicola Aerococcus sp B. brevis E. coli B. sphaericus Alcaligenes sp Leave of O. gratissimum Flower of H.rosa-sinensis 11.9 ± 0.3 c ± 0.4 a Leave of C.ternatea 18.7 ± 0.4 b 10.8 ± 1.0 a 19.1 ± 0.3 b 13.8 ± 0.1 b Herbal hair powder 21.8 ± 0.2 a 10.8 ± 0.1 a 20.9 ±0.1 a 18.0 ± 0.1 a 10.9 ± ± 0.1 Standard 25.4 ± ± ± ± ± ± ± ± 0.1 Control (-)No activity. Values are mean ± SD. Values with different superscript on the same column show significant (P <0.05) difference.zone of inhibition includes the diameter of the well (8mm in diameter) B. coagulans and E. herbicola were inhibited by three extracts among the tested four extracts, and Yersinia sp. and Aerococcus sp. were inhibited by only two extracts. Although B. brevis and Alcaligenes sp. were only sensitive to herbal hair powder, E. coli and B. sphaericus were not inhibited by any of the test extracts (Table 4). The standard commercial shampoo inhibited the growth of all test bacteria, and the diameter of clear zone ranged from 25.4 mm for B. coagulans to 13.0 mm for Alcaligenes sp. Interestingly, even though it produced higher activity against B. coagulans, Yersinia sp., B. brevis and Alcaligenes sp. compared to test extracts, the results produced by the herbal hair powder, C. ternatea and H. rosa-sinensis against E. herbicola and by the herbal hair powder against Aerococcus sp. were higher than the standard. The control, sterile distilled water had no effect on any of the test bacteria at all. The antibacterial activity of plant extracts depends on the available bioactive secondary metabolites in the plant part. It has been reported that H. rosa-sinensis possess flavonoids, saponins, tannins and terpenoids [7]. The important chemical constituents of T. foenum-graecum are saponins, coumarin, fenugreekine, nicotinic acid, phytic acid, scopoletin and trigonelline [8]. C. ternatea consist alkaloids, flavonoids, saponins and tannins [9]. Therefore, the inhibitory effect of these extracts may be due to the presence of these phytochemicals. CONCLUSION Present study clearly demonstrated the antibacterial activity of traditional hair washing substances, Hibiscus rosa-sinensis, Clitoria ternatea and herbal powder on hair borne bacteria. Therefore, present study has enriched the value of traditional knowledge regarding the use of plant based hair washing substances. REFERENCES [1] M.M. Summers, P.F. Lynch, T. Black, J. clin. Path., 1965, 18, [2] J.S. Pastuszka, E. Marchwiñska-Wyrwa, A. Wlaz, Polish Journal of Environmental Studies, 2005, 14, [3] B. Lange-Asschenfeldt, D. Marenbach, C. Lang, A. Patzelt, M. Ulrich, A. Maltusch, D. Terhorst, E. Stockfleth, W. Sterry, J. Lademann, Skin Pharmacol Physiol, 2011, 24,

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