In-vitro Pediculicidal Activity of Juice from Clove of Allium sativum

 

Vivek Shrivastava and U.K. Jain*

Bhopal Institute of Technology and Science -Pharmacy, Bhojpur road, Bhopal (M.P).

*Corresponding Author E-mail: ukjain65@gmail.com

ABSTRACT:

Head lice are a global public-health apprehension affecting primary school age children. The control of lice presents research challenges and prediction for the identification of new, safe insecticides. Allium sativum (commonly known as garlic) is a species belonging to family Alliaceae. Garlic has been used for both culinary and medicinal purposes. In an attempt to assess the lethality, Allium sativum aqueous extracts were tested in In- vitro toxicity model against human adult lice. The lice were observed for lack of response to stimuli over 3 hour period. The results demonstrated that raw juice of Allium sativum clove showed pediculicidal efficacies of 90±10 per cent (P< 0.01 over control). The experimental evidence obtained in the laboratory model could provide a rationale for the use of Allium sativum juice to be included in formulations for controlling head lice.

 

 

INTRODUCTION:

Head lice are a global public-health apprehension affecting primary school age children. They depend on the host for survival and need to eat every three-to-four hours. They feed by injecting saliva with vasodilatory properties into the scalp to draw blood¹.

 

Head lice are tiny six-legged insects that hold to the scalp and neck and nourish on human blood. The head louse is a grey-white animal about 2-3 mm in length. There are three different types of lice infections and each is caused by a unique parasite. A Head louse (Pediculus humanus capitis) is the most common infestation and favors no particular socioeconomic group. Pediculus humanus corporis, is responsible for body lice that is more commonly associated with poverty, and poor hygiene. Pubic lice ("crabs") is caused by Pthirus pubis and is transmitted by intimate and/or sexual contact.

 

The life span of the female louse is about one month. During this time, she will produce between seven to 10 eggs ("nits") per day and attach them firmly to the hair shaft region close to the scalp or body. Following six to 10 days, the nits produce nymphs and grew adults in 10 days. The mode of transmission is most commonly via direct head-to-head contact, Sharing pillows, combs/brushes, caps, headphones.

 

Lice do not often cause significant illness (typhus, relapsing fever and trench fever), but a lice infestation is really a itchy and uneasy experience. Although the head lice are not a main health hazard yet they can cause substantial distress due to erythema, itching and probable secondary infections. The clinical manifestations of head lice include pruritus, excoriation, local erythema, scalp impetigo, papules and cervical and occipital lymphadenopathy. Wet combing is a safe and effective method of eradication, especially when combined with a conventional or natural pediculicide. Many herbal medicines and essential oils appear to be effective at eradicating both the adult lice and their eggs¹.

 

Allium sativum, commonly known as garlic, is a species belonging to family Alliaceae. Garlic has been used for both culinary and medicinal purposes. It has a characteristic pungent, spicy flavor that mellow and sweetens considerably with cooking. A bulb of garlic, the most commonly used part of the plant, is divided into numerous fleshy sections called cloves. The cloves are used as seed, for consumption (raw or cooked), and for medicinal purposes².

 

A wide range of microorganisms have been shown to be sensitive to crushed garlic preparations. The use of garlic has been investigated as antitumor³, antifungal⁴, antibacterial⁵, as well as possessing antithrombotic⁶ properties. Garlic has been reported to reduce blood lipids⁷. Garlic is a very complex compound consisting not only of allyl components but many other components, some of which (kaempferol and quercetin) have been shown to have antigiardial activity in vitro⁸. Chemical analysis of garlic have revealed an unusual concentration of sulfur-containing compounds (1–3%)⁸.

Table 1: Licicidal activity of Allium sativum clove juice

Group	Treatments	Mean death time (In minutes± S.D)	Percent Morbidity after 3 hours	P value Vs Negative control
Negative control group	0.5 ml of Distill water	--------	0
Test Group 1	0.5 ml, 1/10 diluted juice of Allium sativum clove	--------	0
Test Group 2	0.5 ml, 1/2 diluted juice of Allium sativum clove	110 ± 3.19	65± 2.89	P<0.01
Test Group 3	0.5 ml Raw juice of Allium sativum clove	71 ± 2.83	90±10	P<0.01
positive control group	0.5 ml of 1 % w/w Lindane solution	50±2.12	95±5.77	P<0.01

 

The test was done in duplicate and average considered. Values are presented as the mean ± SE (standard error); n = 10 for all groups

One way ANOVA followed by Dunnett compare all vs control test.

 

MATERIALS:

Plant material:

Cloves of Allium sativum (Alliaceae) were purchased from local market and were identified and authenticated at Department of Pharmacognosy, BITS-Pharmacy, Bhopal.

 

Experimental Organism:

Human head lice Pediculus humanus capitis were collected from children between the age of 3-12, with the approval of their parents, residing in Bangrasia near Bhopal (M.P.). The insects were collected by combing the children scalps.

 

METHODS:

Preparation of Garlic juice:

The garlic preparations used in this study were raw garlic juice. It was prepared from raw garlic cloves crushed in a blender for 3min and juice was allowed to stand for 30 min at 25^oC. After filtering through cheese cloth, raw garlic juice was obtained⁹.

 

Test solutions:

Test solutions of Allium sativum extracts were prepared. ½ dilution and 1/10 dilution of aqueous extract were also prepared.

 

Standard solution:

1 % w/w Lindane solution was used as standard

 

Pediculicidal activity:^10-12

The In-vitro tests were started within one hour after collection. A filter paper diffusion bioassay was made. After careful selection of lice under a dissecting microscope, filter paper discs (Whatman No 1) matching with internal diameter of petridish were cut, soaked in respective test solution and placed in petridishes. 0.5 ml of each test solution was spread over the lice and filter paper discs. Negative control lice were placed directly on the filter paper spread with only distill water. The 1 % w/w Lindane solution was simultaneously run as a positive control. The number of lice was 10 in each group. The licicidal activity were investigated with raw Allium sativum juice and 1/2 dilution and 1/10 dilutions of raw juice with distilled water. The lice were judged as dead if there were no vital signs such as movements of antennae or minimal leg movements upon stimulation with or without a forceps. The test was done in triplicate and average considered.

 

Statistical analysis:

The data were subjected to statistical analysis. One-way ANOVA followed by Dunnett post-test was employed to identify pairs of results with significantly different means. Both ANOVA and post-test were performed by Graph- Pad InStat computer package.

 

RESULTS:

The results demonstrate that aqueous extract of Allium sativum clove show pediculicidal efficacies as shown in table 1. The mortality rate evaluated three hours after treatment was 90±10 per cent for the test group 3 (Raw juice of Allium sativum) compared to 65± 5 per cent for the 1/2 diluted juice (test group 2) and 0 per cent for negative control and test group 1. Positive control group, test group 2 and test group 3 showed significant difference over negative control (P< 0.01). Raw juice of Allium sativum showed significant pediculicidal activity.

 

CONCLUSION:

The control of lice presents research challenges and prospects for the identification of new, safe and environmentally acceptable insecticides. The experimental evidence obtained in the laboratory model could provide a rationale for the use of Allium sativum aqueous extracts for controlling head lice which are difficult to control because of their resistance to the currently used anti-louse agents. The overall results of the study indicate that the Allium sativum aqueous extracts is highly effective against adult head lice.

 

REFERENCES:

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9.           Kasuga. 2001. In: Proceedings of “Recent Advances on the Nutritional Benefits Accompanying the Use of Garlic as a Supplement” held during November 15–17 in Newport Beach, CA.1998.

10.     Meinking TL, Taplin D, Kalter DC, Eberle, MW, Comparative efficacy of treatments for pediculosis capitis infection. Arch Dermatol, 1986; 122: 267-71.

11.     Nalamwar VP, Khadabadi SS, Aswar PB, Kosalge SB, Rajurkar RM. In vitro Licicidal activity of different extracts of Acorus calamus Linn. (Araceae) Rhizome. International Journal of Pharm Tech Research, 2009; 1: 96-100.

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Received on 15.04.2010       Modified on 13.05.2010

Accepted on 29.05.2010      © RJPT All right reserved