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Efficacy of Herbal Shampoo and Pesticides for Head Lice

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Efficacy of herbal shampoo of Garcinia dulcis(Roxb.) Kurz ( Clusiaceae) compared with chemical pediculicides for head lice treatment in Thailand   

  • Sirawut Sittichok1 and Mayura Soonwera

Abstract

Currently, alternative pediculicide from plant are need for head lice treatment because of chemical over-the -counter have lost in efficacy due to increasing resistance of head lice( Pediculus humanus capitis ) to chemical over-the-counter worldwide .As the result , this study was to assess the efficacy of herbal shampoo of Garcinia dulcis (Roxb.) Kurz (Clusiaceae) compared with two chemical pediculides ( carbaryl shampoo ; Hafif shampoo®; 0.6% w/v carbaryl and permethrin shampoo ; Scully Anti-Lice shampoo®; 0.5% w/v permethrin) and baby shampoo (Care baby shampoo®) for head lice treatment. In vitro study used the filter paper method with three doses of 0.002, 0.003 and 0.006 ml/cm2 of each treatment for assessing the toxicity of each treatment on mortality of nymph and adult of head ice. The results revealed that as the concentration of treatment increased from the doses of 0.002, 0.003 and 0.006 ml/cm2, the mortality rate of nymphs and adults of head liceincreased. Moreover,the most pediculicidal activity was shown by the dose 0.006 ml/cm2 of G. dulcis shampoo at 10 min,showing 100% mortality of nymphs( LT50 value of 0.2 min and LC50 value of 0.00001 ml/cm2) and adults(LT50 value of 0.7 min and LC50 value of 0.001 ml/cm2). In addition, these results showed more effective on pediculicidal activity than carbaryl shampoo (LT50 values ranged from 6.3 to 10.1 min), permethrin shampoo (LT50 values ranged from 28.9 to 61.8 min) and baby shampoo (LT50 values ranged from 77.3 to 87.1 min). For the results of in vivo test showed that G. dulcis shampoo also exhibited 100 % of cure rate after the 2nd and 3rd treatments and these result was more effective for head lice treatment than two chemical pediculicides and baby shampoo( cure rate ranged from 0 to 96.7%). Consequently, the results of this study pointed that G. dulcis shampoo showed high potential for to be used as alternative pediculicide for head lice treatment of children in Thailand .

Keywords: Pediculus humanus capitis, herbal shampoo, Garcinia dulcis , carbaryl shampoo, permethrin shampoo

Corresponding author: M. Soonwera () and S. Sittichok

Introduction   

Pediculosis capitis (head lice infestation), caused by the wingless insect, Pediculus humanus capitis De Geer (Phthiraptera: Pediculicidae), is one of the most ectoparasitic infestation of human worldwide and affecting mostly children with more than a million per year worldwide. However,Each year ,nearly five million of children and parent are newly infested ( pediculosis) with this ectoparasitic insect (Pilger et al., 2010 and Roberts, 2002). The some researchers reported that, an annual rate of 15 million infestation with head lice and cost of 150 US$ per treatment in the USA, the overall annual cost of head lice treatments are estimated to be US$ 2,250 million . In addition, over-the-counter pediculicides sales in the USA were estimated at >US$ 240 million per year in 1997 and increased to > US$ 350 million per year in 2003 (West , 2004). Furthermore, the American infested children are lost 12-24 million school days per year by " No Nit Policy " (the policy for infested schoolchildren, before they can return to schools ,they to be free of nits ,nymphs and adults of head lice )( National Association of School Nurses,2016) . So, annual economic loss owing to missed workdays and schooldays by parents and infected children who have to stay home estimated of US$ 4-8 billion to the country's economy (Ko and Elston, 2004 ; Leung et al.,2005). However, numerous case of head lice infestations has been reported throughout the world (Eroglu et al., 2016; Doroodgar et al., 2014). Generally , the neurotoxic insecticides (such as malathion, cabaryl, lindane, permethrin) used as pediculicides for head lice treatment worldwide and head lice resistance to neurotoxic pediculicides have occurred in several parts of the world. (Centers for Disease Control and Prevention, 2016,Devore et al.,2015 and Devore and Schutze ,2015). The alternative pediculicides for head lice treatments are critical needed. However ,alternative pediculicides from plants or herbs have attracted a good pediculicides to develop the new potion for head lice treatment, because of their low mammalian toxicity and save for children, mode of action that are not neurotoxic and the possibility that they will have a low potential for the development of head lice resistance (Rassami and Soonwera, 2013; Strycharz et al.,2014). In addition, Gallardo et al.(2012) and Toloza et al.(2006) reported the pediculicidal activities of essential oils from Geranium maculatum ( geranium) ,Myrcianthes cisplatensis ,Eucalyptus cinerea , Eucalyptus viminalis and Eucalyptus saligna .The lotions based on essential oils from lavender ,peppermint and eucalyptus exhibited pediculicidal properties( Audino et al.,2007).Moreover, herbal shampoos of Averrhoa bilimbi ,Clitoria ternatea,Myristica fragrans , Plectranthus amboincus, Tacca chantrieri, Zingiber cassumunar and Zanthoxylum limonella exhibited high pediculicidal activities ( Rassami and Soonwera,2013).

The pediculosis among Thai preschool children(3-5 years old) and Thai primaryschool children(6-12 years old) ,especially the children in the rural area of Thailand were high level with more than 50% and normally the frist option for head lice treatment of them were chemical control. Unfortunately, mostly the chemical pediculicides in Thailand market are neurotoxic pediculicides ( such as pediculicides consist of lindane, malathion ,carbaryl and permethrin ) and several products have lost in efficacy due to increasing resistance of head lice to chemical pediculicides (Rassami and Soonwera, 2013). Moreover ,the high toxic effects of chemical pediculicides to children were recorded in several part of the world.The mode of action of chemical pediculicides to children were toxic to the children's respiratory and nervous systems(Eisenhower and Farrington,2012). Consequently , the infested children in Thailand needed the effective and safety pedicilicides for head lice treatment, the aim of this study was to evaluate the pediculicidal activity of herbal shampoo from Garcinia dulcis (Roxb.) Kurz (Clusiaceae) against head lice and compared pediculicidal activity with two chemical pediculicides (carbaryl shampoo (Hafif shampoo®, 0.6% w/v carbaryl), permethrin shampoo (Scully Anti-Lice shampoo®, 0.5% w/v permethrin)) and one baby shampoo (Care baby Shampoo®) in vitro and in vivo test. In addition, G. dulcis plant is known as "Maphut"in Thailand.This plant specie is an edible plant and fruit and use asmedicinal plant in Thailand. All part of G. dulcis have long been used in traditional Thai medicines such as expectorant , antipyretic, antitoxic, anti-inflammatory and tonic. The mature fruit of G. dulcis also can be used for soft drink , jam or fruit paste (Fig.1, Deachathai et al., 2005 and Lim, 2011). For this reason, herbal shampoo from G. dulcis is the selected herbal shampoo in this study. We will hope that G. dulcis shampoo is the good , effective and safety herbal product for to be used as alternative pediculicide for Thai infested children especially rural children in Thailand.

Materials and Methods

Collection of Garcinia dulcis fruits and herbal shampoo of Garcinia dulcis

The leaves, flowers, raw and mature fruits of this plant were collected from Bangkok and Nakhonratchasima province (North-eastern part of Thailand),Thailand during May-June 2016 (Fig. 1) for identification by the taxonomist of Faculty of Agricultural Technology, King Mongkut's Institute of Technology Ladkrabang (KMITL), Bangkok, Thailand. The extracts from mature fruits of Garcinia dulcis were used for herbal shampoo by medicinal plant scientist of Medicinal Plant Laboratory, Faculty of Agricultural Technology, KMITL. All herbal shampoo of Garcinia dulcis for this research was provided free of charge from Medicinal Plant Laboratory.

Chemical shampoo and baby shampoo:

  1. Carbaryl shampoo (Hafif shampoo®, 0.6% w/v carbaryl) : this chemical product was a common pediculicide in Thailand . This product was purchased from IDS Manufacturing Co. Ltd., Pathumthani province, Thailand and used as positive control.
  2. Permethrin shampoo (Scully shampoo®, 0.5% w/v permethrin) ,It was the chemical product and usually used as pediculicide in Thailand. It was purchased from Sherwood Chemical Manufacturing Co. Ltd., Chacheangsao province, Thailand and used as positive control.
  3. Baby shampoo (Care baby shampoo®) ,this baby shampoo was purchased from Colgate-Palmolive (Thailand) Ltd. 700/362, Bangna-Trad km57, Amphur Muang, Chonburi Province 20000, Thailand and used as negative control.

Collection of head lice

This study was approved by the Ethics committee of the Faculty of Agricultural Technology, King Mongkut's Institute of Technology Ladkrabang (KMITL), Bangkok, Thailand ( 2559-01-04-013) and in collaboration with the directors and teachers of thee primary schools at Bangkok, Thailand. However, the 250 of schoolchildren were infested with head lice and to collaboration in this study. All head lice ( 3rd nymphs and adults) were collected by the anti-head lice combs from collaborating schoolchildren . Moreover, the protocol for head lice treatment of collaborating schoolchildren ,allowed using only the anti-head lice combs during 30 days.Afterward, nymphs and adults of head lice were separated by carefully removing them from teeth of the combs into clean insect boxes (18.0x23.0x5.5 cm) and were transported them to Entomological Laboratory, Faculty of Agricultural Technology, KMITL, within 30 min. All nymph and adults of head lice were identification by Entomologist of our Faculty, as Pediculus humanus capitis De Geer (Pediculidae: Phthiraptera). The P.humanus capitis specimens to be used as the further reference specimens for further research and were kept in the museum of Faculty of Agricultural Technology , KMITL.

Bioassay

The bioassay used filter paper contact to evaluate the pediculicidal activity of four shampoos (G dulcis shampoo, carbaryl shampoo ,permethrin shampoo and baby shampoo) and the distrillation water( negative control) in vitro. This bioassay was adapted from Rssami and Soonwera( 2013) and Soonwera(2016) and 30 min after head lice collection,the bioassay was stared. The doses of 0.002 ,0.003 and 0.006 ml/cm2 of G. dulcis shampoo, carbaryl shampoo, permethrin shampoo, baby shampoo and distillation water ( negative control ) were applied to the filter paper (Whatman® No1, 4.8 cm diameter) and after drying for 30 s, each filter paper was placed on the bottom of a petri dish (5.0 cm diameter). Afterward, selection under the stereomicroscope (Nikon® Type 102) for the10 nymphs (the 3rd stage nymphs) or the adults of head lice and were placed them on each filter paper. The mortalities of nymphs or adults on each filter paper were recorded under dissecting stereomicroscope at 10, 30 and 60 min. In addition, the criteria for mortality of all head louse was defined as the complete absence of any vital signs from the external and the internal structures of head lice such as legs ,antennae ,digestive system, nervous system and muscular system with or without stimulation by micro-paintbrush (Rassami and Soonwera , 2013).The criteria for the pediculicidal activity was adapted from Sonwera(2016) was defined as the LT50 value of treatement was less than 1.0 min. All treatments were replicated 10 times. The mortality data was statistic analyzed with Duncan's multiple range test (DMRT) using SPSS for windows version 16.0. The LT50 and LC50 values were calculated by Probit analysis.

In vivo test, The total number of 120 infested (pediculosis) schoolchildren between the ages 5 to 12 years from three primary schools in Ladkrabang district, Bangkok, Thailand were selected to participate in the experiment. In addition, the criteria for pediculosis was defined as the presence of at least on the alive egg or nit (alive nit was within 0.5-1.0cm of hair from the scalp and tan to coffee colour with an intact operculum) , the living nymph or the living adult. All infested schoolchildren in this study allowed using only the anti-head lice comb for head lice treatment and during the experimental period, they had not been treated head lice with any chemical pediculicides or any herbal shampoos. The 120 infested schoolchildren were randomly and separated into four groups (10 schoolchildren per group per school).However, all treatment was replicated three times, as follows;

  • Group 1 - treated with G. dulcis shampoo
  • Group 2 - treated with carbaryl shampoo (Hafif shampoo®, 0.6% w/v carbaryl)
  • Group 3 - treated with permethrin shampoo (Scully shampoo®, 0.5% w/v permethrin)
  • Group 4 - treated with baby shampoo (Care baby shampoo®)

However, all infested schoolchildren in the groups were treated with G. dulcis shampoo, carbaryl shampoo, permethrin shampoo and baby shampoo by applying 20 ml of each shampoo to a wet hair and scalp per head for 5 min and then rinsed off with the clean water. The cure rate (no pediculosis) was recorded of 1 day after the 1st application . In addition , the second application was started 1 day after the first application for the infested schoolchildren (pidiculosis)in the groups ,after the first application.They were treated with the tested shampoos for cured and recorded the cure rate . The third application was started 1 day after the second application, the infested schoolchildren( pediculosis) in the groups ( after the 2nd application) were treated with the tested shampoos for cured and recorded the cure rate . In addition , the criteria for cure rate was defined as the complete absence of the alive nit, the living nymph and the living adult of head lice on hair and scalp of schoolchildren and criteria for pediculicide was defined as 100% of cure rate.

Results

The efficacy of four tested shampoos ( G. dulcis shampoo, carbaryl shampoo, permethrin shampoo and baby shampoo) on mortality of 3rd stage nymphs of P. humanus capitis as shown in Table 1 . The most effective in pediculicide was shown by the 0.006 ml/cm2 of G. dulcis shampoo caused 100% mortality of nymphs at 10 min, LT50 value of 0.2 min and LC50 value of 0.00001 ml/cm2 . Moreover , this results exhibited significant in pediculicide (LT50value less than 1.0 min) over carbaryl shampoo ( 82.0% mortality,LT50value of 6.3min) ,permethrin shampoo( 42.0%mortality ,LT50value of28.9 min) and baby shampoo ( 35.0% mortality ,LT50 value of 77.3 min). However, at the dose of 0.003 ml/cm2 revealed that G. dulcis shampoo caused 100% mortality at 30 min and LT50 values ranged from 2.2 min , followed by carbaryl shampoo , permethrin shampoo and baby shampoo with 78.0 , 48.0 and 39.0 % mortality and LT50 values of 8.2 , 55.8 and 79.2 min ,respectively. At the dose of 0.002 ml/cm2, G. dulcis shampoo exhibited 100% mortality at 30 min and LT50 value of 3.7 min followed by carbaryl shampoo , permethrin shampoo and baby shampoo with 77.0 , 44.0 and 37.0% mortality and LT50 values of 9.1 , 58.5 and 87.1 min , respectively. However , based on the LC50 values indicated the order of the effective in pediulicide of tested shampoo as G. dulcis shampoo ( 0.00001 ml/cm2) > carbaryl shampoo( 0.002ml/cm2) > permethrin shampoo ( 0.1ml/cm2) > baby shampoo( 1.8 ml/cm2). However , no mortality of all nymph at all dose in distillation water group was observed ,so distillation water was nontoxic to all nymphs during the testing time

The results of in vitro test of P. humanus capitis adults as shown in Table 2 and Fig. 3. At the dose 0.006 ml/cm2 of G. dulcis shampoo was the most toxic to head lice adults with 100% mortality at 10.0 min, LT50 value of 0.7 min , followed by carbaryl shampoo , permethrin shampoo and baby shampoo with72.0 , 41.0 and 32.0 % mortality and LT50 values of 6.7 , 35.2 and 79.6 min, respectively . Moreover, the mortality results of head lice adults of G. dulcis shampoo showed significant pediculicidal activity (LT50 < 1.0 min) over two chemical (LT50 values ranged from 6.7 to 35.2 min) and baby shampoo (LT50 value of 79.6 min). At dose of 0.003 ml/cm2 revealed that of G. dulcis shampoo also showed the most toxic to head lice adults with 100% mortality at 60 min and LT50 value of 3.3 min , followed by carbaryl shampoo , permethrin shampoo and baby shampoo with77.0 , 60.0 and 41.0 % mortality and LT50 values of 9.5 , 38.5 and 80.6 min, respectively . In addition , G. dulcis shampoo at the dose of 0.02 ml/cm2 caused 84.0 % mortality of head lice adults at 60 min and LT50value of 4.4 min ,followed by carbaryl shampoo , permethrin shampoo and baby shampoo with70.0 , 50.0 and 31.0 % mortality and LT50 values of 10.1 , 61.8 and 87.6 min, respectively. Moreover, based on LC50 value pointed that of G. dulcis shampoo was the highest effective in pediculicide to head lice adults showing LC50 value of 0.001ml/cm2, followed by carbaryl shampoo (0.002 ml/cm2), permethrin shampoo (0.2 ml/cm2) and baby shampoo ( 2.2 ml/cm2) ,respectively. Meanwhile , no mortality of adults were observed in distillation water( negative control). All adults in negative control survived and normality throughout the testing periods.In addition, statistical analysis of head lice nymphs and adults showed significant differences between G. dulcis shampoo and two chemical shampoos( carbaryl shampoo and permethrin shampoo). In addition, nymphs(LT50 values 0.2 to 87.1 min) were more susceptibility to all tested shampoo than adults (with LT50 values 0.7 to 87.6 min).

In vivo results as shown in Table 3 and Fig.2 ( the first application) , 4( the second application) and 5( the third application) , the results for the first application revealed that G. dulcis shampoo exhibited the highest cure rate(97.6%) and the lowest pediculosis(3.3%) ,followed by carbaryl shampoo and permethrin shampoo with the cure rate of 73.3 and 16.7 % and pediculosis of 26.7 and 83.3 % ,respectively. Meanwhile,Care baby shampoo®as the negative control showed 0% of cure rate and Pediculusis of 100%(100% of schoolchildren still had head lice on the head ) after the first application. The results for the second application showed that G. dulcis shampoo also exhibited the highest of cure rate (100%) and 0% of pediculosis followed by carbaryl shampoo and permethrin shampoo showed cure rate of 83.3 and 23.3% and pediculosis of 16.7 and 76.7 %, respectively. However , 100% pediculosis of all schoolchildren were recorded in the Care baby shampoo® treatment. Therefore, the Care baby shampoo®( the negative control ) was not effective in pediculicide for head lice treatment. The results of the third application, G. dulcis shampoo also exhibited the most effective in pediculicide to head lice showing 100% of cure rate. Moreover, G. dulcis shampoo showed more effective in pediculicide( 100% of cure rate) than carbanyl shampoo ( 90.0% cure rate,10.0% pediculosis) and permethrin shampoo( 27.8% cure rate , 72.2% pediculicides). However,Care baby shampoo® also showed nonpediculicidal activity with 100% of pediculosis in tested schoolchildren. All the result of this study pointed that G. dulcis shampoo was the highest efective pediculicide( 100% of cure rate,LT50 value less than 1.0 min) for controlling head lice. The G. dulcis shampoo exhibited more effective in pediculicide than carbaryl shampoo , permethrin shampoo and Care baby shampoo®. In addition, after 1st, 2nd and 3rd applications, none of schoolchildren showed negative side effect such as the red spot of the scalp and neck, burning sensation or irritation of the scalp.

Discussion

Our data in this study, G. dulcis shampoo showed the highest toxic to nymph and adult of head lice with 100% mortality at 10 min, LT50 value < 1.0 min and LC50 values ranged from 0.00001 to 0.001 ml/cm2, in vitro test. Moreover, this result showed more effective pediculicide than carbaryl shampoo (Hafif shampoo®), permethrin shampoo (Scully shampoo®) and baby shampoo (Care baby shampoo®). In addition , in vivo test, G. dulcis shampoo also showed toxicity to head lice and was more effective pediculicide for head lice treatment than two chemical and baby shampoos with 100% cure rate after the second application. Meanwhile, two chemical shampoos showed the cure rate of 27.0-90.0% after the third application. However,Care baby shampoo®as negative control in this research showed 0% of cure rate and 100% pediculosis( the tested schoolchildren still had head lice after the third application). In addition, baby shampoo cannot be used as pediculicide for head lice treatment of schoolchildren, it was a mild formulation to gently cleanse baby hair without sting baby's eyes. As the result, baby shampoo is not suitable for controlling head lice. However, Rassami and Soonwera (2013) reported the high effective in pediculicide of herbal shampoos for head lice treatment of schoolchildren in Thailand such as herbal shampoos of Phyllanthus emblica (Euphorbiaceae) and Zanthoxylum limonella (Rutaceae) and Acorus calamus (Acoraceae) and the results for head lice treatments of all herbal shampoo were more effective in pediculicides than carbaryl and malathion shampoos. However, carbaryl shampoo (0.6% w/v carbaryl) in this study was a common pediculicide in Thailand market for head lice treatment. The toxicity of carbaryl shampoo to children ,after application of this chememical shampoo for head lice control also recorded in several parts of the world. Moreover, this chemical shampoo was the hazardous shampoo for infested children , especially the children less than 5 years of age and it high toxic to children's nervous system(Eisenhower and Farrington,2012).

Moreover, carbaryl pediculicide showed decreased efficacy against head lice and head lice resistance to carbaryl pediculicide has been reported in several countries such as Australia, UK and USA (Durand et al., 2012 and Eisenhower and Farrington,2012). Since 1995 In the UK, since 1995 carbaryl pediculicide has been available only on prescription, because of concerns about possible carcinogenic effect ( any substance that produces cancer). However,in the UK, 1% carbaryl product in aqueous formulation is still recommended for UK head lice guidelines (Durand et al., 2012). On the other side, permethrin shampoo (0.5% w/v permethrin; Scully shampoo®) showed 30-75% mortality of nymphs and adults of head lice (LT50 values ranged from 28.9 to 61.8 min) in vitro test and exhibited 16.7-27.8% of cure rate (72.2-83.3% of pediculosis in schoolchildren ) in vivo test and these results were less pediculicidal activities than G. dulcis shampoo. Moreover, in this study, permethrin shampoo showed the low efficacy in pediculicide may be explained by nymphs , adults and nits of head lice were resistance to permethrin shampoo . However, permethrin shampoo is the common and preferred shampoo of Thai infested children especially for the urban children for head lice treatment , because permethrin shampoo exhibited a rapid effect in mortality of head lice. Unfortunately, the toxicity of permethrin shampoo to children and head lice resistance to permethrin shampoo were recored in serveral parts of the world . Moreover, permethrin shampoo was the neurotoxic product for children ,it showed high toxic to nerve cells of children. The side effect after treated with permethrin shampoo were the itching , rash and burning of the children's scalp and corneal damage of children's eyes (Allen and Cox, 2016 and Ko and Elston, 2004 ). However ,permethrin shampoo also toxic to the head lice's nervous system , it destroyed the nerve cells of head lice , resulting in incapability and mortality of head lice (Eisenhower and Farrington, 2012 and Cute et al., 2008). However, pyrethroids products such as permethrin and D-phenothrin have been registered as pediculicides since the 1970s and have been widely common as over-the-counter pediculicides since the 1980s (Devore and Schutz,2015) .The Food and Drug Administration (FDA) of USA approved permethrin pediculicide can be used in patients as young as 2 months of age and older and pregnant women ( American Academy of Pediatrics,2015). Consequently, caution should be used in patients with ragweed allergy, because respiratory difficulty or asthmatic episodes may occur with permethrin use in such patients (Devore et al.,2015 and Eisenhower and Farrington, 2012). Moreover, The clinical and parasitologic resistance to pyrethroids was first reported in France in a randomized controlled trial and followed by additional reports from Europe (Czech Republic, United Kingdom and Denmark), the Middle East (Israel), North (United States), and South (Argentina) America, Asia (Japan), and Australia ( Durand et al.2012, Ko and Elton,2004 and West,2004).

However, G. dulcis shampoo showed the most effective pediculicide in vitro and in vivo in this study. The alternative pediciculicides from herbs such as neem exhibited high potential for head lice treatment ( Abdel-Ghaffar et al., 2016). In addition, herbal shampoo base on 10% of long pepper (Piper retrofactum) fruits of Acorus calamus, Phyllanthus emblica and Zanthoxylum limonella extracts showed highly effective pediculicides for head lice treatment of infested schoolchildren in Ladkrabang area, Bangkok, Thailand, and these results were more effective in pediculicide than carbaryl and permethrin shampooes ( Rassami and Soonwera,2013 ; Soonwera, 2016). However , Audino et al.(2007) reported the lotions containing essential oils from eucalyptus , peppermint and lavender showed more effective pediculicide than the commercial lotion in the Argentina market. Moreover, the commercial pediculicide products based on plants and herbs such as Licatack shampoo® (extract of grapefruit), Aesculo Gel® "L" (active compound noted Cocos nucifera oil), WashAway Laus shampoo® (active compound noted neem extracts), Nopucid Bio Citrus® (active compound noted bergamot essential oil), Paranix® (active compound noted essential oils from coconut, anise, and ylang ylang), Puressntiel® (active compound noted essential oils of lavender, clove, tea tree, geranium, vegetable oils of cocos, calophyllum, jojoba, sunflower, almond, ricin) showed high effectiveness against head lice (Abdel-Ghaffar et al., 2016 and Soonwera, 2016). However, G. dulcis is known as an Asian medicinal plant, belongs to the family Clusiaceae (Guttiferae) and is widely distributed in Thailand and other Southeast Asian countries (Malaysia, Indonesia, the Philippines (Deachathai et al., 2005 and Pinkaew et al., 2012).This plant is locally known as "Maphuut" in Thailand, "Mundu" in Indonesia and Malaysia. The G dulcis is an edidible plant and evergreen tree (size of 5 to 20 m high ,Fig 1). The fruit with yellow and white latexes and fruit latexes were turns to lightly brown on exposure to the air. The bark is dark brown and rough and much branched. The leaves are large and leathery with dark green color and leaves are opposite, simple large, ovate, and oval to belong up to 10-30 cm long by 3-15 cm wide(Fig 1). The fruits are oblate to round and the diameter of fruit size of 5.5-8.5 cm (Fig.1). The fruit peels are smooth, green when immature and tuning yellow or yellow-dark when ripe and usually 1 fruit has 1 to 5 seeds . The fruit peel is thin and soft when ripe while the fresh is pulpy and yellow in color. However, soft yellow flesh has a butter-like consistency and slightly acidic but pleasant to eat. The raw fruit is too sour to be eaten but local Thai people used raw fruit of G. dulcis to improve the taste in sour and spicy soup, Thai curry , spicy soup , pickles and used mature fruits in jam ,butter or sherbert (Abu-Baker et al., 2015 and Lim, 2011). Furthermore,all part of G. dulcis has long been used in traditional Thai medicine to treat human diseases such as fever,cold ,antipyretic and detoxification (Deachathai et al., 2006 and Pinkaew et al., 2012). The stem bark has been used traditionally as anti-inflammatory agent and crushed extract from the bark is used for cleaning wounds while the fruit juice was used as expectorant for coughs, and scurvy. The extracts of G. dulcis roots are used for antipyretic, anti-toxic and detoxification (Deachathai et al., 2005; Lim, 2011 and Lamai et al., 2013). The traditional medicine of Indonesia have been used seeds and leaves of G. dulcis to treat human disase such as thyrocele , lymphatitis and parotitis and the extracts of G . dulcis fruits and leves to treat of anti-HIV activity , antiviral, ant-bacterial, anti-inflammatory, antitumor , anti-cancer and anti-oxidant (Abu-Bakar et al., 2015; Lamai et al., 2013; Hutadilox-Towatana et al., 2007; Pang et al., 2009; Pinkaew et al., 2012 and Tuansulong et al., 2011).

However, G. dulcis is an edible plant in Thailand and also used as the medicinal plant in traditional Thai medicine and G.dulcis shampoo also exhibited the high effective in pediculicide to controlling head lice.Therefore, herbal shampoo from G. dulcis in this study is the effective and safe pediculicide and after application with G dulcis shampoo , the tested children were no-side effect such as the red spot or irritarion on the scalpes of the chidren.This herbal shampoo is suitable to be used as alternative herbal pediculicide for head lice treatment, especially for infested children in rural area of Thailand and may be the good and safe pediculicide for children of rural area of Southeast -Asia.

Acknowledgments

The authors are grateful to the Faculty of Agricultural Technology, King Mongkut's Institute of Technology Ladkrabang (KMITL), Bangkok, Thailand and The National Research Council of Thailand for financial support of this study (Grant No 2559-01-04-013 and Thesis grant for Doctoral Degree Student). The authors would like to thank all Thai infested schoolchildren and the teachers of primary schools and at Bangkok area for in vivo test . The authors wish to thank plant taxonomist, entomologist and Medicinal Plant Laboratory, Faculty of Agricultural Technology, KMITL for classification, identification and provided free of charge of G. dulcis shampoo, respectively

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