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Jörg Heukelbach . Fabíola A. S. Oliveira .
Richard Speare A new shampoo based on neem (Azadirachta indica) is highlyeffective against head lice in vitro Received: 7 February 2006 / Accepted: 10 February 2006 / Published online: 28 March 2006 Abstract Because topical compounds based on insecticid- al chemicals are the mainstay of head lice treatment, butresistance is increasing, alternatives, such as herbs and oils Prevalence of head lice (caused by infection with Pediculus are being sold to treat head lice. To test a commercial capitis) has been increasing worldwide (Burgess shampoo based on seed extract of Azadirachta indica Topical insecticides based on insecticidal chemicals are (neem tree) for its in vitro effect, head lice (n=17) were still the mainstay of head lice treatment (Heukelbach and collected from school children in Australia and immersed Feldmeier ), and resistance to the common over-the- in Wash-Away Louse™ shampoo (Alpha-Biocare GmbH, counter pediculicides has been reported repeatedly by Germany). Vitality was evaluated for more than 3 h by clinicians and confirmed by in vitro studies (Burkhart et al.
examination under a dissecting microscope. Positive and negative controls were a commercially available head lice Elston Hunter and Barker ; Mumcuoglu et al.
treatment containing permethrin 1% (n=19) and no ; Picollo et al. ; Yoon et al. ). Resistance is treatment (n=14). All lice treated with the neem shampoo particularly frequent in countries where head lice are did not show any vital signs from the initial examination common and where chemical insecticides are extensively after immersion at 5–30 min; after 3 h, only a single louse used, such as in the US, UK, Israel, and Australia. Because showed minor signs of life, indicated by gut movements, a the prevalence and degree of insecticide resistance is mortality of 94%. In the permethrin group, mortality was expected to increase, alternative topical therapies for head 20% at 5 min, 50% at 15 min, and 74% after 3 h. All 14 head lice of the negative control group survived during the We tested the shampoo “Wash-Away Louse™” based on observation period. Our data show that Wash-Away seeds of Azadirachta indica (neem tree) for its effect on Louse™ is highly effective in vitro against head lice. The head lice in vitro. The results show that the product is neem shampoo was more effective than the permethrin- highly effective against P. capitis.
based product. We speculate that complex plant-basedcompounds will replace the well-defined chemical pedicu-licides if resistance to the commonly used products further Head lice were collected from two school children and their mothers in Townsville, Australia on two occasions Department of Community Health, School of Medicine, separated by a month. After collection, lice were pooled and held on human hair in 5 cm diameter plastic Petri dishes at 27°C and 50% relative humidity. In vitro tests were started within 1 h after collection of lice, or when this was not possible, lice were fed on the dorsum of the hand of Tropical Medicine and Rehabilitation Sciences, one of the investigators and used for testing within 1 h after A commercially available head lice shampoo based on extracts of neem seeds (Wash-Away Louse™; Alpha- Departamento de Saúde Comunitária, Faculdade de Medicina, Biocare GmbH, Düsseldorf, Germany) was tested.
Wash-Away Louse™ contains a patented oily extract from seeds of the plant A. indica, formulated with the following caprate, cocamidoprophybetaine, glycerol, lauryl gluco- sive fashion or no righting reflex when rolled onto the back side, dicaprylyl ether, and as preservation components benzyl alcohol, benzoic acid, and sorbic acid.
Examinations were performed after 5, 10, 15, 20, 30, 45, As a positive control, a 1%-permethrin-based over-the- 60, 90, and 180 min. All lice were maintained at 27°C counter compound was used (Quellada Head Lice Treat- For the tests, lice clasping hair strands were immersed completely in the product for 1 min, then placed with hairs onto Whatman filter paper in Petri dishes. To prevent licefrom desiccation, the filter paper had been previously Using the strict criteria for mortality, all lice treated with the moistened by 200 μl tap water. After lice were placed on Wash-Away Louse™ shampoo did not show any vital signs the filter paper, obvious pools of the liquid product rests after 5 min (Fig. ). After 30 min, in two lice minimal adherent to the body of lice were wiped from the lice by a internal gut movements were observed, giving a mortality jeweller’s forceps directed under a dissecting microscope.
of 88.2%. After 3 h, mortality was 94.1% with one louse Negative control lice were placed directly on the filter showing persistent gut movements. All 14 head lice in the paper without any treatment. To simulate treatment on an negative control group survived during the observation infested host, head lice were washed in tap water after period. In the permethrin group, the number of vital lice 20 min and placed into a new Petri dish.
decreased steadily from 20% at 5 min to 73.7% at 3 h The number of lice was 17, 19, and 14 in the neem (Fig. Interestingly, the activity of two lice remained group, the permethrin group, and the control group, normal throughout the 3-h observation period.
respectively. Lice were tested in batches of seven to 14 “Mortality” as defined by the usual criteria of loss of righting reflex was 100% in the Wash-Away Louse™ Lice on the filter paper were examined under a dissecting shampoo group during the observation period and ranged microscope by a single observer in all cases to prevent between 68 and 90% in the permethrin group (Fig. ).
The criteria used for survival of lice were extremely strict: if any even minor signs of life, such as internal movements, or movements of antennae or minimal legmovements were observed (with or without stimulation by The usual criteria for “mortality” in head and body lice is a forceps), the lice were categorized as alive. The lice were not physical death, but a lesser state judged by the inability judged as dead if there were no vital signs at all (complete of the insects to walk in a progressive fashion (Mougabure physical death). These data were compared to the usual et al. to show a righting reflex when rolled onto its criteria for “mortality”: the inability to walk in a progres- back (Hunter and Barker or when movement hasceased and appendages fail to move when touched with a Fig. 1 Mortality of head licetreated with Wash-Away Louse™ different points in time, usinghighly strict criteria for mortal- Mortality
Time (min)
different points in time, asdefined by usual criteria for survival (inability to walk in aprogressive fashion, no righting Mortality
Time (min)
needle (Oladimeji et al. However, the use of strict criteria (complete physical death of the insects) is formulation has been shown to be efficacious as a repellent particularly important, as it is known that head lice can against head lice (Mumcuoglu et al. ). In clinical resurrect within a short period of time (Burkhart and studies, a combination of paw paw, thymol, and tea tree oil, Burkhart Using usual and also strict criteria for a combination of coconut oil, anise, and ylang ylang, and a mortality, our data show that Wash-Away Louse™ shampoo preparation from Annona squamosa seed extract were is highly effective in vitro against head lice collected in highly effective against head lice infestations (McCage et Australia, an area where resistance to commonly used al. ; Mumcuoglu et al. ; Tiangda et al. ).
pediculicides is prevalent. After 5 min, all head lice did not In conclusion, it can be speculated that the tested show any vital signs and did not recover after 3 h. The shampoo and similar products will substitute chemical effect of the shampoo was better than a permethrin-based compounds on the market, if resistance to the commonly product, which is considered the first line treatment.
used pediculicides further increases.
First data of a field study from Egypt indicate that Wash- Away Louse™ is also highly effective for treatment of The authors thank Chris Cahill for assistance individuals heavily infested with head lice (Ghaffar and in obtaining head lice. Wash-Away Louse™ neem shampoo was provided free of charge by Alpha Biocare GmbH, Düsseldorf, The neem tree has the widest spectrum of use of all Germany. Jorg Heukelbach was supported by an EndeavourAustralia Research Fellowship.
natural products. The first known use of neem by theHarrappa culture in ancient India dates back 4,500 years(Dasgupta et al. ). Today, neem extracts are used to treat various skin diseases, as an antiseptic substance,against endo- and ectoparasites, or simply as hair Burgess IF (2004) Human lice and their control. Annu Rev Entomol Anecdotal clinical data suggested a good efficacy of Burkhart CG, Burkhart CN (2006) Safety and efficacy of shampoos based on neem oil against pediculosis (Knust pediculicides for head lice. Expert Opin Drug Saf 5:169–179 Burkhart CG, Burkhart CN, Burkhart KM (1998) An assessment of Tross et al. ). Neem extract has also an excellent topical and oral prescription and over-the-counter treatments for effect as a nontoxic repellent, insecticide, and pesticide (Li head lice. J Am Acad Dermatol 38:979–982 et al. ; Mulla and Su The Ayurvedic medicine Dasgupta T, Banerjee S, Yadava PK, Rao AR (2004) Chemopre- uses the neem tree as one of the main ingredients of its ventive potential of Azadirachta indica (Neem) leaf extract in murine carcinogenesis model systems. J Ethnopharmacol92:23–36 It is possible that in the long run, plant extracts will Downs AM, Stafford KA, Hunt LP, Ravenscroft JC, Coles GC substitute chemical compounds. For example, products (2002) Widespread insecticide resistance in head lice to the based on cinnamon, eucalyptus, marjoram, pennyroyal, over-the-counter pediculocides in England, and the emergence Lippia multiflora, and rosemary oils were effective against of carbaryl resistance. Br J Dermatol 146:88–93 Elston DM (2003) Drug-resistant lice. Arch Dermatol 139:1061–1064 head lice in vitro (Oladimeji et al. ; Yang et al. Ghaffar FA, Semmler M (2006) Effects of new neem extracts on Oladimeji FA, Orafidiya OO, Ogunniyi TA, Adewunmi TA (2000) head lice in heavily infested persons. Parasitol Res in press Pediculocidal and scabicidal properties of Lippia multiflora Heukelbach J, Feldmeier H (2004) Ectoparasites—the underesti- essential oil. J Ethnopharmacol 72:305–311 Picollo MI, Vassena CV, Mougabure Cueto GA, Vernetti M, Zerba Hunter JA, Barker SC (2003) Susceptibility of head lice (Pediculus EN (2000) Resistance to insecticides and effect of synergists on humanus capitis) to pediculicides in Australia. Parasitol Res permethrin toxicity in Pediculus capitis (Anoplura: Pedicul- idae) from Buenos Aires. J Med Entomol 37:721–725 Knust FJ (1998) Neem-Therapie der Pediculosis capitis und der Tiangda CH, Gritsanapan W, Sookvanichsilp N, Limchalearn A Scabies im Kindesalter. Arzt Umw 11:319–322 (2000) Anti-headlice activity of a preparation of Annona Li SY, Skinner AC, Rideout T et al (2003) Lethal and sublethal squamosa seed extract. Southeast Asian J Trop Med Public effects of a neem-based insecticide on balsam fir sawfly (Hymenoptera: Diprionidae). J Econ Entomol 96:35–42 Tross R, Bernauer-Jacob V, Hummel E, Kleeberg H (1998) McCage CM, Ward SM, Paling CA et al (2002) Development of a AzadirachtinA-content and bio-efficacy in hair treated with paw paw herbal shampoo for the removal of head lice.
NeemAzal Fomulations. In: H Kleeberg (ed) Practice oriented results on use and production of neem-ingredients and Mougabure CG, Gonzalez AP, Vassena CV, Picollo MI, Zerba EN pheromones VII. Druck und Graphic, p 9–20 (2002) Toxic effect of aliphatic alcohols against susceptible and Yang YC, Choi HY, Choi WS, Clark JM, Ahn YJ (2004a) Ovicidal permethrin-resistant Pediculus humanus capitis (Anoplura: and adulticidal activity of Eucalyptus globulus leaf oil terpenoids against Pediculus humanus capitis (Anoplura: Mulla MS, Su T (1999) Activity and biological effects of neem Pediculidae). J Agric Food Chem 52:2507–2511 products against arthropods of medical and veterinary im- Yang YC, Lee HS, Clark JM, Ahn YJ (2004b) Insecticidal activity of portance. J Am Mosq Control Assoc 15:133–152 plant essential oils against Pediculus humanus capitis (Anoplura: Mumcuoglu KY, Hemingway J, Miller J et al (1995) Permethrin resistance in the head louse Pediculus capitis from Israel. Med Yang YC, Lee HS, Lee SH, Clark JM, Ahn YJ (2005) Ovicidal and adulticidal activities of Cinnamomum zeylanicum bark essential Mumcuoglu KY, Magdassi S, Miller J et al (2004) Repellency of oil compounds and related compounds against Pediculus citronella for head lice: double-blind randomized trial of humanus capitis (Anoplura: Pediculicidae). Int J Parasitol efficacy and safety. Isr Med Assoc J 6:756–759 Mumcuoglu KY, Miller J, Zamir C et al (2002) The in vivo Yoon KS, Gao JR, Lee SH et al (2003) Permethrin-resistant human pediculicidal efficacy of a natural remedy. Isr Med Assoc J head lice, Pediculus capitis, and their treatment. Arch Dermatol


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