Phytomedicine Ovicidal Effect of Essential Oils from Zingiberaceae
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3K\WRPHGLFLQH ² Contents lists available at ScienceDirect Phytomedicine journal homepage: www.elsevier.com/locate/phymed Original Article Ovicidal effect of essential oils from Zingiberaceae plants and Eucalytus 7 globulus on eggs of head lice, Pediculus humanus capitis De Geer Mayura Soonwera⁎, Orawan Wongnet, Sirawut Sittichok Department of Plant Production Technology, Faculty of Agricultural Technology, King Mongkut's Institute of Technology Ladkrabang, Bangkok 10520, Thailand ARTICLE INFO ABSTRACT Keywords: Background: Head lice infestation is an important public health problem worldwide. Chemical pediculicides Ovicidal activity have lost their efficacy because lice have developed resistance to them. Therefore, alternative pediculicides such Head lice egg as essential oils and herbal products have been proposed for treating head lice infestation. Zingiberaceae EOs Study design: To determine the efficacy of essential oils from three Zingiberaceae plants (Curcuma xanthorrhiza, Eucalyptus globulus EO Curcuma zedoaria and Zingiber zerumbet) against head lice eggs and to investigate an augmenting substance (Eucalyptus globulus EO) for improving the efficacy of these essential oils in killing head lice eggs, especially on the inhibition of their hatching process. Permethrin pediculicide, soyabean oil, and drinking water were used as positive, negative, and neutral controls, respectively. Methods: An immersion test was used to evaluate the ovicidal activity of 12 essential oil formulations. Head lice eggs were immersed for 1, 5 and 10 min in the treatments. Mortality rate was observed on day 7 and day 14; mortality was checked under a stereomicroscope. Results: All head lice eggs that were immersed in a combination of 10% C. zedoaria EO and 10% E. globulus EO for 5 min did not hatch at all for 7–14 days of incubation. All head lice eggs that were immersed in soyabean oil and drinking water for 1, 5, and 10 min showed 100% hatching rate in 7–14 days of incubation. All head lice eggs that were immersed in permethrin pediculicide for 1, 5 min, showed 100% hatching rate, but when they were immersed for 10 min, permethrin provided 4.0–6.0% inhibition rate with 94.0–96.0% hatching rate for 7–14 days of incubation. All combinations of Zingiberaceae EOs and E. globulus EO at low and high con- centrations (5 and 10%) exhibited high ovicidal activities against head lice eggs, and the combinations showed a synergistic effect with an increase in the inhibition rate of more than 50%. Conclusion: These results demonstrated that Zingiberaceae EOs augmented with E. globulus EO are promising ovicidal agents for head lice control. Introduction may take between 7 to 10 days to hatch into nymphs. Once hatched, nymphs feed on blood drawn from human scalp and grow into adults. The head louse Pediculus humanus capitis De Geer is one of common Adults are able to reproduce for 14 to 21 days after they have hatched medical insect pests worldwide. This insect has been an ectoparasite of and female head lice may lay up to 8–10 nits a day for a total of 50–300 human beings for a very long time. This pest is a small wingless insect nits during their lifetime. Nits can be gray to white and 3–4 mm in size. (2–4 mm) belonging to the family Pediculicidae and order Phthiraptera. They can be found on hair shafts 1 mm from the scalp, particularly at The history of head lice and pediculosis infection dated back to pre- the back of the head and neck and behind the ears. Adult lice that feed historic time. The oldest fossil of nit (egg of head louse) discovered is on a human host may survive up to 30 days but cannot survive more about 10,000 years old (Araujo et al., 2000). It is a highly host-specific than 2 days away from their human food source (Eisenhower and insect that lives and feeds only on human blood at the scalp and the Farrington, 2012; Frankowski and Bocchini, 2010; Guenther and Cunh, neck area (Feldmeier, 2012; Ko and Elston, 2004; Nutanson et al., 2017; Ko and Elston, 2004). Traditionally, control of these insects has 2008). In general, the male is smaller (2–3 mm) than the female been done by application of neurotoxic synthetic chemical insecticides (3–4 mm). The life cycle of head louse involves three stages. Nits (eggs) such as organochlorine (lindane), organophophate (malathion), Abbreviation: EO, Essential oil; EOs, Essential Oils; IRC, Percentage inhibition rate change ⁎ Corresponding author. E-mail address: [email protected] (M. Soonwera). https://doi.org/10.1016/j.phymed.2018.04.050 Received 19 October 2017; Received in revised form 16 February 2018; Accepted 22 April 2018 (OVHYLHU*PE+$OOULJKWVUHVHUYHG M. Soonwera et al. 3K\WRPHGLFLQH ² carbamate (carbaryl) and permethrin (permethrin). Unfortunately, 2016. All plant specimens were identified by a botanical taxonomist at these insecticides are very persistent, harmful and highly toxic. More- King Monkut's Institute Technology Ladkrabang (KMITL) (Fig. 1). over, head louse population usually develops resistance to them after Rhizomes of Zingiberaceae plants and leaves of eucalyptus were some exposure (Abdel-Ghaffar and Semmler, 2007; Rassami and cleaned, cut into small pieces and put in a 5 L flask. Sterile water was Soonwera, 2013; Soonwera, 2014). On top of that, chemical pediculi- added at a ratio of 1:2 (w/v) and the plant materials were extracted for cides can kill the nymphs and adults but not the eggs (Bowles et al., essential oils (EOs) by a water distillation method in a modified Cle- 2017). There have been reports that neem extract anti-louse shampoo venger type apparatus for 6–8 h. Water was removed from the oils after had a high efficacy against head lice eggs (Abdel-Ghaffar et al., 2012; they were extracted by mixing them with anhydrous sodium sulphate. Al-Quraishy et al., 2015; Di Campli et al., 2012; Mehlhorn et al., 2011). Gas chromatography and gas chromatography/mass spectrometry were Yang et al. (2003, 2005, 2009) summarized that the EOs from Eugenia used to analyse the plant essential oil components. The major com- caryophyllata, Origanum majorana and Cinnamomum zeylanicum were pounds of C. xanthorrhiza EO were xanthorrhizol 45.5%, zingiberene highly toxic to the adults and eggs of P. capitis. Neem seed extract with 18.2%, curcumene 20.7%, bisabolol 8.1% and curcumene 7.5%. The and without azadirachtin were able to kill all stages of head lice (eggs, major compounds of C. zedoaria EO were camphor 45.5%, camphene nymphs and adults) (Abdel-Ghaffar et al., 2010, 2016). 15.3%, zingiberene 14.8%, 1,8-cineole 10.9% and isoborneol 13.5%. ff Gallardo et al. (2012) reported that the most e ective ovicidal agent The major compounds of Z. zerumbet EO were camphene 40.3%, α- against head lice eggs was a commercial pediculicide based on ber- humulene 18.5%, camphor 15.7%, 1,8-cineole 7.8% and zerumbone gamot EO, ciclopentaxiloxane, dimethicone (Nopucid Bio Citrus®) and 17.7%. The major compounds of E. globulus EO were 1,8-cineole 48.5%, ® dimethicone (Nyda ). Yones et al. (2016) presented that spearmint oil α-pinene 20.6%, β-pinene 15.5% and terpineol 15.4%. All prepared was the most effective ovicidal agent that effected a complete inhibition plant essential oils were stored in the laboratory (27.1 ± 1.2 °C; of head lice eggs. 75.0 ± 1.2% RH) for subsequent experiments. The therapeutic prop- Currently, there are considerable concerns about insecticides in erties and chemical constituents of the plant essential oils are presented regard to human health, long chemical persistence in water, the en- in Table 1, while the formulations used in this study are presented in vironment and food supply, and developed insect pest resistance. For Table 2. this reason, ongoing research for safe and effective insecticide alter- natives has been conducted in order to protect children from their Approval of the protocol for head lice egg collection from human beings harmful effects. Natural plant products can perform various functions, and many of them provide useful insecticidal activities. Many of these The protocol for head lice egg collection from human beings was plants have essential oils (EOs) that they produce as a defense against approved by the KMITL Ethics committee, Ladkrabang, Bangkok, insect pests and diseases. Normally, EOs have some insecticidal, fun- Thailand, in March 2016 with a registration number of 2560-1-04-003 gicidal, bactericidal and virucidal properties (Koul et al., 2008; and by the Institute for Development of Human Research Protections Regnault-Roger et al., 2012). Many essential oils (EOs) from Zingiber- (IHRP) Ethic committee, Bangkok, Thailand, with a permit number aceae plants have already been used as alternative insecticides for 76–2558. controlling insect pests including head lice (Burgess, 2004; Regnault- Roger et al., 2012). Their mammalian toxicity is low and their en- Head lice egg collection vironmental persistence is short (Mcallister and Adams, 2010; Regnault-Roger et al., 2012). Curcuma xanthorrhiza (C. xanthorrhiza), Eggs of head lice (P. humanus capitis) were collected from the heads Curcuma zedoaria (C. zedoaria) and Zingiber zerumbet (Z. zerumbet) be- of 20 severely infested persons—10 girls between 13–15 years old and long to the family Zingiberaceae. They are native plants found in all 10 women between 25–30 years old—who were students and parents of regions of Thailand and South East Asia (Chien et al., 2008; some students at several primary schools in Ladkrabang district, Mangunwardoyo and Usia, 2012; Lobo et al., 2009). The rhizomes of Bangkok, Thailand. Their hair was cut with scissors and put in small these three Zingiberaceae plants have essential oils that are toxic to insect boxes (2 ×3×1 cm). More than one thousand head lice eggs insect pests and also have antioxidant, antimicrobial, antifungal, anti- were collected.