Artemisia absinthium-borne compounds as novel larvicides: effectiveness against six mosquito vectors and acute toxicity on non-target aquatic organisms
Parasitol Res (2016) 115:4649–4661
DOI 10.1007/s00436-016-5257-1
ORIGINAL PAPER
Artemisia absinthium-borne compounds as novel larvicides:
effectiveness against six mosquito vectors and acute toxicity
on non-target aquatic organisms
Marimuthu Govindarajan 1 & Giovanni Benelli 2
Received: 30 August 2016 / Accepted: 7 September 2016 / Published online: 15 September 2016
# Springer-Verlag Berlin Heidelberg 2016
Abstract The eco-friendly control of mosquito vectors is a
crucial challenge of public health importance. Here we evaluated the larvicidal potential of Artemisia absinthium essential
oil (EO) and its three major chemical constituents against six
mosquito vectors: Anopheles stephensi, Anopheles subpictus,
Aedes aegypti, Aedes albopictus, Culex quinquefasciatus, and
Culex tritaeniorhynchus. The EO was obtained by leaf hydrodistillation. Its chemical composition was analyzed using gas
chromatography-mass spectrometry. Major components were
(E)-β-farnesene (31.6 %), (Z)-en-yn-dicycloether (11.12 %),
and (Z)-β-ocimene (27.8 %). The EO was toxic effect against
larval populations of An. stephensi, An. subpictus, Ae. aegypti,
Ae. albopictus, Cx. quinquefasciatus, and Cx.
tritaeniorhynchus, with LC50 values of 41.85, 52.02, 46.33,
57.57, 50.57, and 62.16 μg/ml. (E)-β-farnesene, (Z)-en-yndicycloether, and (Z)-β-ocimene were highly effective on
An. stephensi (LC50 = 8.13, 16.24 and 25.84 μg/ml) followed
by An. subpictus (LC50 = 10.18, 20.99, and 30.86 μg/ml), Ae.
aegypti (LC50 = 8.83,17.66, and 28.35 μg/ml), Ae. albopictus
(LC50 = 11.38,23.47, and 33.72 μg/ml), Cx. quinquefasciatus
( L C 5 0 = 9. 6 6, 1 9 .7 6, a n d 31 . 52 μ g / m l ) , a nd C x .
tritaeniorhynchus (LC50 = 12.51,25.88, and 37.13 μg/ml).
Notably, the EO and its major compounds were safer to the
* Marimuthu Govindarajan
* Giovanni Benelli
1
Department of Zoology, Unit of Vector Control, Phytochemistry and
Nanotechnology, Annamalai University, Annamalainagar 608
002, Tamil Nadu, India
2
Department of Agriculture, Food and Environment, University of
Pisa, via del Borghetto 80, 56124 Pisa, Italy
non-target organisms Chironomous circumdatus, Anisops
bouvieri and Gambusia affinis, with LC50 values ranging from
207.22 to 4385 μg/ml. Overall, our results highlight that
(E)-β-farnesene, (Z)-en-yn-dicycloether, and (Z)-β-ocimene
from the A. absinthium EO represent promising eco-friendly
larvicides against six key mosquito vectors with moderate
toxicity against non-target organisms.
Keywords Biosafety . Culcidae . (E)-β-farnesene .
(Z)-en-yn-dicycloether . (Z)-β-ocimene . Non-target
arthropods . Gambusia affinis
Introduction
Arthropods are important vectors of a great number of
pathogens and parasites, which may hit as epidemics or
pandemics in the increasing world populations of humans
and animals (Mehlhorn 2015; Benelli and Mehlhorn 2016;
Benelli et al. 2016a). Mosquitoes (Diptera: Culicidae) represent a key threat for millions of organisms worldwide,
since they act as vectors of the agents of malaria, dengue,
yellow fever, West Nile virus fever, Japanese encephalitis,
filariasis and, more recently, Zika virus (Mehlhorn et al.
2012; Benelli 2015a; Benelli et al. 2016b, c).
According to the latest estimates, there were at least 198
million cases of malaria in 2013 and an estimated 584,000
deaths. Malaria mortality rates have fallen by 47 % globally
since 2000 and by 54 % in the African region, but are still
high. Most deaths occur among children living in Africa,
where a child dies every minute from malaria (Jensen and
Mehlhorn 2009; WHO 2014). Dengue is ranked among the
most important mosquito-borne viral diseases in the world. In
the last 50 years, the incidence has increased 30-fold. An
estimated 2.5 billion people live in over 100 endemic
4650
countries and areas where dengue viruses can be transmitted.
Up to 50 million infections occur annually with 500,000 cases
of dengue hemorrhagic fever and 22,000 deaths, mainly
among children (WHO 2012a). In the past decade, West
Nile virus has emerged in the Americas, becoming endemic
throughout the region. Chikungunya, a formerly obscure arbovirus endemic to East Africa, has also emerged, causing
millions of cases in the Indian Ocean basin and mainland
South and Southeast Asia. The Japanese encephalitis virus
has expanded its range in the Indian subcontinent and
Australasia, where it chiefly affects children (Tolle 2009;
Benelli and Mehlhorn 2016).
Currently, the use of mosquito larvicides faces several serious problems. Besides the negative effects of synthetic insecticides on the environment and non-target organisms, including
man (Hodgson and Levi 1996; WHO 2012b), the development
of resistant mosquito populations in particular is one of the
most serious problems (Hemingway and Ranson 2000;
Naqqash et al. 2016). Insecticide resistance is viewed as an
extremely serious threat to crop protection and vector control,
and is considered by many parties, including industry, the
WHO, regulatory bodies, and the public, to be an issue that
needs a proactive approach (Hemingway and Ranson 2000;
McCaffery and Nauen 2006; Nauen 2007; WHO 2012b).
These problems highlighted the needing of new pest control
alternatives, acceptable for the environment and human health
(Benelli 2016a, b; Pavela and Benelli 2016). Among the
existing alternative tools aimed at decreasing pest populations,
the use of pesticides based on plant extracts is currently one of
the most promising (Amer and Mehlhorn 2006a, b, c, d; Dubey
2011; Benelli 2016c; Govindarajan et al. 2016a, b, c, d).
Essential oils and related main compounds are also an environmentally interesting tool because they are biodegradable and
have minimal side effects on non-target organisms, as well as
on the environment (Govindarajan 2010; Govindarajan et al.
2012, 2013; Pavela 2014, 2015; Benelli 2015b, c Govindarajan
and Benelli 2016a, b).
Essential oils can be used as an alternative to synthetic
larvicides for vector control programs (Pavela 2015). It is
well known that plant-derived natural products are extensively used as biologically active compounds (Zebitz
1984). Among them, essential oils were the first preservatives used by the man (Bakkali et al. 2008). Essential
oils are mainly composed by isoprenoid compounds,
mainly mono- and sesquiterpenes, which are mainly responsible of the smell of many aromatic plants (Franzios
et al. 1997). Commercially, essential oils are used in four
primary ways: as pharmaceuticals, as flavor enhancers in
many food products, as odorants in fragrances, and as
insecticides (Zhu et al. 2001; Pavela 2015; Benelli
2015a).
Artemisia, the largest and most widely distributed genus of
family Asteraceae, comprises over 500 species geographically
Parasitol Res (2016) 115:4649–4661
spread in the temperate zones of Europe, North America,
Asia, and South Africa (He et al. 2009). Many Artemisia
species are popular traditional Chinese medicinal plants
and have been used for the treatment of a variety (...truncated)
