SHORT COMMUNICATION

Antivectorial Activities of Cashew Nut Shell

Extracts from Anacardium occidentale L.
Alain Laurens
1
, Christophe Fourneau
1
, Reynald Hocquemiller
1
, Andr e Cav e
1
, Christian Bories
2
and
Philippe M. Loiseau
2
1
Laboratoire de Pharmacognosie URA 1843 CNRS
2
Laboratoire de Parasitologie, Facult e de Pharmacie 92296 Ch atenay-Malabry, France
Sodium salts of cashew nut shell extracts (CNSL) and anacardic acids isolated from Anacardium occidentale
demonstrate a potent antivectorial activity against Aedes aegypti larvae and Biomphalaria glabrata snails. The
structureactivity relationship is discussed, particularly the hydrogenation of CNSL and anacardic acids which
lower dramatically these properties, showing the importance of the double bonds on the side chains of anacardic
acids. 1997 by John Wiley & Sons, Ltd. Phytother. Res. 11, 145146, 1997
(No. of Figures: 0. No. of Tables: 1. No. of Refs: 10.)
Keywords: Anacardium occidentale; Anacardiaceae; CNSL; anacardic acids; larvicides; molluscicides.
INTRODUCTION
In the tropics, numerous human diseases are transmitted by
means of blood-sucking mosquitoes (e.g. malaria, yellow
fever, lariasis) or aquatic snails (e.g. schistosomiasis,
fasciolasis). The control of the mosquito by destruction of
the aquatic stages is a rapid and efcient means of reducing
and eliminating the transmission of the disease. In the same
way, snail vector control is possible (WHO, 1985; Marston
et al., 1985). Larvicides of plant origin are currently
receiving considerable attention because of their relatively
harmless biodegradable properties. Since the 1920s more
than 2000 plants have been tested for insecticidal activity
(Klocke, 1989) but bioassays with plant extracts against
mosquito larvae are rare. Moreover the use of tropical plants
cultivated near the endemic areas should be of great interest
and cheap for the antivectorial struggle in developing
countries.
In this eld, Anacardium occidentale (Anacardiaceae), a
fruit tree growing widely in tropical and subtropical area is
cultivated in Africa, South America and India for its kernel
(cashew nut). Cashew nut shell liquid (CNSL) showed a
potent molluscicidal activity (Sullivan et al., 1982; Kubo et
al., 1986; Laurens et al., 1987) and various pharmacological
activities (Bhattacharya et al., 1987). The aim of this work
is to study the larvicidal activity of the extract and to
develop a tensioactive extract of Anacardium occidentale.
These antivectorial properties against aquatic pests, i.e. the
larvae of the mosquito Aedes aegypti, a yellow fever and
lariasis vector, are studied compared with the well known
molluscicidal activity against the snail Biom-
phalaria glabrata the intermediate host of intestinal schisto-
somiasis. The presporal toxin of Bacillus thuringiensis
israeli was the reference compound against mosquito larvae
(Nathan, 1993); Phytolacca dodecandra (Endod) spray
dried extract and niclosamide were used as reference
compounds against snails (Goldsmith, 1991).
MATERIALS AND METHODS
Plant material. Cashew nuts of Anacardium occidentale
were collected in MBao area near Dakar (Senegal) in 1993.
A voucher specimen was deposited in Jardin des Plantes
Utiles herbarium, Facult e de M edecine et de Pharmacie,
Dakar, Sngal.
Extracts. CNSL: Cashew nut shells were extracted with n-
hexane in a Soxhlet apparatus. Solvent was removed under
reduced pressure with a yield of 26%.
HPLC. Analysis of CNSL extract was performed using an
ODS Novapack 15 cm column. The mobile phase was
methanol with acetic acid 4% (88:15) with detection at
280 nm. Separation of anacardic acids: 15 g of extract was
chromatographed on a silica gel column (450 g of silica) to
isolate anacardic acids. Compounds are eluted by hexane
ethyl acetateacetic acid (90:10:1).
Hydrogenation of anacardic acids and CNSL. 80 mg of
anacardic acids was dissolved in toluene and hydrogenated
by H
2
. The catalyst was Pd-carbon (8 mg). The solvent was
evaporated to dryness in vacuum and the residue obtained
was constituted by saturated anacardic acid. CNSL was
treated in the same way.
Reference standards. Endod S type 44 spray dried extract of
berries of Phytolacca dodecandra from Ethiopia was
purchased from Dr Lugt. Support Group Endod, Borneo-
straat 124, The Netherland 2585, La Hague. Niclosamide
was a gift from Rh one-Poulenc/Rorer; the bacterial toxin
from Bacillus thuringiensis israeli IPS82 was kindly
provided by Professor H. de Barjac, Institut Pasteur, Paris,
France.
Biological assays. Larvicidal activities: The biological home
test consists of GKep Aedes aegypti laboratory reared strain.

Correspondence to: A. Laurens.

CCC 0951418X/97/02014502
1997 by John Wiley & Sons, Ltd. Accepted 15 July 1996
PHYTOTHERAPY RESEARCH, VOL. 11, 145146 (1997)
24 h old mosquito larvae were exposed to concentrations of
each tested compound for 24 h at 26C. Twenty larvae were
exposed using a small glass cup in 4.5 mL of non-
chlorinated water. Three test cups for each tested
concentration and control were used and three replications
were run over a period of time to ensure sufcient genetic
variability in the colony larvae that were used. After
correction for control mortality with Abbotts formula, the
concentration that produced 50% mortality was determined
by plotting the mortality points on log-probit paper.
Molluscicidal activity was monitored according to the
WHO memorandum Molluscicide screening and Evalua-
tion, 1965. Laboratory reared snails Biomphalaria
glabrata, Puerto Rico strain, were used. Two 100 mL
containers at each concentration with ten young mature
snails were used. The plant extracts or chemical compounds
were dissolved or suspended in dimethylsulphoxide and
diluted with water to solutions of known concentrations.
Assays were carried out in aerated standard reference water
(10% hardness). Third fold series were performed at 26C,
12 h day-night periods. Death was measured after a 24 h
incubation time period by the lack of heart rate. The lethal
concentration 50% was determined using probit/log
10
dose
paper.
RESULTS AND DISCUSSION
The analytical study of CNSL by HPLC showed that
anacardic acids are the main components (73.7%) of CNSL
with 39.5% for the trien, 9.3% for the dien and 24.9% for
the monoen. In addition cardanols, 2-methylcardols and
cardols were identied according to previous work (Shobha
et al., 1992).
Biological activities are shown in Table 1. CNSL and
Endod molluscicidal activities were potent and quite similar
(respectively 5.4 and 5.9 ppm), nevertheless CNSL demon-
strated a more potent larvicidal activity at 2.6 ppm.
Moerover, chemical transformation of CNSL using sodium
hydroxide, into a tensioactive derivative enhanced the
antivectorial properties against mosquito larvae and snails
by 20% (respectively 2.3 and 4.0 ppm).
After the hydrogenation of CNSL, these properties
dropped dramatically for both vectors (18 ppm for larvae
and 34 ppm for snails). Saturated anacardic acids with
LC
50
>100 ppm demonstrating the importance of the dou-
ble-bonds on the side chain for snails. Nevertheless, the
hydrogenated CNSL remained toxic, but at a higher level
(LC
50
=34 ppm) probably because phenolic components
other than anacardic acids are involved in the lethal process.
These chemical products are mainly cardanols and cardols.
For mosquito larvae saturated anacardic acids at
12.5 ppm were more active than the whole extract. Compar-
atively niclosamide was only efcient against snails and the
IPS 82 toxin was only efcient against larvae.
Therefore the CNSL sodium salt is a compound of
interest for controlling mosquito larvae and aquatic snails in
tropical countries. This low-cost natural product could be a
by-product of the agroalimentary industry. The CNSL
sodium salt could be incorporated easily in washing powder
or soap manufactured in developing countries. It could
destroy and control mosquito larvae or snail pollution in
waste water, sewage or open main sewers.
REFERENCES
Bhattacharya, S. K., Mukhopadhyay, M., Mohan Rao, J. R.,
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research: possible self-reliant control of schistosomiasis.
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Table 1. Results of biological assays
Snails Larvae
24 h LC
50
(ppm) 24 h LC
50
(ppm)
CNSL 5.4 2.6
CNSL Na (expressed in free CNSL) 4.0 2.3
Endod 5.9 >100
Niclosamide 0.15 >100
IPS 82 >10 0.03
Unsaturated anacardic acids 3.7 1.5
Saturated anacardic acid >100 12.5
CNSL hydrogenated 34 18
A. LAURENS ET AL. 146