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TOXICITY OF SOME INSECTICIDES AGAINST

MOSQUITO LARVAE OF CULEX PIPIENS

Abdulla A. AL Zarog 1 Mohamed Ali Elssaidi 2


1. Faculty of Science , Sebha University; 2. Faculty of Engineering
and Technology, Sebha University

ABSTRACT

This study was aimed to investigate the efficiency of some


insecticides, (Fenitrothion, Cyphenothrin, Azadirachtin and
Pyriproxyfen) against mosquito larvae of Culex Pipiens. Data indicated
the superior effectiveness of fenitrothion against larvae of Culex
Pipiens followed by azadirachtin, cyphenothrin and pyriproxyfen. The
lethal concentration values (LC50) were 4.1, 4.5, 8.4 and 60.0 ppm for
fenitrothion, azadirachtin, cyphenothrin and pyriproxyfen,
respectively. The relative potencies, based on pyriproxyfen, revealed
the following values, 14.6, 13.3 and 7.1 fold for fenitrothion,
azadirachtin and cyphenothrin, respectively.

KEYWORDS:
Toxicity, Bioassay, Insecticide, Fenitrothion, Cyphenothrin, Azadirachtin,
Pyriproxyfen, Mosquito.
INTRODUCTION

Brogdon and McAllister (1998) stated that the methods for treating
bottles, obtaining baseline data and applying this technique to insects from
the field are described. Sample data are presented from tests run on
different vector species (Anopheles albimanus, A. gambiae, A. dirus, A.
stephensi, A. freeborni, Aedes aegypti, A. albopictus, Culex restuans, C.
pipiens, C. nigripalpus, C. salinarius, C. territans and Lutzomyia youngi)
using a variety of insecticides (including permethrin, lambda-cyhalothrin,
resmethrin, naled, malathion, chlorphoxim and fenitrothion). Time-
mortality data from the bottle bioassay are presented alongside results from
biochemical detection methods applied to the same mosquito population.
The potential role, advantages and limitations of the time-mortality bottle
method are discussed.
Elssaidi (1998) stated that fenitrothion was degraded under lab.
conditions slightly faster than malathion, the residues half-life values (t0.5)
for fenitrothion were 33, 22 & 17 days, while were 22, 20 & 15 days for
malathion in distilled, hard and ground water respectively.
Nazni, et al., (1998) The rates of resistance development were
measured by the LC50 value of a larval bioassay, LT50 value of an adult
bioassay and the frequency of elevated esterase levels. In another set of
experiments using the same batch of Culex, the larvae were not exposed to
any insecticides and the decrease in resistance rate was monitored in each
subsequent 9 generations using similar methods. The heterozygous
standard laboratory strain was selected for susceptibility using the single
raft sib-selection method. The result showed that the field collected F1
generation was 96.0- and 6.3-fold more resistant to malathion and
permethrin, respectively. After selection for about 9 generations, the
resistance ratio to malathion and permethrin was 6.2- and 767.3-fold more
compared to the LC50 values of F1 generations, respectively. Esterase in F1
larvae was 6.0-fold more than the standard laboratory strain.

Bisset-Lazcano, et al., (1991) Resistance to malathion was studied in


the "Quibu" strain of C. quinquefasciatus from Cuba. This strain was
known to be resistant to malathion (with an LD50 500X normal). Adult
insects were kept in the laboratory at 25ٌ2ّC and RH 80ٌ5%. The resulting
larvae were reared on fish food and adults fed on sugar solution and
granular sugar. Esterase analysis was performed on 3-day-old adult
mosquitoes using horizontal gel electrophoresis. Results showed that
mosquitoes were polymorphic for esterases; the esterases Est-1, Est-2 and
Est-3 were the most common morphs. The number of potentially-resistant
female mosquitoes was higher than the number of potentially-resistant

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males. It is suggested that Est-2 was responsible for malathion resistance in
this strain of C. quinquefasciatus, although Est-3 may also play a role in the
resistance of neotropical C. quinquefasciatus.

The present study aimed to investigate the efficiency of some


insecticides, (Fenitrothion, Cyphenothrin, Azadirachtin and Pyriproxyfen)
against mosquito larvae of Culex Pipiens .

MATERIALS AND METHODS

Insecticides used
Sumithion 50% EC. (Fenitrothion):

- O,O, Dimethyl O-(3-methyl-4-nitrophenyl) phsophorethioate,


- The pesticide used was an emulsifiable concentrate having 50% active
ingredient.
- The acute oral LD50 (Rat) = 570-800 mg/kg body weight.

Gokilaht 5% EC. (Cyphenothrin):

- (Rs)-α-cyano-3-phenoxy benzyl (IR)-cis, trans chrysanthemate.


- The preparation used was an emulsifiable concentrate containing 5%
active material.
- The acute oral LD50 (Rat) = 318-2640 mg/kg.

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Agroneem 7% EC. (Azadirachtin):

- Nortriterpenoid botanical insecticide of the liminoid class extracted


from the neem tree Azadirachta indica, yellow-green powder.
- The preparation used was an emulsifiable concentrate containing 75%
active component.
- The acute oral LD50 (Rat) = > 500 mg/kg.

Admiral 10% EC. (Pyriproxyfen):

- 4-Phenoxyphenyl (RS)-2-(2-pyridyloxy) prophyl ether.


- C20H19NO3)
- The preparation used was an emulsifiable concentrate containing
10% active material.
- The acute oral LD50 (Rat) = > 5000 mg/kg.

Toxicological Studies On Culex pipiens (L):

Rearing Techniques:
The strain used in the present investigation was wild larvae of
Culex Pepiens (L) which were collected during their season of abundance
and had been reared, for development of a homogeneous culture under
laboratory conditions of 25 ± 2˚C and 80 ± 5% relative humidity.

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Bioassay evaluation was done in enamel trays. Transformed pupa were
collected from the fore mentioned trays, by means of a wide-mouth glass
dropper, then pipetted into Petri-dishes, that were placed in the adult cages,
the latter known as emerging-adult cages had 33 cm3 size. The emerging
males and females were fed on 10% sucrose solutions and guinea pig blood
meals, respectively. Suitable containers for egg laying, were provided to the
cages 48 hours after the females had their blood meals. Receptacles
containing egg rafts were daily collected from the cages, then left
undisturbed till hatching. The newly hatched larvae were then transferred to
the breeding trays, each containing 2 inch high tab water, such breeding
trays consist each a maximum of 250 larvae of the same age. After 24 hrs
the hatching larvae were fed on fresh yeast which was evenly sprinkled on
the water surface twice daily.

Bioassay Studies
Tests were conducted according to the procedure of Anonymous (1981).
The following concentrations were tested, fenitrothion 50% at 1, 2, 4, 8 and
12 ppm, cyphenothrin 5% at 2, 4, 8, 12 and 16 ppm, azadirachtin 75% at 1,
2, 4, 8 and 10 ppm and pyriproxyfen 10% at 20, 40, 60, 80 and 100 ppm.
To the beakers, which were used for testing the insecticides, a volume of
224 ml of tap water was added, then one ml of different concentrations in
absolute alcohol. Four replicates of each treatment were performed.
However, untreated check was provided with tap water and one ml of
absolute alcohol. Twenty five of early fourth instar larvae to be tested were
first placed in 25 ml tap water in small containers kept undisturbed for few
hours before being added to the previously mentioned beakers.

The total volume of mixture in such beakers would be 250 ml all together.
Mortality counts of larvae were assessed 24 hours following the application
of the insecticides. Individuals which unable to respond to gentle prodding
with a thin probe or failed to rise to the surface and lacked any sign of
activity were considered dead. IF the untreated mortality is ranged between
5 - 20% the percentage mortalities should be corrected by Abbott’s
formula; Abbott, (1925).

RESULTS & DISCUSSION

Toxicity on mosquito larvae:


Data in Table (1) indicated the important role of insecticide performance
and formulation in determining the toxicity responses against mosquito
larvae. Culex Pepiens (L).

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All insecticides tested yielded similar LC50 estimates for mosquito larvae
that were between 7 to 14 fold higher than Pyriproxyfen. The wide ranges
of LC50 values for both insecticides suggest the presence of resistant and
susceptible of the Culex larvae. Based on the LC50’s values, data show the
superior effectiveness of fenitrothion 50% EC followed by azadirachtin
75% EC, cyphenothrin 5% EC and pyriproxyfen 10% EC showing
LC50’s 4.1, 4.5, 8.4 and 60.0 ppm., respectively. The same trend was
obtained with LC90’s representing 7.9, 8.2, 14.9 and 90.05 ppm,
respectively. Taking the LC50 of pyriproxyfen as abase line for calculation,
the relative potencies revealed 14.6, 13.3 and 7.1 fold for fenitrothion
50% EC, azadirachtin 75% EC, cephenothrin 5% EC, respectively.
The 95% confidence intervals around LC50 and LC90 values for the studied
pesticides overlapped, suggesting that there were no significant differences
among the four pesticides tested. Factors such as developmental stage of
the tested larvae, length of the contact period after treatment and the
temperature would affect LC50 values. Furthermore, the toxicity lines
shown in Figure (1) proved these findings and were in harmony with the
conclusion of some authors such as, Pener et al., (1982), Bekhat et al,
(1983), Kamizono and Eto (1983), Metcalf et al., (1984), Kenaway et al.,
(1985), Tanaka et al., (1986), Kamath and renapurkar (1987), March et al.,
(1990), Cheikh and Pasture (1993), Cuany et al., (1993), Kono et al.,
(1993), Brogdon and Bisset-Lazcano, et al., (1991), McAllister (1998) and
Nazni, et al., (1998).

REFERENCES

Abbott, W.S. (1925) : A method of computing the effectiveness of an


insecticide. J.Econ. Entomol., 18 : 265 -267.

Anonymous, (1981) : Instructions for determining the susceptibility or


resistance of mosquito larvae to insecticides. WHO/VBC. 807 : 6.

Bekheit, S.S (1983): Laboratory and field evaluation of synthetic


pyrethroid decamethrin in controlling mosqueto larvae. J.Path.
Bact., 76:597-605.

Bisset-Lazcano, J; Berovides, V; Rodriguez-Coto, M.M; Diaz-Pantoja, C;


Lazcano, J Bisset; Coto, M.M Rodriguez; Pantoja and C. Diaz
(1991): Esterase patterns in Culex (C.) quinquefasciatus Say,
1823, and their relationship with resistance to the
organophosphorus insecticide malathion. Revista Cubana de
Medicina Tropical. 43: 3, 181-185.

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Brogdon, W.G. and McAllister, J.C. (1998): Simplification of adult
mosquito bioassays through use of time-mortality determinations
in glass bottles. Journal of the American Mosquito Control
Association. 14: 2, 159-164.

Cheikh, H.B and Pasture, N. (1993): Resistance to temephos an


organophosphorous insecticide in Culex pipiens from Tunisia,
North Africa. J. of the American mosqueto control association, 9
(3): 335-337.

Cuany, A. Handani, J. Berge, J. Fournier, D. and Pasture, N (1993):


Action of estrase B1 on chlorpyrifos in organophosphate
resistance Culex pipiens mosqueto. Pesticide biochemistry and
pathology. 45: (1) 1-6.

Elssaidi, M. A. (1998): Laboratory & field studies on the fate of


Fentrothion & Malathion on micro-ecosystem models. M.Sc.
Thesis - Envi Sci. dept. Faculty of Engineering & Technology –
Brack, Sebha University -Libya .

Kamath, V. R. and Renapurkar, D. M. (1987): Evaluation of insecticidal


activity of propetamphos against common household insect pests.
Pesticides – Bombay. 21 (12) : 27-28.

Kamizono, H. and Eto, M. (1983): Synthesis and biological activity of


O,O-dialkyl S-[5-aryl -1,3,4-oxadiazo 1-1-2 (3H) – on 3-yl]
methyl phosphorothioates and phosphorodithioates. Agricultural
and Biological chemistry, 47(4) : 701-706.

Kenaway. M. A, Soliman. SA, Kaschef. AH, and Shanbaku. MN. (1985):


Effect of organophosphorus insecticides on cholinesterase.

Kono, Y. and Tomita, Y. (1993): A role of highly active carboxylesterase in


an organophosphorus insecticide. Japanese J. of Sanitary
Zoology., 44 (1) : 15-21.

Marchi, A. and Addis, G. (1990): Resistance to organophosphorus


insecticides in Sardinian populations of Culex pipiens. Margherita
di pula, 7-11 Maggio. Parassitologya, 32 : 177-178.

Metcalf, R.I. and Metcalf, R.A. (1984): Steric, electronic, and polar
parameters that affect the toxic actions of O-alkyl, - o-phenyl
phosphonothionate insecticides. Pesticide-Biochemistry and
physiology, 22 (2) : 169-177.

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Nazni, W.A.; Lee, H.L. and Sa'-diyah, I. (1998): Rate of resistance
development in wild Culex quinquefasciatus (Say) selected by
malathion and permethrin. Southeast Asian Journal of Tropical
Medicine and Public Health. 29: 4, 849-855.

Pener, H. (1982): Organophosphorus multiresistance in culex pipiens


molestus in Israel. Rivista di parasitologia, 43 (3) : 409- 414.

Tanaka, J.;Kuwano, E. and Eto, M. (1986): Synthesis and pesticidal


activities of phosphonate analogs of amino acids. J. of the Fac. Of
Agric. Kyushu Univ., 30 (4) : 209-223.

Table (1): Toxicity response of tested insecticides against mosquito


larvae, Culex Pipiens (L).

Tested Conc. Observed Corrected LC50 Relative LC90 Slope


insecticide (PPM) mortality ppm ppm potency ppm (b)
1 12 8.33
2 28 25.00
Azadirachtin
75% EC.
4 44 41.67 4.5 13.3 8.2 2.533
8 84 83.33
10 96 95.83
1 8 4.17
2 20 16.67
Fenitrothion
50% EC.
4 48 45.83 4.1 14.6 7.9 2.313
8 92 91.67
12 100 100.00
2 12 8.33
4 28 25.00
Cyphenothrin
5% EC.
8 45 48.83 8.4 7.1 14.9 2.621
12 72 70.83
16 92 91.66
20 12 8.33
40 20 16.67
Pyriproxyfen 90.0
10% EC.
60 52 50.00 60.0 1.0 3.818
80 84 83.33 5
100 96 95.83

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Figure (1): The toxicity lines of azadirachtin (A), fenitrothion (B),
cyphenothrin (C) and pyriproxyten (D) on Culex pipiens larvae.

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‫سمية بعض المبيدات الحشرية على يرقات البعوضة‬
‫جنس كيولكس ‪)Culex pipiens (L‬‬

‫‪2‬‬
‫محمد علي السعيدي‬ ‫‪1‬‬
‫عبد ال عبد القادر الزروق‬
‫‪ .1‬كلية العلوم – جامعة سبها ‪ .2 ،‬كلية العلوم الهندسية والتقنية – جامعة سبها‬

‫المستخلص‬

‫استهدفت هذه الدراسة إلقاء الضوء على سمية بعض المبيدات الحشرية على‬
‫يرقات البعوضة من النوع ‪ . Culex Pipiens‬وتشمل المبيدات المختبرة مبيد‬
‫الفينتروثيون ‪ ( EC %50‬مبيد حشري فسفوري ) ‪ ،‬مبيد السيفنوثرين ‪EC %5‬‬
‫( مبيد بيرترويدي ) ‪ ،‬مبيد الزيدراكتين ‪ ( EC %75‬مستخلص نباتي ) ومبيد‬
‫البيروبروكسفين ‪ ( EC %10‬منظم نمو حشري ) ‪ .‬وقد أوضحت النتائج التأتير‬
‫السـمي العالي لمبيد الفينتروثيون بليه مبيد الزيدراكتين فمبيد السيفنوثرين ثم‬
‫البيربروكسفين على الترتيب ‪ .‬وكانت قيم التراكيز القاتلة النصفية (‪ )LC50‬والكافية‬
‫لقتل ‪ %50‬من اليرقات المختبرة ‪ 60.0 ، 8.4 ، 4.5 ، 4.1‬جزء في المليون‬
‫للفينتروثيون والزيدراكتين والسيفنوثرين والبيربروكسفين على الترتيب ‪ .‬وبتقدير‬
‫الكفاءة النسبية للمبيدات المختبرة على أساس مبيد البيروبروكسفين بلغت هذه‬
‫القيم ‪ 7.1 ، 13.3 ، 14.6 :‬مرة قدر كفاءة البيربروكفين مع مبيدات الفينتروتيون‬
‫والزيدراكتين والسيفنوترين على التوالي ‪.‬‬

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