Cyromazine Induced Effects on Larvae and Adults of ... - Iresa

<strong>Cyromazine</strong> <strong>Induced</strong> <strong>Effects</strong> on Larvae and Adults of 

Laboratory Tunisian Strain of the Mediterranean Fruit Fly 

Ceratitis capitata 

Jouda Mediouni-Ben Jemâa and Emna Boushih, Laboratoire de Protection des 

Végétaux, INRAT, Rue Hédi Karray, 2080 Ariana, Université de 7 Novembre à 

Carthage, Tunisia 

__________________________________________________________________________ 

ABSTRACT 

Mediouni-Ben Jemâa, J. and Boushih, E. 2010. <strong>Cyromazine</strong> induced effects on larvae and adults 

of laboratory Tunisian strain of the Mediterranean fruit fly Ceratitis capitata. Tunisian Journal 

of Plant Protection 5: 213-224. 

<strong>Cyromazine</strong> is an insect growth regulator insecticide mainly used to control dipteran insects. This work 

aimed to assess cyromazine-induced effects on the second instar larvae and adults of the Tunisian 

laboratory strain of Mediterranean fruit fly Ceratitis capitata. <strong>Effects</strong> were assessed through larvae and 

adult mortality together with the evaluation of larval and pupal body deformities, female fecundity and 

adult emergence rate. <strong>Cyromazine</strong> was respectively added to the larvae diet and adult nutrient solution 

at three doses 0.05, 0.1 and 0.2 mg/l. Results indicated that cyromazine induced significant mortalities 

to both larvae and adults. After 96 h of exposure and with the dose of 0.2 mg of cyromazine per liter, 

91 and 69% of mortality were recorded respectively for larvae and adults. Moreover, females exposed 

to different cyromazine doses exhibited significant reductions in their fecundity. The mean number of 

eggs laid per female was 55 against 200 for the control at the highest dose. Additionally, a poisoning 

effect expressed by the appearance of body deformities for larvae and pupae was observed. The 

percentage of deformities was positively correlated to cyromazine doses. Furthermore, adult emergence 

rate was strongly reduced compared to the control. Emergence rates were 83, 73.5, 2.5 and 2.02% 

respectively for the control, 0.05, 0.1 and 0.2 mg of cyromazine per liter. 

Keywords: Ceratitis capitata, cyromazine, emergence, fecundity, IGR (for Insect Growth Regulator) 

__________________________________________________________________________ 

The Mediterranean fruit fly, 

Ceratitis capitata (Medfly) was first 

recorded in Tunisia in 1885 (23). Since 

that, it became a serious pest causing 

quantitative and qualitative losses to 

several crops; mainly citrus. Presently, 

chemical control, using broad-spectrum 

insecticides, essentially belonging to 

organophosphates class, plays a major 

role in controlling this insect. However, 

Corresponding Author: Jouda Mediouni-Ben Jemâa 

Email: joudamediouni@lycos.com 

Accepted for publication 29 December 2010 

insect resistance was recorded and control 

failures have been reported in field 

conditions during last years (4, 5). 

Among the most used insecticides, 

malathion has been declared an ozonedepleting 

substance and therefore, is 

being phased out. Consequently, it has 

been banned from annex I of the EU 

directive 91/414/ EEC (26) since 2009. 

In the past few decades, a 

considerable interest has been paid to the 

potential use of insect growth regulators 

known as IGR in controlling insect pests 

(2). Insecticides, acting as insect growth 

regulator, affect growth and development 

Tunisian Journal of Plant Protection 213 Vol. 5, No. 2, 2010

of the pre-adult life stages of insects, and 

are effective against many insects (25). 

<strong>Cyromazine</strong>, N-cyclopropyl-1,3,5- 

triazine-2,4,6-triamine, is an atypically 

substituted s-triazine, which acts as an 

insect growth regulator used as a chitin 

synthesis inhibitor for fly control in cattle 

manure, field crops, vegetables and fruits 

(7, 34). It is an effective larvicide against 

a number of dipteran and lepidopteran 

species (20, 29). <strong>Cyromazine</strong> has a good 

activity against many dipteran species, 

especially when it is orally administered 

to larvae which usually exhibit a wide 

range of different morphological 

abnormalities (3, 15, 27, 35). 

<strong>Cyromazine</strong> is characterized by a 

rapid stiffening of the cuticle, affecting 

mostly larvae of Diptera (9, 14, 18). 

Reynolds and Blakey (29) have shown 

that an early action of cyromazine leads 

the cuticle to become less extensible 

when subjected to simple constant load 

extension tests. They have suggested that 

the cyromazine-induced reduction in 

cuticle extensibility may be responsible 

for the other symptoms of poisoning and 

hence may be the primary effect of the 

insecticide. Moreover, cyromazine 

induces gross deformities manifest in 

larvae and pupae of dipteran insects (3, 

9), and also reduces egg laying and egg 

hatch when fed to Lucilia cuprina adult 

(38). Moreover, Alam et al. (2) clearly 

demonstrated that cyromazine, ingested 

by female flies, is incorporated into eggs 

and inhibits larval development in the 

first generation. 

<strong>Cyromazine</strong> is also effective as 

foliar spray in horticultural crops against 

leafminers (Liriomyza spp.) and various 

other insects, including fleas, thrips, and 

coleoptera (18, 32). It is reported to be a 

main translaminar pesticide used to 

control respectively the pea leafminer 

Liriomyza huidobrensis and the 

serpentine leaf miners (L. trifolii) (13, 17, 

30, 37). <strong>Cyromazine</strong> was used because it 

is harmless to parasitoids (6, 31). 

Recently, there is an increasing trend 

among onion growers in New York to 

manage onion maggot using a 

combination of cyromazine seed 

treatment and chlorpyrifos (Lorsban 4E, 

Dow AgroSciences LLC) in furrow 

treatments (28). The Medfly, C. capitata 

was reported among the susceptible 

species to cyromazine applications (35). 

Larvae when reared on an artificial diet 

supplemented with different 

concentrations of cyromazine, suffer from 

severe developmental disruption 

depending on larval age and duration of 

exposure (11, 36). Symptoms of 

poisoning include body malformations, 

such as swelling of the integument and 

elongation of the larvae, as well as 

inhibition of larval growth, increase of 

larval mortality and a lower pupation and 

adult emergence rate. Furthermore, Budia 

and Viňuela (10) reported that the 

delivery of cyromazine could affect the 

reproduction and larval development of 

C. capitata. 

The present work aims to assess 

cyromazine effects on larvae and adults 

of Tunisian Medfly strain reared under 

laboratory controlled conditions. 

MATERIALS AND METHODS 

Insect. Larvae of C. capitata were 

reared on an artificial diet based on wheat 

bran, sucrose and yeast (Table 1). The 

wheat bran was first sterilized at 120°C 

for two hours. After that, all solid 

ingredients were weighed and mixed until 

a homogenous consistency was reached. 

Liquid yeast and hydrochloric acid were 

then added. To avoid bacterial and fungal 

contaminations, sodium benzoate and 

nipagin were added to the diet. 

The rearing culture was kept in a rearing 

room at 27 o C ± 1 o C, 75 ± 5% R.H and 

under continuous light. 


Insecticide. <strong>Cyromazine</strong> (registered 

trademark Trigard 75, Syngenta 

AgroServices AG, Switzerland) was 

tested against C. capitata. Trials were 

conducted on the second instar larvae and 

new emerged adult flies. Trigard 75 WP 

was dissolved in distilled water at 

respective doses of 0.2, 0.1 and 0.05 mg/l. 

Technical ingredient, cyromazine, was 

orally administered in the drinking water 

and incorporated into the diet respectively 

to newly emerged adult flies and larvae. 

For adult flies, experiments were carried 

out into circular plastic boxes (∅ = 20 

cm). Each box contains 50 adults (25♂ + 

25♀). Trials were replicated 10 times to 

improve precision. Ten cages with 

untreated drinking water served as 

control. For larvae, experiments carried 

on with 20 larvae, were performed in 

Petri dishes (∅ = 9 cm) containing 100 g 

of diet. The experiment was replicated 10 

times. Ten Petri dishes containing 

untreated diet were used as control. 

The effects of three doses of 

cyromazine on larvae and adult 

mortalities were assessed at 24, 48, 72 

and 96 h after treatment. The dose 0.2 

mg/l corresponded to the recommended 

dose of Trigard 75 by the registration 

pesticide service of the Tunisian Ministry 

of Agriculture. Moreover, female 

fecundity was determined at different 

doses. In addition, body deformities of 

larvae and pupae together with adult 

emergence rate were evaluated. 

Table 1. Composition of C. capitata diet: 

quantities per 1000 g of diet (12) 

Ingredient 

Quantité 

Wheat bran 

550.0 g 

Sucrose 

340.0 g 

Sodium Benzoate 

2.5 g 

Nipagin 

2.5 g 

Yeast (liquid) 

1000 ml 

Hydrochloric acid 

20 ml 

Statistical analyses. Adult and 

larval mortalities were calculated at 24, 

48, 72 and 96 h after treatment using 

Abbott formula (1) expressed by 

C − T , where C = rate of Medfly 

Tunisian Journal of Plant Protection 215 Vol. 5, No. 2, 2010 

TR = 

C 

x 100 

mortality in the control field, T = rate of 

Medfly mortality in the treated field and 

TR = rate of population reduction. Results 

were expressed as percentage of 

mortality. 

Biological parameters, like 

fecundity, body deformities of larvae and 

pupae and adult emergence rate, were 

subjected to analysis of variance 

(ANOVA) using Statistica software (33). 

Significant differences were identified by 

least significant difference (LSD) test at 

the probability level 0.05. 

RESULTS 

<strong>Cyromazine</strong> effects on second 

instar larvae adult mortality. Whatever 

cyromazine concentration, this active 

ingredient was toxic both to second instar 

larvae and to adults of C. capitata (Fig. 

1). Moreover, larvae were more 

susceptible than adults. Results showed 

that insect mortality raises with the 

increase of cyromazine concentration and 

time after treatment. 

The lowest dose of cyromazine 

(0.05 mg/l) caused 8% mortality of C. 

capitata larvae and 10% of adults after 24 

h exposure (Fig. 1). At the highest dose 

(0.2 mg/l), 91 and 69% mortality were 

respectively recorded for larvae and 

adults after 96 h of exposure.

Mortality (%) 

100 

80 

60 

40 

20 

72 h of exposure 

Adults 

Larvae 

Mortality (%) 

100 

80 

60 

40 

20 

96 h of exposure 

Adults 

Larvae 

0 

0 

0 0,05 0,1 0,2 

0 0,05 0,1 0,2 

Concentration (mg/l) 

Concentration (mg/l) 

Fig. 1. Percentage of mortality of C. capitata larvae and adults exposed for various periods of time to different 

cyromazine doses. 

Fig. 2. Fecundity of female C. capitata exposed to different cyromazine doses 

Tunisian Journal of Plant Protection 213 

Vol. 5, No. 2, 2010

<strong>Cyromazine</strong> effect on female 

fecundity. Females exposed to different 

cyromazine doses exhibited significant 

reductions in their fecundity (Fig. 2). At 

the highest dose, the mean number of 

eggs laid per female was 55 against 200 

for the control. Moreover, at this dose, no 

more eggs were laid after the seventh day 

after the treatment. At the dose 0.1 mg/l, 

the mean number of eggs obtained at the 

third day of laying was 6 and was 

significantly different from the control. 

Nevertheless, at the dose 0.05 mg/l, no 

significant reductions were obtained 

concerning the mean number of eggs laid 

per female compared to the control. 

<strong>Cyromazine</strong> effects on body 

deformities of larvae and pupae. 

<strong>Cyromazine</strong> incorporated to second instar 

larvae diet manifested a poisoning effect 

expressed by the appearance of body 

deformities on larvae and pupae. The 

percentage of deformities was positively 

correlated to cyromazine doses (Fig. 3). 

Statistical analyses showed significant 

differences in percentage of body 

deformities of larvae and pupae exposed 

to different doses of the insecticide 

compared to the control. 

Fig. 3. Percentage of body malformations of C. capitata larvae and pupae exposed to different cyromazine doses. 

<strong>Cyromazine</strong> effects on adult 

emergence rate. Results showed that 

emergence rate significantly depended on 

cyromazine dose (Fig. 4). The emergence 

rate was 2.02% at the dose 0.2 mg/l 

compared to 2.5% for the dose 0.1 mg/l, 

while it reached 73.5% at the dose 0.05 

mg/l against 83% for the control (0 mg/l). 

No statistical differences were 

observed between the control and the 

lowest dose 0.05 mg/l. Similarly, no 

statistical differences were obtained 

between the two doses 0.1 and 0.2 mg/l. 


Vol. 5, No. 2, 2010

Emergence rate (%) 

100 

80 

60 

40 

20 

a 

a 

b 

b 

0 

0 0,05 0,1 

Dose (mg/l) 

0,2 

Fig. 4. Emergence rate of C. capitata adults exposed to different cyromazine doses 

DISCUSSION 

In Tunisia, C. capitata has been a 

serious pest of several fruit crops mainly 

citrus and is found all around the year. It 

may complete seven or eight generations 

per year, five of them are during the 

summer (16, 19). 

Results reported in this work clearly 

demonstrate that cyromazine ingested by 

larvae and adults of C. capitata induced a 

poisoning syndrome that include 

significant mortality of all developmental 

stages, reduction in female fertility, 

increase of percentage of body 

deformities and decrease of adult 

emergence rate. 

Many studies have been reported on 

the insecticidal activity of cyromazine 

against C. capitata. Indeed, Viňuela et al. 

(36) reported that C. capitata larvae 

reared on an artificial diet supplemented 

with cyromazine undergo severe 

developmental disruption depending on 

their age and exposure duration. 

Moreover, Viňuela and Budia (35) 

indicated that Medfly larvae treated with 

cyromazine (20 mg/Kg of diet) were 

smaller, more elongate and considerably 


less elastic than control. On the contrary, 

when sub-lethal concentrations were 

added to the larval rearing medium, no 

body alterations were recorded. 

Furthermore, at the concentration of 20 

mg/Kg of diet, larvae of C. capitata 

develop severe symptoms, less than 20% 

of them are able to pupate and none 

emerge adults (36). In addition, 

cyromazine is known to affect dipteran 

puparia with lethal outcome (8). These 

authors reported that cyromazine has an 

effect on pupae cuticle of Musca 

domestica. Besides, Kayser and Palivan 

(22) reported that, when larvae of 

dipteran species (M. domestica, Lucilia 

sericata, Calliphora vicina and 

Protophormia terraenovae) were feed 

with diet added with cyromazine (0.25 

mg/l), there is an important insecticidal 

effect. In another context, Reynolds and 

Blakey (29) have shown that an early 

action of cyromazine is to cause the 

cuticle to become less extensible when 

subjected to simple constant load 

extension tests. Additionally, the effect of 

cyromazine on cuticle mechanical 

properties occurred rapidly after exposure 

Vol. 5, No. 2, 2010

of larvae to the treated diet (24). against Medfly larvae and adults. Thus, 

Moreover, Karras et al. (21) indicated cyromazine could be suggested as an 

that the active ingredient of cyromazine is alternative chemical control method in 

more efficiently utilized at lower doses. Tunisian citrus orchards and could have 

After this study, promising results an effective contribution in Integrated 

are obtained with cyromazine treatment Pest Management programs. 

___________________________________________________________________________ 

RESUME 

Mediouni-Ben Jemâa J. et Boushih E. 2010. Effets induits de la cyromazine sur les larves et les 

adultes d’une souche tunisienne de laboratoire de la mouche méditerranéenne des fruits Ceratitis 

capitata. Tunisian Journal of Plant Protection 5: 213-224. 

La substance active, cyromazine, est un régulateur de croissance des insectes principalement utilisé 

pour la lutte contre les diptères. Ce travail a pour objectif d’évaluer les effets induits par la cyromazine 

sur les larves du deuxième stade et sur les adultes d’une souche tunisienne de laboratoire de la mouche 

méditerranéenne des fruits Ceratitis capitata. Les effets ont été évalués à travers la mortalité des larves 

et des adultes, le taux de malformations corporelles des larves et des adultes, la fécondité des femelles 

et le taux d’émergence des adultes. La cyromazine a été respectivement ajoutée au milieu d’élevage des 

larves et à la solution nutritive des adultes à trois doses différentes 0,05, 0,1 et 0,2 mg/l. Les résultats 

ont indiqué que la cyromazine a induit des mortalités significatives des larves et des adultes. A la dose, 

0,2 mg/l, 91 et 69% de mortalité ont été respectivement enregistrés pour les larves et les adultes après 

96 h suivant le traitement. De plus, les femelles exposées aux différentes doses de cyromazine ont 

montré des différences significatives de leur fécondité. A la plus forte dose (0,2 mg/l), le nombre 

moyen d’œufs pondus par femelle était de 55 contre 200 pour le contrôle. En plus, un effet 

d’empoisonnement exprimé par l’apparition de malformations corporelles chez les larves et les adultes 

a été également observé. Le pourcentage de malformations corporelles a été positivement corrélé avec 

les doses de cyromazine. En outre, le taux d’émergence des adultes a été fortement réduit comparé au 

témoin. Les taux d’émergence ont été respectivement 83, 73,5, 2,5 et 2,02% pour le témoin, la dose de 

0,05, 0,1 et 0,2 mg/l. 

Mots clés: Ceratitis capitata, cyromazine, émergence, fécondité, RCI (Régulateur de Croissance des 

Insectes) 

___________________________________________________________________________ 

.2010 

.(Ceratitis capitata) 

ملخص 

مديوني-بن جماعة،‏ جودة و آمنة بوشيح.‏ 

تونسية للذبابة المتوسطية للفواآه 

تأثيرات استعمال سيراموزين ضد يرقات وبالغات من سلالة مخبرية 

Tunisian Journal of Plant Protection 5: 213-224 

يعدّ‏ سيرومازين منظم نمو للحشرات يستخدم بشكل رئيسي في مكافحة حشرات ثنائيات الأجنحة هدف هذا 

العمل إلى تقييم التأثيرات التي يحدثها استعمال سيراموزين في يرقات الطور الثاني وبالغات سلالة مخبرية للذبابة 

تم تقييم هذه التأثيرات باحتساب نسبة موت اليرقات والبالغات مع التشويهات 

المتوسّطية للفواآه 

الجسمية لليرقات والعذارى،‏ ونسبة خصوبة الإناث،‏ ونسبة ظهور البالغات.‏ تمت إضافة سيراموزين للوسط الغذائي 

لليرقات وللمحلول الغذائي للبالغات وفق ثلاث جرعات و و مغ/ل.‏ أشارت النتائج إلى أن سيرومازين 

أحدث موتًا بليغًا لكل من اليرقات والبالغات عند الجرعة مغ/ل،‏ حيث بلغت نسب الموت و بالنسبة لليرقات 

والبالغات،‏ على التوالي بعد ساعة من التعريض.‏ إضافة إلى ذلك،‏ أظهرت الإناث المعرضة للجرعات المختلفة من 

سيرومازين انخفاضا ملحوظا في نسبة الخصوبة.‏ بلغ معدل عدد البيض بيضة للأنثى الواحدة عند الشاهد مقابل 

بيضة للأنثى بالنسبة للجرعة مغ/ل.‏ زيادة على ذلك،‏ لوحظ الأثر السمي المتمثل في ظهور تشوهات جسمية عند 

اليرقات والبالغات.‏ وآانت نسبة التشوهات مرتبطة ايجابيا بجرعات السيرومازين.‏ علاوة على ذلك،‏ لوحظ انخفاض شديد 

55 

.(Diptera) 

%69 

91 

0.2 

200 

0.1 

0.05 

0.2 

.(Ceratitis capitata) 

Tunisian Journal of Plant Protection 214 Vol. 5, No. 2, 2010 

0.2 

96

%2.02 

2.5 

73.5 

في نسبة ظهور الإناث مقارنة بالشاهد.‏ بلغ معدل ظهور البالغات 83 

وللجرعة والجرعة والجرعة مغ/ل،‏ على التوالي.‏ 

و و و 

بالنسبة إلى الشاهد،‏ 

0.2 

0.1 

0.05 

آلمات مفتاحية:‏ خصوبة،‏ ظهور،‏ سيراموزين،‏ منظم نمو الحشرات،‏ 

Ceratitis capitata 

___________________________________________________________________________ 

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