Raúl A. Alzogaray

Insecticidal Activity of Essential Oils from Eleven Eucalyptus spp. and Two Hybrids: Lethal and Sublethal Effects of Their Major Components on Blattella germanica

ABSTRACT The aim of the current study was to evaluate the fumigant activity of the essential oils from 11 species of the genus Eucalyptus and two of their hybrids on first instar of Blattella germanica L. The fumigant activity and repellence of the four major monoterpene components of these essential oils also were tested. Fumigant activity was evaluated by exposing nymphs to the vapors emitted by 50 µl of essential oil or monoterpene in a closed container. The lowest knockdown time 50% (KT50) values, expressed in minutes, were elicited by the essential oils of the Eucalyptus grandis × Eucalyptus tereticornis (57.9) hybrid, Eucalyptus sideroxylon A. Cunn (62.0), E. grandis × Eucalyptus camaldulensis (63.8) hybrid, Eucalyptus viminalis Labill (64.1), Eucalyptus dunnii Maiden (64.5), and Eucalyptus grandis (Hill) ex Maiden (68.7). The KT50 values for the remaining essential oils ranged between 74.5 (E. saligna Smith) and 161.4 min (E. tereticornis Smith). The essential oil from the hybrid E. grandis × E. tereticornis was 3.7 times less toxic than dichlorvos (positive control). The KT50 values of monoterpenes were 38.8 for &agr;-pinene, 55.3 for 1,8-cineole, 175.6 for p-cymene, and 178.3 for &ggr;-terpinene. &agr;-Pinene was 2.5 times less toxic than dichlorvos. There was a strong positive correlation between the fumigant activity of essential oils and their corresponding 1, 8-cineole and &agr;-pinene concentration. Repellency was quantified using a video tracking system. Two concentrations of monoterpenes were studied (7 and 70 µg/cm2). All compounds produced a light repellent effect but only when applied at 70 µg/cm2. In all cases, the repellent effect was less than that produced by the broad-spectrum insect repellent N,N-diethyl-3-methylbenzamide (positive control).

Attractant Volatiles Released by Female and Male Triatoma infestans (Hemiptera: Reduviidae), a Vector of Chagas disease: Chemical Analysis and Behavioral Bioassay

Abstract Volatiles emitted by male and female T. infestans before and during copula were collected on Porapak-Q filters, desorbed with dichloromethane, and analyzed by gas chromotography and gas chromatography-mass spectrometry after confirmation of attractiveness in an arena bioassay. Chemical analysis confirmed the presence of (R,S)-2- and 3-methylbutan-1-ol in a 2:1 ratio; short chain acids (ethanoic to nonanoic acid); long chain acids decanoic to (Z)-9-octadecenoic acid; aliphatic aldehydes (hexanal to nonanal), benzaldehyde and dipropylsulphide from insects in copula. Electroantennographic studies conducted with a homologous series of aliphatic aldehydes on female and male T. infestans showed that, for a given dose, EAG responses elicited from both sexes increased with increased chain length up to nonanal, after which EAG-activity declined. Attractiveness of non-acidic trace components identified in the volatiles were tested on male and female T. infestans, in an arena bioassay using a video tracking method. Aliphatic C6 to C10 aldehydes were tested: hexanal (1–100 μg) and heptanal (10 μg) were attractive to female T. infestans, high doses of octanal and nonanal (1–100 μg) were unattractive to male and female T. infestans but low doses of nonanal (0.01–0.1 μg) were attractive to male T. infestans. Benzaldehyde was highly attractive to female T. infestans at low doses (0.05–0.1 μg). 3-methylbutan-1-ol was attractive to male T. infestans at high dose (1,000 μg). (S) or (S,R) 2-methyl-butan-1-ol were anattractive to males or females (1–1,000 μg). Blends of hexanal and benzaldehyde (20:1 and 40:1) showed an additive effect on attraction compared with hexanal alone, when tested on female T. infestans. The study has demonstrated the presence of a number of electrophysiologically and behaviorally active compounds in volatiles emitted by T. infestans in copula that may have a role in the postulated copulation pheromone.

Repellency of deet to nymphs of Triatoma infestans

The repellency of N,N‐diethyl‐3‐methylbenzamide (deet) to Triatoma infestans (Klug) (Hemiptera: Reduviidae) was evaluated using third‐instar nymphs and a video tracking technique. Three experimental designs were used: (a) the test arena floor was divided into two halves, only one of which was treated with deet; (b) the arena floor was divided into an inner circle and an outer ring, only the latter treated with deet; (c) half of the test arena was covered by a filter paper roof treated with deet (out of reach of the nymphs). In all three types of experiment, a repellent effect was demonstrated proportional to the dose of deet. When a host (pigeon) was shielded by deet‐treated gauze, the rate of blood‐feeding by the nymphs was inhibited. Topical pre‐treatment of the nymphs with N‐ethylmaleimide, to block chemoreception, inhibited the repellency.

Evaluation of hyperactivity produced by pyrethroid treatment on third instar nymphs of Triatoma infestans (Hemiptera: Reduviidae)

The hyperactivity produced in third instar nymphs of Triatoma infestans by their exposure to films of deltamethrin or cis-permethrin was evaluated. Both pyrethroids produced a significant increase in locomotor activity at 26 and 36 degrees C but not at 16 degrees C. At 26 degrees C, only deltamethrin produced hyperactivity when topically applied on the dorsal side of the nymphs abdomen. However, both pyrethroids produced hyperactivity when topically applied on the head of the nymphs. Hyperactivity was not observed when nymphs were treated with N-ethylmaleimide (20 micrograms/insect) before exposure to the pyrethroids. The effect of both insecticides on locomotor activity reversed the inhibitory influence on locomotion elicited by contact with the walls of the experimental arena (thigmotaxis).

Toxicity of pyrethroids and repellency of diethyltoluamide in two deltamethrin-resistant colonies of Triatoma infestans Klug, 1834 (Hemiptera: Reduviidae).

The aim of the currrent investigation was to evaluate (a) the toxicity of three pyrethroids (deltamethrin, lambda-cyhalothrin, and tetramethrin); (b) the effect of these insecticides on the locomotor activity; and (c) the repellent effect of N,N-diethyl-m-toluamide (DEET) on two deltamethrin-resistant strains of Triatoma infestans from Argentina (El Chorro and La Toma), and one susceptible strain. The resistance ratios (RRs) obtained for the La Toma strain were: > 10,769, 50.7, and > 5.2 for deltamethrin, lambda-cyhalothrin, and tetramethrin respectively. The RRs for the El Chorro strain were: > 10,769, 85.8, and > 5.2 for deltamethrin, lambda-cyhalothrin, and tetramethrin respectively. The hyperactivity usually caused by the three pyrethroids was in both the deltamethrin-resistant strains compared to the susceptible reference strain. No differences were observed in the repellent effect of DEET between the three groups. These results indicate that the deltamethrin-resistant insects have a cross resistance to lambda-cyhalothrin and tetramethrin, and are also resistant to the first symptom of pyrethroid poisoning (hyperactivity). However, the sensorial process related to DEET repellency does not appear to be altered.

Incoordination, paralysis and recovery after pyrethroid treatment on nymphs III of Triatoma infestans (Hemiptera: Reduviidae).

Symptoms of poisoning for deltamethrin and cis-permethrin on nymphs III of Triatoma infestans were described. The time required for incoordination and paralysis were determined. Deltamethrin was equal or more rapid in the onset of the first effect (accordingly to dose), and cis-permethrin in the onset of the second one. There were no significant differences between incoordination doses 50% (IncD50s) at different times for the two pyrethroids. They showed equivalent incoordination power, but the nymphs treated with deltamethrin recovered slightly more rapid and in greater amount than the nymphs treated with cis-permethrin. The recovery was inhibited by the simultaneous application of piperonyl butoxide. This result suggests that biotransformation by mixed-function microsomal oxidases are involved in the process of recovery.

Behavioral response of fifth instar nymphs of Triatoma infestans (Hemiptera : Reduviidae) to pyrethroids

The hyperactivity (an increase in locomotor activity) and repellency produced by eight pyrethroids, applied as films on filter paper, were evaluated on fifth instar nymphs of Triatoma infestans (Hemiptera: Reduviidae) using a video tracking technique. All the pyrethroids studied produced hyperactivity. As a trend, hyperactivity produced by cyanopyrethroids was higher than that produced by non-cyanopyrethroids. Hyperactivity was not observed when nymphs were pretreated with the sulphydryl reagent N-ethylmaleimide before exposure to the pyrethroids. The eight pyrethroids failed to produce repellency. No repellency was also observed for the flowable formulation of deltamethrin at the concentration recommended for T. infestans control.

Independent and joint action of cis- and trans-permethrin in Triatoma infestans (Hemiptera: Reduviidae)

The toxicity of pure cis- and trans-permethrin or mixtures of the two isomers topically applied to first, third, and fifth instar nymphs of Triatoma infestans (Klug) at 26°C was determined. The cis-isomer was more active than the trans-isomer in the three stages evaluated. When the two isomers were simultaneously applied to first instar nymphs, an additive effect was observed. Similar treatments of third and fifth instar nymphs resulted in an antagonistic effect. In third instar nymphs, the cis-isomer was more active than trans-isomer at all the three temperatures assayed (16°, 26°, and 36°C). The toxicity of the cis-isomer was lower at 36°C than at either 16° or 26°C. Temperature had no significant effect on the toxicity of the trans-isomer within the temperature range assayed. The toxicity of either isomer to third instar nymphs was not affected by pretreatment of nymphs with PBO (an inhibitor of mixed-function oxidases activity) or TPP (an inhibitor of esterase activity), suggesting that these detoxification pathways are not relevant in the metabolism of cis- or trans-isomers. Arch. Insect Biochem. Physiol. 37:225–230, 1998.

Lethal and sublethal effects of eucalyptol on Triatoma infestans and Rhodnius prolixus, vectors of Chagas disease

Eucalyptol is the common name for a cyclic ether monoterpene found in essential oils from Eucalyptus species and other plants. Several reports showed its insecticidal activity. In this work, visible symptoms of intoxication, effect on locomotor activity, knock‐down, and repellence produced by eucalyptol were evaluated on nymphs of Triatoma infestans Klug and Rhodnius prolixus Ståhl (both Hemiptera: Reduviidae). Both insects are among the main vectors of Chagas disease in Latin America. Visible symptoms of intoxication were similar to those observed for neurotoxic insecticides. A video tracking technique was used to evaluate locomotor activity and repellence by exposing the nymphs to impregnated papers. Hyperactivity (a non‐directional increase in locomotor activity) is a symptom of intoxication that is used to detect triatomines in rural houses, because it causes the insects to leave their refuges. Eucalyptol produced hyperactivity only in T. infestans at a concentration 1 000× higher than the positive control, deltamethrin [(S)‐cyano(3‐phenoxyphenyl)methyl (1R,3R)‐3‐(2,2‐dibromoethenyl)‐2,2‐dimethylcyclopropanecarboxylate]. It also produced repellence on both species at a concentration 10× higher than the positive control, DEET (N,N‐diethyl‐3‐methylbenzamide). Knock‐down effect was evaluated by exposing the nymphs to impregnated papers in closed containers (contact and fumigation simultaneously). Values of knock‐down time for 50% of exposed nymphs (KT50) were calculated for various concentrations of eucalyptol. The onset of knock‐down occurred more rapidly as the concentration increased. In the best cases, eucalyptol was 12–15× less toxic than the positive control dichlorvos (2,2‐dichlorovinyl dimethyl phosphate). After these results, eucalyptol seems discouraged as a hyperactivant agent for monitoring insects in rural houses. Nevertheless, its knock‐down and repellence effect on vectors of Chagas disease deserve further investigation.

Locomotor Behaviour of Blattella germanica Modified by DEET

N,N-diethyl-3-methylbenzamide (DEET) is the active principle of most insect repellents used worldwide. However, its toxicity on insects has not been widely studied. The aim of this work is to study the effects of DEET on the locomotor activity of Blattella germanica. DEET has a dose-dependent repellent activity on B. germanica. Locomotor activity was significantly lower when insects were pre-exposed to 700 µg/cm2 of DEET for 20 or 30 minutes, but it did not change when pre-exposure was shorter. Locomotor activity of insects that were pre-exposed to 2.000 µg/cm2 of DEET for 10 minutes was significantly lower than the movement registered in controls. No differences were observed when insects were pre-exposed to lower concentrations of DEET. A 30-minute pre-exposure to 700 µg/cm2 of DEET caused a significant decrease in locomotor activity. Movement was totally recovered 24 h later. The locomotor activity measured during the exposure to different concentrations of DEET remained unchanged. Insects with decreased locomotor activity were repelled to the same extent than control insects by the same concentration of DEET. We demonstrated that the repellency and modification of locomotor activity elicited by DEET are non-associated phenomena. We also suggested that the reduction in locomotor activity indicates toxicity of DEET, probably to insect nervous system.