We studied the profile of permethrin resistance in populations of head lice infesting children 6-12 yr old in schools and their homes in and around Buenos Aires, Argentina. Five permethrin-resistant populations with different levels of resistance were collected: Hogar Loyola (HL), Republica de Turquia (RT), Hogar Mitre (HM), Guardia de Honor (GH), and Ricardo Guiraldes (RG). One susceptible population, Bandera Argentina (BA), also was collected. Their level of resistance was evaluated, and results showed resistance ratios of 13 for HL, 16 for RT, 22 for HM, 61 for GH, and 69 for RG. To elucidate the possible involvement of the cytochrome P450 monooxygenase system in conferring permethrin resistance, ethoxycoumarin-O-deethylase (ECOD) activity was measured in abdomens of individual third instars and adults by using a fluorometric assay. The ECOD activity was lower in the susceptible BA population (4.7 ng per louse) than in the resistant ones (13.7 ng per louse for RG, 12.3 ng per louse for GH, 8.6 ng per louse for RT, and 8.2 ng per louse for HL). ECOD activity was significantly correlated with the level of resistance in the field populations (r = 0.97, P = 0.0009), suggesting a role for cytochrome monooxygenase P450 system in permethrin resistance by head louse, Pediculus humanus capitis De Geer.

Topical application of 1-dodecanol was significantly more toxic against teneral first nymphs (1-3 h old) than post-teneral first nymphs (24 h old). The lethal dose ratios were 711,500 for Rhodnius prolixus and 3613 for Triatoma infestans. No significative difference between LD50 was found when 1-dodecanol was injected in recently hatched adult R. prolixus (1-4 h old) nor in older adults (24 h old). These values were similar to those calculated for deltamethrin (an effective triatomicide), showing that 1-dodecanol had no insecticidal properties when it was applied by injection. Topical application of high dose of 1-dodecanol (1 microg/i) on teneral first nymphs of R. prolixus, produced an interruption of the darkening process of the cuticle, and probably in the development of its physiological properties.

Deltamethrin and other pyrethroids have been extensively used in Argentina since 1980, for the chemical control of Triatoma infestans Klug (Hemiptera: Reduviidae). Recently, resistance to deltamethrin was detected in field populations by the survival of bugs exposed by topical application to the diagnostic dose estimated on the CIPEIN susceptible strain. Results of the current study showed low resistant ratios (RRs) to deltamethrin for the resistant populations (RR ranged from 2.0 for San Luis colony to 7.9 for Salta colony). Biochemical studies were made on the most resistant colony (Salta) and the susceptible strain (CIPEIN), in order to establish the importance of degradative mechanisms as a cause of the detected resistance. Esterase activity was measured on 3 days old first instars through phenylthioacetate and a-naphtyl acetate activities. The results showed a significant difference in no cholinesterase esterase activity from susceptible (7.6 +/- 0,7 micro M S./i.min.) and Salta resistant colony (9.5 +/- 0.8 microM S./i.min.). Cytochrome p450 mono-oxygenase (p450) activity was measured on individual insects through ethoxycoumarine deethylase (ECOD) activity using a fluorescence microplate reader. The dependence of ECOD activity on age and body region of the nymphs, and pH and time of incubation were studied in order to optimize the measurement. As a result, comparative studies were performed on abdomens of 2 days old first instars at pH 7.2 and 4 h incubation time. ECOD activity of first nymphs was significantly lower in the susceptible colony (61.3 +/- 9.08 pg ECOD/ insect) than in the resistant one (108.1+/- 5.7 pg ECOD/ insect). These results suggest that degradative esterases (no-cholinesterase) and mono-oxygenases cytochrome p450, play an important role in the resistance to deltamethrin in Salta colony from Argentina.

Abstract Permethrin has been used extensively for control of Pediculus humanus capitis De Geer (Anoplura: Pediculidae) in Argentina since 1990, resulting in the development of resistance to this and other pyrethroids. This resistance was first detected in some field populations in 1997. A survey for resistance in Buenos Aires in 2001 revealed significant resistance levels in lice on children at 24 of 26 (92.3%) schools. When compared with a previously unexposed reference population, resistance ratios (RRs) obtained by exposing the insects to filter papers impregnated with permethrin ranged from 2 to 60 in 10 (39%) of the schools. RRs in the remaining 14 (61%) populations were too great to not be measured with the filter paper method (RR > 88.7). As an alternative, we used topical applications of 0.1 μl of acetone solution of permethrin on the dorsal abdomen of adults and third instars. This topical method, which has not been previously reported for head lice, was capable of quantifying higher levels of resistance. Highly resistant populations had RRs from 162.5 to 655.2. When applied to populations with low and intermediate levels of resistance, results from the filter paper and topical application methods were highly correlated, and RRs from topical application were higher than those from the filter paper method. Results from the combination of the two methods indicated that head lice among Buenos Aries school children are highly resistant to permethrin, and the resistance is widespread.

The effectiveness of 1-octanol, 1-nonanol, 1-decanol, 1-undecanol, and 1-dodecanol was evaluated by immersion method against susceptible and permethrin-resistant head lice, Pediculus humanus capitis De Geer, from Buenos Aires, Argentina. All the tested alcohols showed knockdown effect at 10 min and mortality 18 h after treatment. The highest activity was found for the 1-dodecanol (KC50 2.55%, LC50 2.28%) and the lowest for 1-octanol (KC50 8%, LC50 4.46%). The toxicity to the head lice systematically increased with the increase in carbon atoms in the n-aliphatic alcohol moiety, and with the octanol:water coefficient (r2 = 0.94). The pediculicidal activity of 1-dodecanol was not correlated with resistance to permethrin, because no significant difference was observed between toxicity parameters in the susceptible (MAR) and the permethrin-resistant populations which had different resistant levels (RR 5.77 x for E49 population, RR 9.5 x for HL population and RR > 35.3 x for GH population). The pediculicidal effect of aliphatic alcohols demonstrated in this study and the lack of correlation with the permethrin resistance may prove to have a practical value for use in susceptible and permethrin-resistant head lice control.

Biological effects on Tribolium castaneum larvae were evaluated for three withanolides isolated from Salpichroa origanifolia (Solanaceae), (20S,22R,24S,25S,26R)-5alpha,6alpha:22,26:24,25-triepoxy-26-hydroxy-17(13-->18)-abeo-ergosta-2,13,15,17-tetraen-1-one (salpichrolide A, 1), (20S,22R,24S,25S,26R)-22,26:24,25-diepoxy-5alpha,6beta,26-trihydroxy-17(13-->18)-abeo-ergosta-2,13,15,17-tetraen-1-one (salpichrolide C, 2), and (20S,22R,24S,25S,26R)-5alpha,6alpha:22,26:24,25-triepoxy-15,26-dihydroxy-17(13-->18)-abeo-ergosta-2,13,15,17-tetraen-1-one (salpichrolide G, 3), and for several chemically modified analogues. The compounds were incorporated into the larval diet at concentrations of 500 and 2000 ppm. Salpichrolide C (2) produced a significant delay in the development of neonate larvae to adults at the highest concentration (2000 ppm); development delays and lethal effects were produced by salpichrolides A (1) and G (3) at both concentrations assayed. The size of surviving adults was used as a criterion for assessing feedant deterrent effects; the results suggest that these compounds act as feeding inhibitors. Influence of chemical modifications in development delay was analyzed.

Permethrin-resistant colonies of Pediculus capitis (De Geer) from Buenos Aires were used to establish a resistance profile and to examine resistance mechanisms. All permethrin-resistant head lice (resistance ratio from 52.8 to >88.7) were also resistant to d-phenothrin (resistance ratio from 40.86 to >48.39) and deltamethrin (resistance ratio from 16.24, to 38.06). No cross-resistance to carbaryl was found in any of the pyrethroid-resistant P. capitis tested. Otherwise, all resistant colonies showed low to high levels of resistance to beta-cypermethrin. This pyrethroid had never been applied as a pediculicide in Argentina; however, the high level of resistance found in these permethrin-resistant colonies (resistance ratio from 9.74 to 50.97) demonstrated that pyrethroid cross-resistance occurred to this novel insecticide. Treatment with piperonyl butoxide (PBO) or triphenylphosphate (TPP) significantly decreased the toxicity of permethrin in the four colonies tested. The esterase inhibitor TPP produced lower enhancement of toxicity than the multifunction oxidase inhibitor PBO in the colonies having the highest resistance levels. Results presented here concerning the cross-resistance profile and synergism by enzyme inhibitors in permethrin-resistant head lice demonstrated that enhanced metabolism was involved in the pyrethroid resistance. However, the substantial degree of resistance that remained after synergism suggested the presence of another resistance mechanism. Cross-resistance to pyrethroid and susceptibility to the carbamate carbaryl suggested a common action mechanism.

SummaryResistance levels to insecticides used in control of Chagas Disease vectors were assessed in two species of bugs (Hemiptera: Reduviidae): Triatoma infestans (Klug) from Brazil and Rhodnius prolixus Stål from Venezuela. The resistance ratios, compared to susceptible laboratory strains, were determined by topical application bioassays. The T. infestans PA strain exhibited resistance ratios of 7× to deltamethrin, 3.6× to β‐cyfluthrin and 3.3× to cypermethrin, but was susceptible to β‐cypermethrin and lambda‐cyhalothrin. Rhodnius prolixus CA strain showed resistance to all the pyrethroids evaluated, the resistance ratios ranging between 4.5× to lambda‐cyhalothrin and 12.4× to cypermethrin. Deltamethrin resistance in both strains was decreased by piperonyl butoxide, suggesting oxidative metabolism as cause of resistance.

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 degrees C was determined. The cis-isomer was more active than the transisomer 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 degrees, 26 degrees, and 36 degrees C). The toxicity of the cis-isomer was lower at 36 degrees C than at either 16 degrees or 26 degrees 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. (C) 1998 Wiley-Liss, Inc.

The susceptibility of local head lice to permethrin, sumithrin, deltamethrin, and carbaryl was determined by laboratory bioassays in field-collected colonies. Head lice collected from the infested heads of children 6-12 yr old were tested within 3 h of collection. The longest survival of control insects in the laboratory was obtained by keeping them in the dark at 18 degrees C and 70-80% RH. The base line susceptibility data obtained for insects collected from children not treated for lice, the reference colony, showed that deltamethrin caused the highest mortality of the insecticides tested (LC(50), 0.06%). Permethrin, sumithrin, and carbaryl showed no significant difference in mortality (superposition of confidence intervals), being 10 times lower than that caused by deltamethrin All field-collected lice required a higher LC(50) of permethrin than the reference colony. Resistance levels varied from 3 to >100 for colonies that were taken from children treated with anti-lice products. Lice colonies with permethrin resistance showed resistance to sumithrin and deltamethrin, but resistance was not observed to the carbamate carbaryl.

A series of methyl esters of N-substituted (Z)- and (E)-maleamic acids were synthesized and their effect on food intake measured on fifth-instar nymphs of Triatoma infestans. Suppression of food intake was found only for the (Z)-isomers. The initial reaction rate of the synthesized compounds with glutathione (GSH) was calculated from the reaction in vitro of the (Z)-isomers. No reaction was observed with the (E)-isomers. Good correlation between the suppression of food intake, measured by its ED50 (effective dose that inhibited feeding of 50% of the population) and the initial reaction rate with GSH and the hydrophobic parameter π, was found. © of SCI.

We found that feeding behavior of Triatoma infestans (Klug) 5th-instar nymphs was blocked by topical application of the cis-isomers of methyl esters of N-substituted maleamic acids, synthesized in our laboratory. The antifeeding effect of these compounds was determined by exposing 5th-instar nymphs to impregnated filter papers. The greatest antifeeding effect was observed for the shortest alkyl chain of cis-N-substituted methyl maleamate (EC50: 2.47 micrograms/cm2 when the substituent was ethyl and 65.1 micrograms/cm2 when it was octil). The continuous exposure of a T. infestans population to ethyl, buthyl, heptil, or octil maleamates at an initial concentration of 0.1 mg/cm2 produced significant (P < 0.05) decrease in the development of the exposed populations ranging from 220 to 400 d for the different maleamates assayed. These results suggest the antifeeding maleamates as a specific alternative method of T. infestans control.

The butyl ester of buthionine sulfoximine (BBSO) applied topically to the nymph V stage of Triatoma infestans (Klug) caused glutathione depletion which was maintained for four days after treatment. Topical pre-treatment of nymph V with BBSO significantly synergised the toxicity of DDT and fenitrothion to T. infestans.

l-BSO injected at a sublethal dose in nymph V of Triatoma infestans produces a transient GSH depletion in the abdomen. A depletion of the total content of GSH was observed in nymph II fed with a sublethal concentration of l-BSO. Either ingestion or injection pretreatments of T. infestans nymph V with sublethal concentrations of l-BSO produced a slight synergism on the toxicity of fenitrothion. Feeding administration of a sublethal concentration of l-BSO to nymphs II of T. infestans resulted in a weak potentiation of the acute toxicity of DDT.

Topical application of different sulfhydryl (SH) reagents in acetone solution resulted in a deterrence of the feeding activity of Triatoma infestans nymph V. The ED50 for topical application of N-ethylmaleimide (NEM), the more active compound assayed, was 8.9 μg/insect. This effect showed a dose-dependent reversibility. We also observed feeding deterrency when a gauze cloth impregnated with NEM was located between the food source and the nymphs. After topical NEM treatment, mating deterrency was assessed by counting spermatophores dropped and eggs laid. When a population of adult insects, nymphal stages and eggs was reared in a cage containing filter papers treated with 0.4 mg/cm2 of NEM, a significant decrease of population density was seen after 200 days of exposure. Weekly replacement of filter papers impregnated with 0.13 mg/cm2 NEM for a period of one year enabled population control to be maintained. Antifeeding and antimating effects of NEM in Triatoma infestans and the resulting population control could be attributed to a chemoreceptor blockage produced by SH reagents.

Cuticular penetration of lipid soluble substances in Triatoma infestans is very poor (less than 1% in 24 hr), while the respiratory route is particularly efficient (28% in 5 min). Accordingly, several noxious gases killed the insect at short periods of time. CH3Br and SO2 were particularly effective and they also evidenced ovicide action. With the former compound this effect is also observed at low concentrations. The CH3Br hydrolysis products or its metabolites were not similarly effective, while other methylating agent, diazomethane, was very active. The irreversible binding of 14CH3Br to proteins and lipids of T. infestans, as well as the decrease in the sulfhydryl groups content of the proteins of the insect provoked by CH3Br suggest that this compound kills the insect because of its alkylating properties. The use of gaseous insecticides in the chemical control of T. infestans is discussed.