Comparative developmental and susceptibility to insecticide of Bolivian and Brazilian populations of Triatoma infestans

The triatomine bug Triatoma infestans probably originated in Bolivia and dispersed passively over wide areas of South America, where it is the principal vector of Trypanosoma cruzi. In the region of its probable origin this species shows colonization in two different ecotopes, so that it may be encountered in sylvatic as well as in artificial habitats. The sylvatic colonization pattern is not observed in the rest of its range, where T. infestans is exclusive to man-made habitats. The objective of this study was to compare several aspects of two T. infestans populations, one from Minas Gerais (Brazil) and the other from the Cochabamba Valley (Bolivia), with a view to elucidate the factors associated with the different colonization patterns observed for this species. The differences between the developmental cycle, weight, capacity to ingest blood and mortality rate of first instar nymphs should indicate more fragility of Brazilian population that may be related to its elimination possibility.

The London School of Hygiene and Tropical Medicine: a new century of malaria research

The global malaria situation has scarcely improved in the last 100 years, despite major advances in our knowledge of the basic biology, epidemiology and clinical basis of the disease. Effective malaria control, leading to a significant decrease in the morbidity and mortality attributable to malaria, will require a multidisciplinary approach. New tools - drugs, vaccine and insecticides - are needed but there is also much to be gained by better use of existing tools: using drugs in combination in order to slow the development of drug resistance; targeting resources to areas of greatest need; using geographic information systems to map the populations at risk and more sophisticated marketing techniques to distribute bed nets and insecticides. Sustainable malaria control may require the deployment of a highly effective vaccine, but there is much that can be done in the meantime to reduce the burden of disease.

The use of bacterial larvicides in mosquito and black fly control programmes in Brazil

Bacillus spp. based larvides are increasingly replacing, with numerous advantages, chemical insecticides in programmes for controlling black fly and mosquito populations. Brazil was among the pioneers in adopting Bacillus thuringiensis israelensis (B.t.i) to control black flies. However, the major current mosquito control programme in Brazil, the Programme for Eradication of Aedes aegypti launched in 1997, only recently decided to replace temephos by B.t.i based larvicides, in the State of Rio de Janeiro. In the last decade, works developed by research groups in Brazilian institutions have generated a significant contribution to this subject through the isolation of B. sphaericus new strains, the development of new products and the implementation of field trials of Bacillus efficacy against mosquito species under different environmental conditions.

Expression of mosquito active toxin genes by a Colombian native strain of the gram-negative bacterium Asticcacaulis excentricus

Mosquito control with biological insecticides, such as Bacillus sp. toxins, has been used widely in many countries. However, rapid sedimentation away from the mosquito larvae feeding zone causes a low residual effect. In order to overcome this problem, it has been proposed to clone the Bacillus toxin genes in aquatic bacteria which are able to live in the upper part of the water column. Two strains of Asticcacaulis excentricus were chosen to introduce the B. sphaericus binary toxin gene and B. thuringiensis subsp. medellin cry11Bb gene cloned in suitable vectors. In feeding experiments with these aquatic bacteria, it was shown that Culex quinquefasciatus, Aedes aegypti, and Anopheles albimanus larvae were able to survive on a diet based on this wild bacterium. A. excentricus recombinant strains were able to express both genes, but the recombinant strain expressing the B. sphaericus binary toxin was toxic to mosquito larvae. Crude protease A. excentricus extracts did not degrade the Cry11Bb toxin. The flotability studies indicated that the recombinant A. excentricus strains remained in the upper part of the water column longer than the wild type Bacillus strains.

Immunological and protective properties of novel malaria blood stage peptide antigens

ABSTRACT Malaria is a major worldwide public health problem, with transmission occurring throughout Africa, Asia, Oceania and Latin America. Over two billion people live in malarious areas of the world and it is estimated that 300-500 million cases and 1.5-2.7 million deaths occur annually. The increase in multi-drug resistant parasites and insecticide-resistant vectors has made the development of malaria vaccine a public health priority. The published genome offers tremendous opportunity for the identification of new antigens that can befast-tracked for vaccine development. We identified potential protein antigens present on the surface of asexual malaria blood stages through bioinformatics and published transcriptome and proteorné analysis. Amongst the proteins identified, we selected those that contain predicted a-helical coiled-coil regions, which are generally short and structurally stable as isolated fragments. Peptides were synthesized and used to immunize mice. Most peptides tested were immunogenic as demonstrated in ELISA assays, and induced antibodies of varying titres. In immunofluorescence assays, anti-sera from immunized mice reacted with native proteins expressed at different intraerythrocytic developmental stages of the parasite's cycle. In parallel in vitro ADCI functional studies, human antibodies affinity purified on some of these peptides inhibited parasite growth in association with monocytes in magnitudes similar to that seen in semiimmune African adults. Siudies using human immune sera taken from different malaria endemic regions, demonstrated that majority of peptides were recognized at high prevalence. 73 peptides were next tested in longitudinal studies in two cohorts separated in space and time in coastal Kenya. In these longitudinal analyses, antibody responses to peptides were sequentially examined in two cohorts of children at risk of clinical malaria in order to characterize the level of peptide recognition by age, and the role of anti-peptide antibodies in protection from clinical malaria. Ten peptides were associated ?with a significantly reduced odds ratio for an episode of clinical malaria in the first cohort of children and two of these peptides (LR146 and ÁS202.11) were associated with a significantly reduced odds ratio in both cohorts. This study has identified proteins PFB0145c and MAL6P1.37 among others as likely targets of protective antibodies. Our findings support further studies to systematically assess immunogenicity of peptides of interest in order to establish clear criteria for optimal design of potential vaccine constructs to be tested in clinical trials. RESUME La malaria est un problème de santé publique mondial principalement en Afrique, en Asie, en Océanie et en Amérique latine. Plus de 2 milliards de personnes vivent dans des régions endémiques et le nombre de cas par année est estimé entre 300 et 500 millions. 1.5 à 2.7 millions de décès surviennent annuellement dans ces zones. L'augmentation de la résistance aux médicaments et aux insecticides fait du développement d'un vaccin une priorité. Le séquençage complet du génome du parasite offre l'opportunité d'identifier de nouveaux antigènes qui peuvent rapidement mener au développement d'un vaccin. Des protéines antigéniques potentielles présentes à la surface des globules rouges infectés ont été identifiées par bioinformatique et par l'analyse du protéome et du transcriptome. Nous avons sélectionné, parmi ces protéines, celles contenant des motifs dits "a helical coiled-coil" qui sont généralement courts et structurellement stables. Ces régions ont été obtenues par synthèse peptidique et utilisées pour immuniser des souris. La plupart des peptides testés sont immunogéniques et induisent un titre variable d'anticorps déterminé par ELISA. Les résultats de tests d'immunofluorescence indiquent que les sera produits chez la souris reconnaissent les protéines natives exprimées aux différents stades de développement du parasite. En parallèle, des études d'ADCI in vitro montrent qué des anticorps humains purifiés à partir de ces peptides associés à des monocytes inhibent la croissance du parasite aussi bien que celle observée chez des adultes africains protégés. Des études d'antigénicité utilisant des sera de personnes protégées de différents âges vivant dans des régions endémiques montrent que la majorité des peptides sont reconnus avec une haute prévalence. 73 peptides ont été testés dans une étude longitudinale avec 2 cohortes de la côte du Kenya. Ces 2 groupes viennent de zones bien distinctes et les prélèvements n'ont pas été effectués pendant la même période. Dans cette étude, la réponse anticorps contre les peptides synthétiques a été testée dans les 2 cohortes d'enfants à risque de développer un épisode de malaria afin de caractériser le niveau de reconnaissance des peptides en fonction de l'âge et de déterminer le rôle des anticorps anti-peptides dans la protection contre la malaria. Parmi ces peptides, 10 sont associés à une réduction significative des risques de développer un épisode de malaria dans la première cohorte alors qu'un seul (LR146 et AS202.11) l'est dans les 2 cohortes. Cette étude a identifié, parmi d'autres, les protéines PFB0145c et MAL6P1.37 comme pouvant être la cible d'anticorps. Ces résultats sont en faveur de futures études qui évalueraient systématiquement l'immunogénicité des peptides d'intérêt dans le but d'établir des critères de sélection clairs pour le développement d'un vaccin. Résumé pour un large public La malaria est un problème de santé publique mondial principalement en Afrique, en Asie, en Océanie et en Amérique latine. Plus de 2 milliards de personnes vivent dans des régions endémiques et le nombre de cas par année est estimé entre 300 et 500 millions. 1.5 à 2.7 millions de décès surviennent annuellement dans ces zones. La résistance aux médicaments et aux insecticides augmente de plus en plus d'où la nécessité de développer un vaccin. Le séquençage complet du génome (ensemble des gènes) de P. falciparum a conduit au développement de nouvelles .études à large échelle dans le domaine des protéines du parasite (protéome) ; dans l'utilisation d'algorithmes, de techniques informatiques et statistiques pour l'analyse de données biologiques (bioinformatique) et dans les technologies de transcription et de profiles d'expression (transcriptome). Nous avons identifié, en utilisant les outils ci-dessus, des nouvelles protéines antigéniques qui sont présentes au stade sanguin de la malaria. Nous avons sélectionné, parmi ces protéines, celles contenant un motif dit "a-helical coiled-coil" qui sont des domaines impliqués dans un large éventail de fonctions biologiques. Des peptides représentant ces régions structurellement stables ont été synthétisés et utilisés pour immuniser des souris. La plupart des peptides testés sont immunogéniques et induisent un titre variable d'anticorps déterminé par ELISA. Les résultats de tests d'immunofluorescence indiquent que plusieurs sera de souris immunisées avec ces peptides reconnaissent les protéines natives exprimées à la surface des globules rouges infectés. En parallèle, des études d'ADCI in vitro montrent que des anticorps humains purifiés à partir de ces peptides en présence de monocytes inhibent la croissance du parasite de manière similaire à celle observée chez des adultes africains protégés. Des études d'antigénicité utilisant des sera de personnes immunes de différents âges (adultes et enfants) vivant dans des régions endémiques montrent que la majorité des peptides sont reconnus avec une haute prévalence. 73 peptides ont été testés dans des études épidémiologiques dans 2 villages côtiers du Kenya Ces 2 groupes vivent dans des zones bien distinctes et les prélèvements n'ont pas été effectués pendant la même période. Dans ces études, la réponse anticorps dirigée contre les peptides synthétiques a été testée en utilisant 467 échantillons sanguins d'enfants à risque de développer un épisode de malaria afin de caractériser le niveau de reconnaissance des peptides en fonction de l'âge et de déterminer le rôle des anticorps anti-peptides dans la protection contre la malaria cérébrale. Parmi ces peptides, 10 sont associés à une protection contre un épisode de malaria dans le premier village alors qu'un seul l'est dans les 2 villages. Ces résultats sont en faveur de futures études qui évalueraient systématiquement l'immunogénicité des peptides intéressants dans le but d'établir des critères de sélection clairs pour le développement d'un vaccin.

Effect of diflubenzuron on immature stages of Haematobia irritans (L.) (Diptera: Muscidae) in Uberlândia, State of Minas Gerais, Brazil

Horn fly immatures were raised in media containing different concentrations of diflubenzuron in order to verify their susceptibility to this insect growth regulator (IGR). The 50% and 95% lethal concentrations of diflubenzuron for the population (LC50, LC95) were determined as well as the effect of this IGR on the different immature horn fly stages. The tests were performed using the progeny of adults collected in the field. The immatures were maintained in a growth chamber at 25.0 ± 0.5ºC and 12-12 h photoperiod. IGR concentrations of 300 ppb, 100 ppb and 50 ppb were lethal for 100% of the sample. Pupae malformation occurred in the breeding media containing different diflubenzuron concentrations. Values for LC50 , LC95 (± 95% fiducial limits) and the slope of the regression line were respectively, 25.521 ± 1.981 ppb, 34.650 ± 2.001 ppb and 12.720 ± 1.096. The third larval instar was more sensitive to the sub-lethal concentration of the product than the first and second ones were. The results indicate that this IGR can be an important tool for controlling horn fly populations as well as for managing horn fly resistance to conventional insecticides against Haematobia irritans in Uberlândia, State of Minas Gerais.

Preliminary assays indicate that Antonia ovata (Loganiaceae) and Derris amazonica (Papilionaceae), ichthyotoxic plants used for fishing in Roraima, Brazil, have an insecticide effect on Lutzomyia longipalpis (Diptera: Psychodidae: Phlebotominae)

Laboratory-reared Lutzomyia longipalpis (Lutz and Neiva 1912) was tested with extracts of two ichthyotoxic plants, known as timbós, used as fishing poison in the Amazon. Phlebotomines, L. longipalpis, and plants, Antonia ovata and Derris amazonica, were collected in the Raposa-Serra do Sol Indian Reserve, a focus of visceral leishmaniasis in the State of Roraima, Brazil. Extracts were prepared from dried leaves of A. ovata and roots of D. amazonica that were percolated in water, filtered and dried out at 50°C. The solid extract obtained was diluted in water at 150, 200 and 250 mg/ml. The solution was blotted in filter paper placed at the bottom of cylindric glass tubes containing sand flies. For each plant extract and dilution, two series of triplicates with 5 male and 5 female specimens of L. longipalpis were used. Mortality was recorded every 2 h during 72 h of exposure. At 72 h the mortality was as high as 80% for extracts of A. ovata (LD50 = 233 mg/ ml), and 100% for D. amazonica (LD50 = 212 mg/ ml) whereas in the control groups maximum mortality never surpassed 13%. Preliminary assays indicated that A. ovata and D. amazonica displayed significant insecticide effect against L. longipalpis.

Susceptibility to diazinon in populations of the horn fly, Haematobia irritans (Diptera: Muscidae), in Central Brazil

From October 2000 to April 2001, insecticide bioassays were conducted in 18 ranches from 10 counties in the states of Mato Grosso and Mato Grosso do Sul, in Central Brazil. Horn flies from wild populations were exposed to diazinon-impregnated filter papers immediately after collection on cattle, and mortality was recorded after 2 h. A high susceptibility to diazinon was observed in all tested populations. The LC50s ranged from 0.15 to 0.64 µg/cm², and resistance ratios were always lower than one (ranging 0.1-0.6). Pyrethroid products, most applied by backpack sprayers, have been used since the horn fly entered the region, about 10 years ago. The high susceptibility observed to diazinon indicates that this insecticide (as probably other organophosphate insecticides) represents an useful tool for horn fly control and resistance management, particularly in pyrethroid-resistant populations.

Impact of residual spraying on Rhodnius prolixus and Triatoma dimidiata in the department of Zacapa in Guatemala

As a vector control program to control Chagas disease in Guatemala, residual spraying of Rhodnius prolixus and Triatoma dimidiata was performed, and its impact was measured in the department of Zacapa. In order to identify infested villages and determine the degree of infestation, a baseline entomological survey to identify municipalities infested with vectors followed by an additional vector survey in areas known to be infested was conducted. Residual spraying using pyrethroid insecticides was performed at all the villages identified as being infested with the vectors. The residual spraying was shown to be highly effective against both vectors by the decrease in infestation indices after spraying. Analysis of the cost-effectiveness of the spraying showed that the average cost of insecticides per house is high when compared with that in Southern Cone countries.

Comparative evaluation of pyrethroid insecticide formulations against Triatoma infestans (Klug): residual efficacy on four substrates

We investigated the residual efficacy of four insecticide formulations used in Chagas disease vector control campaigns: cyfluthrin 12.5% suspension concentrace (SC), lambda-cyhalothrin 10% wettable powder (WP), deltamethrin 2.5% SC, and 2.5% WP on four types of circular blocks of wood, straw with mud, straw with mud painted with lime, and mud containing 5% of cement. Three concentrations of these insecticides were tested: the LC90 (previously determined on filter paper), the double of the LC90, and the recommended operational dose. For each bioassay test, 15 third-stage nymphs of Triatoma infestans (Klug) (Hemiptera: Reduviidae) were exposed for 120 h to each treatment at 24 h, 30, 60, 90, and 180 days post-spraying. Mortality rates, moulting history and behaviour were recorded at 24, 48, 72, and 120 h of exposure. Mortality rates were highest during the first 30 days post-spraying. Highest mortality rates (above 50%) were observed for deltamethrin 2.5% SC and lambda-cyhalothrin 10% WP on wood blocks up to three months post-spraying. Mud was the substrate on which treatments showed lowest persistence, with the other two substrates showing intermediate residual efficacy of all treatments. During the first 30 days WP formulations were not as effective as SC flowable formulations but, overall in the longer term, WP gave grater mortality rates of T. infestans nymphs exposed at up to six months post-spraying. Porous surfaces, especially mud, showed most variability presumably due to absorption of the insecticide. In contrast the less porous surfaces (i.e. wood and lime-coated mud) kept mortality rates high for longer post-treatment, irrespective of the insecticide concentration used.

The fumigant and repellent activity of aliphatic lactones against Pediculus humanus capitis (Anoplura: Pediculidae)

New alternative insecticides are necessary for the chemical control of head lice. In this study the fumigant knockdown time 50% (KT50) and repellency index (RI) of three aliphatic lactones was compared with two essential oils and DDVP, against permethrin-resistance Pediculus humanus capitis from Argentina. In the fumigant assay, none of the lactones were effective compared to the highest activity of eucalyptus (KT50 15.53 m). In the repellency test, the three lactones were equally or more effective (RI ranging from 60.50 to 76.68) than the positive control (piperonal). These lactones are promising as head lice repellents.

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.

Infestation of rural houses by Triatoma infestans in the region of Los Llanos (La Rioja, Argentina)

Vectorial transmission of Chagas disease has been strongly reduced in most parts of the Southern Cone countries of South America, except in the Gran Chaco region of Argentina, Bolivia, and Paraguay. Given periodical interruptions of the vector control programmes in the endemic region of the Gran Chaco of Argentina, the vectorial transmission of the disease has been increasing during the last years. From the beginning of 2004, the provincial Ministry of Health of La Rioja, Argentina, started a vector control programme to cover the rural houses of the Los Llanos area in the southwestern area of the Gran Chaco region. This article reports the result of a standardized entomological survey and insecticide application against Chagas disease vectors in the intra and peridomestic structures of the rural houses of Los Llanos. A total of 4062 houses were inspected, of which 46.8% were found to be infested by Triatoma infestans. Infestation by vector species other than T. infestans was less than 0.5%(T. eratyrusiformis and T. platensis). Intradomestic infestation was found in 27.2%, whereas peridomestic infestation was found in 39.3% of the houses. The lowest figure of intradomestic infestation was 6.6% (Department F Varela), and the highest value of intradomestic infestation was 45.1% (Department Independencia). In spite of the demonstrated success of vector control elsewhere, this study shows that the vector populations are susceptible to pyrethroid insecticides in the southern area of the Gran Chaco of Argentina, that there still are regions where rural houses show heavy infestation by T. infestans associated with big peridomestic structures and that the vectorial transmission of the Chagas disease will continue, unless a sustained and well organized vector control effort is installed in the region.

Coadaptation and malaria control

Malaria emerges from a disequilibrium of the system 'human-plasmodium-mosquito' (HPM). If the equilibrium is maintained, malaria does not ensue and the result is asymptomatic plasmodium infection. The relationships among the components of the system involve coadaptive linkages that lead to equilibrium. A vast body of evidence supports this assumption, including the strategies involved in the relationships between plasmodium and human and mosquito immune systems, and the emergence of resistance of plasmodia to antimalarial drugs and of mosquitoes to insecticides. Coadaptive strategies for malaria control are based on the following principles: (1) the system HPM is composed of three highly complex and dynamic components, whose interplay involves coadaptive linkages that tend to maintain the equilibrium of the system; (2) human and mosquito immune systems play a central role in the coadaptive interplay with plasmodium, and hence, in the mainten-ance of the system's equilibrium; the under- or overfunction of human immune system may result in malaria and influence its severity; (3) coadaptation depends on genetic and epigenetic phenomena occurring at the interfaces of the components of the system, and may involve exchange of infectrons (genes or gene fragments) between the partners; (4) plasmodia and mosquitoes have been submitted to selective pressures, leading to adaptation, for an extremely long while and are, therefore, endowed with the capacity to circumvent both natural (immunity) and artificial (drugs, insecticides, vaccines) measures aiming at destroying them; (5) since malaria represents disequilibrium of the system HPM, its control should aim at maintaining or restoring this equilibrium; (6) the disequilibrium of integrated systems involves the disequilibrium of their components, therefore the maintenance or restoration of the system's equilibrium depend on the adoption of integrated and coordinated measures acting on all components, that means, panadaptive strategies. Coadaptive strategies for malaria control should consider that: (1) host immune response has to be induced, since without it, no coadaptation is attained; (2) the immune response has to be sustained and efficient enough to avoid plasmodium overgrowth; (3) the immune response should not destroy all parasites; (4) the immune response has to be well controlled in order to not harm the host. These conditions are mostly influenced by antimalarial drugs, and should also be taken into account for the development of coadaptive malaria vaccines.

Association of insecticide use and alteration on Aedes aegypti susceptibility status

Dengue and dengue hemorrhagic fever, vector-borne diseases transmitted by the mosquito Aedes aegypti, are presently important public health problems in Brazil. As the strategy for disease control is based on vector control through the use of insecticides, the development of resistance is a threat to programs efficacy. The objective of this study was to compare the Aedes aegypti susceptibility in nine vector populations from the state of São Paulo and seven from Northeast region of Brazil, since there was a difference on group of insecticide used between the areas. Bioassays with larvae and adult were performed according to the World Health Organization methods.The results showed higher resistance levels to organophosphates group in populations from the Northeast region where this group was used for both larvae and adult control than in São Paulo where organophosphates were used for larvae and pyretroids for adult control. Resistance to pyretroids in adults was widespread in São Paulo after ten years of use of cypermethrin while in vector populations from the Northeast region it was punctual. The difference in resistance profile between the areas is in accordance to the group of insecticide used.