Isolamento de Haemophiliis aegyptius associado à Febre Purpúrica Brasileira, de cloropídeos (Diptera) dos gêneros Hippelates e Liohippelates

O reconhecimento da Febre Purpúrica Brasileira (FPB), em 1984, originou uma série de estudos que revelaram uma correlação desta doença com conjuntivites causadas por Haemophiliis aegyptius. A associação do aumento de conjuntivites em crianças e a maior densidade populacional de cloropídeos do gênero Hippelates já havia sido verificada desde o século passado. Este fenômeno está relacionado ao tropismo que estes insetos apresentam pelos olhos, secreções e feridas de onde se alimentam. Embora haja evidências do papel destes cloropídeos na transmissão mecânica de conjuntivites bacterianas, o isolamento de Haemophilus aegyptius a partir dos mesmos, no seu habitat natural, ainda não havia sido verificado. No presente trabalho obtivemos o isolamento de cepas invasivas de Haemophilus aegyptius, associadas à FPB, de duas coleções de cloropídeos, classificados como Liohippelates peruanus e uma espécie nova, Hippelates neoproboscideus, coletados ao redor dos olhos de crianças com conjuntivite.

Population dynamics and biting rhythm of the anthropophilic sandfly Lutzomyia cruciata (Diptera: Psychdidae) in Southeast, Mexico

Sandflies attracted by human bait were caught in an endemic focus of localized cutaneous leishmaniasis in the state of Campeche, Mexico. Catches were carried out monthly from February 1994 to January 1995 between 18:00 and 22:00 h. Lutzomyia cruciata was the only species caught. The highest population peak of Lu. cruciata was found in March with lesser peaks in February, December 1994, and January 1995. Maximum biting rate of Lu. cruciata was found between 18:00 and 19:00 h. The host-seeking females of Lu. cruciata were directly related to levels of humidity between 88 and 100%. Low and high temperature had a negative effect upon Lu. cruciata activity. The possible role of Lu. cruciata as vector of leishmaniasis in the state of Campeche, Mexico is discussed.

Description of the egg of Anopheles (Anopheles) intermedius (Peryassu, 1908) (Diptera: Culicidae) by scanning electron microscopy

The egg of Anopheles (Anopheles) intermedius (Peryassu, 1908) is described and illustrated with scanning electron micrographs. Literature data on An. (Ano.) maculipes (Theobald, 1903) is provided

POSSIBLE ROLE OF LUTZOMYIA MARANONENSIS AND LUTZOMYIA ROBUSTA (DIPTERA: PSYCHODIDAE) AS VECTORS OF HUMAN BARTONELLOSIS IN THREE PROVINCES OF REGION NOR ORIENTAL DEL MARAÑON, PERU

Human bartonellosis is found predominantly in Perú2, 6, 8, 12, 15, as well as in Ecuador3, 7, 10 and Colombia13, 15. In Peru, the disease is restricted to the valleys of the western-side and a few inter-andean and eastern-slopes of the andean valleys6, 15, 18 at altitudes between 1000 and 3200 masl. Most human cases are reported from the regions of Chavin, Nor Oriental del Marañon and Lima16. Lutzomyia verrucarum is presumed to be the only vector of human bartonellosis in the valleys of Peru1, 2, 8, 11, 17, 19/ Our research objetive was to detect the presence of Lu. verrucarum in various localities known to be endemic for human bartonellosis in three provinces of Region Nor Oriental del Marañon. Sandfly collections were made between 1987 and 1992 during four visits to bartonellosis-endemic provinces: San Ignacio (districts of San José de Lourdes: 1020-1260 m and La Coipa: 1200-1560 m), Jaén (districts of Santa Rosa: 1300-1680 m and Jaén: 1220-1680 m) and Utcubamba (districts of Lonya Grande: 1200 m and El Milagro: 1200-1540 m)

LARVICIDAL ACTIVITY OF Bacillus sphaericus 2362 AGAINST Anopheles nuneztovari, Anopheles darlingi AND Anopheles braziliensis (DIPTERA, CULICIDAE)

In this present study, preliminary data was obtained regarding the mortality rate of the Amazonian anophelines, Anopheles nuneztovari, Anopheles darlingi and Anopheles braziliensis when subjected to treatment with Bacillus sphaericus strain 2362, the WHO standard strain. Initially, experiments were conducted to test the mortality rate of the three species of anopheline larvae. The third larval instar of An. nuneztovari and the second and third larval instars of An. darlingi proved to be the least susceptible. In other experiments, the same three mosquito species were tested with the standard strain 2362, An. nuneztovari was the least susceptible to this insect pathogen, while An. braziliensis was the most susceptible. This latter species showed a difference in the level of LC50 concentration, when compared to the former, of 2.4, 2.5 and 1.8 in readings taken 24, 48 and 72 hours after exposure to the bacillus.

Assessment of PCR in the detection of Leishmania spp in experimentally infected individual phlebotomine sandflies (Diptera: Psychodidae: Phlebotominae)

DNA amplification by the polymerase chain reaction (PCR) was applied in the investigation of the presence of Leishmania (Kinetoplastida: Trypanosomatidae) parasites in single phlebotomine sandflies. Three phlebotomine/parasite pairs were used: Lutzomyia longipalpis/Leishmania chagasi, Lutzomyia migonei/Leishmania amazonensis and Lutzomyia migonei/Leishmania braziliensis, all of them incriminated in the transmission of visceral or cutaneous leishmaniasis. DNA extraction was performed with whole insects, with no need of previous digestive tract dissection or pooling specimens. The presence of either mouse blood in the digestive tract of the sandflies or the digestive tract itself did not interfere in the PCR. Infection by as few as 10 Leishmania sp. per individual were sufficient for DNA amplification with genus-specific primers. Using primers for L. braziliensis and L. mexicana complexes, respectively, it was possible to discriminate between L. braziliensis and L. amazonensis in experimentally infected vectors (L. migonei).

Scanning electron microscopy of third-instar sarcophagid (Diptera: Sarcophagidae) recovered from a mummified human corpse in Thailand

The third-instar of an unidentified sarcophagid, recovered from a mummified body of a 32-yr-old Thai male was examined using scanning electron microscopy (SEM). Although the morphological features of this larva are similar to the other sarcophagid larvae, some features could be helpful for species identification, which is a basic requirement for estimation of postmortem interval in forensic investigation. These features included number and arrangement of papillae on the anterior spiracle, structure of spines, size of circumspiracular tubercles at caudal segment and branching peculiarity of the posterior spiracular hairs. This information could benefit future identification of the sarcophagid larvae that exist in Thailand.

Effects of eucalyptol on house fly (Diptera: Muscidae) and blow fly (Diptera: Calliphoridae)

The effects of eucalyptol were evaluated against the house fly, Musca domestica L., and blow fly, Chrysomya megacephala (F.). The bioassay of adults, using topical application, indicated that M. domestica males were more susceptible than females, with the LD50 being 118 and 177 µg/fly, respectively. A higher LD50 of C. megacephala was obtained; 197 µg/fly for males and 221 µg/fly for females. Living flies of both species yielded a shorter life span after being treated with eucalyptol. The bioassay of larvae, using the dipping method on the third instar, showed that M. domestica was more susceptible than C. megacephala, with their LC50 being 101 and 642 µg/µl, respectively. The emergence of adults, which had been treated with eucalyptol in larvae, decreased only in M. domestica. Having the volatile property, fumigation or impregnated paper test of eucalyptol or the efficacy of repellence or attractiveness merits further investigations to enhance bio-insecticidal efficacy.

Some ultrastructural superficial changes in house fly (Diptera: Muscidae) and blow fly (Diptera: Calliphoridae) larvae induced by eucalyptol oil

The ultrastructural superficial changes in third instar house fly (Musca domestica) and blow fly (Chrysomya megacephala) induced by eucalyptol oil were observed using scanning electron microscopy. Dipped in 0.902 g/ml eucalyptol for 30 sec, the larvae integument of both species showed significant aberrant appearance of the body surface, particularly swelling integument, bleb formation, partial breach and deformation of spines.

Residual effect of two Bacillus thuringiensis var. israelensis products assayed against Aedes aegypti (Diptera: Culicidae) in laboratory and outdoors at Rio de Janeiro, Brazil

Resistance of the dengue vector to temephos stimulated its substitution for Bacillus thuringiensis var. israelensis (Bti) since 2001 in Brazil. The persistence of the two Bti formulations employed at that time by the Health Ministry, Vectobac G and Aquabac G, was assayed under laboratory and outdoor conditions. Both formulations were tested at 0.2 g/10 liters of water, the same concentration applied in the field for vector control. The tests were done against Ae. aegypti third instar larvae (Rockefeller strain). In the laboratory, Vectobac G and Aquabac G caused at least 95% mortality until 101 and 45 days after treatment, respectively. In the outdoor assays, test containers of different materials were treated with either formulation and placed in a shaded area. Larvae were introduced each 3-6 days and mortality was recorded 24 and 48 hours later. In the first set of assays, performed in June 2001, mortality levels of 70% or more were attained for 2-5 weeks for both formulations in all containers. The exception was for the iron one that rusted, resulting in low mortality after seven days. In the second set of assays (August 2001), 70% mortality was attained for just 1-2 weeks for all the containers and both formulations.

Susceptibility of two karyotypic forms of Anopheles aconitus (Diptera: Culicidae) to Plasmodium falciparum and P. vivax

Four laboratory-raised colonies of two karyotypic forms of Anopheles aconitus, i.e., Form B (Chiang Mai and Phet Buri strains) and C (Chiang Mai and Mae Hong Son strains), were experimentally infected with Plasmodium falciparum and P. vivax using an artificial membrane feeding technique and dissected eight and 12 days after feeding for oocyst and sporozoite rates, respectively. The results revealed that An. aconitus Form B and C were susceptible to P. falciparum and P. vivax, i.e., Form B (Chiang Mai and Phet Buri strains/P. falciparum and P. vivax) and Form C (Chiang Mai and Mae Hong Son strains/P. vivax). Comparative statistical analyses of the oocyst rates, average number of oocysts per infected midgut and sporozoite rates among all strains of An. aconitus Form B and C to the ingroup control vectors, An. minimus A and C, exhibited mostly no significant differences, confirming the high potential vector of the two Plasmodium species. The sporozoite-like crystals found in the median lobe of the salivary glands, which could be a misleading factor in the identification of true sporozoites in salivary glands were found in both An. aconitus Form B and C.

Adulticidal activity against Stegomyia aegypti (Diptera: Culicidae) of three Piper spp.

Three Piper species, Piper longum, P. ribesoides and P. sarmentosum, were selected for investigation of adulticidal potential against Stegomyia aegypti, a main vector of dengue and dengue haemorrhagic fever. Successive extraction by maceration with 95% ethanol showed percentage yields of ethanolic extracts, which derived from P. longum, P. ribesoides and P. sarmentosum, of 8.89, 3.21 and 5.30% (w/w), respectively. All Piper extracts illustrated an impressive adulticidal activity when tested against female mosquitoes by topical application. The susceptibility of St. aegypti females to ethanol-extracted Piper was dose dependent and varied among the plant species. The highest adulticidal effect was established from P. sarmentosum, followed by P. ribesoides and P. longum, with LD50 values of 0.14, 0.15 and 0.26 µg/female, respectively. The potential of these Piper species, as possible mosquitocides, established convincing activity for further researches to develop natural substances for combat against adult mosquitoes.

Effect of stalk and leaf extracts from Euphorbiaceae species on Aedes aegypti (Diptera, Culicidae) larvae

The objective of this study was to evaluate the larvicidal activity of essential oil aqueous solutions (hydrolates) obtained by steam distillation of stalks and leaves of Croton argyrophylloides, Croton nepetaefolius, Croton sonderianus and Croton zehntneri against Aedes aegypti larvae. Twenty-five larvae of third instar were placed in plastic beckers, containing the hydrolates (50 mL), in a four repetitions scheme. Water was used as control and the number of dead larvae was counted after 24 hours. The data obtained were submitted to Variance Analysis and Tukey test. Significant differences were observed among the hydrolates from different species and from different parts of each plant (p < 0.001). The hydrolates of stalk and leaf from C. nepetaefolius and C. zehntneri and leaf hydrolate of C. argyrophylloides presented 100% mortality against larvae. The compounds present in C. zenhtneri and C. nepetaefolius are oxygenated phenylpropanoids that are more soluble in water than the monoterpenes and sesquiterpenes detected in the oils of C. argyrophylloides and C. sonderianus. This study showed that all species analyzed presented compounds with larvicidal properties, with differences between each plant parts.

Salivary gland proteins of the human malaria vector, Anopheles dirus B (Diptera: Culicidae)

Salivary gland proteins of the human malaria vector, Anopheles dirus B were determined and analyzed. The amount of salivary gland proteins in mosquitoes aged between 3 - 10 days was approximately 1.08 ± 0.04 µg/female and 0.1 ± 0.05 µg/male. The salivary glands of both sexes displayed the same morphological organization as that of other anopheline mosquitoes. In females, apyrase accumulated in the distal regions, whereas alpha-glucosidase was found in the proximal region of the lateral lobes. This differential distribution of the analyzed enzymes reflects specialization of different regions for sugar and blood feeding. SDS-PAGE analysis revealed that at least seven major proteins were found in the female salivary glands, of which each morphological region contained different major proteins. Similar electrophoretic protein profiles were detected comparing unfed and blood-fed mosquitoes, suggesting that there is no specific protein induced by blood. Two-dimensional polyacrylamide gel analysis showed the most abundant salivary gland protein, with a molecular mass of approximately 35 kilodaltons and an isoelectric point of approximately 4.0. These results provide basic information that would lead to further study on the role of salivary proteins of An. dirus B in disease transmission and hematophagy.