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Biomedical subjects

R G Andre

Publications and source records attributed to R G Andre.

At least 19 recordsLinked to original sources

A probability model of vector behavior: effects of DDT repellency, irritancy, and toxicity in malaria control.

A probability model of how DDT residues may function within a malaria control program is described. A step-wise organization of endophagic behaviors culminates in a vector acquiring a human blood meal inside the house. Different vector behaviors are described, epidemiologically defined, temporally sequenced, and quantified with field data. Components of vector behavior and the repellent, irritant, and toxic actions of insecticide residues are then assembled into a probability model. The sequence of host-seeking behaviors is used to partition the total impact of sprayed walls according to the three chemical actions. Quantitatively, the combined effect of repellency and irritancy exert the dominant actions of DDT residues in reducing man-vector contact inside of houses. These relationships are demonstrated with published and unpublished data for two separate populations of Anopheles darlingi, for Anopheles gambiae and Anopheles funestus in Tanzania, and Anopheles punctulatus in New Guinea.

Animals↗

A comparison study of house entering and exiting behavior of Anopheles vestitipennis (Diptera: Culicidae) using experimental huts sprayed with DDT or deltamethrin in the southern district of Toledo, Belize, C.A.

An investigation of the house entering and exiting behavior of Anopheles vestitipennis Dyar and Knab was undertaken in the Toledo District of Belize, Central America, between March and December of 1998. Three untreated experimental huts were either fitted with exit or entrance interception traps or used as a control for human landing collections. Human landing collections showed that An. vestitipennis exhibited a high level of biting activity shortly after sunset and continued biting at high levels throughout the night. Under unsprayed conditions, the use of exit and entrance interception traps demonstrated that doors, windows, and eaves were the primary mode of entry; whereas, cracks in the walls served a secondary role. The peak entrance time for An. vestitipennis occurred between 6:45 P.M. and 9:45 P.M. and a peak exit time occurred between 11:45 P.M. and 4:45 A.M. Additional trials were conducted after spraying one of the huts with DDT and another with deltamethrin. The excito-repellent properties of deltamethrin did not affect entrance times but did result in a peak exiting behavior that was five hours earlier than under pre-spray conditions. Deltamethrin also exhibited a repellency effect, showing 66% fewer An. vestitipennis entering the hut two weeks post-spray. DDT had an even more powerful repellency effect resulting in a 97% post-spray reduction of An. vestitipennis females entering the hut up to two weeks post-spray. The control hut showed only a 37% reduction in An. vestitipennis as compared to pre-spray conditions.

Animals↗

Plasmodium vivax polymorphs and Plasmodium falciparum circumsporozoite proteins in Anopheles (Diptera: Culicidae) from Belize, Central America.

Eight species of Anopheles mosquitoes from indoor/outdoor human landing collections in Belize, Central America, were examined for human Plasmodium circumsporozoite protein (CSP) using an enzyme-linked immunosorbent assay (ELISA). A total of 14 of 9,104 females tested were positive from general surveys throughout Belize and three of 11,966 were positive from a longitudinal study in Caledonia, northern Belize. ELISA results, using pooled head-thorax preparations and species-specific monoclonal antibodies directed against the circumsporozoite proteins of Plasmodium falciparum and two Plasmodium vivax polymorphs (210 and VK247), found four species reactive: Anopheles vestitipennis (3 pools), Anopheles darlingi (2 pools), Anopheles albimanus (10 pools), and Anopheles gabaldoni (2 pools). The minimum field infection rates (MFIR) for combined Plasmodium species from the general survey were 0.282% for An. vestitipennis, 0.271% for An. darlingi, 0.126% for An. albimanus, and 0.395% for An. gabaldoni. MFIRs for combined Plasmodium species from the longitudinal study in the village of Caledonia were 0.018% for both An. vestitipennis and An. albimanus and 1.66% for An. gabaldoni. Positive CSP pools were collected from the Cayo, Corozal, Orange Walk, Stann Creek, and Toledo political districts. No CSP positive pools were detected from collections in the Belize District. The study provides valuable information on the spatial distribution and species type of Plasmodium positive mosquitoes. This information, in combination with other vector data, suggest that An. vestitipennis and An. darlingi are commonly involved in malaria transmission. Additionally, these species appear to be much more efficient vectors than An. albimanus in Belize.

Animals↗

Pesticide avoidance behavior in Anopheles albimanus, a malaria vector in the Americas.

The behavioral responses of 4 populations of Anopheles albimanus females to DDT, permethrin, and deltamethrin were characterized in excito-repellency tests. One test population (ST) from El Salvador has been maintained as a colony for 20 years. A second population (ES) from Guatemala was colonized in 1992. Third and fourth populations consisted of field-caught specimens from Toledo District (TO) of southern Belize in 1994 and Corozal District (CO) of northern Belize, respectively. Females of ES, TO, and CO populations rapidly escaped from direct contact with treated surfaces for each of the 3 insecticides. Similarities in escape responses of insecticide-resistant (ES) versus insecticide-susceptible populations (TO, CO) suggest that there is no relationship between physiological and behavioral responses of An. albimanus populations to DDT, permethrin, and deltamethrin. Females from all but the ST colony escaped in greater numbers from chambers without direct contact with treated surfaces than from control chambers (P < 0.05). Few females from the ST colony escaped from test chambers, regardless of which insecticide was used or whether contact was allowed, indicating that the ST colony has lost its capability to respond to insecticides. Repellent responses were significant; but they were not pronounced in 30-min exposures, and they were very pronounced in 4-h exposures. We conclude that irritant and repellent responses of malaria vectors to insecticides are important components of malaria control operations.

Americas↗

Characterization of Anopheles darlingi (Diptera: Culicidae) larval habitats in Belize, Central America.

Surveys for larvae of Anopheles darlingi Root were conducted in April, May, and August 1994 in riverine habitats of central Belize (Cayo and Belize districts). An. darlingi was present during both the dry and wet seasons. Larvae were encountered most frequently in patches of floating debris along river margins. The floating mats were often formed by bamboo hanging over the banks and dense submersed bamboo roots. Larvae were found less frequently in lake margins, small lagoons, and ground pools with submersed roots and patches of floating leaves or vegetation. In addition to their association with floating debris, larvae of An. darlingi were associated positively with shade and submersed plants in riverine environments. Samples from river habitats showed the larvae of Anopheles albimanus Wiedemann to be strongly associated with sun-exposed sites containing green or blue-green algae. Unlike An. darlingi, An. albimanus was an ubiquitous mosquito, the immatures of which occurred in a wide variety of riverine and nonriverine aquatic habitats. Based on published reports and our experience, the association of An. darlingi with river systems was verified, and its distribution in Central America and Mexico was mapped.

Animals↗

Aedes albopictus and other Aedes (Stegomyia) species in Fiji.

During an assessment of the dengue situation in Fiji in early 1992, a house-to-house survey of container-breeding Aedes was made. Discarded tires and water drums were identified as key breeding sites for the 4 potential dengue vectors: Aedes aegypti, Aedes albopictus, Aedes pseudoscutellaris, and Aedes polynesiensis. Aedes albopictus were detected on Viti Levu, Vanua Levu, and on Taveuni. Examination of early records and of used tire importation suggests entry into Fiji after July 1985 but well before July 1988 when the species was first detected. It is also possible the Ae. albopictus was introduced via aircraft from Hawaii. In Suva, the 4 Stegomyia species coexist, but in Lautoka, it appears that Ae. albopictus may be displacing Ae. pseudoscutellaris.

Aedes↗

Plasmodium berghei: sensitivity of chloroquine-resistant and chloroquine-sensitive strains to irradiation and the effect of irradiated malaria parasites on cytochrome P450-dependent monooxygenases.

Differences in sensitivities of chloroquine-sensitive and chloroquine-resistant strains of Plasmodium berghei were observed following irradiation of the parasites. A dose of 15 kilorads from a cobalt-60 source killed the erythrocytic stages of the chloroquine-sensitive strain and no parasitemias were observed when mice were injected with these irradiated parasites. In contrast, when the chloroquine-resistant strain was irradiated with the same dose of cobalt-60 and injected into mice, an infection rate of 12.5% was observed, indicating that the latter strain was more resistant to inactivation by irradiation. Following injection of these irradiated strains of P. berghei into mice, significant decreases in mouse hepatic cytochrome P450 and benzo(a)pyrene hydroxylase activity, with no significant effect on N-demethylase activity, were observed. Serum glutamic-oxaloacetic transaminase (SGOT) and glutamic-pyruvic transaminase (SGPT) levels of mice injected with the irradiated parasites fell within the range of the serum enzyme levels in normal laboratory mice.

Alanine Transaminase↗

Insecticide resistance issues in vector-borne disease control.

Vector-borne diseases are an increasing cause of death and suffering worldwide. Efforts to control these diseases have been focused on the use of chemical pesticides, but arthropod resistance (whether physiological, biochemical, or behavioral) to pesticides is now an immense practical problem. The pharmacokinetic interactions of pesticides with arthropods, mechanisms of resistance, and the strengths and shortcomings of different resistance test methods are briefly reviewed. Using malaria control as an example, the differences between the efficacy of insecticide-sprayed houses in reducing malaria transmission, and the actual effect of such treatments on vectors are discussed. Reduced malaria transmission as a result of spraying house walls occurs through some combination of killing vectors that land on sprayed walls (insecticidal effect) and by preventing vectors from entering or remaining inside long enough to bite (behavioral effects). Both insecticidal and behavioral effects of insecticides are important, but the relative importance of one versus the other is controversial. Field studies in Africa, India, Brazil, and Mexico provide persuasive evidence for strong behavioral avoidance of DDT by the primary vector species. This avoidance behavior, exhibited when malaria vectors avoid insecticides by not entering or by rapidly exiting sprayed houses, should raise serious questions about the overall value of current physiological and biochemical resistance tests. The continued efficacy of DDT in Africa, India, Brazil, and Mexico, where 69% of all reported cases of malaria occur and where vectors are physiologically resistant to DDT (excluding Brazil), serves as one indicator that repellency is very important in preventing indoor transmission of malaria. This experience with DDT has implications for future control efforts because pyrethroids also stimulate avoidance behaviors in arthropods. Each chemical should be studied early (before broad-scale use) to define types of action against vector species by geographic area, especially for impregnated bed net applications. The problems for vector control created by use of insecticides in agriculture and the potential for management of resistance in both agriculture and vector-borne disease control are discussed.

Agrochemicals↗

Evaluation of monoclonal antibodies against Plasmodium vivax sporozoites for ELISA development.

Nine monoclonal antibodies (MAbs) developed against Plasmodium vivax (Grassi & Feletti) salivary gland sporozoites were evaluated for use in an enzyme-linked immunosorbent assay (ELISA), using sporozoites developed in Anopheles dirus Peyton & Harrison An. gambiae Giles and An.maculatus Theobald. Four of the antibodies were unsuitable due to the low sensitivity of the resulting assays or the requirement for high concentrations of capture antibody. An additional two MAbs were rejected because they resulted in assays with high background absorbance, attributed to self-binding. Of the three remaining MAbs, the use of Navy vivax sporozoite (NVS) 3 resulted in an ELISA with the highest sensitivity and the lowest concentration requirement for capture antibody. Assay sensitivity varied with sporozoite strain indicating possible quantitative epitope heterogeneity. None of the MAbs cross-reacted with the heterologous sporozoites tested by immunofluorescence antibody assay (IFA). The IFA activity was not an indicator of ELISA sensitivity. The use of MAb NVS 3 in a standardized ELISA method resulted in an assay 10 times more sensitive than reported previously for P. vivax sporozoites, with a detection limit of fewer than 100 sporozoites per mosquito.

Animals↗

Species- and infective stage-specific monoclonal antibodies to Leishmania major produced by an in vitro immunization method.

Monoclonal antibodies specific to the infective-stage promastigotes of Leishmania major are needed for developing rapid diagnostic assays of infected sand flies. An in vitro immunization protocol was applied for the production of monoclonal antibodies using small amounts of L. major. Infective-stage promastigotes were isolated from sand flies (Phlebotomus papatasi) 7-10 days after infection and used as antigen for immunization. Two weeks after a primary immunization, murine splenocytes were removed and immunized in vitro with antigen in murine EL-4 thymoma cell conditioned medium. Three fusions were performed using X63-Ag.653 myeloma cells as fusion partners and two fusions were performed using FOX-NY cells. Antibodies specific to promastigotes were detected using an indirect enzyme-linked immunosorbent assay (ELISA). Initially 56 monoclonal antibodies were selected, and their species and stage specificity were determined using both an ELISA and an indirect fluorescent antibody assay (IFA). Twelve monoclonal antibodies showed species specificity to L. major when tested against four sympatric species of Leishmania. Four other monoclonal antibodies showed species and infective-stage specificity to L. major promastigotes. When tested in immunoblots, all four species- and stage-specific monoclonal antibodies bound to five protein bands that were unique to the infective-stage promastigotes.

Animals↗

Identification of isomorphic malaria vectors using a DNA probe.

About 7,000 recombinant clones, derived from chromosomally-identified families of wild-caught females of Anopheles dirus species D, were screened. The most promising clone was totally specific to species D when tested against single F1 females of all four species of the complex. In fresh specimens the clone was positive for DNA levels 150 times less than the normal DNA content of single individuals. Fresh adult males and females, larvae, and dried specimens have been successfully identified. The clone was sequenced; it is 124 bp long and appears to be repeated in the genome about 1.8 x 10(4) times.

Animals↗

Comparative testing of monoclonal antibodies against Plasmodium falciparum sporozoites for ELISA development.

Ten monoclonal antibodies developed against Plasmodium falciparum sporozoites at four institutions were evaluated for use in an enzyme-linked immunosorbent assay (ELISA). Four of the antibodies were eliminated because of their low sensitivity or requirement for high concentrations of capture antibody, while an additional four were rejected because they exhibited cross-reactivity with P. berghei sporozoites. Of the two remaining monoclonal antibodies, that designated 2A10 had the highest sensitivity, a requirement for lower concentrations of capture antibody, and had been tested successfully against sporozoites from a wider range of geographical areas than the others. Use of this monoclonal antibody in a standardized ELISA method gave a test ten times more sensitive than previously reported for P. falciparum sporozoites and its detection limit was less than 100 sporozoites per mosquito.

Animals↗

Virus isolations from mosquitoes collected during the 1982 Japanese encephalitis epidemic in northern Thailand.

From 16 June to 15 August, 1982 CDC light traps were used to collect mosquitoes in the province of Kamphaengphet, N. Thailand. 353,042 mosquitoes comprising 59 species were collected and identified, and 345,173 were placed in pools for attempted virus isolation by inoculation of C6/36 Aedes albopictus mosquito cell cultures. Viruses were isolated from 63 mosquito pools. These comprised 56 flaviviruses, identified as 35 isolates of Japanese encephalitis (JE) virus strains, 18 strains of Tembusu (TEM) virus and three untyped flaviviruses (FLA); three alphaviruses, identified as the first isolates of Getah (GET) virus to have been made in Thailand; and four viruses which are still unidentified. Most virus isolates were from Culex tritaeniorhynchus mosquitoes collected in carbon dioxide baited light traps. JE virus was isolated only over a ten-day period and the last isolate was obtained one week before the peak of admission of human encephalitis cases at Kamphaengphet Provincial Hospital. Rapid screening of isolates grown on Ae. pseudoscutellaris (LSTM-AP-61) mosquito cells by indirect immunofluorescence using flavivirus group-specific and JE-specific monoclonal antibodies showed a high degree of correlation with plaque reduction neutralization tests. An antigen capture enzyme immunoassay (EIA) test successfully identified about 50% of the JE virus positive pools, but the method saved considerable processing time.

Alphavirus↗

Genetics of esterases and 6-phosphogluconate dehydrogenase in the Anopheles maculatus complex.

Electromorphic variation for some esterases and 6-phosphogluconate dehydrogenase enzymes in the Anopheles maculatus complex is controlled by four loci which are unlinked to sex. Esterase loci are linked to each other: Est-1-36%-Est-4-16.5%-Est-3; but unlinked to Pgd-2. Linkage data were obtained by selfing the F1 generation from selected parents and analysing genotypes in the F2; the classical dihybrid-cross. The analysis consists of testing observed data for goodness of fit to a) ratios expected from Mendelian ratios without linkage and b) if they do not fit then computing a likely degree of linkage and computing expected ratios with such linkage for further tests. Confidence limits are given for the most likely levels of linkage. This method can provide useful information for population-genetic studies on anopheline mosquitoes, whose laboratory rearing is generally difficult. Through indirect evidence, the enzyme loci are correlated to polytene chromosomes. The esterases probably lie on chromosome three (polytene arms 3 and 4) and Pgd-2 on the second chromosome (arms 2 and 5).

Animals↗

Correlation of survival rates of Anopheles dirus A (Diptera: Culicidae) with different infection densities of Plasmodium cynomolgi.

The survival rates are described for 36 paired replicates of Anopheles dirus A mosquitos that had been allowed to engorge themselves on rhesus monkeys that were either infected or non-infected with Plasmodium cynomolgi. The survival rates of infected mosquitos with a mean number of oocysts less than 10 did not differ significantly from those that were non-infected; however, there was a significant difference in the survival rates of non-infected groups and those with a mean number of oocysts in the range 10-40, 41-70, or >/=71.Daily survival rates for non-infected and infected mosquitos did not differ significantly during the first 8 days of extrinsic incubation. In contrast, for the period 9-30 days the survival rates of mosquitos with mean number of oocysts >/=41 were significantly different from those of non-infected mosquitos. The cumulative daily survival rates of mosquitos with mean number of oocysts up to 40, 41-70, or >/=71 decreased with the oocyst count. Mosquitos with a mean number of oocysts >/=71 frequently exhibited excessive numbers of bacteria and deterioration of both their guts and salivary glands.

Animals↗