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A study of the behavioral responses of Aedes aegypti and Anopheles minimus to 3 Cambodian plant extracts at 3 different concentrations (1%, 2.5%, and 5%) was performed using an excito-repellency test system. These 3 plants were Strophanthus scandens, Capparis micracantha, and Dioscorea hispida, selected according to traditional healer's knowledge, bibliographic studies and market surveys. Results showed that S. scandens leaves' hexane extract was the only one to exert repellency against Ae. aegypti with 23.3% of escaped mosquitoes at a concentration of 5%. Capparis micracantha was responsible for an irritant activity against An. minimus with 20.2% of escaped mosquitoes at a concentration of 2.5% and 22.8% escaping at a concentration of 5%. Dioscorea hispida showed an irritant activity on both mosquito species with 23.2% of escaped Ae. aegypti at a concentration of 5% and about 20% of escaped An. minimus at 2.5% and 5%. This is the first report on the irritant and repellent activities of S. scandens , D. hispida , and C. micracantha against mosquito species.

The Anopheles minimus Complex Theobald (Diptera: Culicidae) is composed of the 3 sibling species A, C, and E. The malaria vectors An. minimus A and C are distributed over the Southeast Asian region, whereas species E is restricted to the Ryukyu Japanese Islands. Because species A and C can be sympatric and present specific behaviors and have a role in malaria transmission, it is important to differentiate them. The literature mentioned the presence of a presector pale spot on the wing costa of An. minimus A, whereas species C may exhibit both presector and humeral pale spots. However, the reliability of their diagnostic power has not been established over large temporal and geographic surveys. From the analyses of 9 populations throughout Southeast Asia, including published data and field populations from 2 sites in Thailand, we showed that the wing patterns present spatial and temporal variations that make these two morphological characters unreliable for the precise identification of An. minimus A and C. Therefore, molecular identification remains the most efficient method to obtain an unambiguous differentiation of these 2 species. Correct species identification is essential and mandatory for any relevant study on the Minimus Complex and for the application of successful control strategies.

Before August 1998, in the Khanh Phu commune (central Vietnam), Anopheles minimus s.l. individuals were identified as species A and showed the typical species A wing form. After a significant decrease over the 3 years 1999-2001, an increase in 2002 of An. minimus s.l. possessing a different wing pattern was observed. To determine the specific status of the An. minimus species collected in 2002 and to follow changes in the species composition, an allele-specific polymerase chain reaction was applied to samples collected from 1993 to 2002. This study reports the first record of An. minimus C in central Vietnam and, since 1998, a significant reduction of An. minimus A that coincided with the wide use of permethrin-treated bednets. This change in anopheline composition may have important consequences on malaria transmission. This work shows that the geographic distribution of malaria vectors in southeast Asia is only partially known and highlights the importance of species identification for understanding changes in the vector composition as a result of selective vector control.

Data from mosquito collections made in Belize, Central America, between September 1990 and April 1993 are presented. A total of 537 collections yielding 15,139 specimens are summarized. One genus, 4 subgenera, and 31 species are recorded from Belize for the 1st time. A checklist of the 111 mosquito species now known to occur in Belize is presented.

Field samples of Anopheles gambiae s.s. from Côte d'Ivoire were tested with 5 pyrethroids (cyfluthrin, lambda-cyhalothrin, alpha-cypermethrin, deltamethrin, permethrin), 1 pseudo-pyrethroid (etofenprox), and an organochlorine (DDT). With the use of World Health Organization diagnostic tests, 5 out of 6 samples were found cross-resistant to these insecticides. A strong decrease in knockdown effect and mortality was also observed when testing deltamethrin-impregnated nettings. With a polymerase chain reaction amplification of specific alleles diagnostic test, resistance was found associated with the presence of a kdr mutation. The strong correlation between kdr allelic frequency and resistance to DDT or etofenprox indicated that kdr was the main resistance factor for these 2 insecticides. On the contrary, a lower correlation was observed between kdr frequency and resistance to 4 of the 5 pyrethroids tested, suggesting that another mechanism was also involved, likely a metabolic detoxification. These results point out the necessity to monitor pyrethroid resistance and the presence of kdr before implementation of any impregnated bed-net programs for malaria control.

Different malarial situations in Africa within the past 40 years are discussed in order to evaluate the impact of climatic and human factors on the disease. North of the equator, more droughts and lower rainfall have been recorded since 1972; and in eastern and southern Africa, there have been alternating dry and wet periods in relation to El Niño. Since 1955, the increase in human population from 125 to 450 million has resulted in both expansion of land cultivation and urbanization. In stable malaria areas of West and Central Africa and on the Madagascar coasts, the endemic situation has not changed since 1955. However, in unstable malaria areas such as the highlands and Sahel significant changes have occurred. In Madagascar, cessation of malaria control programs resulted in the deadly epidemic of 1987-88. The same situation was observed in Swaziland in 1984-85. In Uganda, malaria incidence has increased more than 30 times in the highlands (1,500-1,800 m), but its altitudinal limit has not overcome that of the beginning of the century. Cultivation of valley bottoms and extension of settlements are in large part responsible for this increase, along with abnormally heavy rainfall that favored the severe epidemic of 1994. A similar increase in malaria was observed in neighboring highlands of Rwanda and Burundi, and epidemics have been recorded in Ethiopia since 1958. In contrast, in the Sahel (Niayes region, Senegal), stricken by droughts since 1972, endemic malaria decreased drastically after the disappearance of the main vector, Anopheles funestus, due to the destruction of its larval sites by cultivation. Even during the very wet year of 1995. An funestus did not reinvade the region and malaria did not increase. The same situation was observed in the Sahelian zone of Niger. Therefore, the temperature increase of 0.5 degree C during the last 2 decades cannot be incriminated as a major cause for these malaria changes, which are mainly due to the combination of climatic, human, and operational factors.

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.

A survey of Anopheles pseudopunctipennis larval habitats was performed throughout most of its known geographic range. Eleven key environment variables characterized most larval habitats of this important vector of malaria in the Americas. Larval habitats occurred mainly in valley and foothill areas which were often situated in arid regions. Immatures were found primarily during the dry season in sun-exposed freshwater stream pools with clear, shallow, stagnant water containing abundant filamentous green algae and/or aquatic vegetation.

A survey for larval and adult Anopheles mosquitoes was conducted in Toledo District of southern Belize during August-September 1992. We surveyed for larvae in 145 habitats and conducted paired indoor-outdoor collections of adult mosquitoes landing on humans at 6 houses. In 1940-41, Kumm and Ram reported Anopheles darlingi females to be the most common Anopheles mosquitoes inside houses and reported no specimens of Anopheles vestitipennis in southern Belize. In our 1992 survey we found no An. darlingi mosquitoes either as adults or larvae. More An. vestitipennis females were captured indoors than outdoors, whereas most Anopheles albimanus and Anopheles apicimacula females were captured outdoors. All 3 species were represented occasionally in 145 collections of larvae from diverse habitats. Anopheles vestitipennis now appears to be a potentially important vector of malaria during the wet season in Toledo District.

Results of a recent survey of the 3 species of Anopheles mosquitoes, Anopheles (Nyssorhynchus) aquasalis, Anopheles (Nyssorhynchus) argyritarsis, and Anopheles (Anopheles) pseudopunctipennis, on the island of Grenada are contrasted with the only other published survey of these species on Grenada, which was published in 1938. Results suggest that a significant change in the abundance and distribution of each species has occurred over this period. Physical characteristics of the aquatic habitats for each species are described and compared.