Comparative laboratory bioassays of Tolypocladium cylindrosporum, California strain (Kal) was conducted against third instar larvae of four species of mosquito, viz. Aedes aegypti, Anopheles balabacensis, Culex quinquefasciatus and Mansonia uniformis in Malaysia. Of the four mosquito species tested, Ma. uniformis was found to be the most susceptible, followed by Cx. quinquefasciatus, An. balabacensis and Ae. aegypti, in a decreasing order. The LC50 values for Ma. uniformis, Cx. quinquefasciatus, An. balabacensis and Ae. aegypti after four days of exposure were 1.18 X 10(4), 2.02 X 10(5), 4.76 X 10(5) and 1.84 X 10(7) spores per ml test media, respectively. The high sensitivity of Ma. uniformis and its longer life cycle seems to indicate that T. cylindrosporum Kal has good potential as a biocontrol agent for this species of mosquito. But, for Ae. aegypti, this fungus appears to be less effective.
Mark-release-recapture experiments were carried out in Sabah, East Malaysia on the malaria and filariasis vector, Anopheles balabacensis. Samples of wild females were marked with different colours of fluorescent pigments, released in man-baited huts fitted with exit traps. Simultaneous collections and releases were also made in night-biting catches on a water buffalo and on four men. All subsequent recaptures were made in the same situation in which the mosquitoes were marked. The same individual mosquitoes were caught biting men and buffalo on different occasions and the numbers caught showed a strong preference for man over buffalo. The length of the oviposition cycle in the field was found to be 3.0 days. After blood-feeding on man in a hut, An. balabacensis were found to exit on the night or early morning. The same individual mosquitoes were found resting in the hut or exit trap on different occasions. The results indicate that there is strong evidence for the existence of genetic variability in the tendency of An. balabacensis to rest in houses and to bite man and buffalo. The obvious existence of this phenomenon is considered discouraging for the prospects of interruption of malaria transmitted by An. balabacensis in nature.
Recent studies on cytogenetics, behavioral, geographical and distinct morphological characters on adult, pupal and larval stages have revealed that "balabacensis" is a species complex. Anopheles dirus the mainland species, is distributed widely in Thailand and is renowned for its role as primary vector of human malarial parasites. Further, evidence from cytogenetic and taxonomic studies suggests that "An. dirus" is a species complex comprising at least four distinct species provisionally designated: dirus A, B, C and D. These cryptic species are distinguishable only partially morphologically, but can be separated on the basis of metaphase chromosomes using the Giemsa and Hoechst 33258 staining techniques. Apparently, these siblings show distinct patterns of geographic distribution in Thailand and Peninsular Malaysia. The recognition of dirus as a complex of species in Thailand and Peninsular Malaysia requires a re-evaluation of the role that the individual members of this complex have in the transmission of malaria parasites in this region. Cytological analysis of gene rearrangements in ovarian polytene chromosomes has shown that An. maculatus is a sibling-species complex consisting of at least four species in Thailand provisionally designated: maculatus A, B, C and G. These siblings are sympatric in some populations. Furthermore, species B is so highly polymorphic for chromosome rearrangements that four geographic forms can be recognized. It is not known whether these four forms are subspecies or yet further species within the species B complex. These sibling-species must be differentiated in order to understand any differential capabilities in their transmission of human malaria parasites. Anopheles nivipes was elevated from synonymy under An. philippinensis to full species status by Reid, a decision recently confirmed by cross mating experiments. The Thailand Malaria Division does not differentiate these two species and only identifies An. philippinensis, yet, An. nivipes is by far the most common of the two species in Thailand. Furthermore, preliminary surveys of the ovarian polytene chromosomes of several widely separated populations of An. nivipes in Thailand have revealed at least two distinct chromosomal types of nivipes based on fixed inversions on the X chromosomes.
Susceptibilities of two colonies of the taxon An. dirus (one from Perlis and from Thailand) and one colony of An. balabacensis from Sabah to DDT, dieldrin, malathion, fenitrothion and propoxur were determined. DDT and fenitrothion tolerance was found in An. dirus species B and An. balabacensis. No resistant strain was isolated as the two colonies were not either homozygous or nearly so for resistance. Field testing of the susceptibility of the adults of An. balabacensis to DDT was carried out between 1957 to 1976. The results indicated a progressive rise in the LC50 levels greater than 1% in almost all instance. The variation in the number of sprays applied in some districts have resulted in varying sensitivities. Association between the changes in levels of DDT susceptibility and exophilic habit of An. balabacensis has been observed but needs further confirmation. The significance of these findings and the difficulties in distinguishing tolerant from truly resistant individuals are discussed in relation to accurate species identification.
The toxic effects of Abate (temephos) on mosquito larvae and non-target organisms were studied in the rice-field and in the laboratory. In the laboratory tests, Culex tritaeniorhychus larvae and cladoceran zooplanktons (predominantly Diaphanosoma and Moinodaphnia species) were found to be highly susceptible to Abate with LC50 values of 0.27 and less than 0.10 parts per billion respectively. Other non-target species in decreasing degree of susceptibility to Abate were copepods (Tropodiaptomus spp.), Aplocheilus panchax and Tubifex worms. In field study, Abate at concentrations 60, 100 and 200 gm hectare-1 is effective in maintaining the rice-fields free of Anopheles and Culex mosquitoes for at least 2 days. No mortality was observed for Aplocheilus panchax and Tubifex worms at the above concentrations; nevertheless, populations of cladoceran zooplanktons and copepods were reduced up to seven days posttreatment.
Starch-gel electrophoretic studies on nine gene-enzyme systems comprising 14 loci revealed a fair level of genetic variation in two population samples of Anopheles maculatus from Peninsular Malaysia. The proportion of polymorphic loci was 0.36 for the Fort Bertau sample and 0.29 for the Gua Musang sample, while the mean heterozygosity value was 0.09 for Fort Bertau and 0.07 for Gua Musang. The values of genetic similarity (I = 0.98) and genetic distance (D = 0.02) were of the rank of geographical populations.
Until recently, very little was known of Anopheles species complexes and their relationships to epidemiology and malaria transmission in Southeast Asia. During the past eight years, extensive studies on the genetics of natural populations of anopheline mosquitoes in this region, involving the interdisciplinary efforts of taxonomists, operational entomologists and biologists, have revealed groups of cryptic species of Anopheles vectors, particularly the An. leucos phyrus group. This species group comprise seventeen species and two subspecies widely distributed in the forested areas of Southeast Asia. Among these species. An. dirus Peyton and Harrison, has been shown by cytogenetic and morphological studies to be a complex of at least seven isomorphic species, provisionally designated species A, B, C, D, E, F and takasagoensis, on the Southeast Asian mainland. Cytological identification of these species is based on distinct banding patterns of salivary gland polytene chromosomes as well as heterochromatin differences in mitotic karyotypes. The five species found in Thailand (A-D, F) exhibit distinct geographic distributions. Species A is widespread throughout Thailand except in the south. Species B had been found in sympatry with species C in southern Thailand and both seem to show north-south clinal geographic variation. Species D is common on the west side of southern Thailand and along the Thai-Burmese border in sympatry with species A. Species F, An. nemophilous Peyton and Ramalingam, has been found in a population at the Thai-Malaysian border in this study although it was known to be common in southern and western Thailand and Peninsular Malaysia. Species E is known only from western India. The five species found in Thailand also exhibit seasonal variation in relative abundance and different nocturnal biting cycles. Chromosomal polymorphisms have been observed in mitotic and polytene chromosomes of An. dirus A and D. Species B and C also show heterochromatin variation in the sex chromosomes, but are monomorphic for the standard sequence in polytene chromosomes. These biological characteristics of the An. dirus complex may have implications for understanding the epidemiology of malaria in Southeast Asia. Recent cytogenetic studies of wild-caught samples of An. leucosphyrus from Sumatra, Kalimantan and southern Thailand have revealed the presence of two distinct species within this taxon. Species A is widely distributed in southern Thailand, East Malaysia and Kalimantan, while species B is confined to Sumatra. The two isomorphic species are vectors of human malaria within their range of distribution.(ABSTRACT TRUNCATED AT 400 WORDS)
Two separate observations from recent electrophoretic studies of the systematics and population genetics of laboratory-reared populations which had a long history of colonization in various laboratories, were found to be inconsistent with the present study which used wild-caught populations from East Malaysia. Reanalysis of the two data sets generally indicated a low amount of genetic variation in laboratory colonies. The latter is characterized by higher frequency of monomorphic loci, low average heterozygosity values and, in one extreme case, no variability at two loci. However, natural populations of An. balabacensis and An. leucosphyrus showed more protein variability by the use of polyacrylamide gel electrophoresis. Since laboratory-maintained mosquitoes are genetically and phenotypically different from those in the field, results of laboratory studies on the systematics and population genetics of Anopheles species complexes may be biased.
Susceptibility tests were carried between 1987-1989 on Anopheles maculatus female adults collected from twelve different localities in Peninsular Malaysia to DDT. Mosquitoes collected from all localities except those from Kuala Berang, Trengganu state were found to be susceptible to DDT when tested with 4% impregnated paper. Several factors contributing to the lack of development of resistance despite many years of residual spraying in Peninsular Malaysia include exophily, exophagic and excito-repellency behaviour of An. maculatus and the inadequate spraying coverage of houses.
Anopheles (Cellia) litoralis King and Anopheles (Cellia) sundaicus Rodenwaldt, vectors of malaria, were collected from the same brackis and sea-water habitats in six localities in Sabah. They share the same breeding habitats with predominance of one species over the other. The two species although distinct have small morphological differences and are taxonomically separated by certain wing characters. Hybridization between the two species was successful. Reciprocal crosses produced viable progeny which appeared to develop normally to adults. Hybridized females laid fewer viable eggs in comparison with the parents. The F1 hybrids resembled the litoralis parent in most characters. Backcrosses of both litoralis and sundaicus parents with the F1 hybrids yielded no eggs. F1 male hybrids were thus assumed to be sterile. The results obtained from cross matings between the two species suggested something more than subspecific status.
Emulsifiable concentrates of DursbanR (chlorpyrifos) and Dowco 214 (chlorpyrifos-methyl) were tested as mosquito larvicides using Hudson knapsack sprayers on small plots of rice-fields on Penang Island. The mosquitoes found in these rice-fields are predominantly Anopheles campestris and Culex tritaeniorhynchus. At dosages of 14, 28 and 56 gm hectare-1, Dursban is effective in maintaining the rice-fields free of Anopheles larvae for at least 2, 3 and 7 days respectively. Dowco 214 at 56 gm hectare-1 is able to keep the fields free of all mosquito larvae for at least 2 days.