Esterases in the malaria vector mosquito, Anopheles culicifacies. Genetic and linkage analyses of an alpha and beta polymorphism

The genetic and linkage analyses of an alpha and beta esterase polymorphism in Anopheles culicifacies are presented. A survey of laboratory strains uncovered four electrophoretic variants for the alpha esterase and two for the beta esterase. Genetic analyses indicated that the variants of the alpha esterase are under the control of codominant alleles of a single locus and that this locus is linked to the locus controlling the expression of the codominant alleles of the beta esterases. The esterases are not linked either to sex (chromosome 1) or to maroon eye (chromosome 2) but to the chromosome 3 markers, dieldrin resistance and phosphoglucomutase. The gene sequence is Est-alpha--Dl--Pgm--Est-beta dnd the recombination frequencies are as follows: Est-alpha--Dl = 2.9 percent; Dl--Pgm = 5.8 percent; Pgm--Est-beta = 6.4 percent; Est-alpha--Est-beta = 15.1 percent, Est-alpha--Pgm = 8.7 percent and Dl--Est-beta = 12.2 percent.     

Anopheles culicifacies in Baluchistan, Iran

Abstract. Anopheles culicifacies (probably species A) is the main vector of malaria in Baluchistan, southeastern Iran. Adult mosquitoes were collected during 1990-92 by five methods of sampling: knock-down pyrethrum space-spray indoors, human and animal bait (18.00–05.00 hours), pit shelters and CDC light traps, yielding 62%, 3%, 6%, 4% and 25% of specimens, respectively. Whereas spray-catches comprised c. 70% gravid and semi-gravid females, light trap catches were mostly ( c . 60%) unfed females, while females from pit shelters comprised all abdominal stages more equally (13–36%). An. culicifacies populations peaked in April-May and rose again during August-November. Densities of indoor-resting mosquitoes were consistently greater in an unsprayed village than in villages subjected to residual house-spraying with propoxur, malathion or pirimiphos-methyl. Monthly malaria incidence generally followed fluctuations of An. culicifacies density, usually with a peak in May-June.     

X-linkage of malic enzyme in Anopheles culicifacies species B

A survey for malic enzyme (Me) in laboratory strains of species A and species B of Anopheles culicifacies had uncovered two electrophoretic variants, slow and fast, in two strains of species B. Genetic analysis revealed the two variants to be codominant alleles segregating at a locus, Me, which is sex linked. Because of the XX-XY sex determining mechanism, in F1 females, two electromorphs, viz., slow and fast, were observed, whereas in males only one electromorph of maternal origin was seen. Linkage experiments with another X-linked mutant, white eye (w), indicated the map distance between the two loci to be 9.52 +/- 0.86.     

Lactate dehydrogenase allozyme differentiation of species in the Anopheles culicifacies complex

Abstract Genetically controlled enzyme variation exists within and between four sibling species of the Anopheles culicifacies complex of malaria vectors in India. A study on electrophoretic variation of nine enzymes in An.culicifacies sibling species revealed that the lactate dehydrogenase ( Ldh ) locus has Fast (F) and Slow (S) allozymes distinguishing species A+D from species B+C with a probability of c . 95%.     

The Anopheles culicifacies complex and control of malaria

Of the 50 or so species and varieties of anopheline mosquito in India, about 20 are implicated as vectors of human malaria. Of these, perhaps the most important and widespread is Anopheles culicifacies s.l. For the first 10 years of widespread DDT spraying, An. culicifacies remained susceptible to this insecticide - indeed, some thought it lacked resistance genes altogether. By 1960 however, resistance to DDT was apparent. DDT is a cheap insecticide, and its use is still favoured for mosquito control wherever it remains effective. But by the end of the late 1970s it appeared that DDT resistance in An. culicifacies (and other species) was a major barrier to effective vector control in several areas - particularly in parts of the northwestern states. Yet in other areas DDT still seemed to be effective. There was also increasing evidence from other studies suggesting differences in An. culicifacies found in different areas - particularly differences in seasonal prevalence and man-biting activity. We now know - as Sarala Subbarao discusses here - that An. culicifacies s.l. represents a complex of at least four sibling species. But in this case, one of the most important findings is that DDT resistance is mainly associated with species B which proves to be a very poor vector of malaria. Such findings, made possible by careful cytogenetic studies, have very important consequences for malaria epidemiology and control policies.     

Egg development in the mosquito Anopheles albimanus

Summary

In the mosquito, Anopheles albimanus, previtellogenic egg development was completed by 48 h after emergence, and vitellogenic growth was completed by 36 h after a blood meal. Ecdysteroid levels reached a peak of 800 pg/female by 18 h, while vitellin levels rose to their maximum 36–48 h after a blood meal. Most of the ecdysteroids present in the female before 36 h behaved as ‘free’ hormone, while after 42 h the ecdysteroids were ‘conjugated’. Injection of 20-hydroxyecdysone into non-blood-fed females induced degeneration of the resting stage oocytes, but vitellogenin synthesis was detectable by autoradiography. Injection of 5 μ of 20-hy-hroxyecdysone into blood-fed decapitated females induced almost precisely normal levels of vitellin. Detailed analysis of the effect of decapitating blood-fed females suggested that the release of factors from the head (e.g., egg development neurosecretory hormone) occurs as an all-or-none phenomenon, and probably occurs twice.     

Inversion polymorphism in malarial mosquito Anopheles messeae natural populations from the north-eastern region of the range

A cytogenetic study of eight natural populations of Anopheles messeae from the north-eastern part of the species areal was conducted. Complete predominance of homozygotes XL11 and 3R11 in the northern populations was observed. Change in the chromosome 2 inversion frequencies from south northwards was shown. The 2R11 variant, which was not observed in the southern region, was found in the northern populations. These results indicate the maintenance of chromosome frequency of the distribution of inversions XL1 and 3R (in longitude) and 2R1 (in latitude). The inversion frequency distribution in the examined part of the areal have been preserved for a long time.     

ACE inhibitors reduce fecundity in the mosquito,Anopheles stephensi

Angiotensin-converting enzyme (ACE) is a dipeptidyl carboxypeptidase, which cleaves dipeptides and, in some instances, dipeptide or tripeptide amides from the C-terminus of regulatory peptides (e.g. angiotensin I, bradykinin and substance P). The expression of ACE is highly regulated in insects, where it is thought to have a role in the metabolism of peptide hormones involved in regulating reproduction. After a blood meal, ACE activity in the female mosquito Anopheles stephensi, increases four-fold with much of the enzyme finally accumulating in the ovary. In the present study, we have studied the effect on reproduction of adding two selective inhibitors of ACE, captopril and lisinopril, to the blood meal. Both ACE inhibitors reduced the size of the batch of eggs laid by females in a dose-dependent manner, with no observable effects on the behaviour of the adult insect. The almost total failure to lay eggs after feeding on either 1 mM captopril or 1 mM lisinopril, did not result from interference with the development of the primary follicle, but was due to the inhibition of egg-laying. Since very similar effects on the size of the egg-batch were observed with two selective ACE inhibitors, belonging to different chemical classes, we suggest that these effects are mediated by the selective inhibition of the induced mosquito ACE, a peptidase probably involved in the activation/inactivation of a peptide regulating egg-laying activity in A. stephensi.     

Oogenesis in the malaria mosquito Anopheles gambiae

Summary Oogenesis has been followed with the electron microscope in 2 strains of the malaria mosquito Anopheles gambiae, from the emergence of the adult (oocytes at leptonema) till shortly before the oocytes are ready for oviposition. After pachynema the chromosomes form a karyosphere and a fibrous capsule develops around it. Work on other mosquitoes suggests that the capsule may be related to the synaptonemal complexes. Both Anopheles strains contain at some time an extrachromosomal (not DNA-containing) body comparable to the karyosphere in size. Clusters of granules are present at the surface of the nucleolus and free in the nucleoplasm. Tentative results indicate that they may contain DNA. During oogenesis the nucleolus becomes very large, mainly because of proliferation of the nucleolonema. Towards the end of oocyte development the nucleus assumes the large canoe-shape also seen in Aedes and Culex. Nucleolonema traverse the entire nucleus, and modified granular clusters are found throughout.     

Isolation and characterization of microsatellite markers from malaria vector,Anopheles culicifacies

Anopheles culicifacies, an important vector in the Indian subcontinent is a complex of five sibling species of which four are vectors. We describe the isolation of 31 microsatellite markers from the recently recognized isomorphic species A of which 13 were characterized in sympatric populations of Anopheles culicifacies isomorphic species A and B. The allele frequencies ranges from two to 12 in species A and two to seven in species B. Species A being a vector, and that these markers can be used in closely related species, makes the isolation of these markers important to study population structure of all sibling species in this complex.     

Effect of anti-mosquito midgut antibodies on development of malaria parasite, Plasmodium vivax and fecundity in vector mosquito Anopheles culicifacies (Diptera: culicidae)

The effect of anti-mosquito-midgut antibodies on the development of the malaria parasite, P. vivax was studied by feeding the vector mosquito, An. culicifacies with infected blood supplemented with serum from immunized rabbits. In order to get antisera, rabbits were immunized with midgut proteins of three siblings species of Anopheles culicifacies, reported to exhibit differential vectorial capacity. The mosquitoes that ingested anti-midgut antibodies along with infectious parasites had significantly fewer oocysts compared to the control group of mosquitoes. The immunized rabbits generated high titer of antibodies. Their cross reactivity amongst various tissues of the same species and with other sibling species was also determined. Immunogenic polypeptides expressed in the midgut of glucose or blood fed An. culicifacies sibling species were identified by Western blotting. One immunogenic polypeptide of 62 kDa was exclusively present in the midgut of species A. Similarly, three polypeptides of 97, 94 and 58 kDa and one polypeptide of 23 kDa were present exclusively in species B and C respectively. Immunoelectron microscopy revealed the localization of these antigens on baso-lateral membrane and microvilli. The effects of anti-mosquito midgut antibodies on fecundity, longevity, mortality and engorgement of mosquitoes were studied. Fecundity was also reduced significantly. These observations open an avenue for research toward the development of a vector-based malaria parasite transmission-blocking vaccine.     

Four new mutations and a linkage map of species A of Anopheles culicifacies

Four new mutations, brown eye (bw), colorless eye (c), scarlet eye (sca), and abnormal eye (ae) have been mapped on chromosome 2 of species A of Anopheles culicifacies. The gene sequence is bw--c--sca--ae. A linkage map of the genetically analyzed loci in this species is presented.     

Crossing-over in the long arm of the X and Y chromosomes in Anopheles culicifacies

Genetic and cytogenetic investigations of two unstable Y-linked translocations in Anopheles culicifacies indicate that crossing over can occur between the X and Y chromosomes in this species. Moreover, these observations suggest that the long arms of the X and Y chromosomes are homologous and that the sex determining mechanism(s) is located in the short arms.     

Chromosomal polymorphism in the populations of malaria vector mosquito Anopheles messeae at the south of Russian Plain

Zonal features of the geographic distribution of chromosomal inversions in the populations of An. messae at the south of the Russian Plain were examined. The An. messae was identified based on morphological characters and cytogenetically. The chromosomal inversions identified in populations of An. messae comprised XL₁, XL₄, 2R₁, 3R₁, and 3L₁. Inversion XL₄ was endemic and found with low frequency (2%) in Penza oblast and Moscow oblast. Based on the population karyotype structure similarity, on the territory of Russian Plain, three zones (southwestern, southeastern, and central) were identified. Central zone was characterized by higher levels of inversion polymorphism in all chromosomes, except arm 3L. In the two southern zones, high frequency of the XL₀ inversion, along with complete absence of homo- and heterozygotes for the 2R₁ inversion, and high proportion of the individuals with inversion 3L₁ was observed. Specific feature of southeastern zone was the increased frequency of hetero- and homozygotes for the 3R₁ and 3L₁ inversions. Zonal differences reflected adaptive character of chromosomal polymorphism and pointed to hierarchic organization of the species population structure.     

Mating in the mosquito, Anopheles gambiae s.l.

ABSTRACT. Swarming behaviour in the Anopheles gambiae complex was observed in the field, in the Gambia, West Africa, and in the laboratory. Naturally occurring swarms of A.melas were seen in a clearing at the edge of mangrove swamps close to their breeding sites. Males could be induced to swarm over artificial 'markers' within this 'arena' but not outside it. Females were observed entering the swarm and mating. In the laboratory, in an artificial 'dusk', male A.gambiae s.str. swarmed over a black marker on the floor of their 1.2-m cube cage. In contrast to the males, females made only short flights over the marker, performing brief turning movements at its edge. It is proposed that swarming brings about the aggregation necessary before short-range attraction can take place, and that, in nature, anopheline mosquitoes orientate visually first to an arena and then to a marker within the arena. Female behaviour can be interpreted as a process of scanning possible swarm sites until mating is achieved.     

Inversion polymorphism of the malaria mosquito Anopheles messeae. VIII. Distribution of larvae in biotopes under conditions of co-habitation with Anopheles beklemishevi

The role of chromosomal variability was studied in the process of biotopical distribution of larvae of the sibling species Anopheles messeae and A. beklemishevi. It is shown that frequency of the inversions XL1, XL2, 2R1, 3R1 of A. messeae was changed in the lakes with different larval quantity of the sibling species. Mosquito of A. beklemishevi were developed in the suboptimal biotops having low larwal quantity of A. messeae with the analogous chromosomal arrangement 2R1. The larvae with "northeast" genocomplexes of A. messeae prevailed over A. beklemishevi in the lakes with high larval quantity and this prevalence determined dominant state of A. messeae. It is noted that natural selection on the preimaginal stages of development acts in the direction of karyotypic divergence of the sibling species and promotes more effective utilisation of food resources by each species.