Studies on prevalence of anopheline species and community perception of malaria in Jaffna district, Sri Lanka.

BACKGROUND & OBJECTIVES: Over two decades of civil unrest and the conflict situation have had detrimental effects on vector control activities and management of malaria in Jaffna district which is an endemic region for malaria in Sri Lanka. With the background that only a few small-scale studies on malaria and its vectors have been reported from this district, a study was designed to explore the current status of malaria in the Jaffna district in relation to vector and community aspects. METHODS: Adults and larvae of anopheline mosquitoes were collected monthly from selected endemic localities. Species prevalence of the collected mosquitoes was studied while the collected adults of Anopheles subpictus, a potential vector in the district, was screened for sibling species composition based on morphological characteristics and exposed to common insecticides using WHO bioassay kits. Knowledge, attitude and practices (KAP) of the community were tested using a pre-tested structured questionnaire in high-risk and low-risk localities in the district. RESULTS: The anopheline mosquito species distribution in the district was--An. culicifacies (0.5%), An. subpictus (46%), An. varuna (4%), An. nigerrimus (44%) and An. pallidus (5.5%). Among the collected larvae the percent prevalence of An. culicifacies was 13% and other species follows as: An. subpictus (71%), An. varuna (4%), An. nigerrimus (10%) and An. pallidus (2%). Sibling species B, C and D of An. subpictus were present in the district with the predominance of B in both coastal and inland areas, while all members showed both indoor and outdoor resting characteristics, they were highly resistant to DDT (4%) and highly susceptible to malathion (5%). KAP study in the district showed a reasonable level of knowledge, positive attitude and practices towards malaria. CONCLUSION: An. subpictus, the reported major vector of Jaffna and a well-established secondary vector of malaria in the country, continues to be the predominant anopheline species. The distribution of sibling species of An. subpictus complex in the Jaffna district, revealed for the first time, has implications for future studies on its bionomics and malaria transmission pattern in this area and the planning of control strategies for this region. The community perception of disease, which revealed a satisfactory knowledge indicates the potential for better community participation in future malaria control activities in this region. As potential vectors are still present, health authorities need to be vigilant to prevent any future epidemics of malaria.

Mosquito midgut glycoproteins and recognition sites for malaria parasites.

Midgut glycoproteins of the malaria vector Anopheles tessellatus were partially characterised by gel electrophoresis and lectin binding. Specific binding to wheat germ agglutinin (WGA) and Concanavalin A (Con A) indicated the presence of N-linked core oligosaccharides in many proteins. Rabbit antibodies were produced against wheat germ agglutinin binding proteins (WGABP). These antibodies also recognised distinct proteins in the peritrophic membrane which is secreted into the midgut to enclose a bloodmeal. Rabbit anti-WGABP antibodies ingested in a bloodmeal containing infective gametocytes of the human malaria parasites Plasmodium falciparum and P. vivax tended to reduce infectivity of the parasites to vector mosquitoes. Chitotriose added to a bloodmeal also inhibited parasite development in the mosquito. The results are consistent with a hypothesis that N-acetyl glucosamine residues in mosquito midgut glycoproteins and/or midgut chitin and proteoglycan function as recognition sites for malaria parasites.

Interactions of human malaria parasites, Plasmodium vivax and P.falciparum, with the midgut of Anopheles mosquitoes.

Present understanding of the development of sexual stages of the human malaria parasites Plasmodium vivax and P.falciparum in the Anopheles vector is reviewed, with particular reference to the role of the mosquito midgut in establishing an infection. The sexual stages of the parasite, the gametocytes, are formed in human erythrocytes. The changes in temperature and pH encountered by the gametocyte induce gametogenesis in the lumen of the midgut. Macromolecules derived from mosquito tissue and second messenger pathways regulate events leading to fertilization. In An.tessellatus the movement of the ookinete from the lumen to the midgut epithelium is linked to the release of trypsin in the midgut and the peritrophic matrix is not a firm barrier to this movement. The passage of the P.vivax ookinete through the peritrophic matrix may take place before the latter is fully formed. The late ookinete development in P.falciparum requires chitinase to facilitate penetration of the peritrophic matrix. Recognition sites for the ookinetes are present on the midgut epithelial cells. N-acetyl glucosamine residues in the oligosaccharide side chains of An.tessellatus midgut glycoproteins and peritrophic matrix proteoglycan may function as recognition sites for P.vivax and P.falciparum ookinetes. It is possible that ookinetes penetrating epithelial cells produce stress in the vector. Mosquito molecules may be involved in oocyst development in the basal lamina, and encapsulation of the parasite occurs in vectors that are refractory to the parasite. Detailed knowledge of vector-parasite interactions, particularly in the midgut and the identification of critical mosquito molecules offers prospects for manipulating the vector for the control of malaria.

Different effects of modulation of mosquito (Diptera:Culicidae) trypsin activity on the infectivity of two human malaria (Hemosporidia:Plasmodidae) parasites.

Trypsin production in the malaria vector Anopheles tessellatus Theobald peaks between 12 and 21 h after a blood meal. The presence of leupeptin or soybean trypsin inhibitor in a blood meal delayed the onset of maximal trypsin activity. Trypsin inhibitors in an infective blood meal increased the infectivity of Plasmodium vivax Grassi and decreased infectivity of P. falciparum Welch to An tessellatus. The opposite effects of trypsin inhibitors on infectivity of the 2 malaria parasites were attributed to differences in the biology of the parasites within the midgut of the vector, particularly the time of ookinete formation and the requirement for activation of a chitinase.

Antimidgut antibodies inhibit peritrophic membrane formation in the posterior midgut of Anopheles tessellatus (Diptera: Culicidae).

The effect on midgut structure of feeding rabbit antibodies in a blood meal was investigated in Anopheles tessellatus Theobald. No detectable ultrastructural changes in midgut cells were observed. However, antimidgut antibodies inhibited formation of the peritrophic membrane in the posterior midgut. The results indicate that a peritrophic membrane in the posterior midgut is not essential for survival of the mosquito after a blood meal or for establishing a Plasmodium vivax Grassi infection.

Antibodies to Anopheles midgut reduce vector competence for Plasmodium vivax malaria.

Anopheles tessellatus mosquitoes ingested Plasmodium vivax gametocytes in human erythrocytes suspended in rabbit sera with and without anti-mosquito midgut antibodies. When the mosquito bloodmeal contained anti-midgut antibodies, fewer oocysts of P.vivax developed on the mosquito midgut and the proportion of mosquitoes becoming infected was significantly reduced. Complement inactivated serum also reduced the infection rate and load. A second bloodmeal containing anti-midgut antibodies, given 48 or 72 h later, did not enhance the transmission-blocking effect. IgG purified from anti-midgut sera was shown to mediate the transmission-blocking effect.

Population dynamics of anthropophilic mosquitoes during the northeast monsoon season in the malaria epidemic zone of Sri Lanka.

Mosquito-borne diseases are a major health problem in Sri Lanka. Human biting mosquitoes were collected during the night (18.00-06.00 hours) at Nikawehera village, in the malaria endemic intermediate rainfall zone of the country. Collections were made at monthly intervals in the period October 1991 to April 1992, which included the main rainy season due to the northeast monsoon (October-January). Thirteen Anopheles, eleven Culex, three Aedes, three Mansonia and one Armigeres species were identified, including known vectors of malaria, Bancroftian filariasis, Japanese encephalitis and dengue fever. Mosquito human-biting rates were highest in December. The main malaria vector Anopheles culicifacies showed peak biting between 18.00 and 23.00 hours whereas the predominant culicines Culex fuscocephala and Cx quinquefasciatus preferred to bite after midnight. In 1991-92 the prevalence of some species of anophelines at Nikawehera differed markedly from that observed in 1990-91 and the possible reasons are discussed.

Fecundity of Anopheles tessellatus reduced by the ingestion of murine anti-mosquito antibodies.

High titres of antibodies to antigens derived from head/thorax, midgut or abdomen of Anopheles tessellatus were produced in inbred mice. These antibodies, when ingested in a bloodmeal, reduced the fecundity of An. tessellatus by up to 29% in different experiments. It is postulated that antibodies directed against antigens shared between the head/thorax, abdomen and midgut tissues are involved in the reduction of fecundity.

High seasonal malaria transmission rates in the intermediate rainfall zone of Sri Lanka.

Malaria transmission was studied at Nikawehera, a long-established farming village, located in the intermediate rainfall zone of Sri Lanka. Observations were made over a 12-month period (October-September) that included the main rainy season which occurred during the north-east monsoon in November-January. Anolpheles culicifacies, the recognized vector of malaria in Sri Lanka, was the predominant anopheline mosquito collected by human night baiting at Nikawehera. High entomological inoculation rates with An. culcifacies (0.12/hour for Plasmodium vivax) were observed during the height of the transmission season which occurs during, and immediately after, the north-east monsoon. Anolpheles subpictus was identified as a possible additional vector at Nikawehera. Anopheles annularis, a major vector at Weheragala, a site in a new irrigation development (the Mahaweli Scheme) located 70 km away in the dry zone, was not collected by human baiting at Nikawehera. Clinical, entomological and parasitological data suggest that malaria is hyperendemic at Nikawehera, with high seasonal transmission rates.

Host immunity to mosquitoes: effect of antimosquito antibodies on Anopheles tessellatus and Culex quinquefasciatus (Diptera: Culicidae).

Rabbit antibodies to antigens derived from the tissues of sugar-fed Anopheles tessellatus Theobald were fed to An. tessellatus and Culex quinquefasciatus Say in blood meals. These antibodies tended to reduce the number of eggs produced by An. tessellatus, but had no effect on the fecundity of Cx. quinquefasciatus. Ingestion of the rabbit antibodies did not detectably affect the mortality of An. tessellatus but increased the mortality of Cx. quinquefasciatus. The results provide preliminary evidence for the potential of antimosquito antibodies to disrupt the physiology of mosquito vectors and indicate the need for further studies with high-titer, monospecific antibodies against the relevant target antigens.