METHODS: Mosquitoes found landing on humans and resting on leaves over a 5-day period at two sites in the Lawas District of northern Sarawak were collected and identified. DNA samples extracted from salivary glands of Anopheles mosquitoes were subjected to nested PCR malaria-detection assays. The small subunit ribosomal RNA (SSU rRNA) gene of Plasmodium was sequenced, and the internal transcribed spacer 2 (ITS2) and mitochondrial cytochrome c oxidase subunit 1 (cox1) gene of the mosquitoes were sequenced from the Plasmodium-positive samples for phylogenetic analysis.

RESULTS: Totals of 65 anophelines and 127 culicines were collected. By PCR, 6 An. balabacensis and 5 An. donaldi were found to have single P. knowlesi infections while 3 other An. balabacensis had either single, double or triple infections with P. inui, P. fieldi, P. cynomolgi and P. knowlesi. Phylogenetic analysis of the Plasmodium SSU rRNA gene confirmed 3 An. donaldi and 3 An. balabacensis with single P. knowlesi infections, while 3 other An. balabacensis had two or more Plasmodium species of P. inui, P. knowlesi, P. cynomolgi and some species of Plasmodium that could not be conclusively identified. Phylogenies inferred from the ITS2 and/or cox1 sequences of An. balabacensis and An. donaldi indicate that they are genetically indistinguishable from An. balabacensis and An. donaldi, respectively, found in Sabah, Malaysian Borneo.

METHODOLOGY/PRINCIPAL FINDINGS: Anopheles spp. were sampled using human landing catch (HLC) method at Paradason village in Kudat district of Sabah. The collected Anopheles were identified morphologically and then subjected to total DNA extraction and polymerase chain reaction (PCR) to detect Plasmodium parasites in the mosquitoes. Identification of Plasmodium spp. was confirmed by sequencing the SSU rRNA gene with species specific primers. MEGA4 software was then used to analyse the SSU rRNA sequences and bulid the phylogenetic tree for inferring the relationship between simian malaria parasites in Sabah. PCR results showed that only 1.61% (23/1,425) of the screened An. balabacensis were infected with one or two of the five simian Plasmodium spp. found in Sabah, viz. Plasmodium coatneyi, P. inui, P. fieldi, P. cynomolgi and P. knowlesi. Sequence analysis of SSU rRNA of Plasmodium isolates showed high percentage of identity within the same Plasmodium sp. group. The phylogenetic tree based on the consensus sequences of P. knowlesi showed 99.7%-100.0% nucleotide identity among the isolates from An. balabacensis, human patients and a long-tailed macaque from the same locality.

Methods: : Extensive surveys were carried out from November 2017 to January 2018 (post-monsoon season) randomly in the nine inhabited islands of Lakshadweep for conducting faunastic studies on mosquitoes and to know the basic binomics like breeding and resting preference of mosquitoes. The study islands included, Kavaratti, Agatti, Chetlat, Bitra, Amini, Kadmath, Andrott, Kalpeni and Kiltan. Both immature and adult collections were carried out by standard/appropriate sampling techniques. The obtained data were calculated and analysed in terms of different entomological indices.

Results: : A total of 3356 mosquitoes were collected during the study period which comprised of 16 species from nine genera. Out of the 16 species, six belonged to mosquito vectors. The collection included malaria vector, Anopheles stephensi; Japanese encephalitis vector, Culex tritaeniorhynchus; Bancroftian filariasis vector, Cx. quinquefasciatus; Brugian filariasis vector, Mansonia uniformis; and dengue and chikungunya vectors, Stegomya albopicta and St. aegypti. Stegomya albopicta was the most predominant species observed constituting 54% of the catch, followed by Cx. quinquefasciatus, An. stephensi, Cx. tritaeniorhynchus, and St. aegypti constituting 10.5, 6, 3 and 1.2%, respectively. Apart from vector species many non-vectors such as Heizmannia chandi, An. subpictus, An. varuna, Cx. sitiens, Cx. minutissimus, Cx. rubithoracis, Fredwardsius vittatus, Lutzia fuscana, Malaya genurostris and Armigeres subalbatus were also present in the study area. In Kavaratti Island, the capital of Lakshadweep, a non-vector species of sandfly, Sergentomyia (Parrotomyia) babu was observed during the indoor resting collection. The major breeding sites which supported various mosquito species included, discarded plastic containers, tree holes, open sintex tanks (water storage tanks), unused wells, discarded tyres, discarded iron pots, unused and damaged boats, cement tanks, pleated plastic sheets, coral holes, pits and irrigation canals, discarded washing machines, and Colocasia plant leaf axils. Breteau index ranged between 65.3 and 110, CI ranged between 63.64 and 72.41; and HI ranged between 38.46 and 70 among the various islands.