Salivary gland proteome during adult development and after blood feeding of female anopheles dissidens mosquitoes (Diptera Culicidae)

Understanding changes in mosquito salivary proteins during the time that sporozoite maturation occurs and after blood feeding may give information regarding the roles of salivary proteins during the malarial transmission. Anopheles dissidens (formerly Anopheles barbirostris species A1) is a potential vector of Plasmodium vivax in Thailand. In this study, analyses of the proteomic profiles of female An. dissidens salivary glands during adult development and after blood feeding were carried out using two-dimensional gel electrophoresis coupled with nano-liquid chromatography-mass spectrometry. Results showed at least 17 major salivary gland proteins present from day one to day 21 post emergence at 8 different time points sampled. Although there was variation observed, the patterns of protein expression could be placed into one of four groups. Fifteen protein spots showed significant depletion after blood feeding with the percentages of the amount of depletion ranging from 8.5% to 68.11%. The overall results identified various proteins, including a putative mucin-like protein, an anti-platelet protein, a long form D7 salivary protein, a putative gVAG protein precursor, a D7-related 3.2 protein, gSG7 salivary proteins, and a gSG6 protein. These results allow better understanding of the changes of the salivary proteins during the adult mosquito development. They also provide candidate proteins to investigate any possible link or not between sporozoite maturation, or survival of skin stage sporozoites, and salivary proteins

Impact of ivermectin components on Anopheles dirus and Anopheles minimus mosquito survival

Background: Ivermectin mass drug administration to humans or livestock is a potential vector control tool for malaria elimination. Racemic ivermectin is composed of two components, namely a major component (> 80%; ivermectin B1a), which has an ethyl group at C-26, and a minor component (< 20%; ivermectin B1b), which has a methyl group at C-26. There is no difference between the efficacy of ivermectin B1a and ivermectin B1b efficacy in nematodes, but only ivermectin B1b has been reported to be lethal to snails. The ratios of ivermectin B1a and B1b ratios in ivermectin formulations and tablets can vary between manufacturers and batches. The mosquito-lethal effects of ivermectin B1a and ivermectin B1b have never been assessed. As novel ivermectin formulations are being developed for malaria control, it is important that the mosquito-lethal effects of individual ivermectin B1a and ivermectin B1b compounds be evaluated. Methods: Racemic ivermectin, ivermectin B1a or ivermectin B1b, respectively, was mixed with human blood at various concentrations, blood-fed to Anopheles dirus sensu stricto and Anopheles minimus sensu stricto mosquitoes, and mortality was observed for 10 days. The ivermectin B1a and B1b ratios from commercially available racemic ivermectin and marketed tablets were assessed by liquid chromatography-mass spectrometry. Results: The results revealed that neither the lethal concentrations that kills 50% (LC50) nor 90% (LC90) of mosquitoes differed between racemic ivermectin, ivermectin B1a or ivermectin B1b for An. dirus or An. minimus, confirming that the individual ivermectin components have equal mosquito-lethal effects. The relative ratios of ivermectin B1a and B1b derived from sourced racemic ivermectin powder were 98.84% and 1.16%, respectively, and the relative ratios for ivermectin B1a and B1b derived from human oral ivermectin tablets were 98.55% and 1.45%, respectively. Conclusions: The ratio of ivermectin B1a and B1b does not influence the Anopheles mosquito-lethal outcome, an ideal study result as the separation of ivermectin B1a and B1b components at scale is cost prohibitive. Thus, variations in the ratio of ivermectin B1a and B1b between batches and manufacturers, as well as potentially novel formulations for malaria control, should not influence ivermectin mosquito-lethal efficacy

Risk factors, clinical features and treatment of human infection with Plasmodium knowlesi and other Plasmodium species in Sabah, Malaysia

The simian parasite Plasmodium knowlesi is an emergent public health threat in southeast Asia, &nbsp;with human infections now increasingly reported where its macaque hosts and An. Leucosphyrus &nbsp;group vector are present. In Malaysia, P. knowlesi is now the most common cause of human &nbsp;malaria, and has been demonstrated to cause severe and fatal disease. This thesis aimed to provide further understanding of the epidemiology, clinical and laboratory features, and &nbsp;treatment of P.knowlesi malaria in both children and adults, with studies conducted in an area of &nbsp;northwest Sabah, Malaysia. Firstly, the major epidemiological study in this thesis was a case-control analysis of factors &nbsp;associated with acquiring symptomatic P. knowlesi infection. Key findings demonstrated the &nbsp;highest risk in farmers, specific activities such as plantation work and clearing vegetation, &nbsp;sleeping outside, travel, and other environmental and household factors, particularly at the &nbsp;forest-edge, although peri-domestic transmission was also evident. Intrinsic factors such as G6PD &nbsp;deficiency in humans were shown to have a protective benefit, in addition to conventional &nbsp;malaria prevention activities such as insecticide spraying of household walls. The presence of &nbsp;spatio-temporal case-clustering through analysis of P. knowlesi dhfr sequences was demonstrated,&nbsp;however was not found to be related to drug selection pressure from human-to-human transmission. Secondly, a large prospective study in three district hospitals detailed the clinical and laboratory features in children and adults with knowlesi malaria, highlighting the significant morbidity due &nbsp;to anaemia and acute kidney injury in children, despite an absence of severe complications in this &nbsp;group. A lower pyrogenic threshold was seen with lower parasitaemia compared to other &nbsp;Plasmodium species overall. The risk of severe disease was 6.4% in adults, with independent &nbsp;predictors of severe disease including age &ge;45 years and parasitaemia &gt;15,000/&mu;L. Finally, two major randomised controlled trials for the treatment of uncomplicated knowlesi and &nbsp;vivax malaria in Malaysia were conducted comparing an ACT, artesunate-mefloquine, against &nbsp;chloroquine in adults and children. Faster parasite and fever clearance, and decreased anaemia &nbsp;risk at day 28 was seen in the artesunate-mefloquine arm in both studies. There were no&nbsp;treatment failures in the P. knowlesi study. High-grade P. vivax chloroquine resistance was &nbsp;demonstrated in the P. vivax study, with a 61% risk of recurrence at day 28. These studies support the use of ACT as the first-line blood stage treatment for malaria due to all Plasmodium species in this co-endemic area, which is now reflected in national Malaysian treatment &nbsp;guidelines

Antennal morphology and sensilla ultrastructure of the malaria vectors, Anopheles maculatus and An. sawadwongporni (Diptera: Culicidae).

Mosquitoes rely mainly on the olfactory system to track hosts. Sensilla contain olfactory neuron receptors that perceive different kinds of odorants and transfer crucial information regarding the surrounding environment. Anopheles maculatus and An. sawadwongporni, members of the Maculatus Group, are regarded as vectors of malaria in Thailand. The fine structure of their sensilla has yet to be identified. Herein, scanning electron microscopy is used to examine the sensilla located on the antennae of adults An. maculatus and An. sawadwongporni, collected from the Thai-Myanmar border. Four major types of antennal sensilla are discovered in both species: chaetica, coeloconica, basiconica (grooved pegs) and trichodea. The antennae of female An. maculatus have longer lengths (μm, mean&nbsp;±&nbsp;SE) in the long sharp-tipped trichodea (40.62&nbsp;±&nbsp;0.35&nbsp;&gt;&nbsp;38.20&nbsp;±&nbsp;0.36), blunt-tipped trichodea (20.39&nbsp;±&nbsp;0.62&nbsp;&gt;&nbsp;18.62&nbsp;±&nbsp;0.35), and basiconica (7.84&nbsp;±&nbsp;0.15&nbsp;&gt;&nbsp;7.41&nbsp;±&nbsp;0.12) than those of An. sawadwongporni. Using light microscopy, it is found that the mean numbers of large sensilla coeloconica (lco) on both flagella in An. maculatus (left: 32.97&nbsp;±&nbsp;0.48; right: 33.27&nbsp;±&nbsp;0.65) are also greater when compared to An. sawadwongporni (left: 30.40&nbsp;±&nbsp;0.62; right: 29.97&nbsp;±&nbsp;0.49). The mean counts of lco located on flagellomeres 1-3, 6, and 9 in An. maculatus are significantly higher than those of An. sawadwongporni. The data in this study indicate that two closely related Anopheles species exhibit similar morphology of sensilla types, but show variations in length, and likewise in the number of large sensilla coeloconica between them, suggesting they might be causative factors that affect their behaviors driven by the sense of smell

Ivermectin metabolites reduce Anopheles survival

Ivermectin mass drug administration to humans or livestock is a potential vector control tool for malaria elimination. The mosquito-lethal effect of ivermectin in clinical trials exceeds that predicted from in vitro laboratory experiments, suggesting that ivermectin metabolites have mosquito-lethal effect. The three primary ivermectin metabolites in humans (i.e., M1 (3″-O-demethyl ivermectin), M3 (4-hydroxymethyl ivermectin), and M6 (3″-O-demethyl, 4-hydroxymethyl ivermectin) were obtained by chemical synthesis or bacterial modification/metabolism. Ivermectin and its metabolites were mixed in human blood at various concentrations, blood-fed to Anopheles dirus and Anopheles minimus mosquitoes, and mortality was observed daily for fourteen days. Ivermectin and metabolite concentrations were quantified by liquid chromatography linked with tandem mass spectrometry to confirm the concentrations in the blood matrix. Results revealed that neither the LC50 nor LC90 values differed between ivermectin and its major metabolites for An. dirus or An. minimus., Additionally, there was no substantial differences in the time to median mosquito mortality when comparing ivermectin and its metabolites, demonstrating an equal rate of mosquito killing between the compounds evaluated. These results demonstrate that ivermectin metabolites have a mosquito-lethal effect equal to the parent compound, contributing to Anopheles mortality after treatment of humans

An updated checklist of the ants of Thailand (Hymenoptera, Formicidae)

Thailand has a great diversity of ant fauna as a zoogeographical crossroads and biodiversity hotspot. The last publication presenting a Thai ant checklist was published in 2005. In the present paper, based on an examination of museum specimens and published records, a comprehensive and critical species list of Thai ants is synthesized. Currently, 529 valid species and subspecies in 109 genera among ten subfamilies are known from Thailand with their diversity and distribution within 77 provinces presented and assigned to six geographical regions. Furthermore, Thailand is the type locality for 81 ant species. Forty-one species are here newly recorded for Thailand with photographs illustrating these species. The checklist provides information on distribution and a comprehensive bibliography. This study will also serve as a guide for the upper northeast and central Thailand, which are poorly sampled; a comprehensive reference list relating to endemic taxa and localities where conservation is an important priority, thus an essential resource for policy makers and conservation planners concerned with the management of insect diversity in Thailand; and a list of exotic ant species found in Thailand, which could possibly impact the ecological balance

Seasonal dynamics and molecular differentiation of three natural Anopheles species (Diptera: Culicidae) of the Maculatus group (Neocellia series) in malaria hotspot villages of Thailand.

BACKGROUND: Anopheles sawadwongporni Rattanarithikul &amp; Green, Anopheles maculatus Theobald and Anopheles pseudowillmori (Theobald) of the Anopheles maculatus group (Diptera: Culicidae) are recognized as potential malaria vectors in many countries from the Indian subcontinent through Southeast Asia to Taiwan. A number of malaria vectors in malaria hotspot areas along the Thai-Myanmar border belong to this complex. However, the species distribution and dynamic trends remain understudied in this malaria endemic region. METHODS: Mosquitoes of the Maculatus group were collected using CDC light traps every other week from four villages in Tha Song Yang District, Tak Province, Thailand from January to December 2015. Adult female mosquitoes were morphologically identified on site using taxonomic keys. Molecular species identification was performed by multiplex PCR based on the internal transcribed spacer 2 (ITS2) region of ribosomal DNA (rDNA) and sequencing of the cox1 gene at a DNA barcoding region in a subset of 29 specimens. RESULTS: A total of 1328 An. maculatus (sensu lato) female mosquitoes were captured with An. maculatus, An. sawadwongporni and An. pseudowilmori accounting for 75.2, 22.1 and 2.7% respectively. The field captured mosquitoes of the Maculatus group were most abundant in the wet season and had a preferred distribution in villages at higher elevations. The phylogenetic relationships of 29 cox1 sequences showed a clear-cut separation of the three member species of the Maculatus group, with the An. pseudowillmori cluster being separated from An. sawadwongporni and An. maculatus. CONCLUSIONS: This study provides updated information for the species composition, seasonal dynamics and microgeographical distribution of the Maculatus group in malaria-endemic areas of western Thailand. This information can be used to guide the planning and implementation of mosquito control measures in the pursuance of malaria transmission