Vector Mosquito Anopheles Research Articles

Evaluation of Mosquito Magnet and other collection tools for Anopheles mosquito vectors of simian malaria

BackgroundVector surveillance is essential in determining the geographical distribution of mosquito vectors and understanding the dynamics of malaria transmission. With the elimination of human malaria cases, knowlesi malaria cases in humans are increasing in Malaysia. This necessitates intensive vector studies using safer trapping methods which are both field efficient and able to attract the local vector populations. Thus, this study evaluated the potential of Mosquito Magnet as a collection tool for Anopheles mosquito vectors of simian malaria along with other known collection methods.MethodsA randomized 4 × 4 Latin square designed experiment was conducted to compare the efficiency of the Mosquito Magnet against three other common trapping methods: human landing catch (HLC), CDC light trap and human baited trap (HBT). The experiment was conducted over six replicates where sampling within each replicate was carried out for 4 consecutive nights. An additional 4 nights of sampling was used to further evaluate the Mosquito Magnet against the “gold standard” HLC. The abundance of Anopheles sampled by different methods was compared and evaluated with focus on the Anopheles from the Leucosphyrus group, the vectors of knowlesi malaria.ResultsThe Latin square designed experiment showed HLC caught the greatest number of Anopheles mosquitoes (n = 321) compared to the HBT (n = 87), Mosquito Magnet (n = 58) and CDC light trap (n = 13). The GLMM analysis showed that the HLC method caught significantly more Anopheles mosquitoes compared to Mosquito Magnet (P = 0.049). However, there was no significant difference in mean nightly catch of Anopheles mosquitoes between Mosquito Magnet and the other two trapping methods, HBT (P = 0.646) and CDC light traps (P = 0.197). The mean nightly catch for both An. introlatus (9.33 ± 4.341) and An. cracens (4.00 ± 2.273) caught using HLC was higher than that of Mosquito Magnet, though the differences were not statistically significant (P > 0.05). This is in contrast to the mean nightly catch of An. sinensis (15.75 ± 5.640) and An. maculatus (15.78 ± 3.479) where HLC showed significantly more mosquito catches compared to Mosquito Magnet (P < 0.05).ConclusionsMosquito Magnet has a promising ability to catch An. introlatus and An. cracens, the important vectors of knowlesi and other simian malarias in Peninsular Malaysia. The ability of Mosquito Magnet to catch some of the Anopheles mosquito species is comparable to HLC and makes it an ethical and safer alternative.Graphical

Systematic identification of plausible pathways to potential harm via problem formulation for investigational releases of a population suppression gene drive to control the human malaria vector Anopheles gambiae in West Africa

BackgroundPopulation suppression gene drive has been proposed as a strategy for malaria vector control. A CRISPR-Cas9-based transgene homing at the doublesex locus (dsxFCRISPRh) has recently been shown to increase rapidly in frequency in, and suppress, caged laboratory populations of the malaria mosquito vector Anopheles gambiae. Here, problem formulation, an initial step in environmental risk assessment (ERA), was performed for simulated field releases of the dsxFCRISPRh transgene in West Africa.MethodsBuilding on consultative workshops in Africa that previously identified relevant environmental and health protection goals for ERA of gene drive in malaria vector control, 8 potentially harmful effects from these simulated releases were identified. These were stratified into 46 plausible pathways describing the causal chain of events that would be required for potential harms to occur. Risk hypotheses to interrogate critical steps in each pathway, and an analysis plan involving experiments, modelling and literature review to test each of those risk hypotheses, were developed.ResultsMost potential harms involved increased human (n = 13) or animal (n = 13) disease transmission, emphasizing the importance to subsequent stages of ERA of data on vectorial capacity comparing transgenics to non-transgenics. Although some of the pathways (n = 14) were based on known anatomical alterations in dsxFCRISPRh homozygotes, many could also be applicable to field releases of a range of other transgenic strains of mosquito (n = 18). In addition to population suppression of target organisms being an accepted outcome for existing vector control programmes, these investigations also revealed that the efficacy of population suppression caused by the dsxFCRISPRh transgene should itself directly affect most pathways (n = 35).ConclusionsModelling will play an essential role in subsequent stages of ERA by clarifying the dynamics of this relationship between population suppression and reduction in exposure to specific potential harms. This analysis represents a comprehensive identification of plausible pathways to potential harm using problem formulation for a specific gene drive transgene and organism, and a transparent communication tool that could inform future regulatory studies, guide subsequent stages of ERA, and stimulate further, broader engagement on the use of population suppression gene drive to control malaria vectors in West Africa.

Prevalence and seasonal trends of malaria at general hospital, Palanpur Banaskantha, Western part of India

Aim: Malaria is one of the known endemic disease in India. Malaria is transmitted by infected female anopheles mosquito vector transmitting mainly two Plasmodial species named P. falciparum and P. vivax. Prevalence of malaria is affected by atmospheric and socioeconomic condition of geographical area. Aim of the study is to know the prevalence of malaria and the seasonal trend of malaria at general hospital Palanpur, Banaskantha in western part of India. Material and Methods: This study was carried out at Central Laboratory, General Hospital, Palanpur, Banaskantha over period of July 2014 to June 2018. 41660 samples from fever patients were collected. We used Microscopic examination for the diagnosis of malaria by preparing thick and thin smears from these 41660 samples and stained using field stain. Results: We had collected total 41660 samples of fever patients. Out of 41660 samples, 688 samples were detected positive for malaria. So prevalence of malaria in our hospital was 1.65%. Prevalence of P. Vivax, P. Falciparum and mixed (infection of P. Falciparum and P. Vivax at a same time) were found 77.90%, 21.80% and 0.30% respectively. Malaria was seen relatively higher in males and affecting 15-30 years of age group more. Most cases of malaria were found in months of September and October near the end of monsoon and post monsoon season. Conclusion: From our study we concluded that there was high prevalence of P. vivax than P. Falciparum infection. Maximum numbers of cases were after major rainfall, reported in month of September and October. Morbidity and mortality of malaria must be reduced by strengthening Malaria Surveillance, risk factor assessment during pre and postmonsoon period. Premonsoon preventive actions need strengthening and strict monitoring of all these actions are needed. Keywords: Prevalence, Plasmodium vivax, Plasmodium falciparum, Malaria, Thin and Thick smear, Seasonal trends.

Implications of increasing temperature stress for predatory biocontrol of vector mosquitoes

BackgroundPredators play a critical role in regulating larval mosquito prey populations in aquatic habitats. Understanding predator-prey responses to climate change-induced environmental perturbations may foster optimal efficacy in vector reduction. However, organisms may differentially respond to heterogeneous thermal environments, potentially destabilizing predator-prey trophic systems.MethodsHere, we explored the critical thermal limits of activity (CTLs; critical thermal-maxima [CTmax] and minima [CTmin]) of key predator-prey species. We concurrently examined CTL asynchrony of two notonectid predators (Anisops sardea and Enithares chinai) and one copepod predator (Lovenula falcifera) as well as larvae of three vector mosquito species, Aedes aegypti, Anopheles quadriannulatus and Culex pipiens, across instar stages (early, 1st; intermediate, 2nd/3rd; late, 4th).ResultsOverall, predators and prey differed significantly in CTmax and CTmin. Predators generally had lower CTLs than mosquito prey, dependent on prey instar stage and species, with first instars having the lowest CTmax (lowest warm tolerance), but also the lowest CTmin (highest cold tolerance). For predators, L. falcifera exhibited the narrowest CTLs overall, with E. chinai having the widest and A. sardea intermediate CTLs, respectively. Among prey species, the global invader Ae. aegypti consistently exhibited the highest CTmax, whilst differences among CTmin were inconsistent among prey species according to instar stage.ConclusionThese results point to significant predator-prey mismatches under environmental change, potentially adversely affecting natural mosquito biocontrol given projected shifts in temperature fluctuations in the study region. The overall narrower thermal breadth of native predators relative to larval mosquito prey may reduce natural biotic resistance to pests and harmful mosquito species, with implications for population success and potentially vector capacity under global change.Graphical

A study of malaria vector surveillance as part of the Malaria Elimination Demonstration Project in Mandla, Madhya Pradesh

BackgroundUnderstanding of malaria vector density, distribution, insecticide resistance, vector incrimination, infection status, and identification of sibling species are some of the essential components of vector control measures for achieving malaria elimination goals.MethodsAs part of the malaria elimination demonstration project, entomological surveillance was carried out from October 2017 to October 2019 by collecting indoor resting mosquitoes using hand catch method. Susceptibility test was done for determining the insecticide resistance status of vector mosquito Anopheles culicifacies using standard protocols by the World Health Organization. The cone bioassay method was used for determining the efficacy and quality of insecticide sprayed. Mosquitoes collected from different ecotypes were identified and processed for parasite identification, vector incrimination and sibling species determination.ResultsThe two known malaria vector species (Anopheles culicifacies and Anopheles fluviatilis) were found in the study area, which have been previously reported in this and adjoining areas of the State of Madhya Pradesh. The prevalence of An. culicifacies was significantly higher in all study villages with peak in July while lowest number was recorded in May. Proportion of vector density was observed to be low in foothill terrains. The other anopheline species viz, Anopheles subpictus, Anopheles annularis, Anopheles vagus, Anopheles splendidus, Anopheles pallidus, Anopheles nigerrimus and Anopheles barbirostris were also recorded in the study area, although their prevalence was significantly less compared to the An. culicifacies. In 2017, An. culicifacies was found to be resistant to dichloro-diphenyl-trichloroethane (DDT) and malathion, with possible resistance to alphacypermethrin and susceptible to deltamethrin. However, in 2019, the species was found to be resistant to alphacypermethrin, DDT, malathion, with possible resistance to deltamethrin. The bioassays revealed 82 to > 98% corrected % mortality of An. culicifacies on day-one post-spraying and 35 to 62% on follow-up day-30. Anopheles culicifacies sibling species C was most prevalent (38.5%) followed by A/D and E while B was least pre-dominant (11.9%). Anopheles fluviatilis sibling species T was most prevalent (74.6%) followed by U (25.4%) while species S was not recorded. One An.culicifacies (sibling species C) was found positive for Plasmodium falciparum by PCR tests in the mosquitoes sampled from the test areas.ConclusionBased on the nine entomologic investigations conducted between 2017–2019, it was concluded that An. culicifacies was present throughout the year while An. fluviatilis had seasonal presence in the study areas. Anopheles culicifacies was resistant to alphacypermethrin and emerging resistance to deltamethrin was observed in this area. Anopheles culicifacies was confirmed as the malaria vector. This type of information on indigenous malaria vectors and insecticide resistance is important in implementation of vector control through indoor residual spraying (IRS) and use of insecticide-impregnated bed nets for achieving the malaria elimination goals.

Gene editing reveals obligate and modulatory components of the CO2 receptor complex in the malaria vector mosquito, Anopheles coluzzii

The sensitivity to volatile carbon dioxide (CO2) produced by humans and other animals is a critical component in the host preference behaviors of the malaria vector mosquito Anopheles coluzzii. The molecular receptors responsible for the ability to sense CO2 are encoded by three putative gustatory receptor (Gr) genes (Gr22,23,24) which are expressed in a distinctive array of sensory neurons housed in maxillary palp capitate peg sensilla of An. coluzzii. Despite the identification of these components and subsequent studies, there is a paucity of understanding regarding the respective roles of these three GRs in the mosquito's CO2 transduction process. To address this, we have used CRISPR/Cas9-based gene editing technique combined with in vivo electrophysiological recordings to directly examine the role of Gr22,23,24 in detecting CO2 in An. coluzzii. These studies reveal that both Gr23 and Gr24 are absolutely required to maintain in vivo CO2 sensitivity while, in contrast, Gr22 knock out mutants are still able to respond to CO2 stimuli albeit with significantly weaker sensitivity. Our data supports a model in which Gr22 plays a modulatory role to enhance the functionality of Gr23/24 complexes that are responsible for CO2 sensitivity of mosquitoes.

Natural Products: A Potential Source of Malaria Transmission Blocking Drugs?

The ability to block human-to-mosquito and mosquito-to-human transmission of Plasmodium parasites is fundamental to accomplish the ambitious goal of malaria elimination. The WHO currently recommends only primaquine as a transmission-blocking drug but its use is severely restricted by toxicity in some populations. New, safe and clinically effective transmission-blocking drugs therefore need to be discovered. While natural products have been extensively investigated for the development of chemotherapeutic antimalarial agents, their potential use as transmission-blocking drugs is comparatively poorly explored. Here, we provide a comprehensive summary of the activities of natural products (and their derivatives) of plant and microbial origins against sexual stages of Plasmodium parasites and the Anopheles mosquito vector. We identify the prevailing challenges and opportunities and suggest how these can be mitigated and/or exploited in an endeavor to expedite transmission-blocking drug discovery efforts from natural products.

Evaluation of Actin-1 Expression in Wild Caught Wuchereria bancrofti-Infected Mosquito Vectors.

Background Wuchereria bancrofti is the major cause of lymphatic filariasis transmitted by mosquito vectors. In the vector-parasite interaction and among other proteins, actin-1 has been implicated for successful transmission of the pathogen in laboratory-controlled experiments. However, validation of this finding from the pathogen's natural environment is required. Objective This study is aimed at evaluating actin-1 expression upon Wuchereria bancrofti infection in mosquito vectors collected during an epidemiology study in Tsafe Local Government Area of Zamfara State, Nigeria. Methods Mosquitoes were collected and identified using morphological keys, which include length of maxillary palps, pale spots on the wings, and scale patterns on the abdomen. This was followed by detection of the 188 bp SspI marker of Wuchereria bancrofti infection using polymerase chain reaction (PCR). The mRNA levels of the actin-1 gene were evaluated in the infected Anopheles gambiae sl and Culex quinquefasciatus and their controls, which were adult reared from the larvae in the study area. Results The mosquitoes were identified to be Anopheles gambiae sl and Culex quinquefasciatus, while infection by Wuchereria bancrofti was confirmed by amplification of the 188 bp SspI marker. A 4.85 and 4.09 relative fold increase in actin-1 gene expression in Wuchereria bancrofti-infected Anopheles gambiae sl and Culex quinquefasciatus was observed. Thus, for the first time we reported that the actin-1 gene in wild caught mosquito vectors (Anopheles gambiae sl and Culex quinquefasciatus) infected with Wuchereria bancrofti is upregulated. Conclusion The actin-1 gene is upregulated and similarly expressed during W. bancrofti infection in mosquito vectors in the study area and this may likely serve as a biomarker and viable strategy for the control of parasite transmission in endemic areas.

Heterogeneous expression of the ammonium transporter AgAmt in chemosensory appendages of the malaria vector, Anopheles gambiae

Ammonia is one of the principal kairomones originating from human and other animal emanations and in that context, plays an essential role in the host-seeking behaviors of the malaria vector mosquito Anopheles gambiae. Nevertheless, despite its importance in directing host-seeking, the mechanisms underlying ammonia detection in the mosquito olfactory system remains largely unknown. In addition to ongoing efforts to identify and characterize the molecular receptors that underlie ammonia sensitivity, previous studies have revealed a prominent role for ammonium transporters (Amt) in modulating antennal and behavioral responses in Drosophila melanogaster and An. gambiae. In the former, localization of DmAmt in antennal sensilla to auxiliary cells surrounding the ammonia sensory neurons led to the hypothesis that its role was to clear excess ammonium ions in the sensillar lymph. In the latter, RT-PCR and heterologous expression have been used to examine the expression and functional characteristics of the An. gambiae ammonium transporter, AgAmt. We now employ advanced transgenic tools to comprehensively examine AgAmt spatial localization across the peripheral chemosensory appendages in larvae and adult female An. gambiae. In the larval antennae, AgAmt appears localized in both neuronal and auxiliary cells. In contrast to D. melanogaster, in the adult antennae, AgAmt-derived signals are observed in both non-neuronal auxiliary cells and in sensory neurons in ammonia-responsive basiconic and coeloconic sensilla. In the maxillary palps, labella, and tarsi, AgAmt appears restricted to sensory neurons. We have also characterized the responses to ammonia of adult antennal coeloconic sensilla and maxillary palp capitate pegs revealing a correlation between sensillar AgAmt expression and ammonia sensitivity. Taken together, these data suggest that AgAmt may play heterogeneous roles in the adult and larval chemosensory apparatus and potentially broad utility as a supra-receptor target in mosquito control.

Field assessment of 4-hydroxycoumarin as an attractant for anthropophilic Anopheles spp. vectors of malaria in Madagascar

Mosquito-borne diseases like malaria are a major public health problem in tropical countries, such as Madagascar. Female Anopheles mosquito vectors the human malaria parasites (Plasmodium spp.) and is important indicator in malaria surveillance activities. Among the various means of vector control in Madagascar, the use of attractants for mass trapping of target species could be an alternative to insecticides. The aim of this study is to evaluate whether 4-hydroxycoumarin can be used as an attractant for anthropophilic Anopheles spp. vectors of malaria. For this, a field study was conducted using CDC light traps in the village of Ambohidray, Madagascar. 16 days of trapping was conducted and four replicates nights were performed for each product tested. 4-hydroxycoumarin, octenol and two types of blend of these products were tested. The results showed that 4-hydroxycoumarin (2 mg) have a significant attractive effect on Anopheles spp. and significant selectivity towards Anopheles gambiae s.l, and Anopheles mascarensis which are both significant malaria vectors in Madagascar. A synergy of 4-hydroxycoumarin with octenol was found to attract these mosquito vectors. A significant decrease in vector populations was observed during this experiment. These results suggest that 4-hydroxycoumarin could be useful for malaria surveillance and the control of vector populations.

Digital droplet PCR and IDAA for the detection of CRISPR indel edits in the malaria species Anopheles stephensi.

CRISPR/Cas9 technology is a powerful tool for the design of gene-drive systems to control and/or modify mosquito vector populations; however, CRISPR/Cas9-mediated nonhomologous end joining mutations can have an important impact on generating alleles resistant to the drive and thus on drive efficiency. We demonstrate and compare the insertions or deletions (indels) detection capabilities of two techniques in the malaria vector mosquito Anopheles stephensi: Indel Detection by Amplicon Analysis (IDAA™) and Droplet Digital™ PCR (ddPCR™). Both techniques showed accuracy and reproducibility for indel frequencies across mosquito samples containing different ratios of indels of various sizes. Moreover, these techniques have advantages that make them potentially better suited for high-throughput nonhomologous end joining analysis in cage trials and contained field testing of gene-drive mosquitoes.

Thioester-containing Proteins in the Drosophila melanogaster Immune Response against the Pathogen Photorhabdus.

The fruit fly Drosophila melanogaster forms a magnificent model for interpreting conserved host innate immune signaling and functional processes in response to microbial assaults. In the broad research field of host-microbe interactions, model hosts are used in conjunction with a variety of pathogenic microorganisms to disentangle host immune system activities and microbial pathogenicity strategies. The pathogen Photorhabdus is considered an established model for analyzing bacterial virulence and symbiosis due to its unique life cycle that extends between two invertebrate hosts: an insect and a parasitic nematode. In recent years, particular focus has been given to the mechanistic participation of the D. melanogaster thioester-containing proteins (TEPs) in the overall immune capacity of the fly upon response against the pathogen Photorhabdus alone or in combination with its specific nematode vector Heterorhabditis bacteriophora. The original role of certain TEPs in the insect innate immune machinery was linked to the antibacterial and antiparasite reaction of the mosquito malaria vector Anopheles gambiae; however, revamped interest in the immune competence of these molecules has recently emerged from the D. melanogaster-Photorhabdus infection system. Here, we review the latest findings on this topic with the expectation that such information will refine our understanding of the evolutionary immune role of TEPs in host immune surveillance.

Counting and Classification of Malarial Parasite From Giemsa-Stained Thin Film Images

Malaria is a life-threatening disease causing by an infection of the protozoan parasite Plasmodium . Plasmodium falciparum is the deadliest and most common human infected parasites hosted by anopheles mosquito vector. To cure a malaria infected patient and prevent further spreading, malaria diagnosis using microscopy to visualize Giemsa-stained parasites is commonly done. The microscopy diagnosis is somewhat time consuming and requires well-trained malaria experts to interpret what they see under the microscope. To address this limitation, an automated malaria infected diagnosis is needed. This work proposed a computer-aided automated diagnosis system that can perform remote field diagnosis with high accuracy while requiring less computational demands. The proposed framework consists of two main parts that are red blood cell counting and parasite life-cycle stage classification. The counting process is performed by computer vision techniques, namely Hough transform. Different machine learning techniques, i.e., Multilayer Perceptron, Linear Discriminant Analysis, Support Vector Machine, and Weighted Similarity Extreme Learning Machine, are employed in the classification task. We also demonstrated that combining hand-crafted and deep-learned features can enhance the overall performance of the framework. The experimental results showed that the proposed methods could correctly count and classify at 97.94% and 98.12% accuracy, respectively. The overall proposal system can achieve at 96.18% accuracy. This is achieved by WELM in conjunction with deep-learned (AlexNet_FC7) and the hand-crafted (color) features.

Barrier bednets target malaria vectors and expand the range of usable insecticides

Transmission of Plasmodium falciparum malaria parasites occurs when nocturnal Anopheles mosquito vectors feed on human blood. In Africa, where malaria burden is highest, bednets treated with pyrethroid insecticide were highly effective in preventing mosquito bites and reducing transmission, and essential to achieving unprecedented reductions in malaria until 2015 (ref. 1). Since then, progress has stalled2, and with insecticidal bednets losing efficacy against pyrethroid-resistant Anopheles vectors3,4, methods that restore performance are urgently needed to eliminate any risk of malaria returning to the levels seen before their widespread use throughout sub-Saharan Africa5. Here, we show that the primary malaria vector Anopheles gambiae is targeted and killed by small insecticidal net barriers positioned above a standard bednet in a spatial region of high mosquito activity but zero contact with sleepers, opening the way for deploying many more insecticides on bednets than is currently possible. Tested against wild pyrethroid-resistant A. gambiae in Burkina Faso, pyrethroid bednets with organophosphate barriers achieved significantly higher killing rates than bednets alone. Treated barriers on untreated bednets were equally effective, without significant loss of personal protection. Mathematical modelling of transmission dynamics predicted reductions in clinical malaria incidence with barrier bednets that matched those of 'next-generation' nets recommended by the World Health Organization against resistant vectors. Mathematical models of mosquito-barrier interactions identified alternative barrier designs to increase performance. Barrier bednets that overcome insecticide resistance are feasible using existing insecticides and production technology, and early implementation of affordable vector control tools is a realistic prospect.

Gene silencing through RNAi and antisense Vivo-Morpholino increases the efficacy of pyrethroids on larvae of Anopheles stephensi

BackgroundInsecticides are still at the core of insect pest and vector control programmes. Several lines of evidence indicate that ABC transporters are involved in detoxification processes against insecticides, including permethrin and other pyrethroids. In particular, the ABCG4 gene, a member of the G subfamily, has consistently been shown to be up-regulated in response to insecticide treatments in the mosquito malaria vector Anopheles stephensi (both adults and larvae).MethodsTo verify the actual involvement of this transmembrane protein in the detoxification process of permethrin, bioassays on larvae of An. stephensi, combining the insecticide with a siRNA, specifically designed for the inhibition of ABCG4 gene expression were performed. Administration to larvae of the same siRNA, labeled with a fluorescent molecule, was effected to investigate the systemic distribution of the inhibitory RNA into the larval bodies. Based on siRNA results, similar experiments using antisense Vivo-Morpholinos (Vivo-MOs) were effected. These molecules, compared to siRNA, are expected to guarantee a higher stability in environmental conditions and in the insect gut, and present thus a higher potential for future in-field applications.ResultsBioassays using two different concentrations of siRNA, associated with permethrin, led to an increase of larval mortality, compared with results with permethrin alone. These outcomes confirm that ABCG4 transporter plays a role in the detoxification process against the selected insecticide. Moreover, after fluorescent labelling, it was shown the systemic dissemination of siRNA in different body districts of An. stephensi larvae, which suggest a potential systemic effect of the molecule. At the same time, results of Vivo-MO experiments were congruent with those obtained using siRNA, thus confirming the potential of ABCG4 inhibition as a strategy to increase permethrin susceptibility in mosquitoes. For the first time, Vivo-MOs were administered in water to larvae, with evidence for a biological effect.ConclusionsTargeting ABCG4 gene for silencing through both techniques resulted in an increased pyrethroid efficacy. These results open the way toward the possibility to exploit ABCG4 inhibition in the context of integrated programmes for the control An. stephensi mosquitoes and malaria transmission.

Recent Incidence of Human Malaria Caused by Plasmodium knowlesi in the Villages in Kudat Peninsula, Sabah, Malaysia: Mapping of The Infection Risk Using Remote Sensing Data

Plasmodium knowlesi (Pk) is a malaria parasite that naturally infects macaque monkeys in Southeast Asia. Pk malaria, the zoonosis transmitted from the infected monkeys to the humans by Anopheles mosquito vectors, is now a serious health problem in Malaysian Borneo. To create a strategic plan to control Pk malaria, it is important to estimate the occurrence of the disease correctly. The rise of Pk malaria has been explained as being due to ecological changes, especially deforestation. In this research, we analysed the time-series satellite images of MODIS (MODerate-resolution Imaging Spectroradiometer) of the Kudat Peninsula in Sabah and created the “Pk risk map” on which the Land-Use and Land-Cover (LULC) information was visualised. The case number of Pk malaria of a village appeared to have a correlation with the quantity of two specific LULC classes, the mosaic landscape of oil palm groves and the nearby land-use patches of dense forest, surrounding the village. Applying a Poisson multivariate regression with a generalised linear mixture model (GLMM), the occurrence of Pk malaria cases was estimated from the population and the quantified LULC distribution on the map. The obtained estimations explained the real case numbers well, when the contribution of another risk factor, possibly the occupation of the villagers, is considered. This implies that the occurrence of the Pk malaria cases of a village can be predictable from the population of the village and the LULC distribution shown around it on the map. The Pk risk map will help to assess the Pk malaria risk distributions quantitatively and to discover the hidden key factors behind the spread of this zoonosis.

Killer yeasts exert anti-plasmodial activities against the malaria parasite Plasmodium berghei in the vector mosquito Anopheles stephensi and in mice

BackgroundWickerhamomyces anomalus is a yeast associated with different insects including mosquitoes, where it is proposed to be involved in symbiotic relationships with hosts. Different symbiotic strains of W. anomalus display a killer phenotype mediated by protein toxins with broad-spectrum antimicrobial activities. In particular, a killer toxin purified from a W. anomalus strain (WaF17.12), previously isolated from the malaria vector mosquito Anopheles stephensi, has shown strong in vitro anti-plasmodial activity against early sporogonic stages of the murine malaria parasite Plasmodium berghei.ResultsHere, we provide evidence that WaF17.12 cultures, properly stimulated to induce the expression of the killer toxin, can directly affect in vitro P. berghei early sporogonic stages, causing membrane damage and parasite death. Moreover, we demonstrated by in vivo studies that mosquito dietary supplementation with activated WaF17.12 cells interfere with ookinete development in the midgut of An. stephensi. Besides the anti-sporogonic action of WaF17.12, an inhibitory effect of purified WaF17.12-killer toxin was observed on erythrocytic stages of P. berghei, with a consequent reduction of parasitaemia in mice. The preliminary safety tests on murine cell lines showed no side effects.ConclusionsOur findings demonstrate the anti-plasmodial activity of WaF17.12 against different developmental stages of P. berghei. New studies on P. falciparum are needed to evaluate the use of killer yeasts as innovative tools in the symbiotic control of malaria.

A genetic system for biasing the sex ratio in mice.

Biasing the sex ratio of populations of different organisms, including plants, insects, crustacean, and fish, has been demonstrated by genetic and non-genetic approaches. However, biasing the sex ratio of mammalian populations has not been demonstrated genetically. Here, we provide a first proof of concept for such a genetic system in mammals by crossing two genetically engineered mouse lines. The maternal line encodes a functional Cas9 protein on an autosomal chromosome, whereas the paternal line encodes guide RNAs on the Y chromosome targeting vital mouse genes. After fertilization, the presence of both the Y-encoded guide RNAs from the paternal sperm and the Cas9 protein from the maternal egg targets the vital genes in males. We show that these genes are specifically targeted in males and that this breeding consequently self-destructs solely males. Our results pave the way for a genetic system that allows biased sex production of livestock.

Phenotypic Analysis of Rodent Malaria Parasite Asexual and Sexual Blood Stages and Mosquito Stages.

Recent advances in genetics and systems biology technologies have promoted our understanding of the biology of malaria parasites on the molecular level. However, effective malaria parasite targets for vaccine and chemotherapy development are still limited. This is largely due to the unavailability of relevant and practical in vivo infection models for human Plasmodium species, most notably for P.falciparum and P. vivax. Therefore, rodent malaria species have been extensively used as practical alternative in vivo models for malaria vaccine, drug targeting, immune response, and functional characterization studies of conserved Plasmodiumspp. genes. Indeed, rodent malaria models have proven to be invaluable, especially for exploring mosquito transmission and liver stage biology, and were indispensable for immunological studies. However, there are discrepancies in the methods used to evaluate the phenotypes of transgenic and wild-type asexual and sexual blood-stage parasites.Examples of these discrepancies are the choice of an intravenous vs. intraperitoneal infection of rodents with blood-stage parasites and the evaluation of male gamete exflagellation. Herein, we detail standardized experimental methods to evaluate the phenotypes of asexual and sexual blood stages in transgenic parasites expressing reporter-gene or wild-type rodent malaria parasite species. We also detail the methods to evaluate the phenotypes of malaria parasite mosquito stages (gametes, ookinetes, oocysts, and sporozoites) inside Anopheles mosquito vectors. These methods are detailed and simplified here for the lethal and non-lethal strains of P. berghei and P. yoelii but can also be applied with some adjustments to P. chabaudi and P. vinckei rodent malaria species.

Oasis Malaria, Northern Mauritania1.

A malaria survey was conducted in Atar, the northernmost oasis city in Mauritania, during 2015–2016. All febrile patients in whom malaria was suspected were screened for malaria by using rapid diagnostic testing and microscopic examination of blood smears and later confirmed by PCR. Of 453 suspected malaria cases, 108 (23.8%) were positive by rapid diagnostic testing, 154 (34.0%) by microscopic examination, and 162 (35.7%) by PCR. Malaria cases were observed throughout the year and among all age groups. Plasmodium vivax was present in 120/162 (74.1%) cases, P. falciparum in 4/162 (2.4%), and mixed P. falciparum–P. vivax in 38/162 (23.4%). Malaria is endemic in northern Mauritania and could be spreading farther north in the Sahara, possibly because of human-driven environmental changes. Further entomologic and parasitologic studies and monitoring are needed to relate these findings to major Anopheles mosquito vectors and to design and implement strategies for malaria prevention and control.