Higher risk of malaria transmission outdoors than indoors by Nyssorhynchus darlingi in riverine communities in the Peruvian AmazonParasites and Vectors - Tập 12 Số 1 - 2019
Marlon P. Saavedra, Jan E. Conn, Freddy Alava, Gabriel Carrasco-Escobar, Catharine Prussing, Sara A. Bickersmith, Jorge L. Sangama, Carlos Fernández-Miñope, Mitchel Guzmán, Carlos Tong, Carlos Valderrama, Joseph M. Vinetz, Dionicia Gamboa, Marta Moreno
Identification of Aedes mosquitoes by MALDI-TOF MS biotyping using protein signatures from larval and pupal exuviaeParasites and Vectors - Tập 13 Số 1 - 2020
Amira Nebbak, Lionel Alméras
Abstract
Background
Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) biotyping is an innovative strategy, applied successfully for the identification of numerous arthropod families including mosquitoes. The effective mosquito identification using this emerging tool was demonstrated possible at different steps of their life-cycle, including eggs, immature and adult stages. Unfortunately, for species identification by MS, the euthanasia of the mosquito specimen is required.
Methods
To avoid mosquito euthanasia, the present study assessed whether aedine mosquitoes could be identified by MALDI-TOF MS biotyping, using their respective exuviae. In this way, exuviae from the fourth-instar and pupal stages of Aedes albopictus and Aedes aegypti were submitted to MALDI-TOF MS analysis.
Results
Reproducible and specific MS spectra according to aedine species and stage of exuviae were observed which were objectified by cluster analyses, composite correlation index (CCI) tool and principal components analysis (PCA). The query of our reference MS spectra database (DB) upgraded with MS spectra of exuviae from fourth-instar larvae and pupae of both Aedes species revealed that 100% of the samples were correctly classified at the species and stage levels. Among them, 93.8% (135/144) of the MS profiles reached the threshold log score value (LSV > 1.8) for reliable identification.
Conclusions
The extension of reference MS spectra DB to exuviae from fourth-instar and pupal stages made now possible the identification of mosquitoes throughout their life-cycle at aquatic and aerial stages. The exuviae presenting the advantage to avoid specimen euthanasia, allowing to perform complementary analysis on alive mosquitoes.
Development and evaluation of a novel contamination device that targets multiple life-stages of Aedes aegyptiParasites and Vectors - Tập 7 - Trang 1-10 - 2014
Janneke Snetselaar, Rob Andriessen, Remco A Suer, Anne J Osinga, Bart GJ Knols, Marit Farenhorst
The increasing global threat of Dengue demands new and easily applicable vector control methods. Ovitraps provide a low-tech and inexpensive means to combat Dengue vectors. Here we describe the development and optimization process of a novel contamination device that targets multiple life-stages of the Aedes aegypti mosquito. Special focus is directed to the diverse array of control agents deployed in this trap, covering adulticidal, larvicidal and autodissemination impacts. Different trap prototypes and their parts are described, including a floater to contaminate alighting gravid mosquitoes. The attractiveness of the trap, different odor lures and floater design were studied using fluorescent powder adhering to mosquito legs and via choice tests. We demonstrate the mosquitocidal impacts of the control agents: a combination of the larvicide pyriproxyfen and the adulticidal fungus Beauveria bassiana. The impact of pyriproxyfen was determined in free-flight dissemination experiments. The effect on larval development inside the trap and in surrounding breeding sites was measured, as well as survival impacts on recaptured adults. The developmental process resulted in a design that consists of a black 3 Liter water-filled container with a ring-shaped floater supporting vertically placed gauze dusted with the control agents. On average, 90% of the mosquitoes in the fluorescence experiments made contact with the gauze on the floater. Studies on attractants indicated that a yeast-containing tablet was the most attractive odor lure. Furthermore, the fungus Beauveria bassiana was able to significantly increase mortality of the free-flying adults compared to controls. Dissemination of pyriproxyfen led to >90% larval mortality in alternative breeding sites and 100% larval mortality in the trap itself, against a control mortality of around 5%. This ovitrap is a promising new tool in the battle against Dengue. It has proven to be attractive to Aedes aegypti mosquitoes and effective in contaminating these with Beauveria bassiana. Furthermore, we show that the larvicide pyriproxyfen is successfully disseminated to breeding sites close to the trap. Its low production and operating costs enable large scale deployment in Dengue-affected locations.
Simultaneous external and internal marking of Triatoma sordida nymphs: trace element efficacy and microgeographic dispersal in a peridomestic Brazilian Cerrado rural householdParasites and Vectors -
Edson Santos Dantas, Rodrigo Gurgel-Gonçalves, Rafael Maciel‐de‐Freitas, Fernando A. Monteiro
Abstract
Background
Chagas disease (American trypanosomiasis) is an important neglected tropical illness, which has the flagellate protozoan Trypanosoma cruzi as etiological agent and blood-feeding insects of the Triatominae subfamily as vectors. Despite its importance for disease epidemiology, field studies targeting microgeographic dispersal of triatomines in endemic areas are rare. The ability wingless nymphs have to move (crawl) within peridomestic settings is a key component regarding the design and development of rational control strategies.
Methods
We double-marked Triatoma sordida fourth-instar nymphs (N4) with a reliable fluorescent dye and a trace element. This new methodology allowed us to simultaneously evaluate (i) nymph dispersal and (ii) the effectiveness of copper (Cu), chromium (Cr), and cadmium (Cd) trace elements as potential new markers. In the mark-release-recapture (MRR) experiment, 390 T. sordida N4 were released in the peridomicile of a single rural household, 130 individuals at each of three release points, at distances of 2, 5, and 10 m from the chicken coop (CC) and 27, 32, and 35 m away from the horse corral (HC). All specimens were double marked (Cu/blue, Cr/orange, Cd/green). Recaptures occurred in two intervals: 1–3 days and 15–17 days after release.
Results
Specimens were successfully recaptured at all distances up to 10 m. A total of 19, 23, and 10 specimens were able to disperse 2, 5, and 10 m, respectively, to reach the CC. No insects were recaptured at the HC. Of the three analyte/paint combinations tested, Cr/orange gave the most promising results; Cu/blue marker and Cd/green marker performed very poorly with only 4/19 and 0/10 analyte/paint ratios, respectively.
Conclusions
Triatoma sordida N4 could cover a distance of 10 m in 17 days. This indicates that nymphs seem to have a reduced dispersal capability compared to adults. Ninety-one percent of the 22 recaptured orange-marked nymphs were still Cr positive after the 17-day period evaluated. This makes this analyte a good candidate for future investigations that will apply this marking method in MRR studies.
Graphical Abstract
Good performances but short lasting efficacy of Actellic 50 EC Indoor Residual Spraying (IRS) on malaria transmission in Benin, West AfricaParasites and Vectors - Tập 7 - Trang 1-8 - 2014
Rock Aïkpon, Michel Sèzonlin, Filémon Tokponon, Mariam Okè, Olivier Oussou, Frédéric Oké-Agbo, Raymond Beach, Martin Akogbéto
The National Malaria Control Program (NMCP) has been using pirimiphos methyl for the first time for indoor residual spraying (IRS) in Benin. The first round was a success with a significant decrease of entomological indicators of malaria transmission in the treated districts. We present the results of the entomological impact on malaria transmission. Entomologic parameters in the control area were compared with those in intervention sites. Mosquito collections were carried out in three districts in the Atacora-Dongo region of which two were treated with pirimiphos methyl (Actellic 50EC) (Tanguiéta and Kouandé) and the untreated (Copargo) served as control. Anopheles gambiae s.l. populations were sampled monthly by human landing catch. In addition, window exit traps and pyrethrum spray catches were performed to assess exophagic behavior of Anopheles vectors. In the three districts, mosquito collections were organized to follow the impact of pirimiphos methyl IRS on malaria transmission and possible changes in the behavior of mosquitoes. The residual activity of pirimiphos methyl in the treated walls was also assessed using WHO bioassay test. A significant reduction (94.25%) in human biting rate was recorded in treated districts where an inhabitant received less than 1 bite of An. gambiae per night. During this same time, the entomological inoculation rate (EIR) dramatically declined in the treated area (99.24% reduction). We also noted a significant reduction in longevity of the vectors and an increase in exophily induced by pirimiphos methyl on An. gambiae. However, no significant impact was found on the blood feeding rate. Otherwise, the low residual activity of Actellic 50 EC, which is three months, is a disadvantage. Pirimiphos methyl was found to be effective for IRS in Benin. However, because of the low persistence of Actellic 50EC used in this study on the treated walls, the recourse to another more residual formulation of pirimiphos methyl is required.
Assessment of malaria transmission intensity and insecticide resistance mechanisms in three rural areas of the Moyen Ogooué Province of GabonParasites and Vectors - - 2022
Stravensky Térence Boussougou-Sambe, Tamirat Gebru Woldearegai, Ange Gatien Doumba-Ndalembouly, Barclaye Ngossanga, Romuald Beh, Jean Ronald Edoa, Jeannot Fréjus Zinsou, Yabo Josiane Honkpéhèdji, Ulysse Ateba Ngoa, Jean Claude Dejon‐Agobé, Steffen Borrmann, Peter G. Kremsner, Benjamin Mordmüller, Ayôla Akim Adegnika
Abstract
Background
Vector control is considered to be the most successful component of malaria prevention programs and a major contributor to the reduction of malaria incidence over the last two decades. However, the success of this strategy is threatened by the development of resistance to insecticides and behavioural adaptations of vectors. The aim of this study was to monitor malaria transmission and the distribution of insecticide resistance genes in Anopheles populations from three rural areas of the Moyen Ogooué Province of Gabon.
Methods
Anopheles spp. were collected using human landing catches in Bindo, Nombakélé and Zilé, three villages located in the surroundings of Lambaréné, during both the rainy and dry seasons. Mosquitoes were identified morphologically, and DNA was extracted from heads and thoraces. Members of the Anopheles gambiae complex were identified by molecular methods using the PCR SINE200 protocol and by sequencing of the internal transcribed spacer 2 region. Taqman assays were used to determine Plasmodium infection and the presence of resistance alleles.
Results
Anopheles gambiae sensu lato (97.7%), An. moucheti (1.7%) and An. coustani (0.6%) were the three groups of species collected. Anopheles gambiae sensu stricto (98.5%) and An. coluzzii (1.5%) were the only species of the An. gambiae complex present in the collection. Of the 1235 Anopheles collected, 1193 were collected during the rainy season; these exhibited an exophagic behaviour, and consistently more mosquitoes were collected outdoor than indoor in the three study areas. Of the 1166 Anopheles screened, 26 (2.2%) were infected with Plasmodium species, specifically Plasmodium falciparum (66.7%), P. malariae (15.4%), P. ovale curtisi (11.5%) and P. ovale wallikeri (3.8%). Malaria transmission intensity was high in Zilé, with an average annual entomological inoculation rate (aEIR) of 243 infective bites per year, while aEIRs in Bindo and Nombakélé were 80.2 and 17 infective bites per year, respectively. Both the L1014F and L1014S mutations were present at frequencies > 95% but no Ace1G119S mutation was found.
Conclusion
Our results demonstrate that malaria transmission intensity is heterogeneous in these three rural areas of Moyen Ogooué Province, with areas of high transmission, such as Zilé. The exophagic behaviour of the mosquitoes as well as the high frequency of resistance mutations are serious challenges that need to be addressed by the deployment of control measures adapted to the local setting.
Graphical Abstract
Differences in timing of mating swarms in sympatric populations of Anopheles coluzzii and Anopheles gambiae s.s. (formerly An. gambiae M and S molecular forms) in Burkina Faso, West AfricaParasites and Vectors - Tập 6 - Trang 1-14 - 2013
Simon P Sawadogo, Carlo Costantini, Cédric Pennetier, Abdoulaye Diabaté, Gabriella Gibson, Roch K Dabiré
The M and S molecular forms of Anopheles gambiae s.s. Giles appear to have speciated in West Africa and the M form is now formally named An. coluzzii Coetzee & Wilkerson sp.n. and the S form retains the nominotypical name (abbreviated here to An. gambiae). Reproductive isolation is thought to be the main barrier to hybridisation; even though both species are found in the same mating swarms, hybrid fertilisations in copulae have not been found in the study area. The aim of the study, therefore, was to determine whether differences in circadian and/or environmental control over the timing of swarming in the two species contribute to reproductive isolation. The timing of male swarming in these species was recorded four nights per month over four years at five swarming sites in each of two villages. The timing of the start and end of swarming, and the concurrent environmental parameters, temperature, humidity and light intensity, were recorded for n = 20 swarms/month/species. The timing of 'spontaneous’ activity at dusk of individual An. coluzzii and An. gambiae males was video-recorded in an actograph outdoors for 21 nights. Of the environmental parameters considered, swarming was most strongly correlated with sunset (r2 > 0.946). Anopheles gambiae started and stopped swarming earlier than An. coluzzii (3:35 ± 0:68 min:sec and 4:51 ± 1:21, respectively), and the mean duration of swarming was 23:37 ± 0:33 for An. gambiae and 21:39 ± 0:33 for An. coluzzii. Accordingly, in principle, whenever both species swarm over the same marker, a mean of 15.3 ± 3.1% of An. gambiae swarming would occur before An. coluzzii males arrived, and 19.5 ± 4.55% of An. coluzzii swarming would occurred after An. gambiae males had stopped swarming. These results are consistent with the finding that An. gambiae males became active in the actograph 09:35 ± 00:22 min:sec earlier than An. coluzzii males. The timing of swarming and spontaneous activity at dusk are primarily under circadian control, with the phase linked closely to sunset throughout the year. The mating activity of these two species is temporally segregated for 15-20% of the swarming period, which may contribute to the observed reproductive isolation of these species in local sympatric populations.
Insights about the structure of farnesyl diphosphate synthase (FPPS) and the activity of bisphosphonates on the proliferation and ultrastructure of Leishmania and GiardiaParasites and Vectors - Tập 13 - Trang 1-18 - 2020
Ana Paula R. Gadelha, Claudia Maia Brigagao, Martha Barros da Silva, Aline Beatriz Mello Rodrigues, Ana Carolina Ramos Guimarães, Fernando Paiva, Wanderley de Souza, Cristina Henriques
The enzyme farnesyl diphosphate synthase (FPPS) is positioned in the intersection of different sterol biosynthesis pathways such as those producing isoprenoids, dolichols and ergosterol. FPPS is ubiquitous in eukaryotes and is inhibited by nitrogen-containing bisphosphonates (N-BP). N-BP activity and the mechanisms of cell death as well as damage to the ultrastructure due to N-BP has not yet been investigated in Leishmania infantum and Giardia. Thus, we evaluated the effect of N-BP on cell viability and ultrastructure and then performed structural modelling and phylogenetic analysis on the FPPS enzymes of Leishmania and Giardia. We performed multiple sequence alignment with MAFFT, phylogenetic analysis with MEGA7, and 3D structural modelling for FPPS with Modeller 9.18 and on I-Tasser server. We performed concentration curves with N-BP in Leishmania promastigotes and Giardia trophozoites to estimate the IC50via the MTS/PMS viability method. The ultrastructure was evaluated by transmission electron microscopy, and the mechanism of cell death by flow cytometry. The nitrogen-containing bisphosphonate risedronate had stronger anti-proliferative activity in Leishmania compared to other N-BPs with an IC50 of 13.8 µM, followed by ibandronate and alendronate with IC50 values of 85.1 µM and 112.2 µM, respectively. The effect of N-BPs was much lower on trophozoites of Giardia than Leishmania (IC50 of 311 µM for risedronate). Giardia treated with N-BP displayed concentric membranes around the nucleus and nuclear pyknosis. Leishmania had mitochondrial swelling, myelin figures, double membranes, and plasma membrane blebbing. The same population labelled with annexin-V and 7-AAD had a loss of membrane potential (TMRE), indicative of apoptosis. Multiple sequence alignments and structural alignments of FPPS proteins showed that Giardia and Leishmania FPPS display low amino acid identity but possess the conserved aspartate-rich motifs. Giardia and Leishmania FPPS enzymes are phylogenetically distant but display conserved protein signatures. The N-BPs effect on FPPS was more pronounced in Leishmania than Giardia. This might be due to general differences in metabolism and differences in the FPPS catalytic site.
American tegumentary leishmaniasis caused by Leishmania (Viannia) braziliensis: assessment of parasite genetic variability at intra- and inter-patient levelsParasites and Vectors - Tập 6 - Trang 1-8 - 2013
Fernanda S Oliveira, Cláudia M Valete-Rosalino, Sandro JB Pacheco, Filipe A Carvalho Costa, Armando O Schubach, Raquel S Pacheco
The genetic variability of Leishmania (Viannia) braziliensis was assessed at intra and interpatient levels of individuals with different clinical manifestations of American tegumentary leishmaniasis (ATL). Fifty-two samples, of which 13 originated from cutaneous lesions and 39 from mucosal lesions, provided by 35 patients, were examined by low-stringency single-specific-primer PCR (LSSP-PCR) and phenetic analysis. Genetic variability of L. (V.) braziliensis, in kinetoplast DNA (kDNA) signatures, was compared both from different patients and from different lesions of the same patient. Phenetic analysis was performed to evaluate the degree of heterogeneity of the kDNA minicircles. In order to evaluate inter and intrapatient L. (V.) braziliensis genetic variability, the percentage of shared bands and analysis of the coefficients of similarity were analyzed. Different genetic profiles, representing kDNA signatures of the parasite, were obtained by LSSP-PCR analysis of each sample. Phenetic analysis grouped genetic profiles of different levels of differentiation from more similar to most divergent. The percentage of shared bands at the inter and intrapatient levels was 77% and 89%, respectively. Comparison of the average inter and intrapatient coefficients of similarity and their standard deviations were statistically significant (p < 0.001). Genetic variability at the intrapatient level was less pronounced than that between different patients. A conceptual model was proposed to better understand the complexity at both levels.
Experimental hut evaluation of linalool spatial repellent agar gel against Anopheles gambiae sensu stricto mosquitoes in a semi-field system in Bagamoyo, TanzaniaParasites and Vectors - Tập 7 - Trang 1-6 - 2014
Mgeni Mohamed Tambwe, Edgar Mtaki Mbeyela, Brian Migamyo Massinda, Sarah Jane Moore, Marta Ferreira Maia
Malaria vector control is in need of new tools to face its current challenges such as the spread of pyrethroid-resistance and the increase of outdoor feeding mosquitoes. New strategies such as spatial repellents need to be evaluated as supplemental tools to existing control measures such as insecticide treated bed nets and indoor residual spraying. Linalool is a naturally occurring terpene alcohol commonly found in flowers and spices with reportedly repellent properties. Four experimental huts fitted with exit traps and enclosed inside a large screened semi-field system were used for the evaluation. The tested spatial repellent product consisted of an agar gel emanator containing 73% linalool. Two rounds of experiments using a Latin square design were conducted to evaluate the efficacy of the linalool emanators compared to no treatment (negative control) and a transfluthrin coil (positive) against lab-reared disease free Anopheles gambiae s.s.. The emanators were hung inside experimental huts where two volunteers were sleeping unprotected. The outcome measures were repellency, % feeding inhibition, %mortality and post 24 h % mortality. Unlike the mosquito coil, the linalool emanators did not show any feeding inhibition, repellency or induced mortality compared to the negative control. On the other hand mosquitoes kept for 24 h post exposure were 3 times more likely to die after being exposed to two 73% linalool emanators than the negative control. Our results indicate that linalool agar gel emanators are not adequate as a spatial repellent against Anopheles gambiae s.s.. However adding linalool to known repellent formulations could be advantageous, not only because of its pleasant scent but also because of the delayed mortality effect it has on mosquitoes.
