RESUMEN
Plasmodium vivax is now the main cause of malaria outside Africa. The gametocytocidal effects of antimalarial drugs are important to reduce malaria transmissibility, particularly in low-transmission settings, but they are not well characterized for P. vivax. The transmission-blocking effects of chloroquine, artesunate, and methylene blue on P. vivax gametocytes were assessed. Blood specimens were collected from patients presenting with vivax malaria, incubated with or without the tested drugs, and then fed to mosquitos from a laboratory-adapted colony of Anopheles dirus (a major malaria vector in Southeast Asia). The effects on oocyst and sporozoite development were analyzed under a multi-level Bayesian model accounting for assay variability and the heterogeneity of mosquito Plasmodium infection. Artesunate and methylene blue, but not chloroquine, exhibited potent transmission-blocking effects. Gametocyte exposures to artesunate and methylene blue reduced the mean oocyst count 469-fold (95% CI: 345 to 650) and 1,438-fold (95% CI: 970 to 2,064), respectively. The corresponding estimates for the sporozoite stage were a 148-fold reduction (95% CI: 61 to 470) and a 536-fold reduction (95% CI: 246 to 1,311) in the mean counts, respectively. In contrast, high chloroquine exposures reduced the mean oocyst count only 1.40-fold (95% CI: 1.20 to 1.64) and the mean sporozoite count 1.34-fold (95% CI: 1.12 to 1.66). This suggests that patients with vivax malaria often remain infectious to anopheline mosquitos after treatment with chloroquine. Use of artemisinin combination therapies or immediate initiation of primaquine radical cure should reduce the transmissibility of P. vivax infections.
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Anopheles , Antimaláricos , Artesunato , Cloroquina , Malaria Vivax , Azul de Metileno , Plasmodium vivax , Azul de Metileno/farmacología , Azul de Metileno/uso terapéutico , Artesunato/farmacología , Artesunato/uso terapéutico , Cloroquina/farmacología , Cloroquina/uso terapéutico , Plasmodium vivax/efectos de los fármacos , Antimaláricos/farmacología , Antimaláricos/uso terapéutico , Malaria Vivax/tratamiento farmacológico , Malaria Vivax/parasitología , Malaria Vivax/transmisión , Animales , Humanos , Anopheles/parasitología , Anopheles/efectos de los fármacos , Esporozoítos/efectos de los fármacos , Artemisininas/farmacología , Artemisininas/uso terapéutico , Oocistos/efectos de los fármacosRESUMEN
OBJECTIVE: A 15-month longitudinal study was conducted to determine the duration and infectivity of asymptomatic qPCR-detected Plasmodium falciparum and Plasmodium vivax infections in Ethiopia. METHOD: Total parasite and gametocyte kinetics were determined by molecular methods; infectivity to Anopheles arabiensis mosquitoes by repeated membrane feeding assays. Infectivity results were contrasted with passively recruited symptomatic malaria cases. RESULTS: For P. falciparum and P. vivax infections detected at enrolment, median durations of infection were 37 days (95% confidence interval [CI], 15-93) and 60 days (95% CI, 18-213), respectively. P. falciparum and P. vivax parasite densities declined over the course of infections. From 47 feeding assays on 22 asymptomatic P. falciparum infections, 6.4% (3/47) were infectious and these infected 1.8% (29/1579) of mosquitoes. No transmission was observed in feeding assays on asymptomatic P. vivax mono-infections (0/56); one mixed-species infection was highly infectious. Among the symptomatic cases, 4.3% (2/47) of P. falciparum and 73.3% (53/86) of P. vivax patients were infectious to mosquitoes. CONCLUSION: The majority of asymptomatic infections were of short duration and low parasite density. Only a minority of asymptomatic individuals were infectious to mosquitoes. This contrasts with earlier findings and is plausibly due to the low parasite densities in this population.
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Anopheles , Malaria Falciparum , Malaria Vivax , Plasmodium falciparum , Plasmodium vivax , Etiopía/epidemiología , Malaria Vivax/transmisión , Malaria Vivax/epidemiología , Malaria Vivax/parasitología , Humanos , Estudios Longitudinales , Malaria Falciparum/transmisión , Malaria Falciparum/epidemiología , Malaria Falciparum/parasitología , Animales , Plasmodium vivax/aislamiento & purificación , Plasmodium vivax/fisiología , Plasmodium falciparum/aislamiento & purificación , Anopheles/parasitología , Masculino , Femenino , Adulto , Adolescente , Niño , Adulto Joven , Preescolar , Infecciones Asintomáticas/epidemiología , Mosquitos Vectores/parasitología , Persona de Mediana EdadRESUMEN
BACKGROUND: Plasmodium falciparum and Plasmodium vivax account for >90% global malaria burden. Transmission intervention strategies encompassing transmission-blocking vaccines (TBV) and drugs represent ideal public health tools to eliminate malaria at the population level. The availability of mature P. falciparum gametocytes through in vitro culture has facilitated development of a standard membrane feeding assay to assess efficacy of transmission interventions against P. falciparum. The lack of in vitro culture for P. vivax has significantly hampered similar progress on P. vivax and limited studies have been possible using blood from infected patients in endemic areas. The ethical and logistical limitations of on-time access to blood from patients have impeded the development of P. vivax TBVs. METHODS: Transgenic murine malaria parasites (Plasmodium berghei) expressing TBV candidates offer a promising alternative for evaluation of P. vivax TBVs through in vivo studies in mice, and ex vivo membrane feeding assay (MFA). RESULTS: We describe the development of transmission-competent transgenic TgPbvs25 parasites and optimization of parameters to establish an ex vivo MFA to evaluate P. vivax TBV based on Pvs25 antigen. CONCLUSIONS: The MFA is expected to expedite Pvs25-based TBV development without dependence on blood from P. vivax-infected patients in endemic areas for evaluation.
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Vacunas contra la Malaria , Malaria Vivax , Plasmodium berghei , Plasmodium vivax , Animales , Vacunas contra la Malaria/inmunología , Vacunas contra la Malaria/genética , Plasmodium vivax/genética , Plasmodium vivax/inmunología , Malaria Vivax/transmisión , Malaria Vivax/prevención & control , Malaria Vivax/parasitología , Plasmodium berghei/genética , Plasmodium berghei/inmunología , Ratones , Antígenos de Protozoos/inmunología , Antígenos de Protozoos/genética , Humanos , Femenino , Antígenos de SuperficieRESUMEN
Some parasitic diseases, such as malaria, require two hosts to complete their lifecycle: a human and an insect vector. Although most malaria research has focused on parasite development in the human host, the life cycle within the vector is critical for the propagation of the disease. The mosquito stage of the Plasmodium lifecycle represents a major demographic bottleneck, crucial for transmission blocking strategies. Furthermore, it is in the vector, where sexual recombination occurs generating "de novo" genetic diversity, which can favor the spread of drug resistance and hinder effective vaccine development. However, understanding of vector-parasite interactions is hampered by the lack of experimental systems that mimic the natural environment while allowing to control and standardize the complexity of the interactions. The breakthrough in stem cell technologies has provided new insights into human-pathogen interactions, but these advances have not been translated into insect models. Here, we review in vivo and in vitro systems that have been used so far to study malaria in the mosquito. We also highlight the relevance of single-cell technologies to progress understanding of these interactions with higher resolution and depth. Finally, we emphasize the necessity to develop robust and accessible ex vivo systems (tissues and organs) to enable investigation of the molecular mechanisms of parasite-vector interactions providing new targets for malaria control.
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Culicidae , Malaria , Humanos , Animales , Mosquitos Vectores , Ambiente , TecnologíaRESUMEN
BACKGROUND: Measuring risk of malaria transmission is complex, especially in case of Plasmodium vivax. This may be overcome using membrane feeding assays in the field where P. vivax is endemic. However, mosquito-feeding assays are affected by a number of human, parasite and mosquito factors. Here, this study identified the contributions of Duffy blood group status of P. vivax-infected patients as a risk of parasite transmission to mosquitoes. METHODS: A membrane feeding assay was conducted on a total of 44 conveniently recruited P. vivax infected patients in Adama city and its surroundings in East Shewa Zone, Oromia region, Ethiopia from October, 2019 to January, 2021. The assay was performed in Adama City administration. Mosquito infection rates were determined by midgut dissections at seven to 8 days post-infection. Duffy genotyping was defined for each of the 44 P. vivax infected patients. RESULTS: The infection rate of Anopheles mosquitoes was 32.6% (296/907) with 77.3% proportion of infectious participants (34/44). Infectiousness of participants to Anopheles mosquitoes appeared to be higher among individuals with homozygous Duffy positive blood group (TCT/TCT) than heterozygous (TCT/CCT), but the difference was not statistically significant. The mean oocyst density was significantly higher among mosquitoes fed on blood of participants with FY*B/FY*BES than other genotypes (P = 0.001). CONCLUSION: Duffy antigen polymorphisms appears to contribute to transmissibility difference of P. vivax gametocytes to Anopheles mosquitoes, but further studies are required.
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Anopheles , Antígenos de Grupos Sanguíneos , Malaria Vivax , Animales , Humanos , Plasmodium vivax/genética , Anopheles/parasitología , Malaria Vivax/epidemiología , GenotipoRESUMEN
The discovery and development of transmission-blocking therapies challenge malaria elimination and necessitate standard and reproducible bioassays to measure the blocking properties of antimalarial drugs and candidate compounds. Most of the current bioassays evaluating the transmission-blocking activity of compounds rely on laboratory-adapted Plasmodium strains. Transmission-blocking data from clinical gametocyte isolates could help select novel transmission-blocking candidates for further development. Using freshly collected Plasmodium falciparum gametocytes from asymptomatic individuals, we first optimized ex vivo culture conditions to improve gametocyte viability and infectiousness by testing several culture parameters. We next pre-exposed ex vivo field-isolated gametocytes to chloroquine, dihydroartemisinin, primaquine, KDU691, GNF179, and oryzalin for 48 h prior to direct membrane feeding. We measured the activity of the drug on the ability of gametocytes to resume the sexual life cycle in Anopheles after drug exposure. Using 57 blood samples collected from Malian volunteers aged 6 to 15 years, we demonstrate that the infectivity of freshly collected field gametocytes can be preserved and improved ex vivo in a culture medium supplemented with 10% horse serum at 4% hematocrit for 48 h. Moreover, our optimized drug assay displays the weak transmission-blocking activity of chloroquine and dihydroartemisinin, while primaquine and oryzalin exhibited a transmission-blocking activity of ~50% at 1 µM. KDU691 and GNF179 both interrupted Plasmodium transmission at 1 µM and 5 nM, respectively. This new approach, if implemented, has the potential to accelerate the screening of compounds with transmission-blocking activity.
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Antimaláricos , Malaria Falciparum , Humanos , Plasmodium falciparum , Primaquina , Malaria Falciparum/prevención & control , Antimaláricos/farmacología , Antimaláricos/uso terapéutico , Cloroquina/farmacología , Cloroquina/uso terapéuticoRESUMEN
BACKGROUND: Insecticide-based vector control interventions in combination with case management with artemisinin-based combination therapy has reduced malaria incidence and prevalence worldwide. Current control methods focus on the primary malaria vectors, Anopheles gambiae sensu lato (s.l.) and the An. funestus group; however, the impact of secondary and suspected vectors has been either sidelined or received limited attention. Defining the susceptibility of secondary, suspected vector species to different parasites in time and space is essential for efficient malaria control and elimination programs. The aim of this study was to assess the susceptibility of An. gambiae s.l., An. coustani complex and An. pharoensis to Plasmodium vivax and P. falciparum infection in Ethiopia. METHODS: Larvae of Anopheles spp. were collected from different aquatic habitats and reared to adults under laboratory conditions, with the temperature and humidity maintained at 27 ± 1 °C and 75 ± 5%, respectively. Adult female mosquitoes were identified to species as An. gambiae s.l., An. coustani complex and An. pharoensis. Females of these three Anopheles spp. were allowed to feed in parallel feeding assays on infected blood containing the same gametocytes isolated from P. falciparum and P. vivax gametocyte-positive patients by indirect membrane feeding assays. All blood-fed mosquitoes were held under laboratory conditions. After 7 days, all surviving mosquitoes were dissected to detect mid-gut oocyst and enumerated under a microscope. RESULTS: Of 5915 female Anopheles mosquitoes exposed to gametocyte-infected blood, 2106 (35.6%)s fed successfully in the 32 independent infection experiments. There was a significant variation in feeding rates among An. gambiae s.l., An. pharoensis and An. coustani complex (G-test = 48.43, P = 3.049e-11). All three exposed mosquito species were receptive to P. vivax and P. falciparum infection development. The percentage of infected mosquitoes following feeding on an infected blood meal was significantly different among species (G-test = 6.49, P = 0.03886). The median infection intensity (II) for An. coustani complex, An. gambiae s.l. and An. pharoensis was 1.16, 2.00 and 1.25, respectively. Although the proportion of infected mosquitoes significantly differed in terms of II, infection rate (IR) and mean oocyst density among the species, mean oocyst density and IR were highly correlated with gametocyte density in all tests (P < 0.001). CONCLUSION: Primary, secondary and suspected vectors were experimentally susceptible to both P. vivax and P. falciparum infection. An effective malaria elimination program might include surveillance and control tools which target secondary and suspected vectors that might play an outdoor transmission role, possibly resulting in reduced focal malaria transmission. Comparison of the three species' mean infection rates with standard deviation.
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Anopheles , Artemisininas , Insecticidas , Malaria Falciparum , Malaria Vivax , Malaria , Animales , Humanos , Femenino , Plasmodium vivax , Plasmodium falciparum , Mosquitos Vectores/parasitología , Etiopía/epidemiología , Malaria Falciparum/parasitología , Anopheles/parasitología , Malaria Vivax/parasitología , OocistosRESUMEN
The membrane feeding assay is widely used to evaluate the efficacy of transmission-blocking interventions (TBIs) and identify the reservoir of malaria. This study aimed to determine the infectivity of blood meals from symptomatic Plasmodium-infected patients to an Anopheles arabiensis colony in Ethiopia. A membrane feeding assay was conducted on a total of 63 Plasmodium falciparum- and/or Plasmodium vivax-infected clinical patients in East Shoa Zone, Ethiopia. Detection of P. falciparum and P. vivax in blood samples was done using microscopy. Mosquito infection rates were determined by dissection of mosquitoes' midguts, while mosquito infectiousness was observed by dissection of their salivary glands. The proportion of infectious symptomatic patients was 68.3% (43/63). Using the chi-square or Fisher's exact test, the oocyst infection levels were higher among patients infected with P. vivax, females, and rural residents. Nearly 57% (56.7%, 17/30) of assays produced sporozoites in the salivary glands of mosquitoes. Both oocyst and sporozoite infection rates had positive correlations with parasitemia and gametocytemia. High infectiousness of symptomatic patients was observed, with a greater proportion of infectious mosquitoes per assay. Demonstrating oocyst infection in the mosquitoes might confirm estimates of the infectiousness of mosquitoes, although some of the oocyst-infected mosquitoes failed to produce sporozoites. IMPORTANCE Malaria remains one of the most devastating infectious diseases globally, and transmission-blocking activities are needed. Plasmodium transmission from human to mosquitoes is poorly studied, particularly in endemic countries, and the membrane feeding assay allows it to be determined. In this study, we demonstrated human infectious reservoirs of malaria. Moreover, the effect of Plasmodium-infected patients on the infectiousness of mosquitoes was also observed. These findings are therefore important for designing future evaluation of transmission-blocking interventions that will support the malaria elimination program.
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Anopheles , Malaria Falciparum , Malaria Vivax , Malaria , Animales , Femenino , Humanos , Etiopía/epidemiología , Plasmodium vivax , Malaria Vivax/epidemiología , Plasmodium falciparum , Malaria Falciparum/epidemiología , OocistosRESUMEN
Plasmodium falciparum is the parasite responsible for the disease malaria. In vitro cultivation of mature gametocytes of P. falciparum plays a central role in evaluating and developing the transmission-blocking drugs and sexual stage vaccines. These types of preventive molecules are crucial for controlling malaria in the future. Among different Plasmodium species that are involved in human malaria, only P. falciparum is cultivable. Therefore, an efficient method is required for in vitro culture of P. falciparum producing mature and infective gametocytes. This chapter describes a reliable and efficient protocol for the production of adult and infective gametocytes that is suitable for small- and large-scale culture.
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Anopheles , Malaria Falciparum , Plasmodium falciparum , Animales , Bioensayo , Humanos , MalariaRESUMEN
Traditional and modern approaches have been applied to combat the malaria disease. Malaria eradication is a priority in several developing countries. Transmission-blocking vaccines are one of the suggested solutions for malaria eradication. Therefore, there is a demand for introducing the new targets and evaluation methods. Standard membrane feeding assay is the base of the evaluation process of transmission-blocking candidate molecules. Hence, this process is explained in this chapter in detail.
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Vacunas contra la Malaria , Malaria , Bioensayo , Humanos , Malaria/prevención & control , Malaria Falciparum , Membranas , Plasmodium falciparum/inmunologíaRESUMEN
BACKGROUND: Plasmodium vivax transmission-blocking vaccines (TBVs) are receiving increasing attention. Based on excellent transmission-blocking activities of the PbPH (PBANKA_0417200) and PbSOP26 (PBANKA_1457700) antigens in Plasmodium berghei, their orthologs in P. vivax, PVX_098655 (PvPH) and PVX_101120 (PvSOP26), were selected for the evaluation of their potential as TBVs. METHODS: Fragments of PvPH (amino acids 22-304) and PvSOP26 (amino acids 30-272) were expressed in the yeast expression system. The recombinant proteins were used to immunize mice to obtain antisera. The transmission-reducing activities of these antisera were evaluated using the direct membrane feeding assay (DMFA) using Anopheles dirus mosquitoes and P. vivax clinical isolates. RESULTS: The recombinant proteins PvPH and PvSOP26 induced robust antibody responses in mice. The DMFA showed that the anti-PvSOP26 sera significantly reduced oocyst densities by 92.0 and 84.1% in two parasite isolates, respectively, whereas the anti-PvPH sera did not show evident transmission-reducing activity. The variation in the DMFA results was unlikely due to the genetic polymorphisms of the two genes since their respective sequences were identical in the clinical P. vivax isolates. CONCLUSION: PvSOP26 could be a promising TBV candidate for P. vivax, which warrants further evaluation.
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Anticuerpos Antiprotozoarios/sangre , Antígenos de Protozoos/inmunología , Vacunas contra la Malaria/inmunología , Malaria Vivax/prevención & control , Plasmodium vivax/inmunología , Animales , Femenino , Humanos , Inmunogenicidad Vacunal , Vacunas contra la Malaria/genética , Malaria Vivax/parasitología , Malaria Vivax/transmisión , Ratones , Ratones Endogámicos BALB C , Proteínas Protozoarias/genética , Proteínas Protozoarias/inmunología , Proteínas Recombinantes/genética , Proteínas Recombinantes/inmunología , Vacunación/métodos , Vacunas Sintéticas/administración & dosificación , Vacunas Sintéticas/inmunología , Levaduras/genéticaRESUMEN
BACKGROUND: Direct membrane feeding assays (DMFA) are an important tool to study parasite transmission to mosquitoes. Mosquito feeding rates in these artificial systems require optimization, as there are a number of factors that potentially influence the feeding rates and there are no standardized methods that apply to all anopheline species. METHODS: A range of parameters prior to and during direct membrane feeding (DMF) were evaluated for their impact on Anopheles farauti sensu stricto feeding rates, including the starving conditions and duration of starving prior to feeding, membrane type, DMF exposure time, mosquito age, feeding in the light versus the dark, blood volume, mosquito density and temperature of water bath. RESULTS: The average successful DMFA feeding rate for An. farauti s.s. colony mosquitoes increased from 50 to 85% when assay parameters were varied. Overnight starvation and Baudruche membrane yielded the highest feeding rates but rates were also affected by blood volume in the feeder and the mosquito density in the feeding cups. Availability of water during the pre-feed starvation period did not significantly impact feeding rates, nor did the exposure duration to blood in membrane feeders, the age of mosquitoes (3, 5 and 7 days post-emergence), feeding in the light versus the dark, or the temperature (34 °C, 38 °C, 42 °C and 46 °C) of the water bath. CONCLUSION: Optimal feeding conditions in An. farauti s.s. DMFA were to offer 50 female mosquitoes in a cup (with a total surface area of ~ 340 cm2 with 1 mosquito/6.8 cm2) that were starved overnight 350-500 µL of blood (collected in heparin-coated Vacutainer tubes) per feeder in feeders with a surface area ~ 5 cm2 (with a maximum capacity of 1.5 mL of blood) via a Baudruche membrane, for at least 10-20 min.
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Anopheles/fisiología , Bioensayo/métodos , Conducta Alimentaria , Mosquitos Vectores/fisiología , Animales , FemeninoRESUMEN
The control and elimination of malaria caused by Plasmodium vivax both represent a great challenge due to the biological aspects of the species. Gametocytes are the forms responsible for the transmission of the parasite to the vector and the search for new strategies for blocking transmission are essential in a scenario of control and elimination The challenges in this search in regard to P. vivax mainly stem from the lack of a long-term culture and the limitation of studies of gametocytes. This study evaluated the viability and infectivity of P. vivax gametocytes in short-term culture. The samples enriched in gametocytes using Percoll (i), using magnetic-activated cell sorting (MACS®) (ii), and using non-enriched samples (iii) were evaluated. After the procedures, gametocytes were cultured in IMDM medium for up to 48 h. Cultured P. vivax gametocytes were viable and infectious for up to 48 h, however differences in viability and infectivity were observed in the samples after 12 h of culture in relation to 0 h. Percoll-enriched samples were shown to be viable in culture for longer intervals than those purified using MACS®. Gametocyte viability after enrichment procedures and short-term culture may provide new avenues in the development of methods for evaluating P. vivax TB.
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Malaria Vivax , Malaria , Humanos , Plasmodium falciparum , Plasmodium vivaxRESUMEN
Experimental studies for understanding the relationship between Plasmodium vivax and its vector hosts are difficult, because of to the lack of a long-term, in vitro continuous culture system unavailability of infected blood samples, seasonality of the disease, and the concentration of most cases in remote areas. This study evaluates the duration of the infectivity of P. vivax to Anopheles aquasalis after collecting blood from malaria-infected patients. Blood was collected from patients and stored at 4 °C and 37 °C. Every day, for 4 days, the blood was fed to An. aquasalis adult females, and a Giemsa-stained thick blood smear was mounted to account for sexual (gametocytes) and asexual (trophozoites and schizonts) stages and calculate parasitemia. Oocysts in the midgut of the mosquitoes were counted on the seventh day after feeding. Kruskal-Wallis test was used to compare the mean number of oocysts (MO) and the parasite density (PD) in each storage condition and post-infection time-points. The Mann-Whitney test was used to compare the number of oocysts for each day between temperatures. The results show that P. vivax stored at 4 °C and at 37 °C has its infectivity to An. aquasalis preserved for 2 days and 3 days, respectively. Infection rate (IR), PD and MO were higher on the day of blood collection and decreased gradually over time. The parasite density (number of parasites/µL) diminished faster at 4 °C than at 37 °C. In this study, a preservation protocol is shown for long-lasting infectivity of P. vivax in a blood sample taken from malaria-infected patients. These results show that infectivity of P. vivax stored at 4 °C and at 37 °C to An. aquasalis persist until 3 days after blood collection, but parasite density, infection rate, and mean of oocysts decreased 24h after blood collection. Since the malaria cases are increasingly far from the urban areas these results indicate that is possible, losing some infectivity, to realize experimental infections several dozen hours after the blood collection. However, it is necessary to improve the procedures for preserving P. vivax gametocytes for mosquito infection in the laboratory.
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Anopheles/parasitología , Malaria Vivax/parasitología , Mosquitos Vectores/parasitología , Plasmodium vivax/fisiología , Adulto , Anciano , Animales , Brasil , Femenino , Humanos , Malaria Vivax/sangre , Malaria Vivax/transmisión , Masculino , Persona de Mediana Edad , Plasmodium vivax/patogenicidad , Población Rural , Temperatura , Factores de Tiempo , Adulto JovenRESUMEN
Introduction: The development of effective vaccines remains a major health priority to combat the global burden of malaria, a life-threatening disease caused by Plasmodium parasites. Transmission-blocking vaccines (TBVs) elicit antibodies that neutralize the sexual stages of the parasite in blood meals ingested by the Anopheles mosquito, interrupting parasite development in the vector host and preventing disease spread to other individuals.Areas covered: The P. falciparum gametocyte surface antigens Pfs230, Pfs48/45, and Pfs47, the parasite ookinete surface protein Pfs25, and the male gametocyte specific protein PfHAP2 are leading TBV candidates, some of which are in clinical development. The recent expansion of methodology to study monoclonal antibodies isolated directly from humans and animal models, coupled with effective measures for parasite neutralization, has provided unprecedented insight into TBV efficacy and development.Expert opinion: Available structural and functional data on antigen-monoclonal antibody (Ag-mAb) complexes, as well as epitope classification studies, have identified neutralizing epitopes that may aid vaccine development and improve protection. Here, we review the clinical prospects of TBV candidates, progress in the development of novel vaccine strategies for TBVs, and the impact of structural vaccinology in TBV design.
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Vacunas contra la Malaria/administración & dosificación , Malaria Falciparum/prevención & control , Plasmodium falciparum/inmunología , Animales , Anopheles/parasitología , Anticuerpos Monoclonales/inmunología , Anticuerpos Antiprotozoarios/inmunología , Antígenos de Protozoos/inmunología , Humanos , Vacunas contra la Malaria/inmunología , Malaria Falciparum/inmunología , Malaria Falciparum/transmisión , Mosquitos Vectores/inmunología , Mosquitos Vectores/parasitología , Plasmodium falciparum/parasitología , VacunologíaRESUMEN
Plasmodium transmission from humans to mosquitoes is an understudied bottleneck in the transmission of malaria. Direct membrane feeding assays (DMFA) allow detailed malaria transmission studies from humans to mosquitoes. Especially for Plasmodium vivax, which cannot be cultured long-term under laboratory conditions, implementation of DMFAs requires proximity to P. vivax endemic areas. In this study, we investigated the infectivity of symptomatic Plasmodium infections to Anopheles farauti colony mosquitoes in Papua New Guinea (PNG). A total of 182 DMFAs were performed with venous blood collected from rapid diagnostic test (RDT) positive symptomatic malaria patients and subsequently analysed by light microscopy and quantitative real time polymerase chain reaction (qPCR). DMFAs resulted in mosquito infections in 20.9% (38/182) of cases. By light microscopy and qPCR, 10 - 11% of P. falciparum and 32 - 44% of P. vivax positive individuals infected An. farauti. Fifty-eight percent of P. vivax and 15% of P. falciparum gametocytaemic infections infected An farauti.
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Anopheles , Malaria Vivax , Malaria , Animales , Humanos , Malaria Vivax/epidemiología , Papúa Nueva Guinea , Plasmodium falciparum , Plasmodium vivaxRESUMEN
BACKGROUND Different strategies for improvement of malaria control and elimination are based on the blockage of malaria parasite transmission to the mosquito vector. These strategies include the drugs that target the plasmodial sexual stages in humans and the early developmental stages inside mosquitoes. OBJECTIVES Here we tested Malaria Box compounds in order to evaluate their activity against male and female gametocytes in Plasmodium berghei, mosquito infection in P. vivax and ookinete formation in both species. METHODS/FINDINGS The membrane feeding assay and the development of ookinetes by a 24 h ex vivo culture and the ookinete yield per 1000 erythrocytes were used to test transmission-blocking potential of the Malaria Box compounds in P. vivax. For P. berghei we used flow cytometry to evaluate male and female gametocyte time course and fluorescence microscopy to check the ookinete development. The two species used in this study showed similar results concerning the compounds' activity against gametocytes and ookinetes, which were different from those in P. falciparum. In addition, from the eight Malaria Box compounds tested in both species, compounds MMV665830, MMV665878 and MMV665941 were selected as a hit compounds due the high inhibition observed. CONCLUSION Our results showed that P. berghei is suitable as an initial screening system to test compounds against P. vivax.
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Animales , Plasmodium berghei/efectos de los fármacos , Plasmodium vivax/efectos de los fármacos , Malaria Vivax/prevención & control , Mosquitos Vectores/parasitología , Malaria Vivax/tratamiento farmacológico , Malaria Vivax/transmisiónRESUMEN
Spider mites (Acari: Tetranychidae) are pests of a wide range of agricultural crops, vegetables, and ornamental plants. Their ability to rapidly develop resistance to synthetic pesticides has prompted the development of new strategies for their control. Evaluation of synthetic pesticides and bio-pesticides-and more recently the identification of RNA interference (RNAi) target genes-requires an ability to deliver test compounds efficiently. Here we describe a novel method that uses a sheet-like structure mimicking plant leaves and allows for oral delivery of liquid test compounds to a large number of individuals in a limited area simultaneously (~100 mites cm-2). The main component is a fine nylon mesh sheet that holds the liquid within each pore, much like a plant cell, and consequently allows for greater distribution of specific surface area even in small amounts (10 µl cm-2 for 100-µm mesh opening size). The nylon mesh sheet is placed on a solid plane (e.g., the undersurface of a Petri dish), a solution or suspension of test compounds is pipetted into the mesh sheet, and finally a piece of paraffin wax film is gently stretched above the mesh so that the test mites can feed through it. We demonstrate the use of the method for oral delivery of a tracer dye (Brilliant Blue FCF), pesticides (abamectin and bifenazate), dsRNA targeting the Vacuolar-type H+-VATPase gene, or fluorescent nanoparticles to three species of Tetranychus spider mites (Acari: Tetranychidae) and to the cotton aphid, Aphis gossypii Glover (Hemiptera: Aphididae). The method is fast, easy, and highly reproducible and can be adapted to facilitate several aspects of bioassays.
RESUMEN
BACKGROUND: The sterile insect technique (SIT) is a vector control strategy relying on the mass release of sterile males into wild vector populations. Current sex separation techniques are not fully efficient and could lead to the release of a small proportion of females. It is therefore important to evaluate the effect of irradiation on the ability of released females to transmit pathogens. This study aimed to assess the effect of irradiation on the survival and competence of Anopheles arabiensis females for Plasmodium falciparum in laboratory conditions. METHODS: Pupae were irradiated at 95 Gy of gamma-rays, and emerging females were challenged with one of 14 natural isolates of P. falciparum. Seven days post-blood meal (dpbm), irradiated and unirradiated-control females were dissected to assess the presence of oocysts, using 8 parasite isolates. On 14 dpbm, sporozoite dissemination in the head/thorax was also examined, using 10 parasites isolates including 4 in common with the 7 dpbm dissection (oocyst data). The survivorship of irradiated and unirradiated-control mosquitoes was monitored. RESULTS: Overall, irradiation reduced the proportion of mosquitoes infected with the oocyst stages by 17% but this effect was highly inconsistent among parasite isolates. Secondly, there was no significant effect of irradiation on the number of developing oocysts. Thirdly, there was no significant difference in both the sporozoite infection rate and load between the irradiated and unirradiated-control mosquitoes. Fourthly, irradiation had varying effects on female survival with either a negative effect or no effect. CONCLUSIONS: The effect of irradiation on mosquito competence strongly varied among parasite isolates. Because of such isolate variability and, the fact that different parasite isolates were used to collect oocyst and sporozoite data, the irradiation-mediated reduction of oocyst prevalence was not confirmed for the sporozoite stages. Our data indicate that irradiated female An. arabiensis could contribute to malaria transmission, and highlight the need for perfect sexing tools, which would prevent the release of females as part of SIT programmes.
Asunto(s)
Anopheles/parasitología , Anopheles/efectos de la radiación , Rayos gamma , Control de Mosquitos/métodos , Plasmodium falciparum/fisiología , Animales , Anopheles/fisiología , Sangre , Conducta Alimentaria , Femenino , Mosquitos Vectores/parasitología , Mosquitos Vectores/efectos de la radiación , Oocistos/fisiología , Pupa/efectos de la radiaciónRESUMEN
Although malaria is endemic to the Amazon region, little is known about the susceptibility of potential parasite vectors in Brazil. Assessing the vector susceptibility of Anopheles mosquitoes will increase our understanding of parasite-vector interactions and aid the design of vector control strategies. This study assessed the susceptibility of three Anopheles species to midgut infection by Plasmodium vivax, the predominant malaria species in Rondônia State, Brazil. Blood from P. vivax infected patients was fed to Anopheles aquasalis, Anopheles darlingi, and Anopheles deaneorum mosquitoes using a membrane feeding assay (MFA). Gametocytemia was estimated by microscopic examination of blood smears and oocyst prevalence, and infection intensity was assessed. The presence of oocysts was determined by microscopy, and the infection rates and infection intensity were determined for all species. Data from six MFAs showed that An. darlingi and An. deaneorum exhibited the highest infection rates (97% and 90%, respectively) and developed a similar median number of P. vivax oocysts (142 and 123, respectively), while An. aquasalis exhibited the smallest infection rates (77%) and the median number of oocysts (88). Established laboratory colonies of An. darlingi and An. deaneorum and susceptibility to plasmodial infection would be beneficial for modeling P. vivax vector-parasite interactions in Brazil.