Safety and immunogenicity of AGS-v PLUS, a mosquito saliva peptide vaccine against arboviral diseases: A randomized, double-blind, placebo-controlled Phase 1 trial.

BACKGROUND: Immunity to mosquito salivary proteins could provide protection against multiple mosquito-borne diseases and significantly impact public health. We evaluated the safety and immunogenicity of AGS-v PLUS, a mosquito salivary peptide vaccine, in healthy adults 18-50 years old. METHODS: We conducted a randomized, double-blind, placebo-controlled Phase 1 study of AGS-v PLUS administered subcutaneously on Days 1 and 22 at the Center for Vaccine Development and Global Health, Baltimore, MD, USA. Participants were block randomized 1:1:1:1:1 to two doses saline placebo, two doses AGS-v PLUS, AGS-v PLUS/ISA-51 and saline placebo, two doses AGS-v PLUS/ISA-51, or two doses AGS-v PLUS/Alhydrogel. Primary endpoints were safety (all participants receiving ≥1 injection) and antibody and cytokine responses (all participants with day 43 samples), analysed by intention to treat. FINDINGS: Between 26 August 2019 and 25 February 2020, 51 participants were enrolled and randomized, 11 into the single dose AGS-v PLUS/ISA-51 group and ten in other groups. Due to COVID-19, 15 participants did not return for day 43 samplings. Participants experienced no treatment-emergent or serious adverse events. All solicited symptoms in 2/10 placebo recipients and 22/41 AGS-v PLUS recipients after dose one and 1/10 placebo recipients and 22/41 AGS-v PLUS recipients after dose two were mild/moderate except for one severe fever the day after vaccination (placebo group). Only injection site pain was more common in vaccine groups (15/51 after dose 1 and 11/51 after dose 2) versus placebo. Compared to placebo, all vaccine groups had significantly greater fold change in anti-AGS-v PLUS IgG and IFN-É£ from baseline. INTERPRETATION: AGS-v PLUS had favourable safety profile and induced robust immune responses. Next steps will determine if findings translate into clinical efficacy against mosquito-borne diseases. FUNDING: UK Department of Health and Social Care.

Ecological and genetic relationships of the Forest-M form among chromosomal and molecular forms of the malaria vector Anopheles gambiae sensu stricto.

BACKGROUND: Anopheles gambiae sensu stricto, one of the principal vectors of malaria, has been divided into two subspecific groups, known as the M and S molecular forms. Recent studies suggest that the M form found in Cameroon is genetically distinct from the M form found in Mali and elsewhere in West Africa, suggesting further subdivision within that form. METHODS: Chromosomal, microsatellite and geographic/ecological evidence are synthesized to identify sources of genetic polymorphism among chromosomal and molecular forms of the malaria vector Anopheles gambiae s.s. RESULTS: Cytogenetically the Forest M form is characterized as carrying the standard chromosome arrangement for six major chromosomal inversions, namely 2La, 2Rj, 2Rb, 2Rc, 2Rd, and 2Ru. Bayesian clustering analysis based on molecular form and chromosome inversion polymorphisms as well as microsatellites describe the Forest M form as a distinct population relative to the West African M form (Mopti-M form) and the S form. The Forest-M form was the most highly diverged of the An. gambiae s.s. groups based on microsatellite markers. The prevalence of the Forest M form was highly correlated with precipitation, suggesting that this form prefers much wetter environments than the Mopti-M form. CONCLUSION: Chromosome inversions, microsatellite allele frequencies and habitat preference all indicate that the Forest M form of An. gambiae is genetically distinct from the other recognized forms within the taxon Anopheles gambiae sensu stricto. Since this study covers limited regions of Cameroon, the possibility of gene flow between the Forest-M form and Mopti-M form cannot be rejected. However, association studies of important phenotypes, such as insecticide resistance and refractoriness against malaria parasites, should take into consideration this complex population structure.

Desiccation resistance among subpopulations of Anopheles gambiae s.s. from Selinkenyi, Mali.

Certain forms of Anopheles gambiae s.s. actively maintain malaria transmission in the driest areas and months of the year because of considerable drought tolerance. We monitored desiccation resistance of F1 offspring of both the M and S forms of field-collected An. gambiae s.s. Our results indicate that the geographic cline in the distribution of the two forms, as observed in Mali, corresponds to a physiological difference in response to arid environments. In addition, female mosquitoes survived significantly longer than males, enhancing the vector competence for the malaria parasite. Our study supports a genetic link to the drought tolerance phenotype, a phenotype with important consequences to malaria transmission in many places in Africa.

Effects of varying moisture on egg production and longevity of Lutzomyia longipalpis (Diptera: Psychodidae).

The sand fly Lutzomyia longipalpis (Lutz & Neiva) (Diptera: Psychodidae), is the vector of the Neotropical parasite Leishmania chagasi. Designing methods to control the spread of this pathogen involves maintaining large laboratory colonies of these flies. However, transmission studies have been hampered particularly in regard to significant fluctuations in colony productivity. In the process of optimizing our sand fly rearing protocol, we increased moisture levels in the environment of the adult blood-fed female and found that egg development increased, whereas sand fly longevity decreased. Interestingly, continuous moisture from the fourth day after a bloodmeal significantly increased the longevity of the fly, but it did not significantly decrease the number of eggs developed. These results will help guide investigators aiming to manipulate the longevity and productivity of sand fly colonies.

Comparative studies of populations of Lutzomyia umbratilis (Diptera: Psychodidae) in Brazil and Venezuela.

We analyzed 64 quantitative and qualitative morphological characters in different populations of Lutzomyia (Nyssomyia) umbratilis Ward & Fraiha from Venezuela (Gran Sabana, Bolívar/BO) and Brazil (Serra do Navio, Amapá/AP Manaus, Amazonas/AM, Monte Dourado, Pará/ PA, and Peixoto de Azevedo, Mato Grosso/MT). Based on an analysis of qualitative characters, no differences were observed that could distinguish between the populations. Parametric tests were used to verify differences between sample means and variance homogeneity. The analysis-of-variance (ANOVA) test showed the existence of differences between character means in the five populations. In the graphic representation of the Student-Newman-Keuls test (Student-Newman-Keuls), we observed that 77% of the analyzed characters displayed disjunctions between the Venezuelan and Brazilian populations. Considering only the Brazilian population averages, we concluded that they are statistically different. When the Student-Newman-Keuls test was applied, no pattern was found that could separate them. However, in part, the phenetic analysis showed some evidences of heterogeneity in MT population.

Clarification of anomalies in the application of a 2La molecular karyotyping method for the malaria vector Anopheles gambiae.

BACKGROUND: Chromosomal inversions have been considered to be potentially important barriers to gene flow in many groups of animals through their effect on recombination suppression in heterokaryotypic individuals. Inversions can also enhance local adaptation in different groups of organisms and may often represent species-specific differences among closely related taxa. We conducted a study to characterize the 2La inversion karyotypes of An. gambiae sensu stricto mosquitoes sampled from the Kilombero Valley (Tanzania) using a newly designed PCR assay. RESULTS: We frequently encountered a (687 bp) fragment which was only present in the Kilombero Valley populations. Laboratory crossing between An. gambiae s.s. from Njage (Tanzania) and Kisumu (Western Kenya) populations resulted in F1 offspring carrying the observed fragment. Karyotype analysis did not indicate differences in 2La region chromosome morphology between individuals carrying the PCR fragments, the 207 bp fragment, or the 687 bp fragement. CONCLUSION: The observed insertion/deletion polymorphism within the region amplified by the 2La PCR diagnostic test may confound the interpretation of this assay and should be well considered in order to maintain an acceptable level of reliability in studies using this assay to describe the distribution and frequency of the 2La inversion among natural populations of An. gambiae s.s.

Ecology of sand flies (Diptera: psychodidae: phlebotominae) in the north of the state of Mato Grosso, Brazil.

Peixoto de Azevedo is located in the north of State of Mato Grosso, where environmental alterations led to an outbreak of American cutaneous leishmaniasis in the 80s. The parasite from patients was characterized as Leishmania (V.) braziliensis. The aim of this study is to contribute to the sand fly ecology of Central-West Brazil. Captures were carried out monthly using CDC light traps. Twenty-six species of sand fly were characterized; among which Lutzomyia (Lutzomyia) spathotrichia, L. runoides and L. (Psychodopygus) llanosmartinsi were recorded in the State of Mato Grosso for the first time. L. (Nyssomyia) whitmani, L. (N.) antunesi, L. (L.) spathotrichia, L. (P.) c. carrerai, L. (P.) complexa, L. (P.) lainsoni and L. (N.) umbratilis constituted 92.4% of the local fauna, among which L. (N.) whitmani and L. (N.) antunesi, accounting for about 53% of the fauna at the stations of capture. On the vertical distribution of sand flies on the Beira-Rio Farm, L. (N.) whitmani and L. (N.) antunesi prevailed at ground level and in the canopy, respectively, whereas on the BR-080, L. (P.) llanosmartinsi was prevalent on the ground and L. (P.) c. carrerai, in the canopy. It is suggested that L. (N.) umbratilis is the local vector.

Ecology of Sand Flies (Diptera: Psychodidae: Phlebotominae) in the North of the State of Mato Grosso, Brazil

Peixoto de Azevedo is located in the north of State of Mato Grosso, where environmental alterations led to an outbreak of American cutaneous leishmaniasis in the 80s. The parasite from patients was characterized as Leishmania (V.) braziliensis. The aim of this study is to contribute to the sand fly ecology of Central-West Brazil. Captures were carried out monthly using CDC light traps. Twenty-six species of sand fly were characterized; among which Lutzomyia (Lutzomyia) spathotrichia, L. runoides and L. (Psychodopygus) llanosmartinsi were recorded in the State of Mato Grosso for the first time. L. (Nyssomyia) whitmani, L. (N.) antunesi, L. (L.) spathotrichia, L. (P.) c. carrerai, L. (P.) complexa, L. (P.) lainsoni and L. (N.) umbratilis constituted 92.4 percent of the local fauna, among which L. (N.) whitmani and L. (N.) antunesi, accounting for about 53 percent of the fauna at the stations of capture. On the vertical distribution of sand flies on the Beira-Rio Farm, L. (N.) whitmani and L. (N.) antunesi prevailed at ground level and in the canopy, respectively, whereas on the BR-080, L. (P.) llanosmartinsi was prevalent on the ground and L. (P.) c. carrerai, in the canopy. It is suggested that L. (N.) umbratilis is the local vector