Transfected Mosquito Vectors

A method is provided for producing an artificial infection in a Culicidae (mosquito) species. The mosquitoes include species within the subfamilies Culicinae and Anophelinae, and the species include Aedes albopictus, Aedes aegypti and Aedes polynesiensis infected with a Wolbachia infection. The infection may be a strain of Wolbachia which does not normally or naturally infect the selected mosquito species. The artificially infected Aedes mosquito can be introduced into a mosquito population to control the reproduction capability of the population by introducing an incompatible Wolbachia infection. The present method can be used as a novel means to limit mosquito-borne pathogens and thus control or prevent mosquito-borne diseases such as dengue, lymphatic filariasis, etc

Population genetic structure of \u3cem\u3eAedes polynesiensis\u3c/em\u3e in the Society Islands of French Polynesia: implications for control using a \u3cem\u3eWolbachia\u3c/em\u3e-based autocidal strategy

BACKGROUND: Aedes polynesiensis is the primary vector of Wuchereria bancrofti in the South Pacific and an important vector of dengue virus. An improved understanding of the mosquito population genetics is needed for insight into the population dynamics and dispersal, which can aid in understanding the epidemiology of disease transmission and control of the vector. In light of the potential release of a Wolbachia infected strain for vector control, our objectives were to investigate the microgeographical and temporal population genetic structure of A. polynesiensis within the Society Islands of French Polynesia, and to compare the genetic background of a laboratory strain intended for release into its population of origin. METHODS: A panel of eight microsatellite loci were used to genotype A. polynesiensis samples collected in French Polynesia from 2005-2008 and introgressed A. polynesiensis and Aedes riversi laboratory strains. Examination of genetic differentiation was performed using F-statistics, STRUCTURE, and an AMOVA. BAYESASS was used to estimate direction and rates of mosquito movement. RESULTS: FST values, AMOVA, and STRUCTURE analyses suggest low levels of intra-island differentiation from multiple collection sites on Tahiti, Raiatea, and Maupiti. Significant pair-wise FST values translate to relatively minor levels of inter-island genetic differentiation between more isolated islands and little differentiation between islands with greater commercial traffic (i.e., Tahiti, Raiatea, and Moorea). STRUCTURE analyses also indicate two population groups across the Society Islands, and the genetic makeup of Wolbachia infected strains intended for release is similar to that of wild-type populations from its island of origin, and unlike that of A. riversi. CONCLUSIONS: The observed panmictic population on Tahiti, Raiatea, and Moorea is consistent with hypothesized gene flow occurring between islands that have relatively high levels of air and maritime traffic, compared to that of the more isolated Maupiti and Tahaa. Gene flow and potential mosquito movement is discussed in relation to trials of applied autocidal strategies

Integration of irradiation with cytoplasmic incompatibility to facilitate a lymphatic filariasis vector elimination approach

<p>Abstract</p> <p>Background</p> <p>Mass drug administration (MDA) is the emphasis of an ongoing global lymphatic filariasis (LF) elimination program by the World Health Organization, in which the entire 'at risk' human population is treated annually with anti-filarial drugs. However, there is evidence that the MDA strategy may not be equally appropriate in all areas of LF transmission, leading to calls for the augmentation of MDA with anti-vector interventions. One potential augmentative intervention is the elimination of vectors via repeated inundative releases of male mosquitoes made cytoplasmically incompatible via an infection with <it>Wolbachia </it>bacteria. However, with a reduction in the vector population size, there is the risk that an accidental female release would permit the establishment of the incompatible <it>Wolbachia </it>infection type, resulting in population replacement instead of population elimination. To avoid the release of fertile females, we propose the exposure of release individuals to low doses of radiation to sterilize any accidentally released females, reducing the risk of population replacement.</p> <p>Results</p> <p><it>Aedes polynesiensis </it>pupae of differing ages were irradiated to determine a radiation dose that results in sterility but that does not affect the survival and competitiveness of males. Laboratory assays demonstrate that males irradiated at a female sterilizing dosage of 40 Gy are equally competitive with un-irradiated males. No effect of irradiation on the ability of <it>Wolbachia </it>to affect egg hatch was observed.</p> <p>Conclusion</p> <p>An irradiation dose of 40 Gy is sufficient to cause female sterility, but has no observed negative effect on male fitness. The results support further development of this approach as a preventative measure against accidental population replacement.</p

Life-Shortening \u3cem\u3eWolbachia\u3c/em\u3e Infection Reduces Population Growth of \u3cem\u3eAedes aegypti\u3c/em\u3e

Wolbachia bacteria are being introduced into natural populations of vector mosquitoes, with the goal of reducing the transmission of human diseases such as Zika and dengue fever. The successful establishment of Wolbachia infection is largely dependent on the effects of Wolbachia infection to host fitness, but the effects of Wolbachia infection on the individual life-history traits of immature mosquitoes can vary. Here, the effects of life-shortening Wolbachia (wMelPop) on population growth of infected individuals were evaluated by measuring larval survival, developmental time and adult size of Aedes aegypti in intra- (infected or uninfected only) and inter-group (mixed with infected and uninfected) larval competition assays. At low larval density conditions, the population growth of wMelPop infected and uninfected individuals was similar. At high larval densities, wMelPop infected individuals had a significantly reduced population growth rate relative to uninfected individuals, regardless of competition type. We discuss the results in relation to the invasion of the wMelPop Wolbachia infection into naturally uninfected populations

The Effects of Boric Acid Sugar Bait on \u3ci\u3eWolbachia\u3c/i\u3e Trans-Infected Male \u3ci\u3eAedes albopictus\u3c/i\u3e (ZAP Males\u3csup\u3e®\u3c/sup\u3e) in Laboratory Conditions

The field release of Wolbachia trans-infected male mosquitoes, as well as the use of toxic sugar baits, is a novel and promising candidate technique for integrated mosquito management programs. However, the methods of action of the two techniques may not be complementary, because the Wolbachia method releases mosquitoes into the environment expecting a wild population reduction in subsequent generations while the toxic baits are intended to reduce the wild population by killing mosquitoes. This laboratory study was conducted to evaluate the effectiveness of boric acid toxic sugar baits on Wolbachia trans-infected male Aedes albopictus, relative to wild-type Ae. albopictus males. Wolbachia trans-infected (ZAP male®) and the wild-type Ae. albopictus males were exposed separately to 1% boric acid in a 10% sucrose solution in BugDorms. In the control test, the two groups were exposed to 10% sucrose solution without boric acid. Percent mortalities were counted for 24 h, 48 h and 72 h post exposure periods. The results show that 1% boric acid toxic sugar bait can effectively kill ZAP males under laboratory conditions, and the effectiveness was significantly higher after 24 h and 48 h, compared to wild-type male Ae. albopictus. This finding will help in planning and coordinating integrated mosquito management programs, including both Wolbachia trans-infected mosquito releases and the use of toxic sugar baits against Ae. albopictus

Genome-wide analysis of the interaction between the endosymbiotic bacterium Wolbachia and its Drosophila host

BACKGROUND: Intracellular Wolbachia bacteria are obligate, maternally-inherited, endosymbionts found frequently in insects and other invertebrates. The success of Wolbachia can be attributed in part to an ability to alter host reproduction via mechanisms including cytoplasmic incompatibility (CI), parthenogenesis, feminization and male killing. Despite substantial scientific effort, the molecular mechanisms underlying the Wolbachia/host interaction are unknown. RESULTS: Here, an in vitro Wolbachia infection was generated in the Drosophila S2 cell line, and transcription profiles of infected and uninfected cells were compared by microarray. Differentially-expressed patterns related to reproduction, immune response and heat stress response are observed, including multiple genes that have been previously reported to be involved in the Wolbachia/host interaction. Subsequent in vivo characterization of differentially-expressed products in gonads demonstrates that Angiotensin Converting Enzyme (Ance) varies between Wolbachia infected and uninfected flies and that the variation occurs in a sex-specific manner. Consistent with expectations for the conserved CI mechanism, the observed Ance expression pattern is repeatable in different Drosophila species and with different Wolbachia types. To examine Ance involvement in the CI phenotype, compatible and incompatible crosses of Ance mutant flies were conducted. Significant differences are observed in the egg hatch rate resulting from incompatible crosses, providing support for additional experiments examining for an interaction of Ance with the CI mechanism. CONCLUSION: Wolbachia infection is shown to affect the expression of multiple host genes, including Ance. Evidence for potential Ance involvement in the CI mechanism is described, including the prior report of Ance in spermatid differentiation, Wolbachia-induced sex-specific effects on Ance expression and an Ance mutation effect on CI levels. The results support the use of Wolbachia infected cell cultures as an appropriate model for predicting in vivo host/Wolbachia interactions

The Influence of a Slow-Breathing Protocol on Heart Rate and Blood Pressure from Exercise in Moderately Trained Females

International Journal of Exercise Science 12(2): 714-725, 2019. Heart rate recovery (HRR) and blood pressure recovery (BPR) from exercise are both important indicators of health and fitness and are strongly associated with cardiovascular disease. The purpose of this study was to compare the effects of a slow-breathing technique, upright passive recovery (PASS), and active recovery (ACT) on HRR and BPR from exercise. Nine moderately trained, college-aged (20.22 ± 0.97 yrs) female participants cycled three times on an ergometer for 15 mins at 70% of their heart rate maximum (HRmax), each of which was followed by one of three 5 min recovery interventions with heart rate (HR) and blood pressure (BP) objectively measured. Each participant completed all three recovery protocols. One recovery protocol consisted of breathing at a rate of 6 breaths per minute (BRE), another involved PASS and the third was ACT at 60 RPM and 25 W. A repeated measures ANOVA revealed there was a significant effect of protocol (p= 0.00, hp2= 0.67) with HRR. BRE resulted in the fastest HRR of 69 ± 9.31 bpm (40.12%) at the end of the 5 min recovery compared to 63 ± 10.60 bpm (36.57%) and 47 ± 12.54 bpm (27.34%) for PASS and ACT, respectively. A second repeated measures ANOVA indicated there was no effect of protocol (P = 0.43), nor was there a significant interaction with time (p= 0.68), for BPR. The results indicated that BRE increased HRR after exercise more rapidly than PASS or ACT with no influence on BPR. These findings lead to future research needed to explore different breathing protocols following exercise in at-risk populations, such as individuals with cardiovascular disease

Female Adult \u3cem\u3eAedes albopictus\u3c/em\u3e Suppression by \u3cem\u3eWolbachia\u3c/em\u3e-Infected Male Mosquitoes

Dengue, chikungunya and zika viruses are pathogens with an increasing global impact. In the absence of an approved vaccine or therapy, their management relies on controlling the mosquito vectors. But traditional controls are inadequate, and the range of invasive species such as Aedes albopictus (Asian Tiger Mosquito) is expanding. Genetically modified mosquitoes are being tested, but their use has encountered regulatory barriers and public opposition in some countries. Wolbachia bacteria can cause a form of conditional sterility, which can provide an alternative to genetic modification or irradiation. It is unknown however, whether openly released, artificially infected male Ae. albopictus can competitively mate and sterilize females at a level adequate to suppress a field population. Also, the unintended establishment of Wolbachia at the introduction site could result from horizontal transmission or inadvertent female release. In 2014, an Experimental Use Permit from the United States Environmental Protection Agency approved a pilot field trial in Lexington, Kentucky, USA. Here, we present data showing localized reduction of both egg hatch and adult female numbers. The artificial Wolbachia type was not observed to establish in the field. The results are discussed in relation to the applied use of Wolbachia-infected males as a biopesticide to suppress field populations of Ae. albopictus

Population Impacts of \u3cem\u3eWolbachia\u3c/em\u3e on \u3cem\u3eAedes albopictus\u3c/em\u3e

Prior studies have demonstrated that Wolbachia, a commonly occurring bacterium capable of manipulating host reproduction, can affect life history traits in insect hosts, which in turn can have population-level effects. Effects on hosts at the individual level are predicted to impact population dynamics, but the latter has not been examined empirically. Here, we describe a biological model system based on Aedes albopictus (Asian tiger mosquito) that allows for measurement of population dynamics, which has not been accomplished in prior field trials or laboratory designs. The results demonstrate the studied populations to be robust and allow for persistent, closed populations with overlapping generations, which are regulated solely through density-dependent, intraspecific competition for limited resources. Using a novel experimental design, we compare populations that are either uninfected or infected with Wolbachia. The results show differences that include population size, eclosion rates, adult survivorship, and fecundity. The aposymbiotic populations were generally larger and adults longer lived relative to the infected populations. The outcome is discussed in context with naturally occurring Wolbachia invasions, proposed autocidal strategies, and the utility of the developed system as a biological platform for hypothesis testing and improved parameterization

Leishmania sexual reproductive strategies as resolved through computational methods designed for aneuploid genomes

A cryptic sexual reproductive cycle i