The direct and indirect effects of copper on vector-borne disease dynamics.

Metal pollution is a growing concern that affects the health of humans and animals globally. Copper is an essential insect micronutrient required for respiration, pigmentation and oxidative stress protection but can also act as a potentially toxic trace element. While several studies have focused on the negative fitness effects of copper on the aquatic larvae of mosquitoes, the effects of larval copper exposure on adult mosquito fitness (i.e., survival and fecundity) and their ability to transmit parasites (i.e., vector competence) remains unclear. Here, using a well-studied model vector-parasite system, the mosquito Aedes aegypti and parasite Dirofilaria immitis, we show that sublethal copper exposure in larval mosquitoes alters adult female fecundity and vector competence. Specifically, mosquitoes exposed to copper had a hormetic fecundity response and mosquitoes exposed to 600 µg/L of copper had significantly fewer infective parasite larvae than control mosquitoes not exposed to copper. Thus, exposure of mosquito larvae to copper levels far below EPA-mandated safe drinking water limits (1300 µg/L) can impact vector-borne disease dynamics not only by reducing mosquito abundance (through increased larval mortality), but also by reducing parasite transmission risk. Our results also demonstrated that larval copper is retained through metamorphosis to adulthood in mosquitoes, indicating that these insects could transfer copper from aquatic to terrestrial foodwebs, especially in urban areas where they are abundant. To our knowledge this is the first study to directly link metal exposure with vector competence (i.e., ability to transmit parasites) in any vector-parasite system. Additionally, it also demonstrates unequivocally that mosquitoes can transfer contaminants from aquatic to terrestrial ecosystems. These results have broad implications for public health because they directly linking contaminants and vector-borne disease dynamics, as well as linking mosquitoes and contaminant dynamics.

Drug Resistance in Filarial Parasites Does Not Affect Mosquito Vectorial Capacity.

Parasite drug resistance presents a major obstacle to controlling and eliminating vector-borne diseases affecting humans and animals. While vector-borne disease dynamics are affected by factors related to parasite, vertebrate host and vector, research on drug resistance in filarial parasites has primarily focused on the parasite and vertebrate host, rather than the mosquito. However, we expect that the physiological costs associated with drug resistance would reduce the fitness of drug-resistant vs. drug-susceptible parasites in the mosquito wherein parasites are not exposed to drugs. Here we test this hypothesis using four isolates of the dog heartworm (Dirofilaria immitis)-two drug susceptible and two drug resistant-and two vectors-the yellow fever mosquito (Aedes aegypti) and the Asian tiger mosquito (Ae. albopictus)-as our model system. Our data indicated that while vector species had a significant effect on vectorial capacity, there was no significant difference in the vectorial capacity of mosquitoes infected with drug-resistant vs. drug-susceptible parasites. Consequently, contrary to expectations, our data indicate that drug resistance in D. immitis does not appear to reduce the transmission efficiency of these parasites, and thus the spread of drug-resistant parasites in the vertebrate population is unlikely to be mitigated by reduced fitness in the mosquito vector.

Laboratory colonization by Dirofilaria immitis alters the microbiome of female Aedes aegypti mosquitoes.

BACKGROUND: The ability of blood-feeding arthropods to successfully acquire and transmit pathogens of medical and veterinary importance has been shown to be interfered with, or enhanced by, the arthropod's native microbiome. Mosquitoes transmit viruses, protozoan and filarial nematodes, the majority of which contribute to the 17% of infectious disease cases worldwide. Dirofilaria immitis, a mosquito-transmitted filarial nematodes of dogs and cats, is vectored by several mosquito species including Aedes aegypti. METHODS: In this study, we investigated the impact of D. immitis colonization on the microbiome of laboratory reared female Ae. aegypti. Metagenomic analysis of the V3-V4 variable region of the microbial 16S RNA gene was used for identification of the microbial differences down to species level. RESULTS: We generated a total of 1068 OTUs representing 16 phyla, 181 genera and 271 bacterial species. Overall, in order of abundance, Proteobacteria, Bacteroidetes, Actinobacteria and Firmicutes were the most represented phylum with D. immitis-infected mosquitoes having more of Proteobacteria (71%) than uninfected mosquitoes (56.9%). An interesting finding in this study is the detection of Klebsiella oxytoca in relatively similar abundance in infected and uninfected mosquitoes, suggesting a possible endosymbiotic relationship, and has been previously shown to indirectly compete for nutrients with fungi on domestic housefly eggs and larvae. While D. immitis colonization has no effect on the overall species richness, we identified significant differences in the composition of selected bacterial genera and phyla between the two groups. We also reported distinct compositional and phylogenetic differences in the individual bacterial species when commonly identified bacteria were compared. CONCLUSIONS: To the best of our knowledge, this is the first study to understand the impact of a filarial infection on the microbiome of its mosquito vector. Further studies are required to identify bacteria species that could play an important role in the mosquito biology. While the microbiome composition of Ae. aegypti mosquito have been previously reported, our study shows that in an effort to establish itself, a filarial nematode modifies and alters the overall microbial diversity within its mosquito host.

Effects of methylmercury on mosquito oviposition behavior: Maladaptive response to non-toxic exposure.

Animals can modulate their own exposure to environmental contaminants through behavioral plasticity such as diet and habitat choice. However, it remains unclear if behavior also has cascading effects on contaminant exposure across multiple generations. In insects, oviposition site selection is an important behavior females can use to modify offspring contaminant exposure risk. In this study, we use the yellow fever mosquito, Aedes aegypti, to test how methylmercury (MeHg) affects oviposition site selection. We found that mosquito larval development rate and survival were negatively affected at MeHg concentrations ≥100 ppb. Adult females not exposed to MeHg as larvae avoided oviposition sites with high MeHg concentrations (>50 ppb), but MeHg exposure at the larval stage significantly affected this oviposition site selection. Specifically, females raised from larvae exposed to non-toxic MeHg levels (i.e., five-50 ppb) showed a significant increase in preference for oviposition sites contaminated with toxic MeHg concentrations (≥500 ppb), compared to unexposed controls. This maladaptive behavioral response could be because, when conditioned with non-toxic MeHg concentrations, MeHg-associated olfactory cues act as a "supernormal" stimulus during oviposition site selection. Importantly, however, this maladaptive behavioral response is eliminated in female mosquitoes raised from larvae exposed to toxic MeHg concentrations (i.e. 100 ppb), and these mosquitoes showed a significant increase in preference for MeHg uncontaminated oviposition sites, compared to unexposed controls. Thus, in mosquitoes, the magnitude of MeHg exposure in one generation can impact MeHg exposure in subsequent generations by altering oviposition site selection behavior. Our results have broad implications for our understanding of how contaminant-mediated behavioral modifications can feedback on contaminant exposure risk across multiple generations, and consequently how behavior can affect the evolutionary trajectory of organisms inhabiting a heterogeneously contaminated environment.