Characterization of the adult Aedes aegypti early midgut peritrophic matrix proteome using LC-MS.

The Aedes aegypti mosquito is the principal vector of arboviruses such as dengue, chikungunya, yellow fever, and Zika virus. These arboviruses are transmitted during adult female mosquito bloodfeeding. While these viruses must transverse the midgut to replicate, the blood meal must also reach the midgut to be digested, absorbed, or excreted, as aggregation of blood meal metabolites can be toxic to the female mosquito midgut. The midgut peritrophic matrix (PM), a semipermeable extracellular layer comprised of chitin fibrils, glycoproteins, and proteoglycans, is one such mechanism of protection for the mosquito midgut. However, this structure has not been characterized for adult female Ae. aegypti. We conducted a mass spectrometry based proteomic analysis to identify proteins that comprise or are associated with the adult female Ae. aegypti early midgut PM. Altogether, 474 unique proteins were identified, with 115 predicted as secreted. GO-term enrichment analysis revealed an abundance of serine-type proteases and several known and novel intestinal mucins. In addition, approximately 10% of the peptides identified corresponded to known salivary proteins, indicating Ae. aegypti mosquitoes extensively swallow their own salivary secretions. However, the physiological relevance of this remains unclear, and further studies are needed to determine PM proteins integral for midgut protection from blood meal derived toxicity and pathogen protection. Finally, we describe substantial discordance between previously described transcriptionally changes observed in the midgut in response to a bloodmeal and the presence of the corresponding protein in the PM. Data are available via ProteomeXchange with identifier PXD007627.

Acute Toxicity of Permethrin, Deltamethrin, and Etofenprox to the Alfalfa Leafcutting Bee.

Current regulatory requirements for insecticide toxicity to nontarget insects focus on the honey bee, Apis mellifera (L.; Hymenoptera: Apidae), but this species cannot represent all insect pollinator species in terms of response to insecticides. Therefore, we characterized the toxicity of pyrethroid insecticides used for adult mosquito management (permethrin, deltamethrin, and etofenprox) on a nontarget insect, the adult alfalfa leafcutting bee, Megachile rotundata (F.; Hymenoptera: Megachilidae) in two separate studies. In the first study, the doses causing 50 and 90% mortality (LD50 and LD90, respectively) were used as endpoints and 2-d-old adult females were exposed to eight concentrations ranging from 0.0075 to 0.076 µg/bee for permethrin and etofenprox, and 0.0013-0.0075 µg/bee for deltamethrin. For the second study, respiration rates of female M. rotundata were also recorded for 2 h after bees were dosed at the LD50 values to give an indication of stress response. Results indicated a relatively similar LD50 for permethrin and etofenprox, 0.057 and 0.051 µg/bee, respectively, and a more toxic response, 0.0016 µg/bee for deltamethrin. Comparatively, female A. mellifera workers have a LD50 value of 0.024 µg/bee for permethrin and 0.015 µg/bee for etofenprox indicating that female M. rotundata are less susceptible to topical doses of these insecticides, except for deltamethrin, where both A. mellifera and M. rotundata have an identical LD50 of 0.0016 µg/bee. Respiration rates comparing each active ingredient to control groups, as well as rates between each active ingredient, were statistically different (P < 0.0001). The addition of these results to existing information on A. mellifera may provide more insights on how other economically beneficial and nontarget bees respond to pyrethroids.

Ironing out the Details: Exploring the Role of Iron and Heme in Blood-Sucking Arthropods.

Heme and iron are essential molecules for many physiological processes and yet have the ability to cause oxidative damage such as lipid peroxidation, protein degradation, and ultimately cell death if not controlled. Blood-sucking arthropods have evolved diverse methods to protect themselves against iron/heme-related damage, as the act of bloodfeeding itself is high risk, high reward process. Protective mechanisms in medically important arthropods include the midgut peritrophic matrix in mosquitoes, heme aggregation into the crystalline structure hemozoin in kissing bugs and hemosomes in ticks. Once heme and iron pass these protective mechanisms they are presumed to enter the midgut epithelial cells via membrane-bound transporters, though relatively few iron or heme transporters have been identified in bloodsucking arthropods. Upon iron entry into midgut epithelial cells, ferritin serves as the universal storage protein and transport for dietary iron in many organisms including arthropods. In addition to its role as a nutrient, heme is also an important signaling molecule in the midgut epithelial cells for many physiological processes including vitellogenesis. This review article will summarize recent advancements in heme/iron uptake, detoxification and exportation in bloodfeeding arthropods. While initial strides have been made at ironing out the role of dietary iron and heme in arthropods, much still remains to be discovered as these molecules may serve as novel targets for the control of many arthropod pests.

The Influence of Ambient Temperature on the Susceptibility of Aedes aegypti (Diptera: Culicidae) to the Pyrethroid Insecticide Permethrin.

Insecticides are the most common strategy used for the management of mosquitoes. Changes in ambient temperature can alter the toxicity of insecticides to ectothermic organisms. Studies show organophosphate insecticides exhibit a positive correlation between ambient temperature and mortality for many insect species, and carbamate insecticides exhibit a slightly negative correlation between ambient temperature and mortality. Pyrethroid insecticides exhibit a distinctly negative correlation between increasing ambient temperature and mortality for insects. However, this relationship has not been systematically studied for adult mosquitoes. Therefore, we examined the influence of temperature on the susceptibility of adult Aedes aegypti L. (Diptera: Culicidae) when exposed to permethrin. The median lethal concentration, LC50, was estimated for adult Ae. aegypti when exposed to eight concentrations of permethrin (ranging from 0.06­0.58 ng/cm2) at each of the following temperatures­16, 23, 26, 30, 32, and 34C­for 24 h in bottle assays. The estimated LC50 for each temperature was 0.26, 0.36, 0.36, 0.45, 0.27, and 0.31 ng/cm2, respectively. Results indicated a negative correlation between temperature and mortality from 16 to 30C, a positive correlation between temperature and mortality from 30 to 32C, and a negative correlation between temperature and mortality from 32 to 34C. If mosquito populations are expanding in space and time because of increased ambient temperatures and cannot be managed as effectively with pyrethroids, the spread of mosquito-borne diseases may pose considerable additional risk to public health.