The global burden of arboviral diseases and the limited success in controlling them calls for innovative methods to understand arbovirus infections. Metabolomics has been applied to detect alterations in host physiology during infection. This approach relies on mass spectrometry or nuclear magnetic resonance spectroscopy to evaluate how perturbations in biological systems alter metabolic pathways, allowing for differentiation of closely related conditions. Because viruses heavily depend on host resources and pathways, they present unique challenges for characterizing metabolic changes. Here, we review the literature on metabolomics of arboviruses and focus on the interpretation of identified molecular features. Metabolomics has revealed biomarkers that differentiate disease states and outcomes, and has shown similarities in metabolic alterations caused by different viruses (e.g., lipid metabolism). Researchers investigating such metabolomic alterations aim to better understand host–virus dynamics, identify diagnostically useful molecular features, discern perturbed pathways for therapeutics, and guide further biochemical research. This review focuses on lessons derived from metabolomics studies on samples from arbovirus-infected humans.

Chikungunya virus (CHIKV) and the closely related onyong-nyong virus (ONNV) are arthritogenic arboviruses that have caused significant, often debilitating, disease in millions of people. However, despite their kinship, they are vectored by different mosquito subfamilies that diverged 180 million years ago (anopheline versus culicine subfamilies). Previous work indicated that the nonstructural protein 3 (nsP3) of these alphaviruses was partially responsible for this vector specificity. To better understand the cellular components controlling alphavirus vector specificity, a cell culture model system of the anopheline restriction of CHIKV was developed along with a protein expression strategy. Mosquito proteins that differentially interacted with CHIKV nsP3 or ONNV nsP3 were identified. Six proteins were identified that specifically bound ONNV nsP3, ten that bound CHIKV nsP3 and eight that interacted with both. In addition to identifying novel factors that may play a role in virus/vector processing, these lists included host proteins that have been previously implicated as contributing to alphavirus replication. Alphaviruses such as CHIKV and ONNV, cause severely painful human illnesses and are capable of producing large outbreaks with endemic persistence. Half of the alphavirus lifecycle is in mosquitoes, making it important to understand the molecular interactions between the virus and the vector that influence the capacity of these viruses to spread. This work identifies 24 vector proteins that may be responsible for restricting CHIKV from infecting the subfamily of mosquitoes that ONNV, CHIKV’s closest relative, uses for transmission. This is the first study to identify Anopheles host proteins that interact with alphavirus nonstructural protein 3. Understanding these virus/vector interactions is important for identifying risk factors for viral transmission and potential mechanisms for controlling vector infections by human pathogens.

Across the United States, Dirofilaria immitis (Leidy), commonly known as dog heartworm, poses a significant threat to common pets, including dogs, ferrets, and cats, but also to exotic and feral animals. This study focused on surveillance of D. immitis in mosquitoes collected in the Salt Lake Valley of Utah, with an emphasis on animals housed at a local zoo. Mosquitoes (Diptera: Culicidae) from the genera Culex, Culiseta, and Aedes were collected, identified, sorted, and subjected to DNA extraction. In the summer of 2023, extracted DNA from 320 pooled samples of 3,459 mosquitoes was tested by PCR with D. immitis-specific and panfilarial primers. Two samples collected at the zoo were positive for D. immitis, and 2 additional samples tested positive with panfilarial primers, though the precise filarial species could not be determined. In the summer of 2024, an additional 71 samples of 1,999 mosquitoes were collected from trapping at 12 locations across Salt Lake Valley, but none were positive for D. immitis. This study confirmed the presence of D. immitis-infected mosquitoes in the Salt Lake Valley, prompting the zoo to expand prophylaxis for at-risk animals and underscoring the importance of ongoing surveillance efforts. Further monitoring is recommended to assess and manage ongoing threats posed by mosquito-transmitted heartworm.

Mosquito control programs deploy mosquito traps for surveillance and targeting of vector mosquitoes. The primary attractant in these traps is generally carbon dioxide (CO2), which acts as a powerful long-distance lure. Historically, dry ice and compressed gas have been accessible as lures, but they may exhibit logistical hardships (storage, safety, equipment, calibration, consistency, etc.). Additionally, the aforementioned standards can be inaccessible in remote or low-resource environments. Microbial lures have often been proposed to bridge this gap, but prior options, such as yeast, have proven inferior compared to traditional CO2 lures. However, a mycelium-based CO2 generator, the Exhale CO2 Bag, is now commercially available. Initial studies suggested this product may be suitable for mosquito surveillance. To validate this, semi-field cage assays were conducted in Florida by releasing several species of vector mosquito into enclosures to be recollected by either Biogents or United States Centers for Disease Control and Prevention (CDC)-style traps paired with the Exhale, gas cylinders, dry ice, or unbaited. Dry ice and compressed gas performed equivalently, with Exhale collecting 25% to 50% fewer mosquitoes, and unbaited traps collected 10% to 30% fewer than Exhale. Additionally, field testing in Utah, with traps deployed across rural wetlands, industrial transition areas, and urban metropolitan areas, all yielded the same result: the Exhale mycelium and unbaited traps collected an average of 35 or fewer mosquitoes, as compared to regulated gas CO2 traps collecting 1,000 to 3,000 mosquitoes in the same areas. Though some merits exist for a mycelium-based trap, the current data and recent literature fail to support this technology for host-seeking traps in existing mosquito surveillance programs.