The discovery of an odd virus from hematophagous arthropods 40 years ago by Stollar and Thomas described cell fusing agent virus in cells derived from Aedes aegypti mosquitoes. Then came the report of Kamiti River virus from Ae. macintoshi in 1999, followed by worldwide reports of the discovery of other viruses of mosquitoes, ticks, and midges that replicate only in arthropods and not in vertebrates or in vertebrate cells. These viruses (now totaling at least 64 published) have genomes analogous to viruses in various families that include arboviruses and nonarboviruses. It is likely that some of these viruses have been insufficiently studied and may yet be shown to infect vertebrates. However, there is no doubt that the vast majority are restricted to arthropods alone and that they represent a recently recognized clade. Their biology, modes of transmission, worldwide distribution (some have been detected in wild-caught mosquitoes in both Asia and the United States, for example), molecular characteristics of their genomes, and potential for becoming vertebrate pathogens, or at least serving as virus reservoirs, are fascinating and may provide evidence useful in understanding virus evolution. Because metagenomics studies of arthropods have shown that arthropod genomes are the sources of arthropod virus genomes, further studies may also provide insights into the evolution of arthropods. More recently, others have published excellent papers that briefly review discoveries of arthropod viruses and that characterize certain genomic peculiarities, but, to now, there have been no reviews that encompass all these facets. We therefore anticipate that this review is published at a time and in a manner that is helpful for both virologists and entomologists to make more sense and understanding of this recently recognized and obviously important virus group. This review focuses specifically on arthropod viruses in hematophagous arthropods.

The Asian tiger mosquito Aedes albopictus (Skuse), is a highly invasive species that continues to expand its geographic distribution both in the United States and in countries on other continents. Studies have demonstrated its susceptibility to infection with at least 32 viruses, including 13 that are present in the United States. Despite this susceptibility, its role as a significant competent vector in natural transmission cycles of arboviruses, has been limited. However, with the recent introductions of chikungunya and Zika viruses into the Americas, for which Ae. albopictus is a recognized vector, it is possible that the species may contribute to the transmission of these viruses to humans and perhaps other susceptible vertebrates.

Until 2005, chikungunya virus (CHIKV) was a relatively little-studied pathogen restricted to parts of Africa and Asia. Epidemics were sporadic and separated by years of quiescence. In late 2004, the East Central South African genotype of CHIKV moved from Kenya onto the Indian Ocean island of Comoros. The global onslaught of CHIKV had begun. In November of 2005, viral isolates were identified with, what might normally be regarded as an insignificant, single alanine to valine mutation at position 226 of the envelope E1 gene.1 This simple mutation had a remarkable effect; making the virus approximately 100 times more infectious to the Asian tiger mosquito, Aedes albopictus, and it was this species that was transmitting the virus rather than the usual vector, Aedes aegypti.2 Subsequent ‘second-step’ mutations further enhanced the ability of the virus to infect and/or disseminate from the midgut to the salivary glands in Ae. albopictus.3 However, these viruses can still be transmitted by Ae. aegypti. Within a year of the Indian Ocean lineage emerging, over 250 000 people had been infected. An epidemic in Asia began within months and has infected several million people in India and other Asian countries. Chikungunya infections occurred in many countries as a result of people travelling from areas with active transmission. The presence of tiger mosquito vector has been critical to enable localized CHIKV outbreaks in Italy and France. This species continues to invade new territory,4 being identified in the United Kingdom for the first time in 2014.

The study of many arthropod-borne pathogens requires high biosafety considerations, including the use of specialized facilities and equipment for arthropod containment. Mosquito- and tick-borne viruses such as yellow fever, West Nile, and Crimean Congo hemorrhagic fever viruses require facilities that are suitable for housing vertebrates. Multidisciplinary studies that incorporate the vector, vertebrate, and pathogens are essential for a complete understanding of the interactions between these transmission cycle components, especially if they aim to evaluate and model relative susceptibilities of different arthropods and vertebrates to infection and transmission between these. Under laboratory conditions, these studies can be relatively simple, for example, involving colonized arthropods, small animals, and attenuated viruses. Other studies are complex with large animals, high-biocontainment pathogens, and field-collected arthropods. These require a higher level of containment and special design considerations. Both of these types of experiments have their relative merits. A thorough understanding of the issues related to these types of studies and the benefits and drawbacks to using various challenge models will enable the researcher to develop realistic goals for various experiments. This review examines the varied issues that should be considered prior to starting these experiments and covers the basics from the procurement of various arthropods, rearing, high-containment facilities and operational issues specific to work with arthropods, types of infection experiments, and specific issues with arthropod and animal experiments in biosafety levels 3 and 4.