Understanding the ecological and epidemiological roles of pets in the transmission of SARS-CoV-2 is critical for animal and human health, identifying household reservoirs, and predicting the potential enzootic maintenance of the virus. We conducted a longitudinal household transmission study of 76 dogs and cats living with at least one SARS-CoV-2-infected human in Texas and found that 17 pets from 25.6% of 39 households met the national case definition for SARS-CoV-2 infections in animals. This includes three out of seventeen (17.6%) cats and one out of fifty-nine (1.7%) dogs that were positive by RT-PCR and sequencing, with the virus successfully isolated from the respiratory swabs of one cat and one dog. Whole-genome sequences of SARS-CoV-2 obtained from all four PCR-positive animals were unique variants grouping with genomes circulating among people with COVID-19 in Texas. Re-sampling showed persistence of viral RNA for at least 25 d-post initial test. Additionally, seven out of sixteen (43.8%) cats and seven out of fifty-nine (11.9%) dogs harbored SARS-CoV-2 neutralizing antibodies upon initial sampling, with relatively stable or increasing titers over the 2–3 months of follow-up and no evidence of seroreversion. The majority (82.4%) of infected pets were asymptomatic. ‘Reverse zoonotic’ transmission of SARS-CoV-2 from infected people to animals may occur more frequently than recognized.

Rift Valley fever virus (RVFV) is a mosquito-borne virus in the family Bunyaviridiae that has spread throughout continental Africa to Madagascar and the Arabian Peninsula. The establishment of RVFV in North America would have serious consequences for human and animal health in addition to a significant economic impact on the livestock industry. Published and unpublished data on RVFV vector competence, vertebrate host competence, and mosquito feeding patterns from the United States were combined to quantitatively implicate mosquito vectors and vertebrate hosts that may be important to RVFV transmission in the United States. A viremia-vector competence relationship based on published mosquito transmission studies was used to calculate a vertebrate host competence index which was then combined with mosquito blood feeding patterns to approximate the vector and vertebrate amplification fraction, defined as the relative contribution of the mosquito or vertebrate host to pathogen transmission. Results implicate several Aedes spp. mosquitoes and vertebrates in the order Artiodactyla as important hosts for RVFV transmission in the U.S. Moreover, this study identifies critical gaps in knowledge which would be necessary to complete a comprehensive analysis identifying the different contributions of mosquitoes and vertebrates to potential RVFV transmission in the U.S. Future research should focus on (1) the dose-dependent relationship between viremic exposure and the subsequent infectiousness of key mosquito species, (2) evaluation of vertebrate host competence for RVFV among North American mammal species, with particular emphasis on the order Artiodactyla, and (3) identification of areas with a high risk for RVFV introduction so data on local vector and host populations can help generate geographically appropriate amplification fraction estimates. In anticipation of continued pathogen emergence in the U.S. due to globalization climate change, and other factors, the development of proactive management plans and interventions to predict and then intervene is going to be more efficient and effective than retrospective plans developed after pathogen emergence. Effective management of mosquito-borne pathogens like Rift Valley fever virus (RVFV) requires an understanding of the roles that different mosquito species and vertebrate hosts play in transmission. This study combines data on mosquito transmission efficiency, mosquito feeding patterns, and vertebrate infectiousness to quantitatively evaluate the relative importance of different mosquito species and vertebrate hosts to the amplification of RVFV in the U.S. We identify several species of floodwater Aedes spp. mosquitoes that would be the most likely vectors for RVFV, and hoofed ungulates (deer, cows, sheep) would be the most important amplifying vertebrate hosts. Although these data provide public and animal health agencies a priori knowledge on the primary mosquitoes that should be targeted for vector control and the highest priority animals to receive vaccines, this analysis reveals many gaps in knowledge reducing our ability to predict and then manage a potential invasion of RVFV.

Dispersal is a critical life history behavior for mosquitoes and is important for the spread of mosquito-borne disease. We implemented the first stable isotope mark-capture study to measure mosquito dispersal, focusing on Culex pipiens in southwest suburban Chicago, Illinois, a hotspot of West Nile virus (WNV) transmission. We enriched nine catch basins in 2010 and 2011 with ¹⁵N-potassium nitrate and detected dispersal of enriched adult females emerging from these catch basins using CDC light and gravid traps to distances as far as 3 km. We detected 12 isotopically enriched pools of mosquitoes out of 2,442 tested during the two years and calculated a mean dispersal distance of 1.15 km and maximum flight range of 2.48 km. According to a logistic distribution function, 90% of the female Culex mosquitoes stayed within 3 km of their larval habitat, which corresponds with the distance-limited genetic variation of WNV observed in this study region. This study provides new insights on the dispersal of the most important vector of WNV in the eastern United States and demonstrates the utility of stable isotope enrichment for studying the biology of mosquitoes in other disease systems. The distance and direction of adult mosquitoes movement on the landscape are important processes in the spread of mosquito-borne diseases, and are critical to understand to the development of effective intervention programs. Here we present a novel approach to study adult mosquito dispersal by using stable isotope enrichment of natural larval habitats. We apply this technique in a focal hotspot of West Nile virus (WNV) transmission in suburban, Chicago, USA to measure dispersal of Culex spp. mosquitoes. We enriched larval mosquitoes in residential catch basins using ¹⁵N-potassium nitrate and captured adult mosquitoes in traps surrounding these catch basins. Of 10,817 adult female Culex mosquitoes trapped and tested for stable isotopes, 12 individuals were enriched with ¹⁵N, indicating they originated from the catch basins receiving stable isotope amendments. The mean dispersal distance was 1.15 km and maximum flight range was 2.48 km. Ninety percent of the female Culex mosquitoes stayed within 3 km of their larval habitat, which corresponds with the distance-limited genetic variation of WNV observed in this study region. This study provides new insights on the dispersal of the most important vector of WNV in the eastern United States and demonstrates the utility of stable isotope enrichment for studying the biology of mosquitoes in other disease systems.

The dispersal patterns of mosquito vectors are important drivers of vector-borne infectious disease dynamics and understanding movement patterns is pivotal to devise successful intervention strategies. Here, we investigate the dispersal patterns of two globally important mosquito vectors, Aedes albopictus and Culex quinquefasciatus, by marking naturally-occurring larvae with stable isotopes (¹³C or ¹⁵N). Marked individuals were captured with 32 CDC light trap, 32 gravid trap, and 16 BG Sentinel at different locations within two-kilometer radii of six larval habitats enriched with either ¹³C or ¹⁵N. In total, 720 trap nights from July to August 2013 yielded a total of 32,140 Cx. quinquefasciatus and 7,722 Ae. albopictus. Overall, 69 marked female mosquitoes and 24 marked male mosquitoes were captured throughout the study period. The distance that Cx. quinquefasciatus females traveled differed for host-seeking and oviposition-seeking traps, with females seeking oviposition sites traveling further than those seeking hosts. Our analysis suggests that 41% of Cx. quinquefasciatus females that were host-seeking occurred 1–2 kilometer from their respective natal site, while 59% remained within a kilometer of their natal site. In contrast, 59% of Cx. quinquefasciatus females that were seeking oviposition sites occurred between 1–2 kilometer away from their larval habitat, while 15% occurred > 2 kilometer away from their natal site. Our analysis estimated that approximately 100% of Ae. albopictus females remained within 1 km of their respective natal site, with 79% occurring within 250m. In addition, we found that male Ae. albopictus dispersed farther than females, suggesting male-biased dispersal in this Ae. albopictus population. This study provides important insights on the dispersal patterns of two globally relevant vector species, and will be important in planning next generation vector control strategies that mitigate mosquito-borne disease through sterile insect techniques, novel Wolbachia infection, and gene drive strategies. Resolving patterns of mosquito dispersal across landscapes is a critical step toward the development of effective control strategies that mitigate vector-borne disease transmission and its public health burden. Here, we used a recently developed technique involving the enrichment of aquatic larval habitat with stable isotopes to mark male and female mosquitoes of two important vector species, Aedes albopictus and Culex quinquefasciatus. We show that the patterns of dispersal were fundamentally different between these urban vectors. Culex quinquefasciatus dispersed much further than Aedes albopictus. In addition, male Aedes albopictus dispersed further than female mosquitoes. Our study suggests that infectious disease agents transmitted by Culex quinquefasciatus will be more difficult to control locally than those transmitted by Aedes albopictus. Our results on sex-biased dispersal in Aedes albopictus have implications for next-generation vector control strategies that rely on the release of sterile or sterilizing males to control mosquito populations. Finally, our study continues to show the utility of the stable-isotope marking technique to study mosquito movement.

Across the Americas, triatomine insects harbor diverse strains of Trypanosoma cruzi (T. cruzi), agent of Chagas disease. Geographic patterns of vector infection and parasite strain associations, especially in vectors encountered by the public, may be useful in assessing entomological risk, but are largely unknown across the US. We collected Triatoma spp. from across the US (mainly Texas), in part using a citizen science initiative, and amplified T. cruzi DNA to determine infection prevalence and parasite discrete typing units (DTUs). We found 54.4% infection prevalence in 1510 triatomines of 6 species; prevalence in adult T. gerstaeckeri (63.3%; n=897) and T. lecticularia (66.7%; n=66) was greater than in T. sanguisuga (47.6%; n=315), T. indictiva (47.8% n=67), T. rubida (14.1%; n=64), and T. protracta (10.5%; n=19). The odds of infection in adults were 9.73 times higher than in nymphs (95% CI 4.46-25.83). PCR of the spliced leader intergenic region (SL-IR) and/or the putative lathosterol/episterol oxidase TcSC5D gene revealed exclusively T. cruzi DTUs TcI and TcIV; 5.5% of T. cruzi-positive samples were not successfully typed. T. gerstaeckeri (n=548) were more frequently infected with TcI (53.9%) than TcIV (34.4%), and 11.9% showed mixed TcI/TcIV infections. In contrast, T. sanguisuga (n=135) were more frequently infected with TcIV (79.3%) than TcI (15.6%), and 5.2% showed mixed infections. Relative abundance of parasite DTUs varied spatially, with both TcI and TcIV co-circulating in vectors in central Texas, while TcIV predominated in northern Texas. Given prior findings implicating TcI in human disease and TcI and TcIV in animal disease in the US, knowledge of spatial distribution of T. cruzi infection and DTUs in vectors is important to understanding public and veterinary health risk of T. cruzi infection.

Defining spatial and temporal occurrences of triatomine vectors of Trypanosoma cruzi, the agent of Chagas disease, in the US is critical for public health protection. Through a citizen science program and field collections from 2012 to 2016, we collected 3,215 triatomines, mainly from Texas. Using morphological and molecular approaches, we identified seven Triatoma species and report sex, length, and blood engorgement status. Many citizen-collected triatomines (92.9%) were encountered indoors, in peridomestic settings, or in dog kennels and represent spillover transmission risk of T. cruzi to humans and domestic animals. The most commonly collected species were Triatoma gerstaeckeri and Triatoma sanguisuga. Adult T. gerstaeckeri were collected from May to September, peaking from June to July, whereas adult T. sanguisuga were active later, from June to October, peaking from July to September. Based on cross correlation analyses, peaks of captures varied by species and across years. Point pattern analyses revealed unique occurrences of T. sanguisuga in north and east Texas, T. gerstaeckeri in south and west Texas, Triatoma indictiva and Triatoma lecticularia in central Texas, and Triatoma rubida in west Texas. These relatively unique spatial occurrences suggest associations with different suitable habitats and serve as a basis for future models evaluating the ecological niches of different vector species. Understanding the temporal and spatial heterogeneity of triatomines in the southern United States will improve targeted interventions of vector control and will guide public outreach and education to reduce human and animal contact with vectors and reduce the risk of exposure to T. cruzi.