Assessing and managing the risk of Aedes mosquito introductions via the global maritime trade network.

The global shipping network (GSN) has been suggested as a pathway for the establishment and reintroduction of Aedes aegypti and Aedes albopictus primarily via the tire trade. We used historical maritime movement data in combination with an agent-based model to understand invasion risk in the United States Gulf Coast and how the risk of these invasions could be reduced. We found a strong correlation between the total number of cargo ship arrivals at each port and likelihood of arrival by both Ae. aegypti and Ae. albopictus. Additionally, in 2012, 99.2% of the arrivals into target ports had most recently visited ports likely occupied by both Ae. aegypti and Ae. albopictus, increasing risk of Aedes invasion. Our model results indicated that detection and removal of mosquitoes from containers when they are unloaded effectively reduced the probability of mosquito populations establishment even when the connectivity of ports increased. To reduce the risk of invasion and reintroduction of Ae. aegypti and Ae. albopictus, surveillance and control efforts should be employed when containers leave high risk locations and when they arrive in ports at high risk of establishment.

Which trap is best? Alternatives to outdoor human landing catches for malaria vector surveillance: a meta-analysis.

BACKGROUND: Human landing catches (HLC) are an entomological collection technique in which humans are used as attractants to capture medically relevant host-seeking mosquitoes. The use of this method has been a topic of extensive debate for decades mainly due to ethical concerns. Many alternatives to HLC have been proposed; however, no quantitative review and meta-analysis comparing HLC to outdoor alternative trapping methods has been conducted. METHODS: A total of 58 comparisons across 12 countries were identified. We conducted a meta-analysis comparing the standardized mean difference of Anopheles captured by HLC and alternative traps. To explain heterogeneity, three moderators were chosen for analysis: trap type, location of study, and species captured. A meta-regression was fit to understand how the linear combination of moderators helped in explaining heterogeneity. The possibility of biased results due to publication bias was also explored. RESULTS: Random-effects meta-analysis showed no statistically significant difference in the mean difference of Anopheles collected. Moderator analysis was conducted to determine the effects of trap type, geographical location of study, and the species of Anopheles captured. On average, tent-based traps captured significantly more Anopheles than outdoor HLC (95% CI: [- .9065, - 0.0544]), alternative traps in Africa captured on average more mosquitoes than outdoor HLC (95% CI: [- 2.8750, - 0.0294]), and alternative traps overall captured significantly more Anopheles gambiae s.l. than outdoor HLC (95% CI: [- 4.4613, - 0.2473]) on average. Meta-regression showed that up to 55.77% of the total heterogeneity found can be explained by a linear combination of the three moderators and the interaction between trap type and species. Subset analysis on An. gambiae s.l. showed that light traps specifically captured on average more of this species than HLC (95% CI: [- 18.3751, - 1.0629]). Publication bias likely exists. With 59.65% of studies reporting p-values less than 0.025, we believe there is an over representation in the literature of results indicating that alternative traps are superior to outdoor HLC. CONCLUSIONS: Currently, there is no consensus on a single "magic bullet" alternative to outdoor HLC. The diversity of many alternative trap comparisons restricts potential metrics for comparisons to outdoor HLC. Further standardization and specific question-driven trap evaluations that consider target vector species and the vector control landscape are needed to allow for robust meta-analyses with less heterogeneity and to develop data-driven decision-making tools for malaria vector surveillance and control.

Developing a granular scale environmental burden index (EBI) for diverse land cover types across the contiguous United States.

Critical to identifying the risk of environmentally driven disease is an understanding of the cumulative impact of environmental conditions on human health. Here we describe the methodology used to develop an environmental burden index (EBI). The EBI is calculated at U.S. census tract level, a finer scale than many similar national-level tools. EBI scores are also stratified by tract land cover type as per the National Land Cover Database (NLCD), controlling for urbanicity. The EBI was developed over the course of four stages: 1) literature review to identify potential indicators, 2) data source acquisition and indicator variable construction, 3) index creation, and 4) stratification by land cover type. For each potential indicator, data sources were assessed for completeness, update frequency, and availability. These indicators were: (1) particulate matter (PM2.5), (2) ozone, (3) Superfund National Priority List (NPL) locations, (4) Toxics Release Inventory (TRI) facilities, (5) Treatment, Storage, and Disposal (TSD) facilities, (6) recreational parks, (7) railways, (8) highways, (9) airports, and (10) impaired water sources. Indicators were statistically normalized and checked for collinearity. For each indicator, we computed and summed percentile ranking scores to create an overall ranking for each tract. Tracts having the same plurality of land cover type form a 'peer' group. We re-ranked the tracts into percentiles within each peer group for each indicator. The percentile scores were combined for each tract to obtain a stratified EBI. A higher score reveals a tract with increased environmental burden relative to other tracts of the same peer group. We compared our results to those of related indices, finding good convergent validity between the overall EBI and CalEnviroScreen 4.0. The EBI has many potential applications for research and use as a tool to develop public health interventions at a granular scale.

Arboviral diseases and poverty in Alabama, 2007-2017.

Mosquito-borne viruses cause diseases of great public health concern. Arboviral disease case distributions have complex relationships with socioeconomic and environmental factors. We combined information about socio-economic (population, and poverty rate) and environmental (precipitation, and land use) characteristics with reported human cases of arboviral disease in the counties of Alabama, USA, from 2007-2017. We used county level data on West Nile virus (WNV), dengue virus (DENV), chikungunya virus (CHIKV), Zika virus (ZIKV), California serogroup virus, Eastern equine encephalitis virus, and Saint Louis encephalitis virus to provide a detailed description of their spatio-temporal pattern. We found a significant spatial convergence between incidence of WNV and poverty rate clustered in the southern part of Alabama. DENV, CHIKV and ZIKV cases showed a different spatial pattern, being mostly located in the northern part, in areas of high socioeconomic status. The results of our study establish that poverty-driven inequities in arboviral risk exist in the southern USA, and should be taken into account when planning prevention and intervention strategies.

Widespread zoophagy and detection of Plasmodium spp. in Anopheles mosquitoes in southeastern Madagascar.

BACKGROUND: Malaria is a top cause of mortality on the island nation of Madagascar, where many rural communities rely on subsistence agriculture and livestock production. Understanding feeding behaviours of Anopheles in this landscape is crucial for optimizing malaria control and prevention strategies. Previous studies in southeastern Madagascar have shown that Anopheles mosquitoes are more frequently captured within 50 m of livestock. However, it remains unknown whether these mosquitoes preferentially feed on livestock. Here, mosquito blood meal sources and Plasmodium sporozoite rates were determined to evaluate patterns of feeding behaviour in Anopheles spp. and malaria transmission in southeastern Madagascar. METHODS: Across a habitat gradient in southeastern Madagascar 7762 female Anopheles spp. mosquitoes were collected. Of the captured mosquitoes, 492 were visibly blood fed and morphologically identifiable, and a direct enzyme-linked immunosorbent assay (ELISA) was used to test for swine, cattle, chicken, human, and dog blood among these specimens. Host species identification was confirmed for multiple blood meals using PCR along with Sanger sequencing. Additionally, 1,607 Anopheles spp. were screened for the presence of Plasmodium falciparum, P. vivax-210, and P. vivax 247 circumsporozoites (cs) by ELISA. RESULTS: Cattle and swine accounted, respectively, for 51% and 41% of all blood meals, with the remaining 8% split between domesticated animals and humans. Of the 1,607 Anopheles spp. screened for Plasmodium falciparum, Plasmodium vivax 210, and Plasmodium vivax 247 cs-protein, 45 tested positive, the most prevalent being P. vivax 247, followed by P. vivax 210 and P. falciparum. Both variants of P. vivax were observed in secondary vectors, including Anopheles squamosus/cydippis, Anopheles coustani, and unknown Anopheles spp. Furthermore, evidence of coinfection of P. falciparum and P. vivax 210 in Anopheles gambiae sensu lato (s.l.) was found. CONCLUSIONS: Here, feeding behaviour of Anopheles spp. mosquitoes in southeastern Madagascar was evaluated, in a livestock rich landscape. These findings suggest largely zoophagic feeding behaviors of Anopheles spp., including An. gambiae s.l. and presence of both P. vivax and P. falciparum sporozoites in Anopheles spp. A discordance between P. vivax reports in mosquitoes and humans exists, suggesting high prevalence of P. vivax circulating in vectors in the ecosystem despite low reports of clinical vivax malaria in humans in Madagascar. Vector surveillance of P. vivax may be relevant to malaria control and elimination efforts in Madagascar. At present, the high proportion of livestock blood meals in Madagascar may play a role in buffering (zooprophylaxis) or amplifying (zoopotentiation) the impacts of malaria. With malaria vector control efforts focused on indoor feeding behaviours, complementary approaches, such as endectocide-aided vector control in livestock may be an effective strategy for malaria reduction in Madagascar.

An investigation of Dirofilaria immitis infection and its effects on mosquito wingbeat frequencies.

Each mosquito species has a different wingbeat frequency by which they attract mates. With just a brief recording (<1/10th of a second) these acoustic signatures can be analyzed to quickly determine if mosquitoes belong to a species that is known to transmit different pathogens. A recent study has shown that mobile phones are capable of capturing acoustic data from mosquito wingbeats. We examined wingbeat signatures and flight duration patterns of D. immitis infected and non-infected Aedes aegypti to determine if mobile phone recordings of wingbeat frequencies can be used to distinguish infected mosquitoes from non-infected ones. Female mosquitoes were recorded prior to and at various time points after feeding on infected or non-infected dog blood by placing individual mosquitoes into a collection vial and recording for 60 s using the Voice Memo app for iPhone 7 plus and 8. To uniformly analyze audio data, recordings were processed using a previously described automated algorithm in Python 3.0 to determine wingbeat frequency. A total of 1669 recordings were gathered, and mosquitoes were dissected to confirm the presence and number of D. immitis larvae. Our findings indicate that there was a significant effect on wingbeat frequency with an increasing number of L3 larvae. Specifically, as the number of L3, infective stage larvae increases, a decrease in wingbeat frequency is seen. However, there was no significant effect of increasing number of L1 or L2 larvae causing increasing wingbeat frequencies. The detection of a significant difference in wingbeat frequencies between mosquitoes harboring infective stage D. immitis larvae is unique and suggests the possibility of using wingbeat recordings as a tool for vector species and pathogen surveillance and monitoring.

Perceptions and practices of mosquito-borne diseases in Alabama - is concern where it should be?

BACKGROUND: The Gulf Coast of the United States is home to mosquito vectors that may spread disease causing pathogens, and environmental conditions that are ideal for the sustained transmission of mosquito-borne pathogens. Understanding public perceptions of mosquito-borne diseases and mosquito prevention strategies is critical for the development of effective vector control strategies and public health interventions. Here, we present a survey conducted in Mobile, Alabama along the Gulf Coast to better understand public perceptions of mosquito-borne diseases, mosquito control activities, and potential risk factors. METHODS: Using Knowledge, Attitude, and Practice (KAPs) assessments, we surveyed populations living in 12 zip codes in Mobile, Alabama using a 7-point Likert scale and frequency assessments. Survey participants were asked about vector control efforts, knowledge of mosquito-borne diseases, and understanding of mosquito ecology and breeding habitats. RESULTS: One hundred twenty-six surveys were completed in Mobile, Alabama, revealing that 73% of participants reported being bitten by a mosquito in the last 30 days and mosquitoes were frequently seen in their homes. Ninety-four percent of respondents had heard of Zika Virus at the time of the survey, and respondents reported being least familiar with dengue virus and chikungunya virus. CONCLUSIONS: Chikungunya virus, dengue virus, malaria, West Nile virus, and Zika virus have been documented in the Gulf Coast of the United States. The mosquitoes which vector all of these diseases are presently in the Gulf Coast meaning all five diseases pose a potential risk to human health. The results of this survey emphasize knowledge gaps that public health officials can address to empower the population to reduce their risk of these mosquito-borne diseases. Each species of mosquito has specific preferences for breeding and feeding and there is no one size fits all prevention approach, educating people on the need for a variety of approaches in order to address all species will further empower them to control mosquitoes where they live and further reduce their risk of disease.

Aedes Vector Surveillance in the Southeastern United States Reveals Growing Threat of Aedes japonicus japonicus (Diptera: Culicidae) and Aedes albopictus.

Mosquito-borne diseases are on the rise globally, and have the potential to thrive along the Gulf Coast of the United States, where subtropical conditions may facilitate the introduction or movement of mosquito vectors. Despite surveillance efforts, Aedes aegypti (L.) had not been detected in the Gulf state of Alabama for nearly three decades. The detection of Ae. aegypti in Alabama may suggest remnant or reemergent populations of this vector. We conducted adult sampling between May and August of 2018 to capture mosquitoes during a time frame when all species should be active. This was to ensure no species were missed due to overwintering and to identify the distributions of Aedes mosquitoes of medical importance. No Ae. aegypti were detected in Alabama over the period of this study. We detected Aedes albopictus (Skuse) in 65 counties and the recently invasive Aedes japonicus japonicus (Theobald) in 30 counties across the state. These results indicate that while Ae. aegypti was recently reported from parts of Alabama, the state is not experiencing a major resurgence of the species, whereas Ae. albopictus remains ubiquitous. Further, results indicate that a third wave of Aedes invasion may be occurring, that of Ae. japonicus japonicus. All three of these species are medically important vectors and may pose threats to the public health of the Gulf Coast of the United States.

Aedes albopictus is a competent vector of Zika virus: A meta-analysis.

BACKGROUND: Much work has been done in recent years to determine the vector competence of Aedes albopictus (Skuse) for Zika virus (ZIKV). If competent, Ae. albopictus could become an important vector in the spread of ZIKV to areas which until now have been considered safe from the virus. Despite much speculation about Ae. albopictus' competence for ZIKV, there have been, to date, no quantitative syntheses of Ae. albopictus' competence, nor have the potentially confounding differences between studies been addressed. METHODOLOGY/ PRINCIPLE FINDINGS: This study represents a quantitative meta-analysis of the literature surrounding this topic by examining infection rates (IR) and transmission rates (TR) among sample populations of Ae. albopictus at 7 and 14 days post infection (dpi) across 15 journal articles comprising 23 studies. Our analyses examined potentially confounding variables in the studies contained therein, including: geographic origin of viral strain or mosquito population tested, whether sympatry of the tested viral strain and mosquito population was important, and freshness of blood meal. Our results suggest 1) Ae albopictus is a competent vector for ZIKV and 2) that origin of Ae. albopictus population and origin of viral strain had significant effects on the competence of Ae. albopictus to transmit ZIKV. CONCLUSIONS/ SIGNIFICANCE: These results indicate a need to further explore the effects of methodology on vector competence studies and to examine in more detail the geographic variation in the competence of Ae. albopictus for ZIKV as well as the underlying causes of said variation. The ability of Ae. albopictus to carry and transmit ZIKV also points to a need to create new vector control strategies in case of a ZIKV outbreak in an area where Ae. albopictus is prominent. Finally, this study represents a potential template for future meta-analyses in the field of vector competence, where this type of study has been under-utilized despite the abundance of relevant studies.

Rosuvastatin Alters the Proteome of High Density Lipoproteins: Generation of alpha-1-antitrypsin Enriched Particles with Anti-inflammatory Properties.

Statins lower plasma cholesterol by as much as 50%, thus reducing future cardiovascular events. However, the physiological effects of statins are diverse and not all are related to low density lipoprotein cholesterol (LDL-C) lowering. We performed a small clinical pilot study to assess the impact of statins on lipoprotein-associated proteins in healthy individuals (n = 10) with normal LDL-C (<130 mg/dL), who were treated with rosuvastatin (20 mg/day) for 28 days. Proteomic analysis of size-exclusion chromatography isolated LDL, large high density lipoprotein (HDL-L), and small HDL (HDL-S) fractions and spectral counting was used to compare relative protein detection before and after statin therapy. Significant protein changes were found in each lipoprotein pool and included both increases and decreases in several proteins involved in lipoprotein metabolism, complement regulation and acute phase response. The most dramatic effect of the rosuvastatin treatment was an increase in α-1-antirypsin (A1AT) spectral counts associated with HDL-L particles. Quantitative measurement by ELISA confirmed an average 5.7-fold increase in HDL-L associated A1AT. Molecular modeling predictions indicated that the hydrophobic reactive center loop of A1AT, the functional domain responsible for its protease inhibitor activity, is likely involved in lipid binding and association with HDL was found to protect A1AT against oxidative inactivation. Cell culture experiments, using J774 macrophages, demonstrated that the association of A1AT with HDL enhances its antiprotease activity, preventing elastase induced production of tumor necrosis factor α. In conclusion, we show that statins can significantly alter the protein composition of both LDL and HDL and our studies reveal a novel functional relationship between A1AT and HDL. The up-regulation of A1AT on HDL enhances its anti-inflammatory functionality, which may contribute to the non-lipid lowering beneficial effects of statins.