Mosquito surveillance in maritime entry ports in Miami-Dade County, Florida to increase preparedness and allow the early detection of invasive mosquito species.

Invasive mosquito vector species have been inadvertently transported to new areas by humans for decades. Strong evidence supports that monitoring maritime, terrestrial, and aerial points of entry is an essential part of the effort to curb the invasion and establishment of invasive vector mosquito species. Miami-Dade County, Florida is an important operational hub for the cruise ship industry and leisure boats that routinely visit nearby areas in the Caribbean, and freight cargo ships transporting goods from Miami-Dade to Caribbean countries and vice versa. To deal with the increasing public health concern, we hypothesized that mosquito surveillance in small- and medium-sized maritime ports of entry in Miami-Dade is crucial to allow the early detection of invasive mosquito species. Therefore, we have selected 12 small- and medium-sized maritime ports of entry in Miami-Dade County with an increased flow of people and commodities that were not covered by the current mosquito surveillance system. Collection sites were comprised of two distinct environments, four marinas with international traffic of leisure boats, and eight maintenance and commercial freight cargo ship ports. Mosquitoes were collected weekly at each of the 12 collection sites for 24 hours for 6 weeks in the Spring and then for 6 additional weeks in the Summer using BG-Sentinel traps. A total of 32,590 mosquitoes were collected, with Culex quinquefasciatus and Aedes aegypti being the most abundant species totaling 19,987 and 11,247 specimens collected, respectively. Our results show that important mosquito vector species were present in great numbers in all of the 12 maritime ports of entry surveyed during this study. The relative abundance of Cx. quinquefasciatus and Ae. aegypti was substantially higher in the commercial freight cargo ship ports than in the marinas. These results indicate that even though both areas are conducive for the proliferation of vector mosquitoes, the port area in the Miami River is especially suitable for the proliferation of vector mosquitoes. Therefore, this potentially allows the establishment of invasive mosquito species inadvertently brought in by cargo freights.

Diel activity patterns of two distinct populations of Aedes aegypti in Miami, FL and Brownsville, TX.

The diel biting activity of Aedes (Stegomyia) aegypti (L) populations was extensively investigated in the early 1900s to gain more information on the biology of Ae. aegypti, and this information was used to devise effective approaches to controlling populations of this species and protect the human population from widespread arbovirus outbreaks. However, few contemporary studies are available regarding the diel activity patterns of Ae. aegypti. To assess the diel activity patterns of Ae. aegypti in southern Florida and Texas, we conducted 96-h uninterrupted mosquito collections once each month from May through November 2019 in Miami, Florida, and Brownsville, Texas, using BG-Sentinel 2 Traps. The overall diel activity pattern in both cities was bimodal with morning and evening peak activity between 7:00 and 8:00 and between 19:00 and 20:00. There were significant daily, monthly, seasonal, and site-specific differences in activity patterns, but these differences did not affect the overall peak activity times. These differences suggest daily, monthly, seasonal, and site-specific variations in human exposure to Ae. aegypti. Our observations can be used in planning and executing Ae. aegypti vector control activities in southern Florida and southern Texas, specifically those targeting the adult mosquito populations.

Evaluation of the effectiveness of BG-Sentinel and CDC light traps in assessing the abundance, richness, and community composition of mosquitoes in rural and natural areas.

BACKGROUND: Vector-borne diseases are a major burden to public health. Controlling mosquitoes is considered the most effective way to prevent vector-borne disease transmission. Mosquito surveillance is a core component of integrated vector management, as surveillance programs are often the cornerstone for the development of mosquito control operations. Two traps are the most commonly used for the surveillance of adult mosquitoes: Centers for Disease Control and Prevention miniature light trap (CDC light trap) and BG-Sentinel trap (BioGents, Regensburg, Germany). However, despite the importance of the BG-Sentinel trap in surveillance programs in the United States, especially in the Southern states, its effectiveness in consistently and reliably collecting mosquitoes in rural and natural areas is still unknown. We hypothesized that BG-Sentinel and CDC light traps would be more attractive to specific mosquito species present in rural and natural areas. Therefore, our objective was to compare the relative abundance, species richness, and community composition of mosquitoes collected in natural and rural areas by BG-Sentinel and CDC light traps. METHODS: Mosquitoes were collected from October 2020 to March 2021 using BG-Sentinel and CDC light traps baited with dry ice, totaling 105 trap-nights. RESULTS: The BG-Sentinel traps collected 195,115 mosquitoes comprising 23 species from eight genera, and the CDC light traps collected 188,594 mosquitoes comprising 23 species from eight genera. The results from the permutational multivariate analysis of variance (PERMANOVA) and generalized estimating equation model for repeated measures indicate the BG-Sentinel and CDC light traps had similar sampling power. CONCLUSION: Even though BG-Sentinel traps had a slightly better performance, the difference was not statistically significant indicating that both traps are suitable to be used in mosquito surveillance in rural and natural areas.

Urbanization favors the proliferation of Aedes aegypti and Culex quinquefasciatus in urban areas of Miami-Dade County, Florida.

Urbanization processes are increasing globally. Anthropogenic alterations in the environment have profound effects on biodiversity. Decreased biodiversity due to biotic homogenization processes as a consequence of urbanization often result in increased levels of mosquito vector species and vector-borne pathogen transmission. Understanding how anthropogenic alterations in the environment will affect the abundance, richness, and composition of vector mosquito species is crucial for the implementation of effective and targeted mosquito control strategies. We hypothesized that anthropogenic alterations in the environment are responsible for increasing the abundance of mosquito species that are adapted to urban environments such as Aedes aegypti and Culex quinquefasciatus. Therefore, our objective was to survey mosquito relative abundance, richness, and community composition in Miami-Dade County, Florida, in areas with different levels of urbanization. We selected 24 areas, 16 remote areas comprised of natural and rural areas, and 8 urban areas comprised of residential and touristic areas in Miami-Dade County, Florida. Mosquitoes were collected weekly in each area for 24 h for 5 consecutive weeks from August to October 2020 using BG-Sentinel traps baited with dry ice. A total of 36,645 mosquitoes were collected, from which 34,048 were collected in the remote areas and 2,597 in the urban areas. Our results show a clear and well-defined pattern of abundance, richness, and community composition according to anthropogenic modifications in land use and land cover. The more urbanized a given area the fewer species were found and those were primary vectors of arboviruses, Ae. aegypti and Cx. quinquefasciatus.

Invasion, establishment, and spread of invasive mosquitoes from the Culex coronator complex in urban areas of Miami-Dade County, Florida.

Species from the Culex coronator complex are Neotropical species and potential vectors of Saint Louis and West Nile viruses. Culex coronator was first described in Trinidad and Tobago in the early twentieth century and since then it has invaded and has been reported established in most countries of the Americas. Species from the Culex coronator complex were first detected in the United States in the state of Louisiana in 2004 and were subsequently detected in Florida in 2005, reaching Miami-Dade County in 2008. We hypothesize that species from the Cx. coronator complex are adapting to urban environments in Miami-Dade County, Florida, and are becoming more present and abundant in these areas. Therefore, our objective was to investigate the patterns of the presence and abundance of species from the Cx. coronator complex in the urban areas of Miami-Dade County. Here we used weekly data comprised of 32 CDC traps from 2012 to 2020 and 150 BG-Sentinel traps from 2016 to 2020. A total of 34,146 female mosquitoes from the Cx. coronator complex were collected, 26,138 by CDC traps and 8008 by BG-Sentinel traps. While the number of CDC traps that were positive was relatively constant at 26-30 positive traps per year, the number of positive BG-Sentinel traps varied substantially from 50 to 87 positive traps per year. Furthermore, the heat map and logistic general linear model for repeated measures analyses showed a significant increase in both the distribution and abundance of mosquitoes from the Cx. coronator complex, indicating that these species are becoming more common in anthropized habitats being able to thrive in highly urbanized areas. The increase in the distribution and abundance of species from the Cx. coronator complex is a major public health concern. The ability of species from the Cx. coronator complex to benefit from urbanization highlights the need to better understand the mechanisms of how invasive vector mosquito species are adapting and exploiting urban habitats.

Effectiveness of adulticide and larvicide in controlling high densities of Aedes aegypti in urban environments.

Current management and control of Aedes aegypti populations in urban areas are based on the spraying of insecticides. Here, we evaluated the effectiveness of spraying larvicide (Bacillus thuringiensis israelensis) using a truck-mounted Buffalo Turbine and adulticide (Deltamethrin) using a Grizzly ULV Sprayer in an urban area with high densities of Ae. aegypti and many cryptic and difficult to reach aquatic breeding habitats. Experiments were conducted in a tire shop located in Miami-Dade County, Florida with approximately 100,000 used airplane tires. Insecticide interventions were performed after a baseline survey consisting of 3 weeks of collections, followed by two insecticide interventions: (i) application of the adulticide followed by the application of larvicide on the subsequent week; and (ii) application of both adulticide and larvicide on two consecutive weeks. The first insecticide intervention resulted in a non-significant decrease in the relative abundance of Ae. aegypti. On the other hand, the second insecticide intervention significantly reduced the Ae. aegypti relative abundance (P < 0.002). Our results demonstrated that the combined insecticide interventions on two consecutive weeks significantly reduced the relative abundance of Ae. aegypti. This result indicated that the larvicide was successfully propelled reaching cryptical and difficult to reach aquatic habitats. However, even though the number of mosquitoes was greatly reduced, it was still greatly above the 10-mosquito threshold by trap night used by the Miami-Dade Mosquito Control Division to deploy an inspector to survey the area. Considering the lack of new and effective mosquito control tools, efficient and mobile insecticide propellers such as Buffalo Turbine can be of great help to manage mosquito populations in urban areas.

Proliferation of Aedes aegypti in urban environments mediated by the availability of key aquatic habitats.

Aedes aegypti is the main vector of dengue, Zika, chikungunya, and yellow fever viruses. Controlling populations of vector mosquito species in urban environments is a major challenge and being able to determine what aquatic habitats should be prioritized for controlling Ae. aegypti populations is key to the development of more effective mosquito control strategies. Therefore, our objective was to leverage on the Miami-Dade County, Florida immature mosquito surveillance system based on requested by citizen complaints through 311 calls to determine what are the most important aquatic habitats in the proliferation of Ae. aegypti in Miami. We used a tobit model for Ae. aegypti larvae and pupae count data, type and count of aquatic habitats, and daily rainfall. Our results revealed that storm drains had 45% lower percentage of Ae. aegypti larvae over the total of larvae and pupae adjusted for daily rainfall when compared to tires, followed by bromeliads with 33% and garbage cans with 17%. These results are indicating that storm drains, bromeliads and garbage cans had significantly more pupae in relation to larvae when compared to tires, traditionally know as productive aquatic habitats for Ae. aegypti. Ultimately, the methodology and results from this study can be used by mosquito control agencies to identify habitats that should be prioritized in mosquito management and control actions, as well as to guide and improve policies and increase community awareness and engagement. Moreover, by targeting the most productive aquatic habitats this approach will allow the development of critical emergency outbreak responses by directing the control response efforts to the most productive aquatic habitats.

Urban farms in Miami-Dade county, Florida have favorable environments for vector mosquitoes.

The creation of urban farms in complex urban built environments may create suitable local conditions for vector mosquitoes. Urban farms have been implicated in the proliferation of mosquitoes in Africa, but there is a dearth in the knowledge of their role in the proliferation of mosquitoes elsewhere. In this study, we surveyed two urban farms in Miami-Dade County, Florida. Our results show that urban farms provide favorable conditions for populations of vector mosquito species by providing a wide range of essential resources such as larval habitats, suitable outdoor resting sites, sugar-feeding centers, and available hosts for blood-feeding. A total of 2,185 specimens comprising 12 species of mosquitoes were collected over 7 weeks. The results varied greatly according to the urban farm. At the Wynwood urban farm, 1,016 specimens were collected but were distributed only between 3 species; while the total number of specimens collected at the Golden Glades urban farm was 1,168 specimens comprising 12 species. The presence of vector mosquitoes in urban farms may represent a new challenge for the development of effective strategies to control populations of vector mosquito species in urban areas.

Cemeteries in Miami-Dade County, Florida are important areas to be targeted in mosquito management and control efforts.

Definable habitats at the neighborhood level provide a wide range of favorable habitats with optimal conditions and environmental resources for mosquito survival. Problematic habitats for controlling mosquitoes in urban environments such as tire shops, bromeliad patches, and construction sites must be taken into consideration in the development of effective mosquito management and control in urban areas. Cemeteries are often located in highly urbanized areas serving as a haven for populations of vector mosquito species due to the availability of natural resources present in most cemeteries. Even though Miami-Dade County, Florida was the most affected area in the United States during the Zika virus outbreak in 2016 and is currently under a mosquito-borne illness alert after 14 confirmed locally transmitted dengue cases, the role of cemeteries in the proliferation of vector mosquitoes is unknown. Therefore, our objective was to use a cross-sectional experimental design to survey twelve cemeteries across Miami-Dade County to assess if vector mosquitoes in Miami can be found in these areas. Our results are indicating that vector mosquitoes are able to successfully exploit the resources available in the cemeteries. Culex quinquefasciatus was the most abundant species but it was neither as frequent nor present in its immature form as Aedes aegypti and Aedes albopictus. This study revealed that vector mosquitoes, such as Ae. aegypti, Ae. albopictus, and Cx. quinquefasciatus are successfully exploiting the resources available in these areas being able to thrive and reach high numbers. Mosquito control strategies should consider both long-term strategies, based on changing human behavior to reduce the availability of aquatic habitats for vector mosquitoes; as well as short-term strategies such as drilling holes or adding larvicide to the flower vases. Simple practices would greatly help improve the effectiveness of mosquito management and control in these problematic urban habitats.

Simalikalactone D, a Potential Anticancer Compound from Simarouba tulae, an Endemic Plant of Puerto Rico.

Species of the genus Simarouba have been studied because of their antimalarial and antileukemic activities. A group of oxygenated terpenes called quassinoids have been isolated from species of the Simarouba genus, and are responsible for its therapeutic properties. We hypothesized that Simarouba tulae, an endemic plant from Puerto Rico, is a natural source rich in quassinoid compounds with anticancer activity. The leaves were processed and extracted with solvents of different polarities. The extracts were screened for their antiproliferative activity, and it was shown that the chloroform extract was the most active extract. This extract was purified using different chromatographic techniques to afford the quassinoid simalikalactone D (SKD). This compound was further characterized using NMR and X-ray diffraction analysis. A reassessment of original structural assignments for SKD is proposed. SKD showed high cytotoxicity activity, with an IC50 of 55, 58, and 65 nM in A2780CP20 (ovarian), MDA-MB-435 (breast), and MDA-MB-231 (breast) cell lines, respectively. Exposure to SKD led to 15% inhibition of the migration of MDA-MB-231 cells.

Urbanization creates diverse aquatic habitats for immature mosquitoes in urban areas.

Global increases in temperatures and urbanization are impacting the epidemiology of mosquito-borne diseases. Urbanization processes create suitable habitats for vector mosquitoes in which there are a reduced number of predators, and human hosts are widely available. We hypothesize that mosquito vector species, especially Aedes aegypti, are locally concentrated primarily in those specific habitats at the neighborhood levels that provide suitable conditions and environmental resources needed for mosquito survival. Determining how mosquito vector species composition and abundance depend on environmental resources across habitats addresses where different types of vector control need to be applied. Therefore, our goal was to analyze and identify the most productive aquatic habitats for mosquitoes in Miami-Dade County, Florida. Immature mosquito surveys were conducted throughout Miami-Dade County from April 2018 to June 2019, totaling 2,488 inspections. Mosquitoes were collected in 76 different types of aquatic habitats scattered throughout 141 neighborhoods located in the urbanized areas of Miami-Dade County. A total of 44,599 immature mosquitoes were collected and Ae. aegypti was the most common and abundant species, comprising 43% of all specimens collected. Aedes aegypti was primarily found in buckets, bromeliads, and flower pots, concentrated in specific neighborhoods. Our results showed that aquatic habitats created by anthropogenic land-use modifications (e.g., ornamental bromeliads, buckets, etc.) were positively correlated with the abundance of Ae. aegypti. This study serves to identify how vector mosquitoes utilize the resources available in urban environments and to determine the exact role of these specific urban features in supporting populations of vector mosquito species. Ultimately, the identification of modifiable urban features will allow the development of targeted mosquito control strategies optimized to preventatively control vector mosquitoes in urban areas.

Community Composition and Year-round Abundance of Vector Species of Mosquitoes make Miami-Dade County, Florida a Receptive Gateway for Arbovirus entry to the United States.

Vector-borne diseases are a heavy burden to human-kind. Global warming and urbanization have a significant impact on vector-borne disease transmission, resulting in more severe outbreaks, and outbreaks in formerly non-endemic areas. Miami-Dade County, Florida was the most affected area in the continental United States during the 2016 Zika virus outbreak. Miami is an important gateway and has suitable conditions for mosquitoes year-round. Therefore, it was critical to establish and validate a surveillance system to guide and improve mosquito control operations. Here we assess two years of mosquito surveillance in Miami established after the 2016 Zika virus outbreak. Our results show that the most abundant mosquito species are either well adapted to urban environments or are adapting to it. The five most abundant species comprised 85% of all specimens collected, with four of them being primary vectors of arboviruses. Aedes aegypti and Culex quinquefasciatus were found year-round throughout Miami regardless of urbanization level, vegetation, or socioeconomic variations. This study serves as a foundation for future efforts to improve mosquito surveillance and control operations.

Assessment of the effectiveness of BG-Sentinel traps baited with CO2 and BG-Lure for the surveillance of vector mosquitoes in Miami-Dade County, Florida.

Vector-borne diseases are an increasing issue to public health, endangering billions of people worldwide. Controlling vector mosquitoes is widely accepted as the most effective way to prevent vector-borne disease outbreaks. Mosquito surveillance is critical for the development of control strategies under the integrated vector management framework. We hypothesize that the effectiveness and reliability of using BG-Sentinel traps for the surveillance strongly depend on the bait used to attract mosquitoes. The objective of this study was to compare the effectiveness of BG-Sentinel traps baited with CO2 and BG-Lure. A total of 72 traps were deployed for 48 hours once a week for four weeks. For the initial 24-hour period, the traps were baited with CO2, and then for an additional 24 hours using the BG-Lure. Collected mosquitoes were analyzed using the Generalized Estimating Equation for repeated measures analysis. Biodiversity was assessed by the Shannon and Simpson indices and by individual rarefaction curves and SHE profiles. A total of 5,154 mosquitoes were collected, from which 3,514 by traps baited with CO2 and 1,640 mosquitoes by traps baited with BG-Lure. Aedes aegypti and Culex quinquefasciatus were the most abundant and dominant species. Results from the Generalized Estimating Equation models indicated that more than twice as many mosquitoes were attracted CO2 than to the BG-Lure. The comparison of attractiveness of CO2 and BG-Lure to Ae. aegypti and Cx. quinquefasciatus was non-significant, suggesting that both species were equally attracted by the baits. The individual rarefaction curves for Ae. aegypti and Cx. quinquefasciatus imply that traps baited with BG-Lure underestimated mosquito species richness compared to those baited with CO2. BG-Lure were less effective in attracting mosquitoes with low abundances and failed to collect Cx. coronator and Cx. nigripalpus, which were consistently collected by traps baited with CO2. According to our results, CO2 significantly (P<0.05) attracted more mosquitoes (2.67 adjusted odds ratios) than the BG-Lure when adjusted for time and species, being more effective in assessing the relative abundance of vector mosquitoes and yielding more trustworthy results. Traps baited with CO2 collected not only more specimens, but also more species in a more consistent pattern.

Inhibitors of urokinase type plasminogen activator and cytostatic activity from crude plants extracts.

In view of the clear evidence that urokinase type plasminogen activator (uPA) plays an important role in the processes of tumor cell metastasis, aortic aneurysm, and multiple sclerosis, it has become a target of choice for pharmacological intervention. The goal of this study was thus to determine the presence of inhibitors of uPA in plants known traditionally for their anti-tumor properties. Crude methanol extracts were prepared from the leaves of plants (14) collected from the subtropical dry forest (Guanica, Puerto Rico), and tested for the presence of inhibitors of uPA using the fibrin plate assay. The extracts that tested positive (6) were then partitioned with petroleum ether, chloroform, ethyl acetate and n-butanol, in a sequential manner. The resulting fractions were then tested again using the fibrin plate assay. Extracts from leaves of Croton lucidus (C. lucidus) showed the presence of a strong uPA inhibitory activity. Serial dilutions of these C. lucidus partitions were performed to determine the uPA inhibition IC50 values. The chloroform extract showed the lowest IC50 value (3.52 µg/mL) and hence contained the most potent uPA inhibitor. Further investigations revealed that the crude methanol extract and its chloroform and n-butanol partitions did not significantly inhibit closely related proteases such as the tissue type plasminogen activator (tPA) and plasmin, indicating their selectivity for uPA, and hence superior potential for medicinal use with fewer side effects. In a further evaluation of their therapeutic potential for prevention of cancer metastasis, the C. lucidus extracts displayed cytostatic activity against human pancreatic carcinoma (PaCa-2) cells, as determined through an MTS assay. The cytostatic activities recorded for each of the partitions correlated with their relative uPA inhibitory activities. There are no existing reports of uPA inhibitors being present in any of the plants reported in this study.