Aquatic microfauna alter larval food resources and affect development and biomass of West Nile and Saint Louis encephalitis vector Culex nigripalpus (Diptera: Culicidae)

Ciliate protists and rotifers are ubiquitous in aquatic habitats and can comprise a significant portion of the microbial food resources available to larval mosquitoes, often showing substantial declines in abundance in the presence of mosquito larvae. This top‐down regulation of protists is reported to be strong for mosquitoes inhabiting small aquatic containers such as pitcher plants or tree holes, but the nature of these interactions with larval mosquitoes developing in other aquatic habitats is poorly understood. We examined the effects of these two microbial groups on lower trophic level microbial food resources, such as bacteria, small flagellates, and organic particles, in the water column, and on Culex larval development and adult production. In three independent laboratory experiments using two microeukaryote species (one ciliate protist and one rotifer) acquired from field larval mosquito habitats and cultured in the laboratory, we determined the effects of Culex nigripalpus larval grazing on water column microbial dynamics, while simultaneously monitoring larval growth and development. The results revealed previously unknown interactions that were different from the top‐down regulation of microbial groups by mosquito larvae in other systems. Both ciliates and rotifers, singly or in combination, altered other microbial populations and inhibited mosquito growth. It is likely that these microeukaryotes, instead of serving as food resources, competed with early instar mosquito larvae for microbes such as small flagellates and bacteria in a density‐dependent manner. These findings help our understanding of the basic larval biology of Culex mosquitoes, variation in mosquito production among various larval habitats, and may have implications for existing vector control strategies and for developing novel microbial‐based control methods.     

Assessment of Aedes albopictus (Skuse) (Diptera: Culicidae) clutch size in wild and laboratory populations

Aedes albopictus (Skuse) is an invasive mosquito species found across the southern U.S. with range expansion into many northern states. Intra‐ and interspecific larval competition have been evaluated for Ae. albopictus with respect to subsequent adult size, immature and adult survivability, and its capacity to vector pathogens as an adult. However, limited data are available on egg production as related to larval rearing conditions. Because Ae. albopictus is a container‐inhabiting mosquito that oviposits in resource‐limited habitats, it is found under variable density‐dependent conditions. Therefore, we examined the impact of specific rearing conditions on Ae. albopictus clutch size and adult body size; comparing the egg production values and wing lengths from known developmental densities to those from field‐collected populations. Field populations varied significantly among collection sites in mean clutch size (23 to 46). These clutch sizes were comparable to the mean clutch sizes of females reared at the larval densities of nine (20 eggs) and three (53 eggs) larvae per 3 ml of water in the laboratory. Field populations experienced density‐dependent effects impacting adult mosquito size. Mosquitoes from the four sample sites had mean wing lengths of 1.99, 2.47, 2.51, and 2.54 mm, which were less than the mean wing length of mosquitoes reared at larval densities of three larvae per 3 ml of water (2.57 mm).     

Combined sewage overflow accelerates immature development and increases body size in the urban mosquito Culex quinquefasciatus

In urban landscapes the crowding of humans and their waste products may alleviate intra‐specific interactions of common mosquitoes. Here, we present the results of a semi‐natural experiment addressing the effects of water from a sewage overflow stream on density dependent fitness components and phenotypic traits of a common tropical and subtropical urban mosquito, Culex quinquefasciatus Say (Diptera: Culicidae). This semi‐natural experiment was designed to quantify the relative importance of density dependence, weather forces and water quality on larval mortality, sex ratio and size at adult emergence. Results showed that mortality hazards were independent of larval density, decreased in sewage overflow water and increased with minimum temperatures. Under all rearing conditions adult mosquito size decreased with density. Mosquitoes from sewage overflow water emerged faster, were bigger and had an increased ratio of females to males. All these traits may contribute to the regulation of mosquito populations.     

Population dynamics of Aedes aegypti and dengue as influenced by weather and human behavior in San Juan, Puerto Rico

Previous studies on the influence of weather on Aedes aegypti dynamics in Puerto Rico suggested that rainfall was a significant driver of immature mosquito populations and dengue incidence, but mostly in the drier areas of the island. We conducted a longitudinal study of Ae. aegypti in two neighborhoods of the metropolitan area of San Juan city, Puerto Rico where rainfall is more uniformly distributed throughout the year. We assessed the impacts of rainfall, temperature, and human activities on the temporal dynamics of adult Ae. aegypti and oviposition. Changes in adult mosquitoes were monitored with BG-Sentinel traps and oviposition activity with CDC enhanced ovitraps. Pupal surveys were conducted during the drier and wetter parts of the year in both neighborhoods to determine the contribution of humans and rains to mosquito production. Mosquito dynamics in each neighborhood was compared with dengue incidence in their respective municipalities during the study. Our results showed that: 1. Most pupae were produced in containers managed by people, which explains the prevalence of adult mosquitoes at times when rainfall was scant; 2. Water meters were documented for the first time as productive habitats for Ae. aegypti; 3. Even though Puerto Rico has a reliable supply of tap water and an active tire recycling program, water storage containers and discarded tires were important mosquito producers; 4. Peaks in mosquito density preceded maximum dengue incidence; and 5. Ae. aegypti dynamics were driven by weather and human activity and oviposition was significantly correlated with dengue incidence.     

Co-occurrence Patterns of the Dengue Vector Aedes aegypti and Aedes mediovitattus, a Dengue Competent Mosquito in Puerto Rico

Aedes aegypti is implicated in dengue transmission in tropical and subtropical urban areas around the world. Ae. aegypti populations are controlled through integrative vector management. However, the efficacy of vector control may be undermined by the presence of alternative, competent species. In Puerto Rico, a native mosquito, Ae. mediovittatus, is a competent dengue vector in laboratory settings and spatially overlaps with Ae. aegypti. It has been proposed that Ae. mediovittatus may act as a dengue reservoir during inter-epidemic periods, perpetuating endemic dengue transmission in rural Puerto Rico. Dengue transmission dynamics may therefore be influenced by the spatial overlap of Ae. mediovittatus, Ae. aegypti, dengue viruses, and humans. We take a landscape epidemiology approach to examine the association between landscape composition and configuration and the distribution of each of these Aedes species and their co-occurrence. We used remotely sensed imagery from a newly launched satellite to map landscape features at very high spatial resolution. We found that the distribution of Ae. aegypti is positively predicted by urban density and by the number of tree patches, Ae. mediovittatus is positively predicted by the number of tree patches, but negatively predicted by large contiguous urban areas, and both species are predicted by urban density and the number of tree patches. This analysis provides evidence that landscape composition and configuration is a surrogate for mosquito community composition, and suggests that mapping landscape structure can be used to inform vector control efforts as well as to inform urban planning.     

Climate as a driver of tropical insular diversity: comparative phylogeography of two ecologically distinctive frogs in Puerto Rico

The effects of late Quaternary climate on distributions and evolutionary dynamics of insular species are poorly understood in most tropical archipelagoes. We used ecological niche models under past and current climate to derive hypotheses regarding how stable climatic conditions shaped genetic diversity in two ecologically distinctive frogs in Puerto Rico. Whereas the mountain coquí Eleutherodactylus portoricensis is restricted to montane forest in the Cayey and Luquillo Mountains, the red‐eyed coquí E. antillensis is a habitat generalist distributed across the entire Puerto Rican Bank (Puerto Rico and the Virgin Islands, excluding St Croix). To test our hypotheses, we conducted phylogeographic and population genetic analyses based on mitochondrial and nuclear loci of each species across their range in Puerto Rico. Patterns of population differentiation in E. portoricensis, but not in E. antillensis, supported our hypotheses. For E. portoricensis, these patterns include: individuals isolated by long‐term unsuitable climate in the Río Grande de Loíza Basin in eastern Puerto Rico belong to different genetic clusters; past and current climate strongly predicted genetic differentiation; and Cayey and Luquillo Mountains populations split prior to the last interglacial. For E. antillensis, these patterns include: genetic clusters did not fully correspond to predicted long‐term unsuitable climate; and past and current climate weakly predicted patterns of genetic differentiation. Genetic signatures in E. antillensis are consistent with a recent range expansion into western Puerto Rico, possibly resulting from climate change and anthropogenic influences. As predicted, regions with a large area of long‐term suitable climate were associated with higher genetic diversity in both species, suggesting larger and more stable populations. Finally, we discussed the implications of our findings for developing evidence‐based management decisions for E. portoricensis, a taxon of special concern. Our findings illustrate the role of persistent suitable climatic conditions in promoting the persistence and diversification of tropical island organisms.     

Mosquito Population Regulation and Larval Source Management in Heterogeneous Environments

An important question for mosquito population dynamics, mosquito-borne pathogen transmission and vector control is how mosquito populations are regulated. Here we develop simple models with heterogeneity in egg laying patterns and in the responses of larval populations to crowding in aquatic habitats. We use the models to evaluate how such heterogeneity affects mosquito population regulation and the effects of larval source management (LSM). We revisit the notion of a carrying capacity and show how heterogeneity changes our understanding of density dependence and the outcome of LSM. Crowding in and productivity of aquatic habitats is highly uneven unless egg-laying distributions are fine-tuned to match the distribution of habitats’ carrying capacities. LSM reduces mosquito population density linearly with coverage if adult mosquitoes avoid laying eggs in treated habitats, but quadratically if eggs are laid in treated habitats and the effort is therefore wasted (i.e., treating 50% of habitat reduces mosquito density by approximately 75%). Unsurprisingly, targeting (i.e. treating a subset of the most productive pools) gives much larger reductions for similar coverage, but with poor targeting, increasing coverage could increase adult mosquito population densities if eggs are laid in higher capacity habitats. Our analysis suggests that, in some contexts, LSM models that accounts for heterogeneity in production of adult mosquitoes provide theoretical support for pursuing mosquito-borne disease prevention through strategic and repeated application of modern larvicides.     

Density dependence in blowfly populations: experimental evaluation of non‐parametric time‐series modelling

Two approaches for describing density dependence in demographic rates of stage‐structured populations are compared in this study. Time‐series data from laboratory blowfly populations (Lucilia sericata) have been analysed in a separate study, with a statistical modelling approach that incorporated density dependences as unspecified (non‐parametric) functions. In this study, we assessed density‐dependent structures by manipulating densities of larvae and adults in cohorts of blowflies and measuring the demographic rates. We here compare the density‐dependent structures revealed by the cohort experiments with those estimated by the non‐parametric model. This model estimates the demographic rates to have the following density‐dependent structures: (i) larval survival was non‐linearly density‐dependent (a ‘humped’ function), (ii) adult survival was density‐independent, and (iii) reproductive rate decreased with adult density. In the cohort experiments reported here, (i) juvenile survival exhibited a positive density dependence in low densities (facilitation), which became negative at higher densities (competition). Pupal and adult size decreased with initial larval density. (ii) Adult survival was reduced by high initial larval density, but it was independent of adult density. (iii) Reproductive rate was reduced by high initial larval density, and by high adult density in populations of large individuals (from low larval density). Hence, the results from these experiments support the non‐parametric model estimates regarding density‐dependent structures of demographic rates in the blowfly populations. The mean demographic rates, however, were apparently underestimated by the model. We conclude that non‐parametric modelling is a useful first approach for exploratory analysis of ecological time‐series data.     

Climatic stability and genetic divergence in the tropical insular lizard Anolis krugi,the Puerto Rican ‘Lagartijo Jardinero de la Montaña’

Two factors that can lead to geographic structuring in conspecific populations are barriers to dispersal and climatic stability. Populations that occur in different physiographic regions may be restricted to those areas by physical and/or ecological barriers, which may facilitate the formation of phylogeographic clades. Long‐term climatic stability can also promote genetic diversification, because new clades are more likely to evolve in areas that experience lesser climatic shifts. We conducted a phylogeographic study of the Puerto Rican lizard Anolis krugi to assess whether populations of this anole show genetic discontinuities across the species’ range, and if they do, whether these breaks coincide with the boundaries of the five physiographic regions of Puerto Rico. We also assessed whether interpopulation genetic distances in A. krugi are positively correlated with relative climatic stability in the island. Anolis krugi exhibits genetic structuring, but the phylogroups do not correspond to the physiographic regions of Puerto Rico. We used climatic reconstructions of two environmental extremes of the Quaternary period, the present conditions and those during the last glacial maximum (LGM), to quantify the degree of climatic stability between sampling locations. We documented positive correlations between genetic distances and relative climatic stability, although these associations were not significant when corrected for autocorrelation. Principal component analyses indicated the existence of climatic niche differences between some phylogeographic clades of A. krugi. The approach that we employed to assess the relationship between climatic stability and the genetic architecture of A. krugi can also be used to investigate the impact of factors such as the spatial distribution of food sources, parasites, predators or competitors on the genetic landscape of a species.     

Direct negative density‐dependence in a pond‐breeding frog population

Understanding population dynamics is critical for the management of animal populations. Comparatively little is known about the relative importance of endogenous (i.e. density‐dependent) and exogenous (i.e. density‐independent) factors on the population dynamics of amphibians with complex life cycles. We examined the potential effects of density‐dependent and ‐independent (i.e. climatic) factors on population dynamics by analyzing a 15‐yr time series data of the agile frog Rana dalmatina population from Târnava Mare Valley, Romania. We used two statistical models: 1) the partial rate correlation function to identify the feedback structure and the potential time lags in the time series data and 2) a Gompertz state‐space model to simultaneously investigate direct and delayed density dependence as well as climatic effects on population growth rate. We found evidence for direct negative density dependence, whereas delayed density dependence and climate did not show a strong influence on population growth rate. Here we demonstrated that direct density dependence rather than delayed density dependence or climate determined the dynamics of our study population. Our results confirm the findings of many experimental studies and suggest that density dependence may buffer amphibian populations against environmental stress. Consequently, it may not be easy to scale up from individual‐level effects to population‐level effects.     

A comparison of mosquito densities,weather and infection rates of Aedes aegypti during the first epidemics of Chikungunya (2014) and Zika (2016) in areas with and without vector control in Puerto Rico

In Puerto Rico, the first records of the transmission of Chikungunya (CHIKV) and Zika (ZIKV) viruses were confirmed in May 2014 and December 2015, respectively. Transmission of CHIKV peaked in September 2014, whereas that of ZIKV peaked in August 2016. The emergence of these mosquito‐transmitted arboviruses in the context of a lack of human population immunity allowed observations of whether the outbreaks were associated with Aedes aegypti (Diptera: Culicidae) densities and weather. Mosquito density was monitored weekly in four communities using sentinel autocidal gravid ovitraps (AGO traps) during 2016 in order to provide data to be compared with the findings of a previous study carried out during the 2014 CHIKV epidemic. Findings in two communities protected against Ae. aegypti using mass AGO trapping (three traps per house in most houses) were compared with those in two nearby communities without vector control. Mosquito pools were collected to detect viral RNA of ZIKV, CHIKV and dengue virus. In areas without vector control, mosquito densities and rates of ZIKV detection in 2016 were significantly higher, similarly to those observed for CHIKV in 2014. The density of Ae. aegypti in treated sites was less than two females/trap/week, which is similar to the putative adult female threshold for CHIKV transmission. No significant differences in mosquito density or infection rates with ZIKV and CHIKV at the same sites between years were observed. Although 2016 was significantly wetter, mosquito densities were similar.     

Body size variation of the floodwater mosquito Aedes albifasciatus in Central Argentina

An inverse relationship between larval density and adult body size has been reported for several mosquito species, affecting their survival and vector competence, response to repellents and other factors. Larvae of the floodwater mosquito Aedes (Ochlerotatus) albifasciatus (Macquart) (Diptera: Culicidae) develop quickly in temporary pools, so intraspecific competition (for food or space) might regulate population abundance and affect the size of adult mosquitoes. We investigated the temporal variation of adult female wing-length (an index of body-size) in natural populations of Ae. albifasciatus, using adults collected during each phase of the rainy season. The relationships between adult mosquito abundance, female wing-length, rainfall and temperature were analysed through simple regressions. Skewness of the frequency distribution of wing-lengths showed a strong negative relationship with mean wing-length. The distribution of wing-lengths varied seasonally and was correlated with rainfall 7-15 days previously as the major consequence of breeding site volume. Thus temporal variation of body size in natural populations of Ae. albifasciatus reflected density-dependent changes in the aquatic habitat where immature stages develop, influenced more by rainfall than by temperature or other environmental variables.     

Modeling the effects of Aedes aegypti’s larval environment on adult body mass at emergence

Mosquitoes vector harmful pathogens that infect millions of people every year, and developing approaches to effectively control mosquitoes is a topic of great interest. However, the success of many control measures is highly dependent upon ecological, physiological, and life history traits of mosquito species. The behavior of mosquitoes and their potential to vector pathogens can also be impacted by these traits. One trait of interest is mosquito body mass, which depends upon many factors associated with the environment in which juvenile mosquitoes develop. Our experiments examined the impact of larval density on the body mass of Aedes aegypti mosquitoes, which are important vectors of dengue, Zika, yellow fever, and other pathogens. To investigate the interactions between the larval environment and mosquito body mass, we built a discrete time mathematical model that incorporates body mass, larval density, and food availability and fit the model to our experimental data. We considered three categories of model complexity informed by data, and selected the best model within each category using Akaike’s Information Criterion. We found that the larval environment is an important determinant of the body mass of mosquitoes upon emergence. Furthermore, we found that larval density has greater impact on body mass of adults at emergence than on development time, and that inclusion of density dependence in the survival of female aquatic stages in models is important. We discuss the implications of our results for the control of Aedes mosquitoes and on their potential to spread disease.     

Sublethal metal stress response of larvae of Aedes aegypti

Aedes aegypti (Diptera: Culicidae) has adapted to urban environments. The urbanization process provides suitable habitats for this disease vector, subsequently increasing the probability of the transmission of pathogens in high‐density environments. Urban environments provide metal‐stressed larval habitats. However, little is known about the physiological cost of metal stress or how this might affect the performance of this mosquito species. The present study aims to characterize the sublethal physiological consequences of metal stress in Aedes aegypti. Various parameters of mosquito physiology under larval metal stress are assessed, including larval metallothionein expression and the effects of larval metal stress on adult performance and their progeny. The results show that environmentally relevant larval metal stress compromises larval and adult development and performance, and also results in larval metal tolerance along with an increase in lipid consumption. These performance costs are coupled with a dramatic increase in metallothionein expression in the midgut. Metal stress results in a lowered adult body mass and neutral storage lipids at emergence, starvation tolerance, fecundity and starvation tolerance of offspring compared with non‐metal‐stressed individuals. Ironically, larval metal stress results in increased adult longevity. Taken together, these findings indicate that even low levels of environmentally relevant larval metal stress have considerable physiological consequences for this important disease vector.     

Sea level,topography and island diversity: phylogeography of the Puerto Rican Red‐eyed Coquí, Eleutherodactylus antillensis

Quaternary climatic oscillations caused changes in sea level that altered the size, number and degree of isolation of islands, particularly in land‐bridge archipelagoes. Elucidating the demographic effects of these oscillations increases our understanding of the role of climate change in shaping evolutionary processes in archipelagoes. The Puerto Rican Bank (PRB) (Puerto Rico and the Eastern Islands, which comprise Vieques, Culebra, the Virgin Islands and associated islets) in the eastern Caribbean Sea periodically coalesced during glaciations and fragmented during interglacial periods of the quaternary. To explore population‐level consequences of sea level changes, we studied the phylogeography of the frog Eleutherodactylus antillensis across the archipelago. We tested hypotheses encompassing vicariance and dispersal narratives by sequencing mtDNA (c. 552 bp) of 285 individuals from 58 localities, and four nuDNA introns (totalling c. 1633 bp) from 173 of these individuals. We found low support for a hypothesis of divergence of the Eastern Islands populations prior to the start of the penultimate interglacial c. 250 kya, and higher support for a hypothesis of colonization of the Eastern Islands from sources in eastern Puerto Rico during the penultimate and last glacial period, when a land bridge united the PRB. The Río Grande de Loíza Basin in eastern Puerto Rico delineates a phylogeographic break. Haplotypes shared between the PRB and St. Croix (an island c. 105 km south‐east of this archipelago) likely represent human‐mediated introductions. Our findings illustrate how varying degrees of connectivity and isolation influence the evolution of tropical island organisms.     

Single‐ or multistage regulation in complex life cycles: does it make a difference?

Data on the different stages of complex life cycles are often rather unbalanced, especially those concerning the effects of density. How does this affect our understanding of a species’ population dynamics? Two discrete three‐stage models with overlapping generations and delayed maturation are constructed to address this question. They assume that survival or emigration in any life stage and/or reproduction can be density dependent. A typical pond‐breeding amphibian species with a well‐studied larval stage serves as an example. Numerical results show that the population dynamics resulting from density dependence at a single (e.g. the larval) stage can be decisively and unpredictably modified by density dependence in additional stages. Superposition of density‐dependent processes could thus be one reason for the difficulties in identifying density dependence in the field. Moreover, in a simulated source‐refuge system with habitat‐specific density‐dependent dispersal of juveniles density dependence in multiple stages can stabilize or destabilize the dynamics and produce misleading age structures. From an applied perspective this model shows that excluding multistage regulation prematurely clearly affects our ability to predict consequences of human impacts.     

Demographic response by a small epiphytic orchid

? Premise of the study: Biotic changes are an inevitable consequence of climate change. Epiphytes may be more susceptible to changes in climate variation, but data regarding responses to climate variability under field conditions are limited. We evaluated whether the abundance of demographic stages in the epiphytic orchid Lepanthes rupestris at the Luquillo Experimental Forest in Puerto Rico was associated with short-term changes in climate variation over an 8-yr period. ? Methods: We used cross-correlation analyses to evaluate associations between the abundance of seedlings, juveniles, adults, and fruits per subpopulation, population growth, colonization and extinction rates in L. rupestris with variables related to precipitation and temperature, with and without lag- responses. ? Key results: We detected significant negative correlations between the average number of seedlings and the number of dry days, between the average number of fruits and minimum average temperature with a 6-mo response lag, and between the average number of adults and the maximum temperature with a 1-yr response lag. Neither population growth rate nor probability of colonization and extinction were directly related to climatic variation between 2000 and 2007. ? Conclusions: Associations between climatic variables and demographic stages could have negative implications for this orchid within the context of expected drying trends in the Caribbean. Results argue for the establishment of long-term monitoring studies of orchid populations, because only long-term studies would provide the appropriate temporal scale to detect and predict climate change effects and adaptive management of orchid populations.     

Conservation status assessment of the Sharp‐shinned Hawk,an endangered insular raptor in Puerto Rico

Sharp‐shinned Hawks (Accipiter striatus) are forest raptors that are widely distributed in the Americas. A subspecies endemic to Puerto Rico (A. s. venator) is listed as endangered and restricted to mature and old secondary montane forests and shade coffee plantations. However, recent information about the population status and distribution of Puerto Rican Sharp‐shinned Hawks is lacking. We developed a spatial geographic distribution model for Sharp‐shinned Hawks in Puerto Rico from 33 locations collected during four breeding seasons (2013–2016) using biologically relevant landscape variables (aspect, canopy closure, elevation, rainfall, slope, and terrain roughness). Elevation accounted for 89.8% of the model fit and predicted that the greatest probability of occurrence of Sharp‐shinned Hawks in Puerto Rico (> 60%) was at elevations above 900 m. Based on our model, an estimated 56.1 km² of habitat exists in Puerto Rico with a high probability of occurrence. This total represents ~0.6% of the island's area. Public lands included 43.8% of habitat with high probability of occurrence (24.6 km²), 96% of which was located within four protected areas. Our results suggest that Sharp‐shinned Hawks are rare in Puerto Rico and restricted to the higher elevations of the Cordillera Central. Additional research is needed to identify and address ecological limiting factors, and recovery actions are needed to avoid the extinction of this endemic island raptor.     

Altered climate leads to positive density‐dependent feedbacks in a tropical wet forest

Climate change is predicted to result in warmer and drier Neotropical forests relative to current conditions. Negative density‐dependent feedbacks, mediated by natural enemies, are key to maintaining the high diversity of tree species found in the tropics, yet we have little understanding of how projected changes in climate are likely to affect these critical controls. Over 3 years, we evaluated the effects of a natural drought and in situ experimental warming on density‐dependent feedbacks on seedling demography in a wet tropical forest in Puerto Rico. In the +4°C warming treatment, we found that seedling survival increased with increasing density of the same species (conspecific). These positive density‐dependent feedbacks were not associated with a decrease in aboveground natural enemy pressure. If positive density‐dependent feedbacks are not transient, the diversity of tropical wet forests, which may rely on negative density dependence to drive diversity, could decline in a future warmer, drier world.