Childhood parasitic infections and gastrointestinal illness in indigenous communities at Lake Atitlán, Guatemala.

Lake Atitlán has experienced a decline in water quality resulting from cultural eutrophication. Indigenous Mayans who already face disproportionate health challenges rely directly on the lake water. Our objectives were to: (1) estimate prevalence of shedding of water-borne fecal parasites among children 5 years of age and younger, (2) assess household-reported incidence of gastrointestinal illness in children, and (3) characterize water sources, treatment, and sanitation conditions in households. We hypothesized that household use of untreated lake water results in increased risk of shedding of parasites and gastrointestinal symptoms. A cross-sectional fecal sampling and physical exam of 401 children were conducted along with WASH surveys in partnership with healthcare providers in seven communities. Fecal samples were screened for Giardia lamblia and Cryptosporidium parvum, using a rapid ELISA, with a portion examined by microscope. The prevalence of parasite shedding was 12.2% (9.7% for Giardia; 2.5% for Cryptosporidium). Risk factors for Giardia shedding included age 3 years or older (3.4 odds ratio, z-stat = 2.781 p = 0.0054), low height-for-age z-score (2.3 odds ratio, z-stat = 2.225, p = 0.0216), lack of any household water treatment (2.5 odds ratio, z-stat = 2.492, p < 0.0012), and open access to household latrine (2.04 odds ratio, z-stat = 1.992, p = 0.0464). The majority (77.3%) of households reported water treatment, boiling and gravity fed filters as the most widespread practices. The vast majority of households (92%) reported usage of a latrine, while 40% reported open and shared access beyond their household. An overwhelming majority of households reported diarrhea and fever several times per year or greater, with approximately half reporting vomiting at that frequency. Lake water use was identified as a risk factor for households reporting frequent gastrointestinal symptoms (odds ratio of 2.5, 4.4, and 2.6; z-stat of 3.10, 3.65, and 3.0; p-values of 0.0021, 0.0003, and 0.0028, for diarrhea, vomiting, and fever, respectively) in children 5 years of age and younger. The frequency of gastrointestinal illness with a strong link to lake drinking water cannot be explained by the prevalence of protozoa, and risk from other enteropathogens must be explored. Improving access to water treatment and sanitation practices could substantially reduce the parasite burden faced by developing children in the region.

Harmful Algal Blooms Threaten the Health of Peri-Urban Fisher Communities: A case study in Kisumu Bay, Lake Victoria, Kenya.

Available guidance to mitigate health risks from exposure to freshwater harmful algal blooms (HABs) is largely derived from temperate ecosystems. Yet in tropical ecosystems, HABs can occur year-round, and resource-dependent populations face multiple routes of exposure to toxic components. Along Winam Gulf, Lake Victoria, Kenya, fisher communities rely on lake water contaminated with microcystins (MCs) from HABs. In these peri-urban communities near Kisumu, we tested hypotheses that MCs exceed exposure guidelines across seasons, and persistent HABs present a chronic risk to fisher communities through ingestion with minimal water treatment and frequent, direct contact. We tested source waters at eleven communities across dry and rainy seasons from September 2015 through May 2016. We measured MCs, other metabolites, physicochemical parameters, chlorophyll a, phytoplankton abundance and diversity, and fecal indicators. We then selected four communities for interviews about water sources, usage, and treatment. Greater than 30% of source water samples exceeded WHO drinking water guidelines for MCs (1µg/L), and over 60% of source water samples exceeded USEPA guidelines for children and immunocompromised individuals. 50% of households reported sole use of raw lake water for drinking and household use, with alternate sources including rain and boreholes. Household chlorination was the most widespread treatment utilized. At this tropical, eutrophic lake, HABs pose a year-round health risk for fisher communities in resource -limited settings. Community-based solutions and site-specific guidance for Kisumu Bay and similarly impacted regions is needed to address a chronic health exposure likely to increase in severity and duration with global climate change.

Heterotrophic N2-fixation contributes to nitrogen economy of a common wetland sedge, Schoenoplectus californicus.

A survey of the ecological variability within 52 populations of Schoenoplectus californicus (C.A. Mey.) Soják across its distributional range revealed that it is commonly found in nitrogen (N) limited areas, but rarely in phosphorus limited soils. We explored the hypothesis that S. californicus supplements its nitrogen demand by bacterial N2-fixation processes associated with its roots and rhizomes. We estimated N2-fixation of diazotrophs associated with plant rhizomes and roots from several locations throughout the species' range and conducted an experiment growing plants in zero, low, and high N additions. Nitrogenase activity in rhizomes and roots was measured using the acetylene reduction assay. The presence of diazotrophs was verified by the detection of the nifH gene. Nitrogenase activity was restricted to rhizomes and roots and it was two orders of magnitude higher in the latter plant organs (81 and 2032 nmol C2H4 g DW-1 d-1, respectively). Correspondingly, 40x more nifH gene copies were found on roots compared to rhizomes. The proportion of the nifH gene copies in total bacterial DNA was positively correlated with the nitrogenase activity. In the experiment, the contribution of fixed N to the plant N content ranged from 13.8% to 32.5% among clones from different locations. These are relatively high values for a non-cultivated plant and justify future research on the link between N-fixing bacteria and S. californicus production.

Employing CBPR to investigate function, utility, and longevity of household filters to improve potable water quality for indigenous peoples at Lake Atitlán, Guatemala: a pilot study with San Pedro de La Laguna.

Cyanobacterial blooms at Lake Atitlán in Guatemala threaten and compromise the livelihood and health of local residents. Indigenous Tz'utujil, Kaqchikel, and K'iche' rely directly on lake water for drinking, bathing, cleaning, cooking, and fishing. Nonpoint source runoff and untreated wastewater pumped directly into the lake contribute to high fecal pathogen loads into source waters. Concurrent nutrient loading results in cyanobacterial blooms further compromising water quality. A lakeside municipality facing high rates of childhood gastrointestinal illness volunteered to engage in community-based participatory research (CBPR) to evaluate efficacy, utility, and longevity of filters in households. The filters consistently reduced the risk of coliforms and E. coli in household water drawn from the lake based on World Health Organization guidelines. Household surveys were simultaneously administered through a student leadership group regarding water usage, water quality, and community health. Filters demonstrated ability to reduce high loads of fecal indicators from source waters and ability to remove a cyanobacterial toxin (microcystin) at 10 µg/L in deionized water. Further studies are imperative to determine longevity of use in households and CBPR provides a powerful avenue to test efficacy of a possible intervention while engaging stakeholders and empowering community members with sustainable solutions.

Fatty acids in anopheline mosquito larvae and their habitats.

Larvae of the three important Central American malaria vectors, Anopheles albimanus, An. vestitipennis, and An. darlingi, are found in distinctly different habitats broadly defined by hydrology and aquatic vegetation, but little is known about the actual food quality and quantity of these habitats. Polyunsaturated fatty acids (PUFA) are of special interest, because mosquitoes require 20:5ω3 (EPA), 20:4ω6 (ARA), and 22:6ω3 (DHA) and without an adequate supply of these PUFAs they are not able to complete their life cycle. We collected samples of larvae and their corresponding habitats and analyzed their fatty acid (FA) composition to reveal if there are any species-specific and habitat-specific differences in FA composition, and if habitat FA differences can be linked to differences in the mosquito FA pattern and, ultimately, mosquito performance. We also assessed how FA of wild larvae compare to the laboratory-reared larvae. Habitats were generally low in essential PUFAs and there were no significant differences among the FA composition of habitat samples. There were significant differences in FA composition of larvae. An. darlingi contained significantly higher amounts of FA, specifically a higher content of ω-6 PUFA, represented mainly by the linoleic acid (18:2ω-6). Large differences were found between field-collected and laboratory-reared An. vestitipennis larvae, especially in the content of PUFAs. The laboratory-reared larvae contained significantly more of the total FA, ω3 PUFA, and MUFA. The laboratory-reared larvae contained three to five times more essential PUFAs, EPA, and DHA. However, there were no differences in the total dry weight of the 4(th) instar larvae between the wild vs laboratory-reared larvae. Total FA in both larvae and habitats of An. albimanus and An. darlingi were positively correlated with the concentration of particulate organic carbon and nitrogen (POC, PON) in their respective habitats, but no such correlation was found for An. vestitipennis. PUFA are a good indicator of nutritional quality, although factors controlling the success of anopheline development from larval habitats are likely to be more complex and would include the presence of predators, pathogens, and toxins as interacting factors.

Patterns of activities of root phosphomonoesterase and phosphodiesterase in wetland plants as a function of macrophyte species and ambient phosphorus regime.

Phosphorus (P)-limited plants produce higher amounts of root phosphatases, but research has mostly focused on phosphomonoesterases (PMEs). Because phosphate diesters can form a significant proportion of organic P in wetlands, we aimed to determine whether wetland plants produce both root PMEs and root phosphodiesterases (PDEs), and, if so, what factors influence activities of these enzymes. We measured the activities of root PMEs and PDEs colorimetrically in a wide range of macrophytes from natural and P-enriched wetlands. Hydrolyzable P in sediments was analyzed using commercially available PMEs and PDEs. In all species, both root PMEs and PDEs were always present, and their activities were closely correlated. Sedges and broadleaved emergents had the highest activity of both enzymes, while those of floating-leaved plants were the lowest. Redundancy analysis revealed close association between root enzymes and the proportion of monoesterase- and diesterase-hydrolyzable dissolved unreactive P. Both enzymes were positively correlated with root tissue N : P ratio. Both plant and sediment traits were important when explaining differences in enzyme activities. Although the activities are related to ambient P regime, the relationship was not close enough to use root enzymes as reliable predictors of dissolved unreactive P that is hydrolyzed by sediment phosphomono- and diesterases.

Linking environmental nutrient enrichment and disease emergence in humans and wildlife.

Worldwide increases in human and wildlife diseases have challenged ecologists to understand how large-scale environmental changes affect host-parasite interactions. One of the most profound changes to Earth's ecosystems is the alteration of global nutrient cycles, including those of phosphorus (P) and especially nitrogen (N). Along with the obvious direct benefits of nutrient application for food production, anthropogenic inputs of N and P can indirectly affect the abundance of infectious and noninfectious pathogens. The mechanisms underpinning observed correlations, however, and how such patterns vary with disease type, have long remained conjectural. Here, we highlight recent experimental advances to critically evaluate the relationship between environmental nutrient enrichment and disease. Given the interrelated nature of human and wildlife disease emergence, we include a broad range of human and wildlife examples from terrestrial, marine, and freshwater ecosystems. We examine the consequences of nutrient pollution on directly transmitted, vector-borne, complex life cycle, and noninfectious pathogens, including West Nile virus, malaria, harmful algal blooms, coral reef diseases, and amphibian malformations. Our synthetic examination suggests that the effects of environmental nutrient enrichment on disease are complex and multifaceted, varying with the type of pathogen, host species and condition, attributes of the ecosystem, and the degree of enrichment; some pathogens increase in abundance whereas others decline or disappear. Nevertheless, available evidence indicates that ecological changes associated with nutrient enrichment often exacerbate infection and disease caused by generalist parasites with direct or simple life cycles. Observed mechanisms include changes in host/vector density, host distribution, infection resistance, pathogen virulence or toxicity, and the direct supplementation of pathogens. Collectively, these pathogens may be particularly dangerous because they can continue to cause mortality even as their hosts decline, potentially leading to sustained epidemics or chronic pathology. We suggest that interactions between nutrient enrichment and disease will become increasingly important in tropical and subtropical regions, where forecasted increases in nutrient application will occur in an environment rich with infectious pathogens. We emphasize the importance of careful disease management in conjunction with continued intensification of global nutrient cycles.

Natural hybridization in tropical spikerushes of Eleocharis subgenus Limnochloa (Cyperaceae): Evidence from morphology and DNA markers.

PREMISE OF THE STUDY: Natural hybridization represents an important force driving plant evolution and affecting community structure and functioning. Hybridization may be overlooked, however, among morphologically highly uniform congeners. An excellent example of such a group is Eleocharis subgenus Limnochloa, which has no reliably proven hybrids. Does this reflect biological barriers to interspecific crosses or difficulties in detecting the hybrids? We tested the hypothesis that hybridization occurs among sympatric Eleocharis cellulosa, E. interstincta, and E. mutata in northern Belize, Central America. • METHODS: Morphometric study (407 plants) was followed by examination of inter-simple sequence repeat (ISSR) polymorphisms (44 plants) and ITS sequence variation (33 plants). • KEY RESULTS: Two putatively hybrid morphotypes were discerned-E. cellulosa-resembling and E. interstincta-resembling. DNA markers of E. cellulosa and E. interstincta displayed additive constitution in plants from one E. cellulosa-resembling population only. The other putatively hybrid populations contained ISSR and ITS markers of the species they resembled morphologically, several unique ISSR markers, and ITS sequences of an undescribed South American Limnochloa entity. DNA markers of E. mutata were absent in the putative hybrids. • CONCLUSIONS: Simultaneous use of various types of molecular markers can overcome many pitfalls of investigations concerning hybridization among closely related and morphologically similar species. Northern Belize represents a hybrid zone of E. cellulosa and E. interstincta. A third participant in the hybridization events occurring in this zone is an unknown Limnochloa lineage but is not E. mutata. Interspecific hybridization may play a significant role in the diversification of Eleocharis.

A mark release-recapture study to define the flight behaviors of Anopheles vestitipennis and Anopheles albimanus in Belize, Central America.

The recapture rates of wild-caught, unengorged Anopheles vestitipennis and Anopheles albimanus females were determined at 0, 400, and 800 m from a fixed release point in Belize, Central America. Three sampling trials, each consisting of two 12-hour collections, were performed at each distance during September-October 2003. A total of 1,621 An. vestitipennis and 1,326 An. albimanus were marked and released during the course of the study. The recapture rate of An. vestitipennis was greatest at 0 m (7.9%; 44/ 556) and declined from 3.0% (16/531) at 400 m to 0.2% (1/534) at 800 m. Anopheles albimanus females were recaptured only at the 0-m distance and in extremely low numbers (1.1%; 5/446). Biting patterns for the unmarked natural populations were similar to those previously described for Belize, and recaptures for both species occurred during these normal biting times. The overall recapture rates for An. vestitipennis (3.76%; 61/ 1,621) and An. albimanus (0.38%; 5/1,326) indicate that An. vestitipennis has a higher probability of being attracted to a human habitation.

Habitat suitability for three species of Anopheles mosquitoes: larval growth and survival in reciprocal placement experiments.

Larval habitats of the main malaria vectors in Belize are associated with three distinctly different aquatic environments: marshes with sparse macrophytes and cyanobacterial mats (Anopheles albimanus), tall dense macrophyte marshes (An. vestitipennis), and floating detritus assemblages within freshwater rivers (An. darlingi). We assessed species-specific habitat suitability based upon nutrient characteristics using larval survival rates (SR) and wing lengths (WL) from floating habitat enclosures. Anopheles albimanus showed a high SR (81%) in all three habitats, while An. vestitipennis had a similarly high SR in its own habitat (82%) and An. darlingi's habitat (81%). Anopheles darlingi only showed high SR (85%) in its own habitat. Both An. vestitipennis and An. darlingi showed very low SR in the An. albimanus habitat. There were no significant WL differences among field-caught, laboratory-reared, and experimental populations of An. vestitipennis and An. albimanus, with the exception of An. vestitipennis experimental populations and An. vestitipennis field populations placed in the An. albimanus habitat. Habitat quality indicators, particulate organic carbon (POC), dissolved organic carbon (DOC), and particulate organic nitrogen (PON), were consistently higher in An. vestitipennis habitats than in the habitats of the other two species. Correspondingly, An. vestitipennis adults were larger when measured both as dry mass and from WL. There were no differences in dry mass, lipids, or protein content among the same species reared at different locations. We compared SR and WL among mosquitoes from shaded and unshaded containers to test whether the high mortality rates for An. vestitipennis and An. darlingi in the An. albimanus habitat were due to intense sun exposure. There were no significant differences among developmental times, survivorship, or adult size for shaded versus sun-exposed populations. This indicates that other factors such as larval toxins, predator avoidance, interspecific species competition, etc. may be responsible for the higher mortality rates in those species not adapted to this particular habitat.

A mark-release-recapture study to define the flight behaviors of Anopheles vestitipennis and Anopheles albimanus in Belize, Central America

The recapture rates of wild-caught, unengorged Anopheles vestitipennis and Anopheles albimanus females were determined at 0, 400, and 800 m from a fixed release point in Belize, Central America. Three sampling trials, each consisting of two 12-hour collections, were performed at each distance during September­October 2003. A total of 1,621 An. vestitipennis and 1,326 An. albimanus were marked and released during the course of the study. The recapture rate of An. vestitipennis was greatest at 0 m (7.9%; 44/ 556) and declined from 3.0% (16/531) at 400 m to 0.2% (1/534) at 800 m. Anopheles albimanus females were recaptured only at the 0-m distance and in extremely low numbers (1.1%; 5/446). Biting patterns for the unmarked natural populations were similar to those previously described for Belize, and recaptures for both species occurred during these normal biting times. The overall recapture rates for An. vestitipennis (3.76%; 61/ 1,621) and An. albimanus (0.38%; 5/1,326) indicate that An. vestitipennis has a higher probability of being attracted to a human habitation.

A delayed release mechanism for mark-release-recapture studies.

A delayed release mechanism was designed for a mark-recapture study that evaluated the flight behavior of anopheline mosquitoes in Belize, Central America. The design prevents marked mosquitoes from being released until after a set time. The time lag allows researchers to return to collecting posts before release and reduces the potential of marked females tracking an odor plume generated during the release procedure. This is especially important in study designs evaluating temporal host-seeking behavior patterns. Detailed descriptions of the design are presented.

Biting patterns and seasonal densities of Anopheles mosquitoes in the Cayo District, Belize, Central America with emphasis on Anopheles darlingi.

The present study utilized an experimental hut to conduct human-baited landing collections for characterizing the all-night biting patterns and seasonal densities of adult Anopheles darlingi in the centrally located Cayo District of Belize, Central America. A total of 25 all-night collections (i.e., sunset to sunrise) were conducted from January 2002 to May 2003, capturing a total of 18,878 An. darlingi females. Anopheles darlingi exhibited a bimodal nightly biting pattern with one predominate peak occurring three h after sunset and a smaller peak occurring one h prior to sunrise. Biting females were collected throughout the night in higher densities indoors (9,611) than outside (9,267) the experimental hut (O:I=1.00:1.04). Seasonal adult collections show An. darlingi densities were highest during the transitional months between the end of the wet and beginning of the dry season (January) and the end of the dry season and beginning of the wet season (May). A total of 2,010 An. darlingi females was captured in 31 two-h, human-baited landing collections performed from January to October 2002. Anopheles darlingi monthly population densities were found to have no significant associations with high or low temperatures, precipitation, or river level. However, qualitative data examination indicates an inverse relationship between river level and An. darlingi adult collections suggesting a disturbance of larval habitats. All-night biting and seasonal distribution patterns for other anopheline species are also described. None of the adult specimens collected throughout the entire study tested positive for Plasmodium spp. infection using the VecTest rapid diagnostic kit.

Experimental evaluation of overhanging bamboo in Anopheles darlingi larval habitat selection in Belize, Central America.

Previous studies in Belize have shown the preferred breeding habitats of the malaria vector Anopheles darlingi were floating detritus patches within riverine systems that were associated with overhanging bamboo. The present study focused on an experimental evaluation of overhanging bamboo in An. darlingi habitat selection using larval counts as an indicator of attractiveness. Four sets of 1-m2 floating screened enclosures were placed at a location with a documented presence of both larval and adult An. darlingi populations. Each enclosure set comprised a control (i.e., open water) and three other experimental treatments consisting of: 1) detritus, 2) detritus with overhanging bamboo, and 3) overhanging bamboo alone. Larvae were sampled from all treatments on Day 5, Day 11, and Day 17 post-setup. A total of 2,461 An. darlingi larvae were collected and identified from three trials conducted from March-May 2002. Of these, 1,997 larvae were sampled from detritus treatments, 256 from enclosures with bamboo and detritus, 139 from bamboo treatments, and 69 from control enclosures. The detritus treatment had a significantly higher average count of An. darlingi larvae than the other treatments (P<0.01), and no difference existed between the control treatment and the treatment containing overhanging bamboo alone (P=0.423). More importantly, enclosures containing the overhanging bamboo with detritus treatments did not have greater larval populations than the enclosures with only detritus. These data suggest that bamboo does not contribute to An. darlingi larval habitat attractiveness but may function as a barrier to surface water flow causing the lodging of debris, the preferred habitat, that will then attract gravid females for oviposition.

Distribution of Anopheles albimanus, Anopheles vestitipennis, and Anopheles crucians associated with land use in northern Belize.

Anthropogenic land use changes often alter natural patterns of disease transmission. The goal of this study was to determine whether phosphorus input from sugarcane, Saccharum officinarum L., cultivation in northern Belize could pose a significant environmental impact on malaria transmission by changing vegetation structure and composition of wetlands and associated larval habitats. Our primary focus was on the increased dominance of cattail, Typha domingensis Pers., a favored habitat for Anopheles vestitipennis Dyar & Knab. A land cover classification based on satellite imagery was used to select 20 marshes impacted by agricultural runoff and 20 marshes surrounded by forest (nonimpacted). A 100-m transect was established into each of the 40 marshes. Water, vegetation, and larval sampling were conducted at the 0-, 10-, 25-, 50-, and 100-m locations along the transect. Analyses of larval density data indicated that Anopheles albimanus Wiedemann was negatively correlated with percentage of cover of Typha (R2 = 0.39, P < 0.001) but positively correlated with sparse Eleocharis cellulosa Torr. (rush) cover (R2 = 0.19, P < 0.05) and presence of cyanobacterial mats (CBM) (R2 = 0.33, P < 0.0001). An. vestitipennis was found to be positively correlated with percentage of cover of Typha (R2 = 0.19, P < 0.001). Canonical correspondence analysis identified CBM and light as the variables associated with the presence of An. albimanuts larvae, Typha cover with An. vestitipennis larvae, and Eleocharis and absence of light with Anopheles crucians (Wiedemann). A positive correlation also existed between marshes adjacent to agricultural activities and presence of An. vestitipennis (R2 = 0.37, P < 0.05). These results indicate that marshes in proximity to agricultural fields are conducive for Typha growth, thereby providing habitat for the more efficient malaria vector

Use of remote sensing and geographic information systems to predict locations of Anopheles darlingi-positive breeding sites within the Sibun River in Belize, Central America.

Previous studies have identified several anopheline species integral to the transmission of malaria in Belize. The highly efficient vector, Anopheles darlingi Root, is currently considered the most important. The preferred larval habitat of An. darlingi has been described as floating detritus patches, which are commonly associated with overhanging spiny bamboo, Guadua longifolia (E. Fourn.), along river margins. The objectives of this study were to use remote sensing and geographic information system (GIS) tools to 1) define the landscape features (i.e., river curvature, land cover, and house locations) associated with An. darlingi-positive breeding habitats and 2) determine the association between cleared land cover and the growth of spiny bamboo. A systematic survey was conducted in which all detritus patches of at least 1 m2 were sampled, mapped using GPS, and characterized by cause of habitat lodging. Bamboo stretches growing along the river margins also were mapped. Spatial analyses of satellite imagery found no associations between river characteristics or land cover with positive An. darlingi habitats. In addition, there was no significant difference in cleared versus forested land cover in relation to the presence or absence of bamboo. Results indicate that the average distance from homes to negative habitats was significantly greater than from positive detritus mats. Based on the land cover and river characteristics used, our results do not support the use of remote sensing as a predictive tool to locate specific areas within rivers positive for An. darlingi habitats.

Nutrient resorption in wetland macrophytes: comparison across several regions of different nutrient status.

This study explored patterns of nutrient resorption in wetland macrophytes to test the prediction that plants from regions with a strong nutrient limitation will show higher resorption of the limiting nutrient. Nitrogen and phosphorus resorption was assessed in macrophytes from marshes of different nutrient status in tropical and temperate regions, and expressed as resorption efficiency (NRE, PRE) and proficiency (NRP, PRP). Macrophytes were grouped into three categories: Typha, graminoids and broadleaved plants. Nitrogen was less limiting than P, consequently N availability varied less than P availability, NRP and NRE were lower, and N resorption was mostly incomplete. NRP was determined more by growth form than by local conditions. The large range of soil P concentrations allowed an exploration of relationships between P availability and resorption along a wide gradient. P-limited macrophytes (N : P > 16) had significantly higher PRP and PRE. Resorption proficiency was found to be a more sensitive indicator of changes in nutrient availability than resorption efficiency. The results confirmed that resorption in wetland macrophytes depends on nutrient availability, and is higher at nutrient-limited sites. A particularly strong relationship was found between resorption indicators and P limitation expressed either as live tissue N : P or soil P.

Volatile substances from larval habitats mediate species-specific oviposition in Anopheles mosquitoes.

Oviposition site selection has been recognized as critical both for the survival and population dynamics of mosquitoes. Volatile substances released from larval habitats have been implicated as potential olfactory cues mediating oviposition. In our continuing studies of cues involved in oviposition site selection, we collected material from the larval habitats of Anopheles albimanus Wiedemann and Anopheles vestitipennis Dyar & Knab, i.e., cyanobacterial mats and Typha domingensis Pers. litter, respectively. The volatile compounds were extracted by freeze-drying the material and trapping the volatilized material on a -55 degrees C titanium condenser. For oviposition trials conducted with wild-caught females, the tested volatile materials were pipetted onto filters floating on the surface of distilled water in Teflon beakers that were placed within oviposition cages. For both species, volatile materials in low concentrations increased oviposition, assessed as egg density, whereas there was a shift to reduced oviposition at higher concentrations. Volatile effect was strongly habitat/species-specific as shown by reciprocal treatment tests.

A mark-release-recapture study using a novel portable hut design to define the flight behavior of Anopheles darlingi in Belize, Central America.

Knowledge of the flight behavior of local vectors is of paramount importance in mosquito control programs. The following study defined the recapture rate of wild-caught, unengorged Anopheles darlingi females at 0, 400, and 800 m from a fixed release point in Belize, Central America, using a newly designed portable experimental hut. Three sampling trials, each consisting of 2 12-h collections, were performed at all distances from July 2002 to June 2003. A total of 1,185 An. darlingi were marked and released during the course of the study. The recapture rate was greatest at 0 m (29.0%; 124/428) and declined from 11.6% (37/318) at 400 m to 5.8% (21/361) at the 800-m site. There was no difference between the average number of marked mosquitoes recaptured inside the experimental hut versus outside the hut at any distance location. Recapture rates of each trial were highest during the first night's collection at all locations. Further examination of the first night data revealed a variation in the peak time of recapture by distances from the release point. The peak in nightly recapture at both the 0- and 400-m sites occurred within the first 2 h after sunset, and the peak recapture at the 800-m site occurred during the 7th h after sunset. Information from the present study is the first to describe the flight behavior of An. darlingi in Belize and will benefit in the development of adult-density risk assessments at the house level based on distances from potential vector breeding sites.

Evaluation of habitat management strategies for the reduction of malaria vectors in northern Belize.

Mowing and burning of emergent vegetation were evaluated as potential management strategies for the control of the malaria vector, Anopheles vestitipennis, in northern Belize, Central America. The primary aim was reduction of tall dense macrophytes (dominated by Typha domingensis) as preferred larval habitat for An. vestitipennis. Nine experimental plots were established in a Typha marsh in Orange Walk District, Belize. Three plots were burned, three were treated by subaquatic mowing, and three were unaltered controls. After treatment, Typha height was most dramatically affected by the mow treatment. Plant heights at 21 and 95 days post-treatment reflected an 89% and 48% decrease, respectively, compared to pretreatment conditions. The Typha height in the burn plots was not as severely affected. Heights at 21 days post-treatment were 39% lower than those of pre-treatment vegetation, with a return to near pre-test heights by 95 days post-treatment. Both treatments resulted in a significant reduction in the number of An. vestitipennis larvae collected as compared to control plots. Conversely, the treatments resulted in increased larval densities of several other vector and pest mosquito species. Larval population densities ofAn. albimanus, Ochlerotatus taeniorhynchus, and Culex coronator were significantly higher in burn plots. In mow plots, there were significant increases in An. albimanus and Oc. taeniorhynchus larval populations. Non-target invertebrate species affected by the treatments were adult Tropisternus collaris, larval Corythrella, and adult Parapleapuella.