Identification, distribution, and probability of infection in sympatric gastropods by digenean parasites at Pa-hay-okee, Everglades National Park using molecules and morphology

In this study, I determined the identity, taxonomic placement, and distribution of digenetic trematodes parasitizing the snails Pomacea paludosa and Planorbella duryi at Pa-hay-okee, Everglades National Park. I also characterized temporal and geographic variation in the probability of parasite infection for these snails based on two years of sampling. Although studies indicate that digenean parasites may have important effects both on individual species and the structure of communities, there have been no studies of digenean parasitism on snails within the Everglades ecosystem. For example, the endangered Everglade Snail Kite, a specialist that feeds almost exclusively on Pomacea paludosa, and is known to be a definitive host of digenean parasites, may suffer direct and indirect effects from consumption of parasitized apple snails. Therefore, information on the diversity and abundance of parasites harbored in snail populations in the Everglades should be of considerable interest for management and conservation of wildlife. Juvenile digeneans (cercariae) representing 20 species were isolated from these two snails, representing a quadrupling of the number of species known. Species were characterized based on morphological, morphometric, and sequence data (18S rDNA, COI, and ITS). Species richness of shed cercariae from P. duryi was greater than P. paludosa, with 13 and 7 species respectively. These species represented 14 families. P. paludosa and P. duryi had no digenean species in common. Probability of digenean infection was higher for P. duryi than P. paludosa and adults showed a greater risk of infection than juveniles for both of these snails. Planorbella duryi showed variation in probability of infection between sampling sites and hydrological seasons. The number of unique combinations of multi-species infections was greatest among P. duryi individuals, while the overall percentage of multi-species infections was greatest in P. paludosa. Analyses of six frequently-observed multiple infections from P. duryi suggest the presence of negative interactions, positive interactions, and neutral associations between larval digeneans. These results should contribute to an understanding of the factors controlling the abundance and distribution of key species in the Everglades ecosystem and may in particular help in the management and recovery planning for the Everglade Snail Kite.

Leaf Gas Exchange and Nutrient Use Efficiency Help Explain the Distribution of Two Neotropical Mangroves under Contrasting Flooding and Salinity

Rhizophora mangle and Laguncularia racemosa cooccur along many intertidal floodplains in the Neotropics. Their patterns of dominance shift along various gradients, coincident with salinity, soil fertility, and tidal flooding. We used leaf gas exchange metrics to investigate the strategies of these two species in mixed culture to simulate competition under different salinity concentrations and hydroperiods. Semidiurnal tidal and permanent flooding hydroperiods at two constant salinity regimes (10 g L−1 and 40 g L−1) were simulated over 10 months. Assimilation ( ), stomatal conductance ( ), intercellular CO2 concentration ( ), instantaneous photosynthetic water use efficiency (PWUE), and photosynthetic nitrogen use efficiency (PNUE) were determined at the leaf level for both species over two time periods. Rhizophora mangle had significantly higher PWUE than did L. racemosa seedlings at low salinities; however, L. racemosa had higher PNUE and and, accordingly, had greater intercellular CO2 (calculated) during measurements. Both species maintained similar capacities for A at 10 and 40 g L−1 salinity and during both permanent and tidal hydroperiod treatments. Hydroperiod alone had no detectable effect on leaf gas exchange. However, PWUE increased and PNUE decreased for both species at 40 g L−1 salinity compared to 10 g L−1. At 40 g L−1 salinity, PNUE was higher for L. racemosa than R. mangle with tidal flooding. These treatments indicated that salinity influences gas exchange efficiency, might affect how gases are apportioned intercellularly, and accentuates different strategies for distributing leaf nitrogen to photosynthesis for these two species while growing competitively.

Phylogenetic and ontogenetic influences on the distribution of anthocyanins and betacyanins in leaves of tropical plants

We examined the anatomy of expanding, mature, and senescing leaves of tropical plants for the presence of red pigments: anthocyanins and betacyanins. We studied 463 species in total, 370 genera, belonging to 94 families. This included 21 species from five families in the Caryophyllales, where betacyanins are the basis for red color. We also included 14 species of ferns and gymnosperms in seven families and 29 species with undersurface coloration at maturity. We analyzed 399 angiosperm species (74 families) for factors (especially developmental and evolutionary) influencing anthocyanin production during expansion and senescence. During expansion, 44.9% produced anthocyanins and only 13.5% during senescence. At both stages, relatively few patterns of tissue distributions developed, primarily in the mesophyll, and very few taxa produced anthocyanins in dermal and ground tissue simultaneously. Of the 35 species producing anthocyanins both in development and senescence, most had similar cellular distributions. Anthocyanin distributions were identical in different developing leaves of three heteroblastic taxa. Phylogeny has influenced the distribution of anthocyanins in the epidermis and mesophyll of expanding leaves and the palisade parenchyma during senescence, although these influences are not strong. Betacyanins appear to have similar distributions in leaves of taxa within the Caryophyllales and, perhaps, similar functions. The presence of anthocyanins in the mesophyll of so many species is inconsistent with the hypothesis of protection against UV damage or fungal pathogens, and the differing tissue distributions indicate that the pigments may function in different ways, as in photoprotection and freeradical scavenging.

Physiological performance measures and tolerance limits of estuarine indicator species in south Florida

Current water management practices in South Florida have negatively impacted many species inhabiting Florida Bay. Variable and high salinity has been identified as a key stressor in these estuaries. The Comprehensive Everglades Restoration Plan (CERP) includes water redistribution projects that will restore natural freshwater flows to northeastern Florida Bay. My studies focused on the following central theme and hypotheses: Biological performance measures (i.e., growth, reproduction, survival), behavior (i.e., habitat preference and locomotor behavior) and diversity of estuarine fish will be controlled by changes in salinity and water quality that will occur as a result of the restoration of freshwater flow to the bay. A series of acute and subchronic physiological toxicity studies were conducted to determine the effects of salinity changes on the life stages (embryo/larval, juvenile, adult) and fecundity of four native estuarine fish (Cyprinodon variegatus, Floridichthys carpio, Poecilia latipinna, and Gambusia holbrooki). Fish were exposed to a range of salinity concentrations (freshwater to hypersaline) based on salinity profiles in the study areas. Growth (length, weight) and survival were measured. Salinity trials included both rapid and gradual change events. Results show negative effects of acute, abrupt salinity changes on fish survival, development and reproductive success as a result of salinity stress. Other studies targeted reproduction and critical embryo-larval/neonate development as key areas for detecting long-term population effects of salinity change in Florida Bay. Adults of C. variegates and P. latipinna were also examined for behavioral responses to pulsed salinity changes. These responses include changes in swimming performance, locomotor behavior and zone preference. Finally, an ecological risk assessment was conducted for adverse salinity conditions in northeastern Florida Bay. Using the U.S. EPA's framework, the risk to estuarine fish species diversity was assessed against regional salinity profiles from a 17-year database. Based on the risk assessment, target salinity profiles for these areas are recommended for managers.^

Interactions of different arsenic species with thols: Chemical and biological implications

Arsenic has been classified as a group I carcinogen. It has been ranked number one in the CERCLA priority list of hazardous substances due to its frequency, toxicity and potential for human exposure. Paradoxically, arsenic has been employed as a successful chemotherapeutic agent for acute promyelocytic leukemia and has found some success in multiple myeloma. Since arsenic toxicity and efficacy is species dependent, a speciation method, based on the complementary use of reverse phase and cation exchange chromatography, was developed. Inductively coupled plasma mass spectrometer (ICP-MS), as an element specific detector, and electrospray ionization mass spectrometer (ESI-MS), as a molecule specific detector, were employed. Low detection limits in the µg. L−1 range on the ICP-MS and mg. L−1 range on the ESI-MS were obtained. The developed methods were validated against each other through the use of a Deming plot. With the developed speciation method, the effects of both pH on the stability of As species and reduced glutathione (GSH) concentration on the formation and stability of arsenic glutathione complexes were studied. To identify arsenicals in multiple myeloma (MM) cell lines post arsenic trioxide (ATO) and darinaparsin (DAR) incubation, an extraction method based on the use of ultrasonic probe was developed. Extraction tools and solvents were evaluated and the effect of GSH concentration on the quantitation of arsenic glutathione (As-GSH) complexes in MM cell extracts was studied. The developed method was employed for the identification of metabolites in DAR incubated cell lines where the effect of extraction pH, DAR incubation concentration and incubation time on the relative distribution of the As metabolites was assessed. A new arsenic species, dimethyarsinothioyl glutathione (DMMTA V-GS), a pentavalent thiolated arsenical, was identified in the cell extracts through the use of liquid chromatography tandem mass spectrometry. The formation of the new metabolite in the extracts was dependent on the decomposition of s-dimethylarsino glutathione (DMA(GS)). These results have major implications in both the medical and toxicological fields of As because they involve the metabolism of a chemotherapeutic agent and the role sulfur compounds play in this mechanism.

Segregation of Palaemonid Shrimp Along an Everglades Estuarine Gradient: Do Multiple Species Have Similar Trophic Function?

Variation in physical gradients and production along estuaries can alter species compositions. Spatiotemporal variation in abundance and distribution of palaemonid shrimp species was investigated in relation to seasonal freshwater inputs and salinity in the Shark River Estuary, Everglades National Park, Florida, USA. Using trapping techniques, multiple sites were sampled repeatedly extending from the headwaters to the Gulf of Mexico. Stable isotope analyses were also performed on a subset of samples. Five palaemonid species occurred in the samples: Palaemonetes paludosus (Gibbes, 1850), Palaemonetes pugio (Holthuis, 1949), Palaemonetes intermedius (Holthuis, 1949), Palaemon floridanus (Chace, 1942), and Leander paulensis(Ortmann, 1897). Overall, shrimp catches in traps doubled in the dry season. Catches in the upper estuary were dominated by P.paludosus, particularly in the wet season, while catch per unit effort at the most downstream and highest salinity sites were dominated by P. floridanus. At mid-estuary, several species co-occurred. δ15n analyses revealed that most species filled similar roles in the community, with the exception of P. paludosus, which shifted from enrichment in the dry season to depletion in the wet season as it expanded downstream in the estuary. Palaemonid δ13C values varied between sites and seasons, with shrimp in upstream sites being more depleted. These data suggest that changes in salinity regimes resulting from Everglades restoration efforts may result in species replacement, with potential implications for trophic dynamics.

Taxonomy and distribution of taxa in the genus Gomphonema from the Florida Everglades, U.S.A.

Located at a subtropical latitude, the expansive Florida Everglades contains a mixture of tropical and temperate diatom taxa, as well as a unique flora adapted to the calcareous, often excessively hot, seasonally flooded wetland conditions. This flora has been poorly documented taxonomically, although diatoms are recognized as important indicators of environmental change in this threatened ecosystem. Gomphonema is a dominant genus in the freshwater marsh, and is represented by highly variable species complexes, including Gomphonema gracile Ehrenberg, Gomphonema intricatum var. vibrio Ehrenberg sensu Fricke, Gomphonema vibrioides Reichardt & Lange-Bertalot and Gomphonema parvulum (Kützing) Grunow. These taxa have been shown to exhibit wide morphological variation in other regions, resulting in considerable nomenclatural confusion. We collected Gomphonema from 237 sites distributed throughout the freshwater Everglades and used qualitative and quantitative morphological data to identify 20 distinguishable populations. Taxonomie assignments were based on descriptions and/or observations of type material of relevant taxa when possible, but deviations from original morphological range descriptions were common. We then compared morphological variation in Everglades Gomphonema taxa to that reported for the same taxa in other regions and suggest revisions of taxonomie concepts when necessary.

Distribution of Diatoms and Development of Diatom-Based Models for Inferring Salinity and Nutrient Concentrations in Florida Bay and Adjacent Coastal Wetlands of South Florida (USA)

The composition and distribution of diatom algae inhabiting estuaries and coasts of the subtropical Americas are poorly documented, especially relative to the central role diatoms play in coastal food webs and to their potential utility as sentinels of environmental change in these threatened ecosystems. Here, we document the distribution of diatoms among the diverse habitat types and long environmental gradients represented by the shallow topographic relief of the South Florida, USA, coastline. A total of 592 species were encountered from 38 freshwater, mangrove, and marine locations in the Everglades wetland and Florida Bay during two seasonal collections, with the highest diversity occurring at sites of high salinity and low water column organic carbon concentration (WTOC). Freshwater, mangrove, and estuarine assemblages were compositionally distinct, but seasonal differences were only detected in mangrove and estuarine sites where solute concentration differed greatly between wet and dry seasons. Epiphytic, planktonic, and sediment assemblages were compositionally similar, implying a high degree of mixing along the shallow, tidal, and storm-prone coast. The relationships between diatom taxa and salinity, water total phosphorus (WTP), water total nitrogen (WTN), and WTOC concentrations were determined and incorporated into weighted averaging partial least squares regression models. Salinity was the most influential variable, resulting in a highly predictive model (r apparent 2  = 0.97, r jackknife 2  = 0.95) that can be used in the future to infer changes in coastal freshwater delivery or sea-level rise in South Florida and compositionally similar environments. Models predicting WTN (r apparent 2  = 0.75, r jackknife 2  = 0.46), WTP (r apparent 2  = 0.75, r jackknife 2  = 0.49), and WTOC (r apparent 2  = 0.79, r jackknife 2  = 0.57) were also strong, suggesting that diatoms can provide reliable inferences of changes in solute delivery to the coastal ecosystem.

Ecology and distribution of diatoms in Biscayne Bay, Florida (USA): Implications for bioassessment and paleoenvironmental studies

The spatial and temporal distribution of planktonic, sediment-associated and epiphytic diatoms among 58 sites in Biscayne Bay, Florida was examined in order to identify diatom taxa indicative of different salinity and water quality conditions, geographic locations and habitat types. Assessments were made in contrasting wet and dry seasons in order to develop robust assessment models for salinity and water quality for this region. We found that diatom assemblages differed between nearshore and offshore locations, especially during the wet season when salinity and nutrient gradients were steepest. In the dry season, habitat structure was primary determinant of diatom assemblage composition. Among a suite of physicochemical variables, water depth and sediment total phosphorus (STP) were most strongly associated with diatom assemblage composition in the dry season, while salinity and water total phosphorus (TP) were more important in the wet season. We used indicator species analysis (ISA) to identify taxa that were most abundant and frequent at nearshore and offshore locations, in planktonic, epiphytic and benthic habitats and in contrasting salinity and water quality regimes. Because surface water concentrations of salts, total phosphorus, nitrogen (TN) and organic carbon (TOC) are partly controlled by water management in this region, diatom-based models were produced to infer these variables in modern and retrospective assessments of management-driven changes. Weighted averaging (WA) and weighted averaging partial least squares (WA-PLS) regressions produced reliable estimates of salinity, TP, TN and TOC from diatoms (r2 = 0.92, 0.77, 0.77 and 0.71, respectively). Because of their sensitivity to salinity, nutrient and TOC concentrations diatom assemblages should be useful in developing protective nutrient criteria for estuaries and coastal waters of Florida.

Distribution of Diatoms Along Environmental Gradients in the Charlotte Harbor, Florida (USA), Estuary and Its Watershed: Implications for Bioassessment of Salinity and Nutrient Concentrations

The relative abundance of diatom species in different habitats can be used as a tool to infer prior environmental conditions and evaluate management decisions that influence habitat quality. Diatom distribution patterns were examined to characterize relationships between assemblage composition and environmental gradients in a subtropical estuarine watershed. We identified environmental correlates of diatom distribution patterns across the Charlotte Harbor, Florida, watershed; evaluated differences among three major river drainages; and determined how accurately local environmental conditions can be predicted using inference models based on diatom assemblages. Sampling locations ranged from freshwater to marine (0.1–37.2 ppt salinity) and spanned broad nutrient concentration gradients. Salinity was the predominant driver of difference among diatom assemblages across the watershed, but other environmental variables had stronger correlations with assemblages within the subregions of the three rivers and harbor. Eighteen indicator taxa were significantly affiliated with subregions. Relationships between diatom taxon distributions and salinity, distance from the harbor, total phosphorus (TP), and total nitrogen (TN) were evaluated to determine the utility of diatom assemblages to predict environmental values using a weighted averaging-regression approach. Diatom-based inferences of these variables were strong (salinity R 2 = 0.96; distance R 2 = 0.93; TN R 2 = 0.83; TP R 2 = 0.83). Diatom assemblages provide reliable estimates of environmental parameters on different spatial scales across the watershed. Because many coastal diatom taxa are ubiquitous, the diatom training sets provided here should enable diatom-based environmental reconstructions in subtropical estuaries that are being rapidly altered by land and water use changes and sea level rise.

Physiological performance measures and tolerance limits of estuarine indicator species in South Florida

Current water management practices in South Florida have negatively impacted many species inhabiting Florida Bay. Variable and high salinity has been identified as a key stressor in these estuaries. The comprehensive Everglades Restoration Plan (CERP) includes water redistribution projects that will restore natural freshwater flows to northeastern Florida Bay. My studies focused on the following central theme and hypotheses: Biological performance measures (i.e., growth, reproduction, survival), behavior (i.e., habitat preference and locomotor behavior) and diversity of estuarine fish will be controlled by changes in salinity and water quality that will occur as a result of the restoration of freshwater flow to the bay. A series of acute and subchronic physiological toxicity studies were conducted to determine the effects of salinity changes on the life stages (embryo/larval, juvenile, adult) and fecundity of four native estuarine fish (Cyprinodon variegatus, Floridichthys carpio, Poecilia latipinna, and Gambusia holbrooki). Fishe were exposed to a range of salinity concentrations (freshwater to hypersaline) based on salinity profiles in the study areas. Growth (length, weight) and survival were measured. Salinity trials included both rapid and gradual change events. Results show negative effects of acute, abrupt salinity changes on fish survival, development and reproductive success as a result of salinity stress. Other studies targeted reproduction and critical embryo-larval/neonate development as key areas for detecting long-term population effects of salinity change in Florida Bay. Adults of C. variegatus and P. latipinna were also examined for behavioral responses to pulsed salinity changes. These responses include changes in swimming performance, locomotor behavior and zone preference. Finally, an ecological risk assessment was conducted for adverse salinity conditions in northeastern Florida Bay. Using the U.S. EPA's framework, the risk to estuarine fish species diversity was assessed against regional salinity profiles from a 17-year database. Based on the risk assessment, target salinity profiles for these areas are recommended for managers.