A review of the potential role of tumour-promoting compounds produced by Lyngbya majuscula in marine turtle fibropapillomatosis

Harmful algal blooms (HABs) have increased in abundance and severity in recent decades. Whereas the implications for human impacts and intoxication resulting from blooms have been extensively studied, the ecological implications of these microalgae are less well understood. Many HAB species produce biologically active, secondary metabolites and the fate of these toxins through the foodweb is generally not well understood unless it culminates in extensive fish mortalities or human poisonings. This review focusses on one HAB species, the cyanobacterium Lyngbya majuscula, and presents a hypothetical role for its involvement in fibro-papillornatosis (FP), a neoplastic disease of marine turtles. FP is expressed as benign tumours that grow both internally and externally on marine turtles, preventing vision, movement and organ function. The aetiology of FP is currently not conclusively understood, but virus material has been associated with tumours and previous studies have suggested a role for naturally produced tumour promoters. In this review, we present a hypothesis regarding the involvement of L. majuscula in FP, either through direct intoxication and action of tumour-promoting compounds or indirectly by causing seagrass loss and compromised immune function, thus leaving the turtles more susceptible to disease.

Feeding preference and deterrence in rabbitfish Siganus fuscescens for the cyanobacterium Lyngbya majuscula in Moreton Bay, south-east Queensland, Australia

Rabbitfish Siganus fuscescens preferences for Lyngbya majuscula collected from three bloom locations in Moreton Bay, Queensland, Australia, were tested along with a range of local plant species in the laboratory. Consumption of L. majuscula by fish did not differ between wild and captive-bred fish (P = 0.152) but did differ between bloom location (P = 0.039). No relationship was found between consumption rates and lyngbyatoxin-a concentration (r(2) = 0.035, P = 0.814). No correlation existed between C : N and proportion of food consumed when all food types were analysed statistically, whereas a clear correlation was observed when L. majuscula was removed from the calculations. In simulated bloom conditions, fish avoided ingestion of L. majuscula by feeding through gaps in the L. majuscula coverage. Both wild and captive-bred S. fuscescens showed a distinct feeding pattern in 10 day no-choice feeding assays, with less L. majuscula being consumed than the preferred red alga Acanthophora spicifera. Lyngbya majuscula however, was consumed in equal quantities to A. spicifera by wild S. fuscescens when lyngbyatoxin-a was not detectable. Wild fish probably do not preferentially feed on L. majuscula when secondary metabolites are present and are not severely impacted by large L. majuscula blooms in Moreton Bay. Furthermore, poor feeding performance in both captive-bred and wild S. fuscescens suggests that they would exert little pressure as a top-down control agent of toxic L. majuscula blooms within Moreton Bay. (c) 2006 The Fisheries Society of the British Isles.

Phototrophic microendoliths bloom during coral "white syndrome"

Following rapid lesion progression of white syndrome in tabular Acropora spp., the white bare skeleton gradually changes to green, a result of endolithic algae blooms (primarily Ostreobium spp.). Endolithic algal biomass and chlorophyll concentration were found to be an order of magnitude higher in the green zone compared with healthy appearing parts of each colony. Chl b to Chl a ratio increased from 1:1.6 in the healthy area to 1:2 and 1:3.5 in the white exposed skeleton and green zones, respectively. These observations together with pulse amplitude modulated (PAM) fluorometry suggest photoacclimation of the endoliths in the green zone. Histopathological microscopy revealed that the endolithic algal filaments penetrate the coral tissue. This study highlights the interaction of endolithic algae with both the skeleton and host tissue. This may have a critical role in the processes that accompany the post-disease state in reef-building corals.

Dietary selectivity for the toxic cyanobacterium Lyngbya majuscula and resultant growth rates in two species of opisthobranch mollusc

Trophodynamics of blooms of the toxic marine cyanobacterium Lyngkya majuscula were investigated to determine dietary specificity in two putative grazers: the opisthobranch molluscs, Stylocheilus striatus and Bursatella leachii. S. striatus is associated with L. majuscula blooms and is known to sequester L. majuscula metabolites. The dietary specificity and toxicodynamics of B. leachii in relation to L. majuscula is less well documented. In this study we found diet history had no significant effect upon dietary selectivity of S. striatus when offered a range of plant species. However, L. majuscula chemotype may alter S. striatus' selectivity for this cyanobacterium. Daily biomass increases between small and large size groups of both species were recorded in no-choice consumption trials using L. majuscula. Both S. striatus and B. leachii preferentially consumed L. majuscula containing lyngbyatoxin-a. Increase in mass over a 10-day period in B. leachii (915%) was significantly greater than S. striatus (150%), yet S. striatus consumed greater quantities of L. majuscula (g day(-1)) and thus had a lower conversion efficiency (0.038) than B. leachii (0.081) based on sea hare weight per gram of L. majuscula consumed day(-1). Our findings suggest that growth rates and conversion efficiencies may be influenced by sea hare maximum growth potential, acquisition of secondary metabolites or diet type. (C) 2005 Elsevier B.V. All rights reserved.

Effects of iron additions on filament growth and productivity of the cyanobacterium Lyngbya majuscula

The bioavailability of iron, in combination with essential macronutrients such as phosphorus, has been hypothesised to be linked to nuisance blooms of the toxic cyanobacterium Lyngbya majuscula. The present laboratory study used two biological assay techniques to test whether various concentrations of added iron (inorganic and organically chelated) enhanced L. majuscula filament growth and productivity (C-14-bicarbonate uptake rate). Organically chelated iron (FeEDTA) with adequate background concentrations of phosphorus and molybdenum caused the largest increases (up to 4.5 times the control) in L. majuscula productivity and filament growth. The addition of inorganic iron (without added phosphorus or molybdenum) also stimulated L. majuscula filament growth. However, overall the FeEDTA was substantially and significantly more effective in promoting L. majuscula growth than inorganic iron (FeCl3). The organic chelator (EDTA) alone and molybdenum alone also enhanced L. majuscula growth but to a lesser extent than the chelated iron. The results of the present laboratory study support the hypothesis that iron and chelating organic compounds may be important in promoting blooms of L. majuscula in coastal waters of Queensland, Australia.

Chemical deterrence of a marine cyanobacterium against sympatric and non-sympatric consumers

This study investigates the influence of mesograzer prior exposure to toxic metabolites on palatability of the marine cyanobacterium, Lyngbya majuscula. We examined the palatability of L. majuscula crude extract obtained from a bloom in Moreton Bay, South East Queensland, Australia, containing lyngbyatoxin-a (LTA) and debromoaplysiatoxin (DAT), to two groups: (1) mesograzers of L. majuscula from Guam where LTA and DAT production is rare; and (2) macro- and mesograzers found feeding on L. majuscula blooms in Moreton Bay where LTA and DAT are often prevalent secondary metabolites. Pair-wise feeding assays using artificial diets consisting of Ulva clathrata suspended in agar (control) or coated with Moreton Bay L. majuscula crude extracts (treatment) were used to determine palatability to a variety of consumers. In Guam, the amphipods, Parhyale hawaiensis and Cymadusa imbroglio; the majid crab Menaethius monoceros; and the urchin Echinometra mathaei were significantly deterred by the Moreton Bay crude extract. The sea hares, Stylocheilus striatus, from Guam were stimulated to feed by treatment food whereas S. striatus collected from Moreton Bay showed no discrimination between food types. In Moreton Bay, the cephalaspidean Diniatys dentifer and wild caught rabbitfish Siganus fuscescens were significantly deterred by the crude extract. However, captive-bred S. fuscescens with no known experience with L. majuscula did not clearly discriminate between food choices. Lyngbya majuscula crude extract deters feeding by most mesograzers regardless of prior contact or association with blooms.

Cognitive-didactic Model for Making Testing Items in Physics

Report published in the Proceedings of the National Conference on "Education and Research in the Information Society", Plovdiv, May, 2015

A molecular basis for polyketide biosynthesis in marine dinoflagellates with emphasis on Karenia brevis

Polyketides derived from dinoflagellates are among the most complex and unique structures identified to date. The carbon framework of all polyketides is assembled by a polyketide synthase (PKS). No studies of the biosynthesis of dinoflagellate derived polyketides at the genomic level have been reported to date. Nine strains representing seven different species of dinoflagellates were screened for the presence of type I and type II polyketide synthases (PKS) by PCR and RT-PCR. Seven of the nine strains yielded products that were homologous with known and putative type I polyketide synthases. In each case, the presence of a PKS gene was correlated with the presence of bacteria in the cultures as identified by amplification of the bacterial 16S rRNA gene. However, residual phylogenetic signals, resistance to methylation sensitive restriction enzymes and the lack of hybridization to bacterial isolates support a dinoflagellate origin for most of these genes. ^ A more detailed analysis of Karenia brevis, a toxic marine dinoflagellate endemic to the Gulf of Mexico, also supports the hypothesis that dinoflagellates have polyketide synthase genes. Blooms of this harmful alga cause fish kills, marine mammal mortalities and neurotoxic shellfish poisonings. These harmful effects are attributed to a suite of polyketide secondary metabolites known as the brevetoxins. PKS encoding genes amplified from K. brevis culture were found to be similar to PKS genes from the closely related protist, Cryptosporidium parvum. This suggested that these genes originate from the dinoflagellate. However, K. brevis has not been grown axenically. The associated bacteria might be the source of the toxins or the PKS genes. This dissertation reports the localization of these PKS encoding genes by a combination of flow cytometry/PCR and fluorescence in situ hybridization (FISH). Two genes localized exclusively to K. brevis cells while a third localized to both K. brevis and associated bacteria. While these genes have not yet been linked to toxin production, the work describes the first definitive evidence of resident PKS genes in any dinoflagellate. ^

Paleoenvironmental implications of the Indidura Formation (Cenomanian/Turonian), northeastern Mexico: A high resolution stratigraphic study

High-resolution lithostratigraphic data from rock sequences known as the Indidura Formation near Parras de La Fuente, Coahuila, NE Mexico, led to achieve a significant improvement of our knowledge of that Formation. The results of this study indicate for the first time that the sequence at Parras de La Fuente developed from the deposition of calcareous cyanobacterial microspheroids that accumulated under perennial blooms during the Late Cenomanian through the Middle Turonian. Multi-proxy analyses included sedimentological, petrographical, scanning electron microscopy, stable isotope, trace element geochemistry, and paleontological data. The combined results allowed the correlation of δ13C and anomalies in Mo, V, and Cr with the abundance and predominance of calcareous cyanobacterial microspheroids, which were the main suppliers of the carbonate components and the organic matter throughout deposition of the Indidura Formation in the Parras de la Fuente area, under dysoxic/anoxic conditions. Conspicuous interbeds of dark and light-gray laminated marly calcilutites, and dark-gray marlstones that characterize the stratigraphic sequence formed in response to external forcing climatic factors of millennial-scale Milankovitch cycles (ca. 20 ka precession). At the microscopic level, the prominent dark and light-gray laminae were formed during cycles similar to the 10 to 15 years solar irradiance maximum, and represent alternating periods of high and low calcareous cyanobacterial microspheroids productivity.

Mechanistic studies on the advanced oxidation and photochemical transformation of cyanotoxins (microcystins)

The increased occurrence of cyanobacteria (blue-green algae) blooms and the production of associated cyanotoxins have presented a threat to drinking water sources. Among the most common types of cyanotoxins found in potable water are microcystins (MCs), a family of cyclic heptapeptides containing substrates. MCs are strongly hepatotoxic and known to initiate tumor promoting activity. The presence of sub-lethal doses of MCs in drinking water is implicated as one of the key risk factors for an unusually high occurrence of primary liver cancer. ^ A variety of traditional water treatment methods have been attempted for the removal of cyanotoxins, but with limited success. Advanced Oxidation Technologies (AOTs) are attractive alternatives to traditional water treatments. We have demonstrated ultrasonic irradiation and UV/H2O2 lead to the degradation of cyanotoxins in drinking water. These studies demonstrate AOTs can effectively degrade MCs and their associated toxicity is dramatically reduced. We have conducted detailed studies of different degradation pathways of MCs and conclude that the hydroxyl radical is responsible for a significant fraction of the observed degradation. Results indicate preliminary products of the sonolysis of MCs are due to the hydroxyl radical attack on the benzene ring and substitution and cleavage of the diene of the Adda peptide residue. AOTs are attractive methods for treatment of cyanotoxins in potable water supplies. ^ The photochemical transformation of MCs is important in the environmental degradation of MCs. Previous studies implicated singlet oxygen as a primary oxidant in the photochemical transformation of MCs. Our results indicate that singlet oxygen predominantly leads to degradation of the phycocyanin, pigments of blue green algae, hence reducing the degradation of MCs. The predominant process involves isomerization of the diene (6E to 6Z) in the Adda side chain via photosensitized isomerization involving the photoexcited phycocyanin. Our results indicate that photosensitized processes play a key role in the environmental fate and elimination of MCs in the natural waters. ^

Phytoplankton bloom status: Chlorophyll a biomass as an indicator of water quality condition in the southern estuaries of Florida, USA

Altered freshwater inflows have affected circulation, salinity, and water quality patterns of Florida Bay, in turn altering the structure and function of this estuary. Changes in water quality and salinity and associated loss of dense turtle grass and other submerged aquatic vegetation (SAV) in Florida Bay have created a condition in the bay where sediments and nutrients have been regularly disturbed, frequently causing large and dense phytoplankton blooms. These algal and cyanobacterial blooms in turn often cause further loss of more recently established SAV, exacerbating the conditions causing the blooms. Chlorophyll a (CHLA) was selected as an indicator of water quality because it is an indicator of phytoplankton biomass, with concentrations reflecting the integrated effect of many of the water quality factors that may be altered by restoration activities. Overall, we assessed the CHLA indicator as being (1) relevant and reflecting the state of the Florida Bay ecosystem, (2) sensitive to ecosystem drivers (stressors, especially nutrient loading), (3) feasible to monitor, and (4) scientifically defensible. Distinct zones within the bay were defined according to statistical and consensual information. Threshold levels of CHLA for each zone were defined using historical data and scientific consensus. A presentation template of condition of the bay using these thresholds is shown as an example of an outreach product.

Florida Bay: Water quality status and trends, historic and emerging algal bloom problems

Florida Bay is a unique subtropical estuary that while historically oligotrophic, has been subjected to both natural and anthropogenic stressors, including hurricanes, coastal eutrophication and other impacts. These stressors have resulted in degradation of water quality in the past several decades, most evidenced by reoccurring blooms of the picocyanobacterium Synechococcus spp. Major nutrient inputs consist of freshwater flows to the eastern region from runoff and regulated canal releases, inputs from the Everglades to the central region via Taylor Slough, exchanges with the Gulf of Mexico, which include intermittent Shark River inputs to the western region, stormwater and wastewater from the Florida Keys, and atmospheric deposition. These nutrient inputs have resulted in a transition from strong phosphorus (P) limitation of phytoplankton in the eastern bay to nitrogen (N) limitation in the western bay. Large blooms of Synechococcus were most pronounced in the central bay region, in the area of transition between P and N limitation, in the mid-1990s. Although non-toxic, these blooms, which have continued intermittently through the early 2000s, resulted in significant sea-grass and benthic organism mortalities. A new suite of stressors in 2005, including the passages of Hurricanes Katrina, Rita, and Wilma, additional canal releases, and the initiation of road construction to widen the main roadway leading to the Keys, were correlated with a large Synechococcus bloom in the previously clear, strongly P- limited, northeastern region of the bay. Sustained for 3 years, this bloom was accompanied by a shift from P limitation to N limitation during its course. Nutrient bioassay experiments suggest that this bloom persisted due to the ability of Synechococcus to access organic N and P sources, microbial and geochemical cycling of organic and inorganic nutrients in the water column and between the water column and sediments (both suspended particles and benthos), and decreased grazing by benthic fauna due to their die-off.

Application of excitation emission matrix fluorescence monitoring in the assessment of spatial and seasonal drivers of dissolved organic matter composition: Sources and physical disturbance controls

The environmental dynamics of dissolved organic matter (DOM) were characterized for a shallow, subtropical, seagrass-dominated estuarine bay, namely Florida Bay, USA. Large spatial and seasonal variations in DOM quantity and quality were assessed using dissolved organic C (DOC) measurements and spectrophotometric properties including excitation emission matrix (EEM) fluorescence with parallel factor analysis (PARAFAC). Surface water samples were collected monthly for 2 years across the bay. DOM characteristics were statistically different across the bay, and the bay was spatially characterized into four basins based on chemical characteristics of DOM as determined by EEM-PARAFAC. Differences between zones were explained based on hydrology, geomorphology, and primary productivity of the local seagrass community. In addition, potential disturbance effects from a very active hurricane season were identified. Although the overall seasonal patterns of DOM variations were not significantly affected on a bay-wide scale by this disturbance, enhanced freshwater delivery and associated P and DOM inputs (both quantity and quality) were suggested as potential drivers for the appearance of algal blooms in high impact areas. The application of EEM-PARAFAC proved to be ideally suited for studies requiring high sample throughput methods to assess spatial and temporal ecological drivers and to determine disturbance-induced impacts in aquatic ecosystems.