Morphological changes during akinete germination in Cylindrospermopsis raciborskii (Nostocales, Cyanobacteria)

The occurrence of the cyanobacterium Cylindrospermopsis raciborskii (Woloszynska) Seenayya and Subba Raju is a global water quality issue. The misidentification of C. raciborskii in the past is a major concern for water quality users, considering the reported cases of human and livestock poisonings associated with the cyanobacterium. Many of the available taxonomic descriptions for this species provide little or no detail of the morphology of early developmental phases that may assist with identification. Therefore, typifying the morphological changes throughout the entire life cycle for such a species requires urgent attention. In this study, five distinct morphological phases identified using a new culturing technique are reported for the process of akinete germination in C. raciborskii. Before the terminal emergence of three to four cell germlings through a ruptured akinete envelope (phase 3), mature akinetes (phase 1) elongated and the endospore separated from the akinete envelope (phase 2). After the association with the envelope was lost, four-cell germlings (phase 4a) matured into young trichomes of more than four cells (phase 4b). Throughout the process of germination, internal granular structures decreased in size and were irregular in shape in germlings and young trichomes. The culturing technique, which used a Sedgwick-Rafter cell, was successful in its application but was limiting in that the development of young trichomes after phase 4b could not be monitored.

The toxins of Lyngbya majuscula and their human and ecological health effects

Lyngbya majuscula is a benthic filamentous marine cyanobacterium, which in recent years appears to have been increasing in frequency and size of blooms in Moreton Bay, Queensland. It has a worldwide distribution throughout the tropics and subtropics in water to 30m. It has been found to contain a variety of chemicals that exert a range of biological effects, including skin, eye and respiratory irritation. The toxins lyngbyatoxin A and debromoaplysiatoxin appear to give the most widely witnessed biological effects in relation to humans, and experiments involving these two toxins show the formation of acute dermal lesions. Studies into the epidemiology of the dermatitic, respiratory and eye effects of the toxins of this organism are reviewed and show that Lyngbya induced dermatitis has occurred in a number of locations. The effects of aerosolised Lyngbya in relation to health outcomes were also reported. Differential effects of bathing behaviour after Lyngbya exposure were examined in relation to the severity of health outcomes. The potential for Lyngbya to exhibit differential toxicologies due to the presence of varying proportions of a range of toxins is also examined. This paper reviews the present state of knowledge on the effects of Lyngbya majuscula on human health, ecosystems and human populations during a toxic cyanobacterial bloom. The potential exists for toxins from Lyngbya majuscula affecting ecological health and in particular marine reptiles. (C) 2001 Elsevier Science Ltd. All rights reserved.

Isolation and identification of the cyanotoxin cylindrospermopsin and deoxy-cylindrospermopsin from a Thailand strain of Cylindrospermopsis raciborskii (Cyanobacteria)

A strain of Cylindrospermopsis (Cyanobacteria) isolated from a fishpond in Thailand was examined for its taxonomy based upon morphology and 16S rRNA gene sequence. It was also examined for production of the hepatotoxic cyanotoxin called cylindrospermopsin (CYN) and deoxycylindrospermopsin (deoxy-CYN). The strain (CY-Thai) was identified as C. raciborskii (Woloszynska) Seenaya and Subba Raju based upon morphological examination which was confirmed by 16S rRNA gene sequences and phylogenetic comparisons based upon its 16S rRNA gene. The alkaloid heptatotoxin CYN was confirmed using mouse bioassay, HPLC and HPLC-MS/MS while deoxy-CYN was confirmed using HPLC-MS/MS. The mouse bioassay gave a minimum lethal dose at 250 mg dry weight cells/kg body weight within 24 h and 125 mg/kg at 72 h, with signs of poisoning the same as in literature reports for CYN. HPLC chromatographic comparison of the CY-Thai toxin with standard CYN gave the same retention time and an absorbance maximum at 262 nm. HPLC-MS/MS confirmed the presence of CYN (M + H 416) and deoxy-CYN (M + H 400). The CYN content in strain CY-Thai was estimated at 1.02 mg/g and approximately 1/10 of this amount for deoxy-CYN. This is the first report from Asia of a CYN, deoxy-CYN producing Cylindrospermopsis raciborskii. (C) 2001 Elsevier Science Ltd. All rights reserved.

Human Fatalities from Cyanobacteria: Chemical and Biological Evidence for Cyanotoxins

An outbreak of acute liver failure occurred at a dialysis center in Caruaru, Brazil (8 degrees 17 'S, 35 degrees 58 'W), 134 km from Recife, the state capital of Pernambuco. At the clinic, 116 (89%) of 131 patients experienced visual disturbances, nausea, and vomiting after routine hemodialysis treatment on 13-20 February 1996. Subsequently, 100 patients developed acute liver failure, and of these 76 died. As of December 1996, 52 of the deaths could be attributed to a common syndrome now called Caruaru syndrome. Examination of phytoplankton from the dialysis clinic's water source, analyses of the clinic's water treatment system, plus serum and liver tissue of clinic patients led to the identification of two groups of cyanobacterial toxins, the hepatotoxic cyclic peptide microcystins and the hepatotoxic alkaloid cylindrospermopsin. Comparison of victims' symptoms and pathology using animal studies of these two cyanotoxins leads us to conclude that the major contributing factor to death of the dialyses patients was intravenous exposure to microcystins, specifically microcystin-YR, -LR, and -AR. From liver concentrations and exposure volumes, it was estimated that 19.5 mug/L microcystin was in the water used for dialysis treatments. This is 19.5 times the level set as a guideline for safe drinking water supplies by the World. Health Organization.

Factors affecting akinete differentiation in Cylindrospermopsis raciborskii (Nostocals, Cyanobacteria)

1. Cylindrospermopsis raciborskii is a potentially toxic freshwater cyanobacterium which can produce akinetes (reproductive spores) that on germinating can contribute to future populations. To further understand factors controlling the formation of these specialised cells, the effects of diurnal temperature fluctuations (magnitude and frequency), in combination with different light intensities and phosphorus concentrations were investigated under laboratory conditions. 2. Akinete differentiation was affected by the frequency of temperature fluctuations. Maximum akinete concentrations were observed in cultures that experienced multiple diurnal temperature fluctuations. 3. Akinete concentrations increased with increasing magnitude of temperature fluctuation. A maximum akinete concentration was achieved under multiple diurnal temperature fluctuations with a magnitude of 10degreesC (25degreesC to 15degreesC). 4. A fourfold increase in light intensity (25-100 mumol m(-2) s(-1)) resulted in an approximate 14-fold increase in akinete concentration. 5. High filterable reactive phosphorus (FRP) concentrations (> 70 mug L-1) in the medium, combined with a multiple diurnal temperature fluctuation of 10degreesC, supported the development of the highest akinete concentration.

Ecophysiology of the marine cyanobacterium, Lyngbya majuscula (Oscillatoriaceae) in Moreton Bay, Australia

Large blooms of the marine cyanobacterium Lyngbya majuscula in Moreton Bay, Australia (27 degrees 05'S, 153 degrees 08'E) have been re-occurring for several years. A bloom was studied in Deception Bay (Northern Moreton Bay) in detail over the period January-March 2000. In situ data loggers and field sampling characterised various environmental parameters before and during the L. majuscula bloom. Various ecophysiological experiments were conducted on L. majuscula collected in the field and transported to the laboratory, including short-term (2h) C-14 incorporation rates and long-term (7 days) pulse amplitude modulated (PAM) fluorometry assessments of photosynthetic capacity. The effects of L. majuscula on various seagrasses in the bloom region were also assessed with repeated biomass sampling. The bloom commenced in January 2000 following usual December rainfall events, water temperatures in excess of 24 degrees C and high light conditions. This bloom expanded rapidly from 0 to a maximum extent of 8 km(2) over 55 days with an average biomass of 210 g(dw)(-1) m(-2) in late February, followed by a rapid decline in early April. Seagrass biomass, especially Syringodium isoetifolium, was found to decline in areas of dense L. majuscula accumulation. Dissolved and total nutrient concentrations did not differ significantly (P > 0.05) preceding or during the bloom. However, water samples from creeks discharging into the study region indicated elevated concentrations of total iron (2.7-80.6 mu M) and dissolved organic carbon (2.5-24.7 mg L-1), associated with low pH values (3.8-6.7). C-14 incorporation rates by L. majuscula were significantly (P < 0.05) elevated by additions of iron (5 mu M Fe), an organic chelator, ethylenediaminetetra-acetic acid (5 mu M EDTA) and phosphorus (5 mu M PO4-3). Photosynthetic capacity measured with PAM fluorometry was also stimulated by various nutrient additions, but not significantly (P > 0.05). These results suggest that the L. majuscula bloom may have been stimulated by bioavailable iron, perhaps complexed by dissolved organic carbon. The rapid bloom expansion observed may then have been sustained by additional inputs of nutrients (N and P) and iron through sediment efflux, stimulated by redox changes due to decomposing L. majuscula mats. (c) 2004 Elsevier B.V. All rights reserved.

Blooms of the cyanobacterium Lyngbya majuscula in coastal Queensland, Australia: disparate sites, common factors

During the last decade there has been a significant rise in observations of blooms of the toxic cyanobacterium, Lyngbya majuscula along the east coast of Queensland, Australia. Whether the increase in cyanobacterial abundance is a biological indicator of widespread water quality degradation or also a function of other environmental change is unknown. A bioassay approach was used to assesses the potential for runoff from various land uses to stimulate productivity of L. majuscula. In Moreton Bay, L. majuscula productivity was significantly (p < 0.05) stimulated by soil extracts, which were high in phosphorus, iron and organic carbon. Productivity of L. majuscula from the Great Barrier Reef was also significantly (p < 0.05) elevated by iron and phosphorus rich extracts, in this case seabird guano adjacent to the bloom site. Hence, it is possible that other L. majuscula blooms are a result of similar stimulating factors (iron, phosphorus and organic carbon), delivered through different mechanisms. (c) 2004 Elsevier Ltd. All rights reserved.

Phosphate uptake and growth kinetics of Trichodesmium (Cyanobacteria) isolates from the North Atlantic Ocean and the Great Barrier Reef, Australia

We compared inorganic phosphate (P-i) uptake and growth kinetics of two cultures of the diazotrophic cyanobacterium Trichodesmium isolated from the North Atlantic Ocean (IMS101) and from the Great Barrier Reef, Australia (GBRTRLI101). Phosphate-limited cultures had up to six times higher maximum P-i uptake rates than P-replete cultures in both strains. For strain GBRTRLI101, cell-specific P-i uptake rates were nearly twice as high, due to larger cell size, but P-specific maximum uptake rates were similar for both isolates. Half saturation constants were 0.4 and 0.6 muM for P-i uptake and 0.1 and 0.2 muM for growth in IMS101 and GBRTRLI101, respectively. Phosphate uptake in both strains was correlated to growth rates rather than to light or temperature. The cellular phosphorus quota for both strains increased with increasing P-i up to 1.0 muM. The C:P ratios were 340-390 and N:P ratios were 40-45 for both strains under severely P-limited growth conditions, similar to reported values for natural populations from the tropical Atlantic and Pacific Oceans. The C:P and N:P ratios were near Redfield values in medium with >1.0 muM P-i. The North Atlantic strain IMS101 is better adapted to growing on P-i at low concentrations than is GBRTRLI101 from the more P-i-enriched Great Barrier Reef. However, neither strain can achieve appreciable growth at the very low (nanomolar) P-i concentrations found in most oligotrophic regimes. Phosphate could be an important source of phosphorus for Trichodesmium on the Great Barrier Reef, but populations growing in the oligotrophic open ocean must rely primarily on dissolved organic phosphorus sources.

Epidemiology of recreational exposure to freshwater cyanobacteria - an international prospective cohort study

Background: Case studies and anecdotal reports have documented a range of acute illnesses associated with exposure to cyanobacteria and their toxins in recreational waters. The epidemiological data to date are limited; we sought to improve on the design of some previously conducted studies in order to facilitate revision and refinement of guidelines for exposure to cyanobacteria in recreational waters. Methods: A prospective cohort study was conducted to investigate the incidence of acute symptoms in individuals exposed, through recreational activities, to low ( cell surface area < 2.4 mm(2)/mL), medium ( 2.4 - 12.0 mm(2)/mL) and high (> 12.0 mm(2)/mL) levels of cyanobacteria in lakes and rivers in southeast Queensland, the central coast area of New South Wales, and northeast and central Florida. Multivariable logistic regression analyses were employed; models adjusted for region, age, smoking, prior history of asthma, hay fever or skin disease ( eczema or dermatitis) and clustering by household. Results: Of individuals approached, 3,595 met the eligibility criteria, 3,193 (89%) agreed to participate and 1,331 (37%) completed both the questionnaire and follow-up interview. Respiratory symptoms were 2.1 (95% CI: 1.1 - 4.0) times more likely to be reported by subjects exposed to high levels of cyanobacteria than by those exposed to low levels. Similarly, when grouping all reported symptoms, individuals exposed to high levels of cyanobacteria were 1.7 ( 95% CI: 1.0 - 2.8) times more likely to report symptoms than their low-level cyanobacteria-exposed counterparts. Conclusion: A significant increase in reporting of minor self-limiting symptoms, particularly respiratory symptoms, was associated with exposure to higher levels of cyanobacteria of mixed genera. We suggest that exposure to cyanobacteria based on total cell surface area above 12 mm(2)/mL could result in increased incidence of symptoms. The potential for severe, life-threatening cyanobacteria-related illness is likely to be greater in recreational waters that have significant levels of cyanobacterial toxins, so future epidemiological investigations should be directed towards recreational exposure to cyanotoxins.