CYLINDROSPERMOPSIS RACIBORSKII (CYANOBACTERIA) INVASION AT MID‐LATITUDES: SELECTION,WIDE PHYSIOLOGICAL TOLERANCE,ORGLOBALWARMING?1

The tropical bloom‐forming cyanobacterium Cylindrospermopsis raciborskii (Woloszynska) Seenaya et Subba Raju is causing increasing concern because of its potential toxicity and invasive behavior at mid‐latitudes. This species has recently been identified in several temperate areas and was first reported in France in 1994, but the mechanisms underlying this acclimation process remain to be elucidated. We performed a range of laboratory experiments in an attempt to identify the physiological characteristics that could account for this behavior. We investigated the three following hypotheses: 1) clones of C. raciborskii adapted to temperate climate have been selected as it advanced north, 2) C. raciborskii has high physiological tolerance that allows it to prosper in a wide range of conditions, and 3) changes inclimate (global warming) have favored the development of C. raciborskii in mid‐latitudes. Ten strains of C. raciborskii from Australia n=1), Brazil (2), France (2), Germany (1), Hungary (1), Mexico (1), and Senegal (2) were cultured at different temperatures and light intensities. The in vitro growth parameters (μ and T_opt) were the same for the tropical and temperate strains. All 10 strains displayed positive net growth in a wide range of temperatures (from 20 to 35°C) and light intensities (from 30 to 400 μmol photons·m^?2·s^?1), with maximum growth rates at around 30°C and 80 μmol photons·m^?2·s^?1. This suggests that the colonization of mid‐latitudes by C. raciborskii may result from a combination of its ability to tolerate a rather wide range of climatic conditions and the global warming phenomenon, which provides this species with better environmental conditions for its growth.     

Seasonal dynamics and toxicity of Cylindrospermopsis raciborskii in Lake Guiers (Senegal, West Africa)

Cylindrospermopsis raciborskii is a toxic bloom-forming cyanobacterium that occurs at tropical and temperate latitudes. Despite several reports from Africa, no data were previously available about its dynamics or toxic potential there. We therefore carried out a 1-year survey of the dynamics of C. raciborskii in the main water reservoir in Senegal, Lake Guiers. Cylindrospermopsis raciborskii never formed a bloom in this lake during the period studied, but was dominant during the dry season. The only observed bloom-forming species was a diatom, Fragilaria sp., which displayed a seasonal pattern contrary to that exhibited by C. raciborskii. Principal component analysis applied to environmental and phytoplankton data showed that high C. raciborskii biomasses were mainly related to high temperature and water column stability. Tests for C. raciborskii species-related toxicity and/or toxin synthesis were performed on 21 isolated clones. All the strains isolated tested negative in mouse toxicity bioassays, toxin analysis (MS/MS) and tests for known cylindrospermopsin genes (ps, pks). The limited number of isolates studied, and the occurrence of toxic and nontoxic clones in natural cyanobacterial populations, mean that we cannot conclude that there is no C. raciborskii-associated health risk in this drinking water reservoir.     

Acclimation at high temperatures increases the ability of Raphidiopsis raciborskii (Cyanobacteria) to withstand phosphate deficiency and reveals distinct strain responses

ABSTRACT

Raphidiopsis (Cylindrospermopsis) raciborskii is one of the most studied potentially harmful cyanobacteria. Single environmental factors such as increased temperature or light are reported to be promoters of R. raciborskii growth, but the interaction of two or more promoting factors is less understood. The performance of two strains of R. raciborskii (MVCC19, Uruguayan and LP1, Brazilian) were evaluated under acclimation and temperature shifts (25–32°C) in combination with two transitions from phosphorus (P) sufficiency to limiting growth conditions. When subjected to transition from high P sufficiency to a P-limiting state, strains were able to grow only at the warmer temperature if previously acclimated. The MVCC19 strain showed higher specific growth rates and a shorter growth phase than LP1. Morphological differences were also found: the MVCC19 strain produced shorter filaments, while the LP1 strain increased in length and the number of cells per filament. The results show the positive effect of high temperature on the ability of R. raciborskii to withstand P-limiting conditions, which may confer resilience of populations to periods of severe nutrient limitation in warm lakes. This finding contributes to the understanding of the success of this species in diverse environmental conditions. The effect of temperature on the tolerance of nutrient deficiency and the performance of strains under lake conditions suggests the need for a drastic reduction in nutrient loads to avoid R. raciborskii dominance in warmer lakes.     

Phytoplankton seasonal variation in a shallow stratified eutrophic reservoir (Garças Pond, Brazil)

This study aimed at describing the phytoplankton dynamics and structure in a shallow eutrophic reservoir, the Garças Pond, located in the Parque Estadual das Fontes do Ipiranga (23°38′40.6′′ S, 46°37′28.0′′ W), in the Municipality of São Paulo, southeast Brazil. Samples were collected monthly from January to December 1997 in five depths (subsurface, 1 m, 2 m, 3 m, and 20 cm above the bottom) in the pelagic zone (Z _max = 4.7 m). Abiotic variables studied were: water temperature, turbidity, transparency, conductivity, pH, dissolved oxygen, alkalinity, inorganic carbon, and N and P dissolved and total forms. Altogether 236 phytoplankton taxa distributed among 10 classes were identified. Phytoplankton seasonal and vertical variation was related to shifts in the water chemical features as a consequence of a warm-wet season with stratified water column (phase 1, January–March and September–December) alternating with a cool-dry season with mixed water column (phase 2, April–August). There were shifts in cyanobacterial dominance over the entire year. During phase 1, Raphidiopsis/Cylindrospermopsis was one of the most important taxon. During phase 2, Raphidiopsis/Cylindrospermopsis biomass decreased, whereas richness and diversity increased and diatoms were relatively abundant. In September, when the water column was markedly stratified, a cyanobacterial bloom (Sphaerocavum brasiliense) occurred. Changes in water chemical variables caused by the bloom allowed recognition of a phase 3, in which pH and chlorophyll a, TP and CO₃ ^2? concentration reached their highest values. According to Reynolds and collaborators’ functional groups approach, phase 1 was marked by groups S/W1/W2/H1/Y, phase 2 by groups K/L _M /L _O /D/P/X1/F, and phase 3 by group M. This sequence was corroborated by canonical correspondence analysis (CCA) results.     

Sublethal responses in Melanoides tuberculata following exposure to Cylindrospermopsis raciborskii containing cylindrospermopsin

Sublethal effects in the aquatic snail Melanoides tuberculata were examined during exposure to whole cell extracts of Cylindrospermopsis raciborskii and live C. raciborskii cultures, containing varying concentrations of algal cells, cellular debris, and the blue-green algal toxin, cylindrospermopsin (CYN). Exposure to whole cell extracts or live algal cultures did not result in significant changes in adult snail behaviour or relative growth rates. However, clear changes in the number of hatchlings released from parent snails were observed. Exposure to whole cell extracts containing ≥200 μg L⁻¹ extracellular CYN resulted in an increase in the number of hatchlings. In contrast, decreases in hatchling number were recorded from treatments containing ≥200 μg L⁻¹ CYN during exposures to live C. raciborskii cultures, compared with controls. This suggests that CYN may be more toxic to grazing invertebrates if present in the intracellular form. Since CYN is a protein synthesis inhibitor, it is possible that CYN may be especially toxic to rapidly developing tissues such as snail embryos. This may also explain the lack of effects observed in adult snails.     

Ontogeny of Anabaenopsis raciboskii Wolosz. and Its Taxonomical Relevance

An annual life cycle, including 4 developmental stages, is discovered in Anabaenopsis raciborskii Wolosz of the West Lake, Hangzhou. The progress of these developmental stages is influenced by the enviroment. The third stage with heterocysts expressed distinctly as Anabaenopsis raciborskii Wolosz., and the fourth stage was formerly recognized as Raphidiopsis sinensis and R. curvata. It is considered based on these facts that Anabaenopsis raciborskii Wolosz., R. curvata Frit. et Rich. and R. sinensis Jao are different forms at dif-ferent stages of ontogeny, and ought be treated as a single species.     

Comparative studies on photosynthesis and phosphate metabolism of Cylindrospermopsis raciborskii with Microcystis aeruginosa and Aphanizomenon flos-aquae

The physiological differences for three bloom-forming cyanobacteria (Cylindrospermopsis raciborskii, Microcystis aeruginosa, and Aphanizomenon flos-aquae) were investigated. In comparison with M. aeruginosa and A. flos-aquae, C. raciborskii exhibited a significantly higher concentration of carotenoids, higher values in maximum photosynthesis rate (P_m), apparent photosynthetic efficieny (a), and maximum electron transport rate (ETR_max) during the growth period. In addition, higher extracellular alkaline phosphatase activities and lower light compensation point (I_c) were also detected in C. raciborskii (p < 0.05, ANOVA). Therefore, it is suggested that the higher photosynthetic activities, more effective uptake and utilization to phosphate, and low light requirements might play important roles in the occurrence and invasive behavior of C. raciborskii.     

Genetic characterisation of Cylindrospermopsis raciborskii (Nostocales, Cyanobacteria) isolates from Africa and Europe

The invasive cyanobacterium Cylindrospermopsis raciborskii is increasingly spreading in temperate freshwater habitats worldwide and is of major concern due to its ability to produce potent toxins. It is, therefore, important to understand the mechanisms behind the dispersal of this species. Different hypotheses have been proposed to explain the phylogeography and mechanisms underlying the recent expansion of C. raciborskii into temperate latitudes, but there is still no conclusive evidence whether the obvious ecological success of C. raciborskii is due to selection mechanisms, physiological tolerance, climatic change or radiation after the last ice age. In the present study, new isolates of C. raciborskii from Europe and Africa were genetically characterised by sequencing the ITS1, PC-IGS, nifH and rpoC1 genes and compared to corresponding sequences of C. raciborskii available in GenBank in order to test different phylogeographical hypotheses. The strains were also morphologically examined and screened for production of the hepatotoxic cylindrospermopsin (CYN). We clearly demonstrate a variation among the populations of C. raciborskii from different geographical regions. The phylogenetic analyses revealed a clustering of the strains due to geographic origin. The ITS1 and nifH genes separated into American, European and Australian–African groups, whereas the PC-IGS and rpoC1 separated into American and European/Australian/African groups. An analysis of concatenated data supported the division into American, European and African/Australian groups, and even indicated a subdivision into an African and an Australian group. Our findings do not strongly support any of the existing hypotheses on the phylogeography of C. raciborskii, and most likely a combination of these hypotheses is the best approach to understand the evolution and dispersal of this species.