Assessing the status of Ichkeul Lake and its catchment through identifying potentially toxic cyanobacteria, simultaneously extracted metals and use of acid-volatile-sulfide model.

Lake Ichkeul is considered one of the most significant wetlands in the Mediterranean basin. It serves as a crucial wintering area for numerous western Palearctic birds. A notable decline in species diversity has been observed in the past decade, attributed to excessive water usage for irrigation and the effects of climate change. This study aimed to assess the status of Ichkeul Lake and its catchment through identifying potentially toxic cyanobacteria, and sediment quality. Our first striking finding was that Lyngbya majuscula the dominant potentially toxic cyanobacterium in the lake originated from the Tinja channel. Trace element concentrations in lake sediments exceeded SQG standards which is indicative of rare detrimental effects to biological life. However, the sediment in front of the Tinja channel exhibited high contamination levels of Zn and Cd. These findings call for an urgent need to ensure the ongoing management and conservation of this world heritage site.

Harmful epiphytic dinoflagellate assemblages on macrophytes in the Gulf of Tunis.

The spatio-temporal distribution of epiphytic and planktonic microalgae coupled with environmental factors was investigated for a one-year period in the Gulf of Tunis (northeastern Tunisia). Harmful microalgae assemblages were dominated by three toxic epiphytic dinoflagellates: Ostreopsis sp., Prorocentrum lima and Coolia monotis. They were observed, both on macrophytes (1.03 × 105 cells g-1 FW ; 1.3 × 104 cells g-1 FW and 865 cells g-1 FW, respectively) and in the water column (2.35 × 104 cells L-1; 3.72 × 103 cells L-1; 1.04 × 103 cells L-1, respectively). Species abundances decreased with depth and maximum concentrations were found in shallow waters (0.5-1 m). The highest species abundance was registered both on macroalgae and seagrass with no special preference observed for either of these substrates. Redundancy analyses (RDA) show significant changes in these species abundances according to sites and seasons. The proliferation of Ostreopsis sp. is widespread in summer, when water temperature is warm, and especially in bay zones. The occurrence of P. lima and C. monotis blooms was mainly correlated to nutrients. In this study, macrophyte beds in the Gulf of Tunis were a reservoir of potentially toxic species that could pose a real threat, both to ecosystems and to public health.

Alexandrium pacificum Litaker sp. nov (Group IV): Resting cyst distribution and toxin profile of vegetative cells in Bizerte Lagoon (Tunisia, Southern Mediterranean Sea).

A high spatial resolution sampling of Alexandrium pacificum cysts, along with sediment characteristics (% H2O, % organic matter (OM), granulometry), vegetative cell abundance and environmental factors were investigated at 123 study stations in Bizerte Lagoon (Tunisia). Morphological examination and ribotyping of cells obtained from a culture called ABZ1 obtained from a cyst isolated in lagoon sediment confirmed that the species was A. pacificum. The toxin profile from the ABZ1 culture harvested during exponential growth phase was simple and composed of the N-sulfocarbamoyl toxins C1 (9.82pgtoxincell-1), the GTX6 (3.26pgtoxincell-1) and the carbamoyl toxin Neo-STX (0.38pgtoxincell-1). The latter represented only 2.8% of the total toxins in this strain. High abundance of A. pacificum cysts correlated with enhanced percentages of water and organic matter in the sediment. In addition, sediment fractions of less than 63µm were examined as a favorable potential seedbed for initiation of future blooms and outbreaks of A. pacificum in the lagoon. A significant difference in the cyst distribution pattern was recorded among the lagoon's different zones, with the higher cyst abundance occurring in the inner waters. Also, no correlation due to the specific hydrodynamics of the lagoon was observed in the spatial distribution of A. pacificum cysts and vegetative cells.

Phylogeny and biogeography of the invasive cyanobacterium Cylindrospermopsis raciborskii.

Cylindrospermopsis raciborskii is a toxic cyanobacterium with an invasive nature. The species is found in all the main continents but its origin and dispersal routes on a worldwide perspective remain yet mostly unknown. In this study, 27 isolates of C. raciborskii gathered worldwide have been used for an in-deep phylogenetic analyses with a concatenated system of three genetic markers (16 rRNA, 16S-23S ITS larger subunit, and RNA polymerase rpoC1) comprehending 3,188 bp. Our results provide support for an origin of C. raciborskii in the American continent. Dispersal routes included afterward a spread into the African continent and then Asia and Australia, being Europe the last continent to be colonized by this species. Our phylogenetic inferences suggest that C. raciborskii seem to have a well-defined dispersal behavior with a well-established population structure around the world.

African origin and europe-mediated global dispersal of the cyanobacterium Microcystis aeruginosa.

Microcystis aeruginosa is a bloom-forming cyanobacteria, which currently has a cosmopolitan distribution. Since M. aeruginosa can produce toxic compounds across all continents that it inhabits, it is of major public health relevance to assess its origin and dispersal. Thus, we conducted a worldwide study using 29 isolates representative of all the main continents, and used a concatenated genetic system for phylogenetic analyses consisting of four genetic markers (spanning ca. 3,485 bp). Our results support an early origin of M. aeruginosa in the African continent, with a subsequent dispersal to establish a second genetic pool in the European continent, from where M. aeruginosa then colonized the remaining continental regions. Our findings indicate that the European population has a cosmopolitan distribution, and is genetically closer to populations from Africa and North America. Our study also highlights the utility of using a concatenated dataset for phylogenetic inferences in cyanobacteria.

Genetic variability of the invasive cyanobacteria Cylindrospermopsis raciborskii from Bir M'cherga reservoir (Tunisia).

Cylindrospermopsis raciborskii is an invasive freshwater cyanobacteria of tropical origin, also found in temperate regions. Due to its known ability to produce potent toxins, such as cylindrospermopsin and the paralytic shellfish poisoning, this species is of major concern from a water quality perspective. This study presents a genetic characterization of four C. raciborskii strains isolated from the Bir M'cherga Tunisian reservoir. The toxicity assessment was investigated via molecular biology tools, which suggested that all the isolated strains were not producing cylindrospermopsin, saxitoxin, or microcystin. This result was further confirmed by HPLC and MALDI-TOF analyses. However, we report for the first time in C. raciborskii the presence of mcyA and mcyE, two segments of the microcystin synthetase mcy cluster. All the strains were identified taxonomically based on the 16S rRNA sequences, and their phylogenetic relationships were assessed using the rpoC1 region. Tunisian strains formed a distinct clade separated from the other African strains.

Molecular and phylogenetic characterization of potentially toxic cyanobacteria in Tunisian freshwaters.

This study presents a genetic characterization of 27 potentially toxic cyanobacterial strains isolated from seven reservoirs located in the north and centre of Tunisia. These strains belonged mainly to Microcystis aeruginosa, Cylindrospermopsis raciborskii and Planktothrix agardhii species. Their toxicological potential was evaluated by molecular biology tools, which showed that none of the isolated strains carried segments of the gene cluster responsible for the production of cylindrospermopsin and saxitoxin. The majority of Microcystis isolates were able to synthesize microcystin, since they presented the six characteristic segments of the microcystin synthetase mcy cluster (mcyA, -B, -C, -D, -E and -G). This was further confirmed by MALDI-TOF analysis that showed the presence of eight microcystin variants, including microcystin-LR. The taxonomic identification of the strains was assessed based on the variability of the 16S rRNA gene sequences. Furthermore, the 16S-23S rRNA ITS sequences of Microcystis isolates and rpoC1 sequences of Cylindrospermopsis strains were also used in the phylogenetic analysis.

Genetic diversity and structure of the invasive toxic cyanobacterium Cylindrospermopsis raciborskii.

Strains of the invasive toxic cyanobacteria Cylindrospermopsis raciborskii were genetically evaluated with four genetic markers encompassing in total 2.9 kb (16S rRNA, ITS longer spacer, ITS shorter spacer and rpoC1) to assess the phylogenetic relationships, genetic variation and population differentiation of the species across all five continents. The phylogenetic analysis showed that the C. raciborskii strains grouped into three well-supported distinct clusters: (I) European (II) African/American, and (III) Asian/Australian. The European group presented a high genetic similarity with the Asian and the Australian isolates than with the African and American isolates. Several Portuguese isolates were analyzed (n = 7) and revealed a low genetic differentiation with little geographical structure. The genetic distance among groups and phylogenetic relationships obtained in this study suggest that the recent invasion of C. raciborskii in Portuguese and other European temperate environments could have had its origin in the Asian and/or Australian continents.

Seasonal occurrence and toxicity of Microcystis spp. and Oscillatoria tenuis in the Lebna Dam, Tunisia.

Physicochemical and biological water quality, including the total microcystin concentrations, was investigated for the first time from January to December 2005 in the Lebna Dam, Tunisia. Microcystin levels and characterization of the different microcystin variants present were measured by protein phosphatase (PP2A) inhibition assays and by LC/MS/MS, respectively. Nutrient values were high, with total inorganic nitrogen and phosphorus concentrations ranging from 0.05 to 8.4 mg L(-1) and from 0.03 to 1.37 mg L(-1), respectively. However, the chlorophyll-a concentrations were very low with a peak (5.32 microg L(-1)) on 20 September 2005 at 9 m depth water samples. Microscopic examination of the phytoplankton samples showed the dominance in the autumn of three morphospecies of the genus Microcystis and the species Oscillatoria tenuis. The total (particulate and dissolved) microcystin concentrations at the surface and at 9 m depth water samples ranged between 0.008 and 1.73, and 0.005 and 5.57 microg microcystin (MC)-LR equivalent L(-1), respectively, with a peak on 20 September. The presence of the microcystin synthetase genes (mcyA, -B, and -C) in the lysates of the three morphospecies of the genus Microcystis and the species O. tenuis indicated that these species were responsible for the microcystin production in this system. The analysis of the field cyanobacterial sample extract containing these species by LC/MS/MS revealed the presence of two microcystin variants: microcystin-LR (MC-LR) and microcystin-YR (MC-YR).