Cytotoxic and Genotoxic Effects of Cyanobacterial and Algal Extracts—Microcystin and Retinoic Acid Content

In the last decade, it has become evident that complex mixtures of cyanobacterial bioactive substances, simultaneously present in blooms, often exert adverse effects that are different from those of pure cyanotoxins, and awareness has been raised on the importance of studying complex mixtures and chemical interactions. We aimed to investigate cytotoxic and genotoxic effects of complex extracts from laboratory cultures of cyanobacterial species from different orders (Cylindrospermopsis raciborskii, Aphanizomenon gracile, Microcystis aeruginosa, M. viridis, M. ichtyoblabe, Planktothrix agardhii, Limnothrix redekei) and algae (Desmodesmus quadricauda), and examine possible relationships between the observed effects and toxin and retinoic acid (RA) content in the extracts. The cytotoxic and genotoxic effects of the extracts were studied in the human hepatocellular carcinoma HepG2 cell line, using the MTT assay, and the comet and cytokinesis-block micronucleus (cytome) assays, respectively. Liquid chromatography electrospray ionization mass spectrometry (LC/ESI-MS) was used to detect toxins (microcystins (MC-LR, MC-RR, MC-YR) and cylindrospermopsin) and RAs (ATRA and 9cis-RA) in the extracts. Six out of eight extracts were cytotoxic (0.04–2 mgDM/mL), and five induced DNA strand breaks at non-cytotoxic concentrations (0.2–2 mgDM/mL). The extracts with genotoxic activity also had the highest content of RAs and there was a linear association between RA content and genotoxicity, indicating their possible involvement; however further research is needed to identify and confirm the compounds involved and to elucidate possible genotoxic effects of RAs.

Functional Groups of Phytoplankton and Their Relationship with Environmental Factors in the Restored Uzarzewskie Lake

Uzarzewskie Lake is a small, postglacial lake, located in western Poland. The lake is under restoration treatment since 2006. At first, iron treatment was done for 2 years. In the second stage, spring water was directed into the hypolimnion in order to improve water oxygenation near the bottom sediments. The purpose of our research was to determine changes in the contribution of functional groups to the total number of taxa and total biomass of phytoplankton due to changes in the physical and chemical characteristics of the restored lake. Phytoplankton composition was analyzed in three periods: (1) before restoration; (2) during the first method of restoration; and (3) when the second method was implemented in the lake. Epilimnetic phytoplankton was sampled every year monthly from March to November. The relationship between phytoplankton groups and environmental factors (water temperature, ammonium nitrogen, nitrate nitrogen, dissolved phosphorus, conductivity and pH) was examined, using the canonical analyses. The redundancy analysis indicated that the temperature, dissolved phosphates concentration, ammonium nitrogen and pH were the main determining factors of the phytoplankton community dynamics. During the study, 13 coda dominated the phytoplankton biomass. Cyanobacteria of the codon H1 with such species as Aphanizomenon gracile, Dolichospermum planctonicum, D. viguieri dominated the phytoplankton community before restoration. S1 group consisting of Planktolyngbya limnetica, Limnothrix redekei and Planktothrix agardhii mostly dominated during the period in which the first method was used. Improvement of water quality due to restoration efforts in the third period caused dominance of other groups, especially J (Actinastrum hantzschii and other Chlorococcales), C (Asterionella formosa and other diatoms), Y (Cryptomonas marssonii and other cryptophytes), Lo (Peridiniopsis cunningtonii and other dinophytes) and X2 (Rhodomonas lacustris).

ОЦЕНКА ЭКОЛОГИЧЕСКОГО СОСТОЯНИЯ ОЗЕРА КАРПОВСКОГО В 2019 Г.

Озеро Карповское относится к категории малых водоемов Калининграда, активно используемых в рекреационных целях. Малые водоемы наиболее уязвимы к загрязнениям и в течение короткого периода времени могут утрачивать способность к самоочищению. Цель данной работы – оценка экологического состояния оз. Карповского по гидрохимическим, гидрологическим, микробиологическим и гидробиологическим параметрам. Гидрохимические показатели озера, за исключением содержания железа, соответствовали значениям рыбохозяйственных нормативов. Превышение нормативов по содержанию общего железа в водоеме характерно для поверхностных вод Калининградской области. Количество сапрофитных бактерий в воде увеличивалось к летне-осеннему сезону, что связано с повышением температуры воды летом в среднем до 22–26 °С и усилением деструктивных процессов в осенний сезон. В бактериопланктоне оз. Карповского преобладали аэробные бактерии, развитие которых определялось наличием в воде легкодоступного органического вещества, достаточного количества кислорода, благоприятного pH среды. Патогенных бактерий, опасных для здоровья человека, в микрофлоре воды не обнаруживали. Альгофлора озера по таксономическим показателям характеризовалась как «зелено-эвгленово-диатомово-синезеленая», что характерно для многих урбанизированных водоемов. В количественном отношении преобладали водоросли из отдела Cyanoprokaryota при доминировании Limnothrix redekei и Planktothrix agardhii. Вегетация водорослей в течение всего периода способствовала поддержанию стабильно высокого содержания кислорода и, возможно, оказывала влияние на снижение содержания биогенных элементов в воде. Результаты проведенного комплексного анализа показали, что оз. Карпов-ское относится к водоемам β-мезосапробной зоны – это соответствует эвтрофному уровню.

Protected Freshwater Ecosystem with Incessant Cyanobacterial Blooming Awaiting a Resolution

For 50 years persistent cyanobacterial blooms have been observed in Lake Ludoš (Serbia), a wetland area of international significance listed as a Ramsar site. Cyanobacteria and cyanotoxins can affect many organisms, including valuable flora and fauna, such as rare and endangered bird species living or visiting the lake. The aim was to carry out monitoring, estimate the current status of the lake, and discuss potential resolutions. Results obtained showed: (a) the poor chemical state of the lake; (b) the presence of potentially toxic (genera Dolichospermum, Microcystis, Planktothrix, Chroococcus, Oscillatoria, Woronichinia and dominant species Limnothrix redekei and Pseudanabaena limnetica) and invasive cyanobacterial species Raphidiopsis raciborskii; (c) the detection of microcystin (MC) and saxitoxin (STX) coding genes in biomass samples; (d) the detection of several microcystin variants (MC-LR, MC-dmLR, MC-RR, MC-dmRR, MC-LF) in water samples; (e) histopathological alterations in fish liver, kidney and gills. The potential health risk to all organisms in the ecosystem and the ecosystem itself is thus still real and present. Although there is still no resolution in sight, urgent remediation measures are needed to alleviate the incessant cyanobacterial problem in Lake Ludoš to break this ecosystem out of the perpetual state of limbo in which it has been trapped for quite some time.

Seasonal changes in phytoplankton and bioindices in the southern part of Lake Jeziorak (NE Poland)

The structure of phytoplankton communities of Lake Jeziorak was presented for the first time. The objective of our research was qualitative and quantitative analysis and bioindices of phytoplankton during and outside the tourist season. Analysis of phytoplankton assemblages were performed in 2011-2012. A total of 96 taxa were identified in Lake Jeziorak, mainly: Cyanobacteria - 20, Bacillariophyceae - 49, and Chlorophyta − 19. Biomass of the phytoplankton varied from 10 mg l−1 in October to 29 mg l−1 in May. In the whole research period, Cyanobacteria dominated and represented up to 68% of the total biomass. The cyanobacterial blooms were constantly observed. Biomass in the summer period was composed of filamentous Aphanizomenon gracile, Limnothrix redekei, Planktothrix agardhii and Pseudanabaena limnetica. Outside the holiday season, i.e. in autumn and spring, filamentous Cyanobacteria accompanied cryptomonads in phytoplankton. The species composition, the biomass of phytoplankton, and TSI indicate the hypertrophic conditions of the lake. Phytoplankton in Lake Jeziorak was in a state of equilibrium for almost the entire study period. S1 was a dominant group and its abundance and biomass did not change by more than 10%. There was no significant direct effect of the seasonal tourism impact on the development of phytoplankton in Lake Jeziorak.

Ecological characteristics of plankton and aquatic vegetation in Lake Qiluhu

Investigations of the phytoplankton, zooplankton, zoobenthos and aquatic vegetation in Lake Qiluhu were carried out in February, 2009. Over the whole lake, 13 sampling sites were set up for the analysis of phytoplankton and zooplankton, and 22 profiles for the collection of macrophytes and zoobenthos. In the survey, 7 phyla, 65 algae species were identified. The average abundance of phytoplankton was 7.16 × 108 cells/L, and the dominant specie was Limnothrix redekei. No obvious surface accumulation of algae was detected. The concentration of Chlorophyll a ranged from 85 to 101 μg/L, and the average value was 93 μg/L. Nineteen species of zooplankton were observed, including 4 species of rotifers, 6 species of cladocerans and 9 species of copepods. Copepods were the dominant species, their abundance reaching 68%, whilst Cladocerans took second place with an abundance proportion of 28%. Six species of submerged vegetation were identified: Potamogeton Pectinatus, Myriophyllum, Elodea Canadensis, Ceratophyllum demersum and Potamogeton crispus. Amongst them, the dominant vegetation was P. Pectinatus, the biomass of which was up to 63% of the total biomass. Emerged macrophytes were cluster distributed across the whole lake, mainly consisting of Scirpus tabernaemontani, phragmites communis and cane shoots. Unfortunately, no living zoobenthos were found at the sites. The results indicated that, in Lake Qiluhu, the abundance of phytoplankton was maintained at a high level. The ecological function of submerged vegetation was gradually being lost because of its low standing crop and coverage, and the benthic animal habitat was severely damaged.

Winter phytoplankton communities in different depths of three mesotrophic lakes (Łęczna-Włodawa Lakeland, Eastern Poland)

Phytoplankton samples were collected from three mesotrophic lakes: Piaseczno, Rogóźno and Krasne during winter seasons (from January to March). The samples were analyzed for species analysis and abundance of planktonic algae in relation to different depths of water column (0–7 m). Selected water physical-chemical parameters were also measured. Abundance of phytoplankton depended strongly on the thickness of snow and ice cover or mixing conditions. The maximal phytoplankton total number reached about 5 × 106 ind. L−1 beneath the clear ice in the Krasne Lake, minimal numbers were recorded under the thick snow and ice layers in the Piaseczno Lake (2 × 103 ind. L−1). The winter phytoplankton communities were dominated by flagellates principally cryptomonads (Cryptomonas spp. Rhodomonas minuta), euglenophytes (Trachelomonas volvocina, T. volvocinopsis), dinoflagellates (Peridinium bipes, Gymnodinium helveticum) and chrysophytes (Mallomonas elongata, M. akrokomos, Dinobryon sociale) or non-motile small species of blue-green algae (e.g. Rhabdoderma lineare, Limnothrix redekei), diatoms (Stephanodiscus spp., Asterionella formosa), and green algae (e.g. Scenedesmus spp., Monoraphidium spp.). Phytoplankton abundance and structure showed differentiation during the winter season and along the water column as well.