Identification of compounds from terrestrial dissolved organic matter toxic to cyanobacteria

Authors

Amanda D. Neilen, Anthony R. Carroll, Darryl W. Hawker, Katherine R. O’Brien, Michele A. Burford

There is emerging evidence for the phytotoxicity of terrestrial dissolved organic matter (DOM), however its sources, transformations and ecological effects in aquatic ecosystems are poorly understood. DOM characterization by Nuclear Magnetic Resonance (NMR) spectroscopy has typically involved solid-state techniques, but poor resolution has often precluded identification of individual components. This study is the first to directly identify individual phytotoxic components using a novel combined approach of preparative HPLC fractionation of DOM (obtained from leaves of two common riparian trees, Casuarina cunninghamiana and Eucalyptus tereticornis). This was followed by chemical characterization of fractions, using one-dimensional (1D) and two-dimensional (2D) solution-state ¹H NMR analyses. Additionally, the phytotoxic effect of the fractions was determined using cultures of the cyanobacteria Raphidiopsis (Cylindrospermopsis) raciborskii. The amino acid, proline, from Casuarina leachate was identified as phytotoxic, while for Eucalyptus leachate, it was gallic acid and polyphenols. These phytotoxicants remained in the leachates when they were incubated in sunlight or the dark conditions over 5 days. Our study identifies phytotoxic compounds with the potential to affect algal species composition, and potentially control nuisance R. raciborskii blooms.

Temporal heterogeneity of microbial communities and metabolic activities during a natural algal bloom

Authors

Jin Zhou, Yong-min Lao, Jun-ting Song, Hui Jin, Jian-ming Zhu, Zhong-hua Cai

Elucidating the interactions between algae and associated microbial communities is critical for understanding the mechanisms that mediate the dynamic of harmful algal blooms (HABs) in marine environment. However, the microbial functional profiles and their biogeochemical potential in HABs process remains elusive, especially during a complete natural HAB cycle. Here, we used pyrosequencing and functional gene array (GeoChip) to investigate microbial community dynamics and metabolic potential during a natural dinoflagellate (Noctiluca scintillans) bloom. The results shown that bacterioplankton exhibited significant temporal heterogeneity over the course of the bloom stages. Microbial succession was co-driven by environmental parameters and biotic interactions. The functional analysis revealed significant variations in microbial metabolism during matter cycling. At bloom onset-stage, metabolic potential associated with iron oxidation and transport was elevated. Carbon fixation and degradation, denitrification, phosphorus acquisition, and sulfur transfer/oxidation were significantly enhanced at the plateau stage. During the decline and terminal stages, oxidative stress, lysis of compounds, and toxin degradation & protease synthesis increased. This work reveal phycosphere microorganisms can enhanced organic C decomposition capacity, altered N assimilation rate and S/P turnover efficiency, and balancing of the Fe budget during HAB process. The ecological linkage analysis has further shown that microbial composition and functional potential were significantly linked to algal blooms occurrence. It suggest that structural variability and functional plasticity of microbial communities influence HAB trajectory.

Effects of lake warming on the seasonal risk of toxic cyanobacteria exposure

Authors

Nicole M. Hayes Heather A. Haig Gavin L. Simpson Peter R. Leavitt

Incidence of elevated harmful algal blooms and concentrations of microcystin are increasing globally as a result of human‐mediated changes in land use and climate. However, few studies document changes in the seasonal and interannual concentrations of microcystin in lakes. Here, we modeled 11 yr of biweekly microcystin data from six lakes to characterize the seasonal patterns in microcystin concentration and to ascertain if there were pronounced changes in the patterns of potential human exposure to microcystin in lakes of central North America. Bayesian time series analysis with generalized additive models found evidence for a regional increase in microcystin maxima and duration but recorded high variation among lakes. During the past decade, warmer temperatures, but not nutrient levels, led to a marked increase in the number of days when concentrations exceeded drinking and recreational water thresholds set by the World Health Organization and United States Environmental Protection Agency.

 

Co-occurrence of microcystin- and geosmin- producing cyanobacteria in the Tri An Reservoir, a drinking-water supply in Vietnam

 

Authors

Pham, Thanh-Luu; Tran, Thi Hoang Yen; Hoang, Nghia Son; Ngo, Xuan Quang; Tran, Thai Thanh

The present study aimed to investigate the occurrence and distribution of microcystin-(MC) and geosmin-producing cyanobacteria in the Tri An Reservoir, a drinking-water supply in Southern Vietnam. The MC concentration was measured using high-performance liquid chromatography (HPLC). Geosmin production was screened using headspace solid-phase microextraction (HPME) and gas chromatography/mass spectrometry (GC/ MS). The ability of cyanobacteria to produce MC and geosmin was determined by the presence of two molecular markers: mcyA and geoA, respectively. A maximum MC concentration of 207.6 ± 14.2 μg g ^–1 dry weight (DW) was detected from an isolated Microcystis aeruginosa (Kützing) (strains TAMA1), and the maximum geosmin concentration of 114.2 ± 6.1 ng g ^–1 wet weight (WW) was detected from an isolated Dolichospermum circinale (Rabenhorst) (strains TADC4); MC and geosmin were also observed in water-bloom samples, with the highest concentrations being 463.6 ± 34.7 μg g ^–1 DW and 148.5 ± 9.8 ng g ^–1 WW, respectively. These results represent the first molecular evidence of the co-occurrence of MC- and geosmin-producing cyanobacteria in a drinking-water-supply reservoir in Vietnam. Given the success of the molecular techniques and chemical analysis used, we propose that multiplex polymerase chain reaction assays, HPLC, and HPME GC/MS can be reliable tools for investigating MC and geosmin in water-bloom and cyanobacterial samples. Further, we propose that they can be used to create early-warning systems for harmful secondary metabolites produced by cyanobacteria in drinking-water supplies.

 

Physiological responses of Raphidiopsis raciborskii (Cyanobacteria) strains to water conductivity: effect of sodium and magnesium ions

Authors

Daniel Vinícius Neves de Lima, Ana Beatriz Furlanetto Pacheco, Carolina Lage Goulart & Sandra Maria Feliciano de Oliveira e Azevedo

Cyanobacterial blooms dominated by Raphidiopsis raciborskii, a potential producer of saxitoxin (STX), are commonly reported in continental water bodies of semiarid regions with high conductivity (> 1000 µS cm⁻¹). STX production is proposed as a cellular protection mechanism against salt stress. Here, we test the hypothesis that high conductivity favors R. raciborskii growth and that STX is advantageous under this condition. We included four tropical strains (two STX⁺ and two STX⁻) cultured for 12 days in control (450 µS cm⁻¹) or in media with NaCl (10 mM) or MgCl₂ (5 mM) (1500 µS cm⁻¹). High conductivity (either NaCl or MgCl₂) had neutral or positive effects on growth for all strains. Total STX content did not differ in response to salt concentrations. All strains could tolerate higher conductivity without signals of stress, as noted for stable photosynthetic parameters and protein expression profiles. For the strain with higher growth in salt, accumulation of intracellular carbohydrates occurred in early times (3 days). In conclusion, high conductivity is a favorable condition for R. raciborskii, with no clear relation with the ability to produce STX. Pronounced intraspecific variability was observed, a fact that must be considered for a better understanding of this species.

Harmful Algal Bloom Toxins in Aerosol Generated from Inland Lake Water

Authors

Nicole E. Olson, Madeline E. Cooke, Jia H. Shi,Johnna A. Birbeck,Judy A. Westrick, Andrew P. Ault

Harmful algal blooms (HABs) caused by cyanobacteria in freshwater environments produce toxins (e.g., microcystin) that are harmful to human and animal health. HAB frequency and intensity are increasing with greater nutrient runoff and a warming climate. Lake spray aerosol (LSA) released from freshwater lakes has been identified on lakeshores and after transport inland, including from lakes with HABs, but little is known about the potential for HAB toxins to be incorporated into LSA. In this study, freshwater samples were collected from two lakes in Michigan: Mona Lake during a severe HAB with microcystin concentrations (>200 μg/L) well above the Environmental Protection Agency (EPA) recommended “do not drink” level (1.6 μg/L) and Muskegon Lake without a HAB (<1 μg/L microcystin). Microcystin toxins were identified in freshwater, as well as aerosol particles generated in the laboratory from Mona Lake water by liquid chromatography–tandem mass spectrometry (LC–MS/MS) at atmospheric concentrations up to 50 ± 20 ng/m³. Enrichment of hydrophobic microcystin congeners (e.g., microcystin-LR) was observed in aerosol particles relative to bulk freshwater, while enrichment of hydrophilic microcystin (e.g., microcystin-RR) was lower. As HABs increase in a warming climate, understanding and quantifying the emissions of toxins into the atmosphere is crucial for evaluating the health consequences of HABs.

Cyanotoxin occurrence in large rivers of the United States

Authors

Jennifer L. Graham, Neil M. Dubrovsky, Guy M. Foster, Lindsey R. King, Keith A. Loftin, Barry H. Rosen & Erin A. Stelzer

Cyanotoxins occur in rivers worldwide but are understudied in lotic ecosystems relative to lakes and reservoirs. We sampled 11 large river sites located throughout the United States during June–September 2017 to determine the occurrence of cyanobacteria with known cyanotoxin-producing strains, cyanotoxin synthetase genes, and cyanotoxins. Chlorophyll a concentrations ranged from oligotrophic to eutrophic (0.5–64.4 µg L⁻¹). Cyanobacteria were present in the algal communities of all rivers (82% of samples, n = 50) but rarely dominated the phytoplankton (0–52% of total abundance; mean = 8.8%). Pseudanabaena and Planktothrix occurred most often, and many (64%) of the cyanobacterial genera identified (n = 25) have known cyanotoxin-producing strains. Cyanotoxin synthetase genes occurred in all but one river. The mcyE and sxtA genes were most common, present in 73% of rivers and 44% and 40% of samples, respectively. The cyrA gene was less common (22% of samples) but occurred in 64% of rivers. The anaC gene was detected in one river (4% of samples). Anatoxin-a and microcystins were detected at low levels (0.10–0.38 µg L⁻¹) in 2 midcontinent rivers. Cylindrospermopsins and saxitoxins were not detected. Cyanobacteria, cyanotoxin synthetase genes, and cyanotoxins were present at low concentrations throughout this subset of US rivers. Eutrophic rivers located in the midcontinent region of the United States had the highest algal biomass, abundance of cyanotoxin synthetase genes, and cyanotoxin occurrence.

Improved detection of mcyA genes and their phylogenetic origins in harmful algal blooms

Authors

Jaejin Lee, Jinlyung Choi, Micah Fatka, Elizabeth Swanner, Kaoru Ikuma, Xuewei Liang, Tania Leung, Adina Howe

Microcystins, a group of cyanotoxins produced by cyanobacterial strains, have become a significant microbial hazard to human and animal health due to increases in the frequency and intensity of cyanobacterial harmful algal blooms (CyanoHABs). Many studies have explored the correlation between microcystin concentrations and abundances of toxin-producing genes (e.g., mcyA genes) measured using quantitative PCR, and discrepancies between toxin concentrations and gene abundances are often observed. In this study, the results show that these discrepancies are at least partially due to primer sets that do not capture the phylogenetic diversity of naturally present toxin-producers. We designed three novel primer gene probes based on known mcyA genes to improve the detection and quantification of these genes in environmental samples. These primers were shown to improve the identification of mcyA genes compared to previously published primers in freshwater metagenomes, cyanobacterial isolates, and lake water samples. Unlike previously published primers, our primer sets could selectively amplify and resolve Microcystis, Anabaena, and Planktothrix mcyA genes. In lake water samples, abundance estimations of mcyA genes were found to correlate strongly with microcystin concentrations. Based on our results, these primers offer significant improvements over previously published probes to accurately identify and quantify mcyA genes in the environment. There is an increasing need to develop models based on microbial information and environmental factors to predict CyanoHABs, and improved primers will play an important role in aiding monitoring efforts to collect reliable and consistent data on toxicity risks.

Environmental behavior of methamphetamine and ketamine in aquatic ecosystem: Degradation, bioaccumulation, distribution, and associated shift in toxicity and bacterial community

Authors

Zhenglu Wang, Sheng Han, Min Cai, Peng Du, Zhaobin Zhang, Xiqing Li

Ketamine (KET) and methamphetamine (METH) have been recognized as emerging contaminants in aquatic ecosystems. This paper aimed to investigate the environmental behaviour, including the degradation, distribution, and bioaccumulation, of METH, KET, and their main metabolites (amphetamine (AMP) and norketamine (NorKET)). The changes in acute toxicity in the aqueous phase and in the bacterial community in sediment were determined to assess the associated eco-risk of the drug exposure. Five types of lab-scale aquatic ecosystems were established and exposed to KET or METH for 40 days: a water- sediment- organisms- KET system (K), a water- sediment- organisms- METH system (M), a water- sediment- organism- METH- KET system (M + K), a water-sediment- KET- METH system (control), and a water- sediment- organisms system (biocontrol). The results demonstrated that much faster degradation occurred for both METH (t_1/2 = 3.89 and 2.37 days in the M and M + K group, respectively) and KET(t_1/2 = 5.69 days 5.39 days in the K group and M + K group, respectively) than in the control group (t_1/2 = 7.83 and 86.71days for METH and KET, respectively). Rapid adsorption of KET, METH, and their metabolites was observed in the sediment, which had clay and silt as the main particle sizes. KET was observed to be absorbed by shallow-water fish (Chinese medaka, rosy bitterling and mosquito fish), while METH was dominantly ingested by bottom-dwellers (loach). Duckweed might play a crucial role in the dissipation process of METH and KET, which were mainly adsorbed by duckweed roots. During incubation, the acute toxic levels in the K and M + K groups changed from non-toxic to medium toxicity levels, and the toxicity in the M and control groups changed from non-toxic to low toxicity levels. Moreover, marked changes in the bacterial community in the sediment induced by METH or KET exposure were observed, and the most significant change in the bacterial community was observed in the group spiked with both METH and KET. This work for the first time elucidated the environmental behaviors of METH and KET in aquatic ecosystem and associated the impact on ecological system equilibrium.

Nanoplastics Promote Microcystin Synthesis and Release from Cyanobacterial Microcystis aeruginosa

Authors

Li-Juan Feng, Xiao-Dong Sun, Fan-Ping Zhu, Yue Feng, Jian-Lu Duan, Fu Xiao, Xiang-Yu Li, Yi Shi, Qian Wang, Jia-Wen Sun, Xiao-Yu Liu, Jia-Qi Liu, Lin-Lin Zhou, Shu-Guang Wang, Zhaojun Ding, Huiyu Tian, Tamara Susan Galloway, Xian-Zheng Yuan

Although the fate of nanoplastics (<100 nm) in freshwater systems is increasingly well studied, much less is known about its potential threats to cyanobacterial blooms, the ultimate phenomenon of eutrophication occurrence worldwide. Previous studies have evaluated the consequences of nanoplastics increasing the membrane permeability of microbes, however, there is no direct evidence for interactions between nanoplastics and microcystin; intracellular hepatotoxins are produced by some genera of cyanobacteria. Here, we show that the amino-modified polystyrene nanoplastics (PS-NH₂) promote microcystin synthesis and release from Microcystis aeruginosa, a dominant species causing cyanobacterial blooms, even without the change of coloration. We demonstrate that PS-NH₂ inhibits photosystem II efficiency, reduces organic substance synthesis, and induces oxidative stress, enhancing the synthesis of microcystin. Furthermore, PS-NH₂ promotes the extracellular release of microcystin from M. aeruginosa via transporter protein upregulation and impaired cell membrane integrity. Our findings propose that the presence of nanoplastics in freshwater ecosystems might enhance the threat of eutrophication to aquatic ecology and human health.