Promoting heterotrophic denitrification of Pseudomonas hunanensis strain PAD-1 using pyrite: A mechanistic study.

Denitrification is critical for removing nitrate from wastewater, but it typically requires large amounts of organic carbon, which can lead to high operating costs and secondary environmental pollution. To address this issue, this study proposes a novel method to reduce the demand for organic carbon in denitrification. In this study, a new denitrifier, Pseudomonas hunanensis strain PAD-1, was obtained with properties for high efficiency nitrogen removal and trace N2O emission. It was also used to explore the feasibility of pyrite-enhanced denitrification to reduce organic carbon demand. The results showed that pyrite significantly improved the heterotrophic denitrification of strain PAD-1, and optimal addition amount was 0.8-1.6 g/L. The strengthening effect of pyrite was positively correlated with carbon to nitrogen ratio, and it could effectively reduce demand for organic carbon sources and enhance carbon metabolism of strain PAD-1. Meanwhile, the pyrite significantly up-regulated electron transport system activity (ETSA) of strain PAD-1 by 80%, nitrate reductase activity by 16%, Complex III activity by 28%, and napA expression by 5.21 times. Overall, the addition of pyrite presents a new avenue for reducing carbon source demand and improving the nitrate harmless rate in the nitrogen removal process.

Multi-residue analysis of 206 pesticides in grass forage by the one-step quick, easy, cheap, effective, rugged, and safe method combined with ultrahigh-performance liquid chromatography quadrupole orbitrap mass spectrometry.

A novel method for detecting pesticide multi-residue in grass forage (alfalfa and oat) was established based on the one-step automatic extraction and purification technology of quick, easy, cheap, effective, rugged, and safe combined with ultrahigh-performance liquid chromatography quadrupole Orbitrap high-resolution mass spectrometry. The crushed sample was extracted with acetonitrile with 1% acetate, followed by a cleanup step with a primary-secondary amine, octadecylsilane, and graphitized carbon black. The extraction and purification were carried out using the one-step automatic pretreatment equipment. The target pesticides were acquired in positive ion electrospray ionization mode and full scan/data dependent secondary scan mode. The calibration curve shows good linearity over the corresponding concentration range, with the coefficient of determination greater than 0.99. The screening detection limits were 0.5-50 µg/kg, and the limit of quantification for the 206 pesticides was set at 1-50 µg/kg. At the spiking levels of one, two, and 10 times of limit of quantification, more than 95% of pesticides had recovery between 70-120%, with a relative standard deviation ≤20%. The method was proved to be simple, rapid, high-sensitivity, and could be routinely used for the high throughput screening and quantitative analysis of pesticide residues in alfalfa and oat.

Development and Validation of a Method for Determination of 43 Antimicrobial Drugs in Western-Style Pork Products by UPLC-MS/MS with the Aid of Experimental Design.

Western-style pork products have attracted many modern urban consumers, and these products have rapidly entered the Chinese market. The current hazard analysis of processed meat products mainly focuses on processing hazards (PAHs, microorganisms, and food additives), with less attention to veterinary drug residues. According to the survey results, the residues of antimicrobial drugs (sulfonamides and quinolones) in pork and its products in China are a severe problem, which may cause metabolic reactions, toxic effects, or enhance drug resistance. This study applied a modified QuEChERS method combined with ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MSMS) to develop a rapid and sensitive method for determining antimicrobial drugs in bacon and ham was successfully evaluated methodologically by EU 2002/657/EC. This study used a three-level, three-factor Box-Behnken design (BBD) to optimize the QuEChERS method by response surface methodology. The excellent linearity of the calibration curve was shown in the corresponding concentration range with a coefficient of determination greater than 0.99. The values of decision limit (CCα) and detection capability (CCß) were in the range of 10.9-31.3 µg/kg and 11.8-52.5 µg/kg, respectively. The method successfully detected two trace levels of antimicrobial drugs in commercially available samples, including sulfadiazine and moxifloxacin.

Spatiotemporal dynamics of cell abundance, colony size and intracellular toxin concentrations of pelagic and benthic Microcystis in Lake Caohai, China.

Lake Caohai has experienced extensive Microcystis blooms in recent years, and to improve its water quality, the local government carried out a series of water control measures. To better understand the dynamics of both pelagic and benthic Microcystis and their characteristics in Lake Caohai, we conducted a 1-year investigation from December 2015 to December 2016 to gain a seasonal outlook on the distribution and dynamics of cell abundance, colony size and intracellular microcystins (MCs) of Microcystis. The results indicated that the Microcystis bloom occupied primarily the northeastern region and then moved gradually from lakeshore to lake center. The perennial southwesterly winds and the water inflow from northeast to southwest in Lake Caohai determined the spatiotemporal distribution of pelagic Microcystis. Benthic Microcystis was mainly distributed in the northeastern region in summer, occupied the lake center in autumn and then occupied the southeastern region in winter, determined by the sedimentation of pelagic Microcystis and the death of benthic Microcystis. Small colonies (20-60 µm) overwintered more easily in both water column and sediment. The concentrations of intracellular toxin of benthic Microcystis were observed to be significantly higher than those of pelagic Microcystis. This might be because Microcystis synthesized large amount of MCs to acclimate to an unfavorable benthic environment. This knowledge on the dynamics of Microcystis expands our understanding of mechanisms underpinning the formation of Microcystis blooms.

Residue investigation of some phenylureas and tebuthiuron herbicides in vegetables by ultra-performance liquid chromatography coupled with integrated selective accelerated solvent extraction-clean up in situ.

BACKGROUND: Some trace amounts of urea herbicide residues can be transferred to humans via the food chain, thereby being potentially harmful to human health. The development of a robust analytical methodology for effective sample preparation and simultaneous determination of herbicide residues in vegetable samples is required for achieving food safety. RESULTS: The diuron-molecularly imprinted polymers (MIPs) synthesized have excellent affinity and high selectivity to phenylureas (monolinuron, isoproturon, diuron and linuron) and tebuthiuron. A novel automated procedure with better selectivity for vegetable sample treatment was developed by integrated matrix solid-phase dispersion-accelerated solvent extraction clean-up in situ. Five herbicides can be baseline separated with runtime down to 5 min by ultra-performance liquid chromatography, and good linearity was obtained with a correlation coefficient (r) of 0.9999. The limit of quantification of the method was in the range of 0.8-2.3 µg kg-1 . Diuron residue in cherry tomato sample was found to be 40 µg kg-1 . CONCLUSION: The developed method has satisfactory selectivity, good linearity, high sensitivity and accuracy as well as speediness, and can ensure rapid selective extraction and sensitive multi-residue analysis at low microgram per kilogram levels of the herbicides in vegetable food. © 2018 Society of Chemical Industry.

Rapid selective accelerated solvent extraction and simultaneous determination of herbicide atrazine and its metabolites in fruit by ultra high performance liquid chromatography.

A selective accelerated solvent extraction procedure achieved one step extraction and cleanup for analysis of herbicide atrazine and its metabolites in fruit. Using a BEH C18 analytical column and the gradient mode with 2 mM ammonium acetate aqueous solution/acetonitrile as a mobile phase achieved effective chromatographic separation of the five analytes within 4 min. The calibration curves were linear over two orders of magnitude of concentration with correlation coefficients (r) of 0.9996-0.9999. The method limit of quantification was 1, 2, 1.5, 3, and 2 µg/kg for atrazine, desethylatrazine, desisopropylatrazine, desethyldesisopropylatrazine, and hydroxyatrazine, respectively, in the case of atrazine it is at least two orders of magnitude lower than the maximum residue limit (0.25 mg/kg). The intra-day and inter-day precisions of the five analytes were in the range of 2.1-3.5 and 3.1-4.8 %, respectively. The recoveries of the five analytes at three spiked levels varied from 85.9 to 107% with a relative standard deviation of 1.8-4.9% for pear and apple samples. The ultra high performance liquid chromatography with diode array detection method was proved to be fast, inexpensive, selective, sensitive, and accurate for the quantification of the analytes in pear and apple samples.

Synthesis and evaluation of dummy molecularly imprinted microspheres for the specific solid-phase extraction of six anthraquinones from slimming tea.

Dummy molecularly imprinted microspheres with danthron as template were synthesized and their performance was evaluated. Accelerated solvent extraction can rapidly and effectively remove template molecules from the microspheres. The microspheres were applied as a specific sorbent for solid-phase extraction of six anthraquinones from slimming tea, showing excellent affinity and high selectivity to danthron and the target analytes. The molecular recognition mechanisms were discussed by the experimental validation with IR spectroscopy. The sample was treated using accelerated solvent extraction followed by dummy molecularly imprinted microspheres solid-phase extraction. Under the optimized ultra high performance liquid chromatographic conditions, the six target analytes can be baseline separated in 8 min, and good linearity was obtained in a range of 0.1-40 µg/mL with the correlation coefficient (r(2)) of ≥0.9998. The method limit of quantification was in a range of 1-2 mg/kg, it can ensure analysis of anthraquinones at mg/kg level. The intra- and interday precision (RSD, n = 6) for the analysis of the six analytes in a slimming tea was less than 4.5 and 5.4%, respectively. The developed method can be applied for the selective extraction, effective separation, and rapid determination of six anthraquinones in slimming tea.

Vertical distribution of Fe and Fe(III)-reducing bacteria in the sediments of Lake Donghu, China.

In lake sediments, iron (Fe) is the most versatile element, and the redox cycling of Fe has a wide influence on the biogeochemical cycling of organic and inorganic substances. The aim of the present study was to analyze the vertical distribution of Fe and Fe(III)-reducing bacteria (FeRB) in the surface sediment (30 cm) of Lake Donghu, China. At the 3 sites we surveyed, FeRB and Fe(II)-oxidizing bacteria (FeOB) coexisted in anoxic sediments. Geobacter-related FeRB accounted for 5%-31% of the total Bacteria, while Gallionella-related FeOB accounted for only 0.1%-1.3%. A significant correlation between the relative abundance of poorly crystalline Fe and Geobacter spp. suggested that poorly crystalline Fe favored microbial Fe(III) reduction. Poorly crystalline Fe and Geobacter spp. were significantly associated with solid-phase Fe(II) and total inorganic phosphorus levels. Pore water Fe(II) concentrations negatively correlated with NO3(-) at all sites. We concluded that Geobacter spp. were abundant in the sediments of Lake Donghu, and the redox of Fe might participate in the cycling of nitrogen and phosphorus in sediments. These observations provided insight into the roles of microbial Fe cycling in lake sediments.

Highly sensitive determination of cyromazine, melamine, and their metabolites in milk by molecularly imprinted solid-phase extraction combined with ultra-performance liquid chromatography.

A novel molecularly imprinted solid-phase extraction-ultra-performance liquid chromatography (MISPE-UPLC) method for effective separation and simultaneous determination of cyromazine, melamine, and their metabolites (ammeline and ammelide) in milk samples was developed. Molecularly imprinted polymers (MIP) were synthesized in an ethanol-water system, with melamine as the template and methacrylic acid as the organic functional monomer. The MIP were applied as a specific sorbent for the selective solid phase extraction of cyromazine, ammelide, melamine and ammeline. The molecular recognition mechanism was investigated by molecular simulation and the experiment was validate by using Fourier transform infrared spectroscopy and (1)H nuclear magnetic resonance spectroscopy. A new mechanism based on the formation of both an amido group and hydrogen bonds was developed. A binding study demonstrated that the MIP showed excellent affinity to and high selectivity for melamine and related compounds. Under optimized conditions, we achieved good linearity of the calibration curves with correlation coefficients >0.999. Low limits of quantification (LOQ) for the method were determined to be 1.25, 1.25, 2.59, and 6.42 µg/kg for cyromazine, ammelide, melamine, and ammeline, respectively, which were 3 orders of magnitude smaller than the maximum residue limit (MRL). The high sensitivity of this method allows detection at the microgram per kilogram level. The proposed MISPE-UPLC method is a highly selective and sensitive method for determination of cyromazine, melamine, and their metabolites (ammeline and ammelide) for use in the control and quality assurance of milk.

Dynamics of Microcystis surface scum formation under different wind conditions: the role of hydrodynamic processes at the air-water interface.

Due to climate change, Microcystis blooms occur at increasing frequencies in aquatic ecosystems worldwide. Wind-generated turbulence is a crucial environmental stressor that can vertically disperse the Microcystis surface scum, reducing its light availability. Yet, the interactions of Microcystis scum with the wind-generated hydrodynamic processes, particularly those at the air-water interface, remain poorly understood. Here, we explore the response of Microcystis (including colony size and migration dynamics) to varying magnitudes and durations of intermittent wind disturbances in a mesocosm system. The flow velocities, size of Microcystis colonies, and the areal coverage of the water surface by scum were measured through video observations. Our results demonstrate that low wind speeds increase colony size by providing a stable condition where Microcystis forms a scum layer and aggregates into large colonies at the air-water interface. In contrast, wind disturbances disperse scum and generate turbulence, resulting in smaller colonies with higher magnitudes of wind disturbance. We observed that surface scum can form rapidly following a long period (6 h) of high-magnitude (4.5 m s-1) wind disturbance. Furthermore, our results indicate reduced water surface tension caused by the presence of Microcystis, which can decrease surface flow velocity and counteract wind-driven mixing. The reduced surface tension may also drive lateral convection at the water surface. These findings suggest that Microcystis reduces surface tension, likely by releasing surface-active materials, as an adaptive response to various wind conditions. This could result in an increased rate of surface scum re-formation under wind conditions and potentially facilitate the lateral expansion of scum patches during weak wind periods. This study reveals new insights into how Microcystis copes with different wind conditions and highlights the importance of the air-water interface for Microcystis scum dynamics.

Selection of photosynthetic traits by turbulent mixing governs formation of cyanobacterial blooms in shallow eutrophic lakes.

Prediction of the complex cyanobacteria-environment interactions is vital for understanding harmful bloom formation. Most previous studies on these interactions considered specific properties of cyanobacterial cells as representative for the entire population (e.g. growth rate, mortality, and photosynthetic capacity (Pmax)), and assumed that they remained spatiotemporally unchanged. Although, at the population level, the alteration of such traits can be driven by intraspecific competition, little is known about how traits and their plasticity change in response to environmental conditions and affect the bloom formation. Here we test the hypothesis that intraspecific variations in Pmax of cyanobacteria (Microcystis spp.) play an important role in its population dynamics. We coupled a one-dimensional hydrodynamic model with a trait-based phytoplankton model to simulate the effects of physical drivers (turbulence and turbidity) on the Pmax of Microcystis populations for a range of dynamic conditions typical for shallow eutrophic lakes. Our results revealed that turbulence acts as a directional selective driver for changes in Pmax. Depending on the intensity of daily-periodic turbulence, representing wind-driven mixing, a shift in population-averaged phenotypes occurred toward either low Pmax, allowing the population to capture additional light in the upper layers, or high Pmax, enhancing the efficiency of light utilization. Moreover, we observed that a high intraspecific diversity in Pmax accelerated the formation of surface scum by up to more than four times compared to a lower diversity. This study offers insights into mechanisms by which cyanobacteria populations respond to turbulence and underscores the significance of intraspecific variations in cyanobacterial bloom formation.

Effect of extracellular polymeric substances on the colony size and morphological changes of Microcystis.

Surface blooms of colony-forming Microcystis are increasingly occurring in aquatic ecosystems on a global scale. Recent studies have found that the Microcystis colonial morphology is a crucial factor in the occurrence, persistence, and dominance of Microcystis blooms, yet the mechanism driving its morphological dynamics has remained unknown. This study conducted a laboratory experiment to test the effect of extracellular polymeric substances on the morphological dynamics of Microcystis. Ultrasound was used to disaggregate colonies, isolating the cells and of the Microcystis suspension. The single cells were then re-cultured under three homologous EPS concentrations: group CK, group Low, and group High. The size, morphology, and EPS [including tightly bound EPS (TB-EPS), loosely bound EPS (LB-EPS), bound polysaccharides (B-polysaccharides), and bound proteins (B-proteins)] changes of colonies were closely monitored over a period of 2 months. It was observed that colonies were rapidly formed in group CK, with median colony size (D50) reaching 183 µm on day 12. The proportion of colonies with a size of 150-500 µm increased from 1% to more than 50%. Colony formation was also observed in both groups Low and High, but their D50 increased at a slower rate and remained around 130 µm after day 17. Colonies with a size of 50-150 µm account for more than 50%. Groups CK and Low successively recovered the initial Microcystis morphology, which is a ring structure formed of several small colonies with a D50 of 130 µm. During the recovery of the colony morphology, the EPS per cell increased and then decreased, with TB-EPS and B-polysaccharides constituting the primary components. The results suggest that colony formation transitioned from adhesion driven to being division driven over time. It is suggested that the homologous EPS released into the ambient environment due to the disaggregation of the colony is a chemical cue that can affect the formation of a colony. This plays an important but largely ignored role in the dynamics of Microcystis and surface blooms.

Rapid determination of volatile benzene derivatives and chlorobenzenes in goat's milk by HS-SPME-GC-MS/MS.

A method for the determination of eight benzenes (BTEXs) and twelve chlorobenzenes (CBs) in goat's milk by headspace solid-phase microextraction-gas chromatography-mass spectrometry (HS-SPME-GC-MS/MS) was developed. The study investigated the impact of various factors such as extraction fiber type, salt amount, equilibrium conditions, and desorption conditions on the outcomes. Target analytes were separated on a DB-HeavyWAX column and quantified using the external standard method. The results showed that the target compounds had a good linear relationship in the range of 0.01 ∼ 50 µg/L (R2 > 0.997), the limit of detection (LOD) was 0.003 ∼ 0.150 µg/L, and the limit of quantification (LOQ) was 0.01 ∼ 0.50 µg/L. The average recoveries were 82%-116% and the relative standard deviation (RSD) was 0.8%-17.3% under the three addition levels of 1×, 2×, and 10 × LOQ. In a survey of twenty goat's milk samples, only ethylbenzene, xylenes, cumene, chlorobenzene, and 1,4-dichlorobenzene were detected at levels exceeding their respective limits of quantification. The method was evaluated using two ecological scales (Eco-Scale), GAPI and AGREEN, to verify its environmental friendliness and applicability. This method is simple, green, and efficient, which provides a certain theoretical basis for the production and quality safety evaluation of dairy products.

Simultaneous determination of 78 pesticide residues and 16 mycotoxins in tsampa by an improved QuEChERS method coupled with ultra performance liquid chromatography-tandem mass spectrometry.

Tsampa may contain pesticide residues and mycotoxins, which may pose a risk to human health. Currently, pesticide detection and mycotoxin detection are two independent experiments. To improve the efficiency of the analysis, a method based on QuEChERS combined with ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) for the simultaneous determination of 78 pesticides and 16 mycotoxins in tsampa was developed. All the target compounds showed good linear correlation with correlation coefficients (R2) greater than 0.9990. The limits of detection (LODs) and limits of quantification (LOQs) were in the ranges of 0.10-3.00 µg kg-1 and 0.40-10.00 µg kg-1, respectively. The average recoveries of the pesticides and mycotoxins spiked at the 1, 2, and 10-fold LOQ were in the range of 73.0-115.2%, and the relative standard deviations (RSDs) were lower than 11.7%. This method was applied to 19 batches of real samples in which 32% of samples exceeded the maximum residue limits of the European Union involving aflatoxin G2, ochratoxin A, and hexaconazole. It proved to be excellent, efficient, greatly simplified, and highly applicable, which could reduce the workload and time significantly for the daily monitoring of the pesticides and mycotoxins in tsampa.

Rapid flotation of Microcystis wesenbergii mediated by high light exposure: implications for surface scum formation and cyanobacterial species succession.

Increasing occurrences of Microcystis surface scum have been observed in the context of global climate change and the increase in anthropogenic pollution, causing deteriorating water quality in aquatic ecosystems. Previous studies on scum formation mainly focus on the buoyancy-driven floating process of larger Microcystis colonies, neglecting other potential mechanisms. To study the non-buoyancy-driven rapid flotation of Microcystis, we here investigate the floating processes of two strains of single-cell species (Microcystis aeruginosa and Microcystis wesenbergii), which are typically buoyant, under light conditions (150 µmol photons s-1 m-2). Our results showed that M. wesenbergii exhibited fast upward migration and formed surface scum within 4 hours, while M. aeruginosa did not form visible scum throughout the experiments. To further explore the underlying mechanism of these processes, we compared the dissolved oxygen (DO), extracellular polymeric substance (EPS) content, and colony size of Microcystis in different treatments. We found supersaturated DO and the formation of micro-bubbles (50-200 µm in diameter) in M. wesenbergii treatments. M. aeruginosa produces bubbles in small quantities and small sizes. Additionally, M. wesenbergii produced more EPS and tended to aggregate into larger colonies. M. wesenbergii had much more derived-soluble extracellular proteins and polysaccharides compared to M. aeruginosa. At the same time, M. wesenbergii contains abundant functional groups, which was beneficial to the formation of agglomerates. The surface scum observed in M. wesenbergii is likely due to micro-bubbles attaching to the surface of cell aggregates or becoming trapped within the colony. Our study reveals a species-specific mechanism for the rapid floatation of Microcystis, providing novel insights into surface scum formation as well as succession of cyanobacterial species.

[Rapid determination of 15 N-nitrosamines in air-dried yak meat using one-step QuEChERS-gas chromatography-triple quadrupole mass spectrometry].

A method based on gas chromatography-triple quadrupole mass spectrometry (GC-MS/MS) coupled with one-step QuEChERS technique was developed for the simultaneous determination of 15 N-nitrosamines in air-dried yak meat. The hydration volume, extraction solvent, extracting salt, and cleaning material were optimized according to the characteristics of the N-nitrosamines and sample matrix. The optimized conditions were as follows: 10 mL of purified water for sample hydration, acetonitrile as the extraction solvent for the sample after hydration, 4.0 g of anhydrous MgSO4 and 1.0 g of NaCl as extracting salts, 500 mg of MgSO4+25 mg of C18+50 mg of PSA as cleaning materials. Favorable recoveries of the 15 N-nitrosamines were obtained when the extraction solution was incompletely dried. Thus, the final extract was dried to below 0.5 mL under a mild nitrogen stream and then redissolved to 0.5 mL with acetonitrile. After filtration, 200 µL of the sample was transferred to an autosampler vial for GC-MS/MS analysis. The 15 N-nitrosamines were determined using GC-MS/MS on a DB-HeavyWAX column (30 m×0.25 mm×0.25 µm) with an electron impact ion source in multiple-reaction monitoring (MRM) mode, and quantified using an external standard method. Under the optimized experimental conditions, the results showed that the calibration curves exhibited good linearities for the 15 N-nitrosamines, with correlation coefficients (r2) greater than 0.9990. The limits of detection (LODs) and the limits of quantification (LOQs) ranged from 0.05 to 0.20 µg/kg and from 0.10 to 0.50 µg/kg, respectively. At spiked levels of 1LOQ, 2LOQ, and 10LOQ, the average recoveries were 79.4%-102.1%, 80.6%-109.5%, and 83.0%-110.6%, respectively, and the relative standard deviations were in the range of 0.8%-16.0%. The low matrix effects of the 15 N-nitrosamines indicated the high sensitivity of the proposed method. The method was applied to detect representative commercial air-dried yak meat samples obtained using different processing techniques. Seven N-nitrosamines, including N-nitrosodimethylamine, N-nitrosodiisobutylamine, N-nitrosodibutylamine, N-methyl-N-phenylnitrous amide, N-ethyl-N-nitrosoaniline, N-nitrosopyrrolidine, and N-nitrosodiphenylamine were detected in all samples. The average contents of the seven N-nitrosamines was 0.08-20.18 µg/kg. The detection rates and average contents of the N-nitrosamines in cooked air-dried yak meat samples were higher than those in traditional raw air-dried yak meat samples. Compared with the manual QuEChERS method, the one-step QuEChERS method developed integrated the extraction and clean-up procedures into one single run, and the detection efficiency was considerably improved. The developed method is simple, rapid, highly sensitive, and insusceptible to human errors. Thus, it is useful for the determination of N-nitrosamines in air-dried yak meat and can be extended to the qualitative and quantitative analysis of N-nitrosamines in other meat products. It also provides method support and a data reference for the general determination of N-nitrosamines, which is of great significance for food safety.

Assessment of in-situ monitoring and tracking the vertical migration of cyanobacterial blooms using LISST-HAB.

Cyanobacterial harmful algal blooms (cyanoHABs) are becoming increasingly common in aquatic ecosystems worldwide. However, their heterogeneous distributions make it difficult to accurately estimate the total algae biomass and forecast the occurrence of surface cyanoHABs by using traditional monitoring methods. Although various optical instruments and remote sensing methods have been employed to monitor the dynamics of cyanoHABs at the water surface (i.e., bloom area, chlorophyll a), there is no effective in-situ methodology to monitor the dynamic change of cell density and integrated biovolume of algae throughout the water column. In this study, we propose a quantitative protocol for simultaneously measurements of multiple indicators (i.e., biovolume concentration, size distribution, cell density, and column-integrated biovolume) of cyanoHABs in water bodies by using the laser in-situ scattering and transmissometry (LISST) instrument. The accuracy of measurements of the biovolume and colony size of algae was evaluated and exceeded 95% when the water bloom was dominated by cyanobacteria. Furthermore, the cell density of cyanobacteria was well estimated based on total biovolume and mean cell volume measured by the instrument. Therefore, this methodology has the potential to be used for broader applications, not only to monitor the spatial and temporal distribution of algal biovolume concentration but also monitor the vertical distribution of cell density, biomass and their relationship with size distribution patterns. This provides new technical means for the monitoring and analysis of algae migration and early warning of the formation of cyanoHABs in lakes and reservoirs.

Combined effect of freshwater salinization and harmful algae on the benthic invertebrate Chironomus pallidivittatus.

Global climate change as well as human activities have been reported to increase the frequency and severity of both salinization and harmful algal blooms (HABs) in many freshwater systems, but their co-effect on benthic invertebrates has rarely been studied. This study simultaneously examined the joint toxicity of salinity and different cyanobacterial diets on the behavior, development, select biomarkers, and partial life cycle of Chironomus pallidivittatus (Diptera). High concentrations of salts (e.g., 1 g/L Ca2+ and Mg2+) and toxic Microcystis had synergistic toxicity, inhibiting development, burrowing ability and causing high mortality of C. pallidivittatus, especially for the Mg2+ treatment, which caused around 90% death. Low Ca2+ concentration (e.g., 0.01 g/L) promoted larval burrowing ability and inhibited toxin accumulation, which increased the tolerance of Chironomus to toxic Microcystis. However, low Mg2+ concentration (e.g., 0.01 g/L) was shown to inhibit the behavior, development and increase algal toxicity to Chironomus. Toxic Microcystis resulted in microcystin (MC) accumulation, inhibited the burrowing ability of larvae, and increased the proportion of male adults (>50%). The combined toxicity level from low to high was verified by the weight of evidence and the grey TOPSIS model, which integrated five lines of evidence to increase the risk assessment accuracy and efficiency. This is the first study that provided insights into ecological risk arising from the joint effect of salinity and harmful algae on benthic organisms. We suggest that freshwater salinization and HABs should be considered together when assessing ecological threats that arise from external stress.

Differences in survivability and toxic potential among Microcystis colonies of different sizes in sediment.

Microcystis colonies have the ability to persist for extended periods in sediment and function as a "seed bank" for the succeeding summer bloom in water column. The colonial morphology and toxin production ability of Microcystis are important for their population maintenance and life history. However, it is unclear about the influence of the colony morphology and toxic potential of Microcystis colonies on their benthic process. To address this question, we classified field Microcystis samples into three groups based on their size (< 150 µm, 150-300 µm, and > 300 µm) and compared their survivability and toxic potential during culturing in sediment. The results showed that Microcystis colonies in sediments disappeared quickly at 25℃ but survived for long periods at 5℃. The survivability of smaller Microcystis colonies (< 300 µm) was significantly higher than that of larger ones (> 300 µm). The activities of catalase (CAT) were significantly increased in large colonies compared to small colonies at 15℃ and 25℃. Real-time PCR indicated that smaller colonies had higher proportion of potential toxic genotype, and Microcystis colonies cultured at 15℃ and 25℃ showed higher percentage of microcystin-producing genotype. These results indicate that Microcystis colonies survived longer at low temperature and that larger Microcystis colonies are more susceptible to oxidative stress in sediments. The difference of toxic potential of Microcystis colonies of different sizes in sediments may be related to their survival ability in sediments.

Instantaneous and reversible flocculation of Scenedesmus via Chitosan and Xanthan Gum complexation.

An instantaneous and reversible flocculation method for Scenedesmus harvesting was developed, based on the complexation of Chitosan (CTS) and Xanthan Gum (XG). Under rapid stirring, Scenedesmus cells formed centimeter-sized flocs within 20 s using binary flocculants of 4 mg/L CTS and 16 mg/L XG. These flocs exhibited a remarkable harvest efficiency exceeding 95 % when filtered through 500-µm-pore-sized sieves. Furthermore, the flocs could be completely disintegrated by using alkaline or NaCl solutions (pH > 11 or NaCl concentration > 1.5 mol/L). Adjusting pH allowed recovery of 50 % CTS and 75 % XG, resulting in microalgae biomass with lower flocculant content and reducing reagent costs. Electrostatic interaction of -COO- of XG and -NH3+ of CTS deduced the formation of polyelectrolyte complexes (PECs), which shrink and wrap the coexisting algal cells to form the flocs under stirring. CTS and XG complexation was instantaneous and reversible, explaining quick flocculation and disintegration.