Single-Cell RNA Sequencing Profiling Cellular Heterogeneity and Specific Responses of Fish Gills to Microplastics and Nanoplastics.

Fish gills are highly sensitive organs for microplastic (MP) and nanoplastic (NP) invasions, but the cellular heterogeneity of fish gills to MPs and NPs remains largely unknown. We employed single-cell RNA sequencing to investigate the responses of individual cell populations in tilapia Oreochromis niloticus gills to MP and NP exposure at an environmentally relevant concentration. Based on the detected differentially expressed gene (DEG) numbers, the most affected immune cells by MP exposure were macrophages, while the stimulus of NPs primarily targeted T cells. In response to MPs and NPs, H+-ATPase-rich cells exhibited distinct changes as compared with Na+/K+-ATPase-rich cells and pavement cells. Fibroblasts were identified as a potential sensitive cell-type biomarker for MP interaction with O. niloticus gills, as evidenced by the largely reduced cell counts and the mostly detected DEGs among the 12 identified cell populations. The most MP-sensitive fibroblast subpopulation in O. niloticus gills was lipofibroblasts. Cell-cell communications between fibroblasts and H+-ATPase-rich cells, neurons, macrophages, neuroepithelial cells, and Na+/K+-ATPase-rich cells in O. niloticus gills were significantly inhibited by MP exposure. Collectively, our study demonstrated the cellular heterogeneity of O. niloticus gills to MPs and NPs and provided sensitive markers for their toxicological mechanisms at single-cell resolution.

Physiological and immune profiling of tilapia Oreochromis niloticus gills by high-throughput single-cell transcriptome sequencing.

The physiological and immune functions of fish gills are largely recognized, but their following functional heterogeneity at the single cell scale has been rarely reported. Here, we performed single cell RNA sequencing (scRNA-seq) on the gills of tilapia fish Oreochromis niloticus. We identified a total of 12 cell populations and analyzed their functional heterogeneity. To investigate the physiological function of O. niloticus gills, expression patterns of genes encoding ion transporters were selected from the identified H+-ATPase-rich cells (HR cells), Na+/K+-ATPase-rich cells (NaR cells), and pavement cells. Specific enrichment of ca4a, slc9a1a, and LOC100692482 in the HR cells of O. niloticus gills explained their functions in acid-base regulation. Genes encoding Ca2+ transporters, including atp2b1, LOC100696627, and LOC 100706765, were specifically expressed in the NaR cells. Pavement cells were presumably the main sites responsible for ammonia and urea transports in O. niloticus gills with specific enrichment of Rhbg and LOC100693008, respectively. The expression patterns of the four immune cell subtypes varied greatly, with B cells being enriched with the most immune-related GO terms. KEGG enrichment analysis showed that MAPK signaling pathway was the most enriched pathway among the four types of immune cells in O. niloticus gills. Our results are important in understanding the physiological and immune responses of fish gills at the cellular resolution.

Unveiling the Microfiber Release Footprint: Guiding Control Strategies in the Textile Production Industry.

Microplastic fibers from textiles have been known to significantly contribute to marine microplastic pollution. However, little is known about the microfiber formation and discharge during textile production. In this study, we have quantified microfiber emissions from one large and representative textile factory during different stages, spanning seven different materials, including cotton, polyester, and blended fabrics, to further guide control strategies. Wet-processing steps released up to 25 times more microfibers than home laundering, with dyeing contributing to 95.0% of the total emissions. Microfiber release could be reduced by using white coloring, a lower dyeing temperature, and a shorter dyeing duration. Thinner, denser yarns increased microfiber pollution, whereas using tightly twisted fibers mitigated release. Globally, wet textile processing potentially produced 6.4 kt of microfibers in 2020, with China, India, and the US as significant contributors. The study underlined the environmental impact of textile production and the need for mitigation strategies, particularly in dyeing processes and fiber choice. In addition, no significant difference was observed between the virgin polyesters and the used ones. Replacing virgin fibers with recycled fibers in polyester fabrics, due to their increasing consumption, might offer another potential solution. The findings highlighted the substantial impact of textile production on microfiber released into the environment, and optimization of material selection, knitting technologies, production processing, and recycled materials could be effective mitigation strategies.

Maltol mitigates cisplatin-evoked cardiotoxicity via inhibiting the PI3K/Akt signaling pathway in rodents in vivo and in vitro.

Our current research aims to evaluate the efficiency of a flavor enhancer, maltol (produced by heating ginseng) against cisplatin-evoked cardiotoxicity by establishing cisplatin-induced heart injury in vivo and H9C2 rat cardiomyocyte model. The cisplatin-treated mice at 3 mg/kg for four times on the 7th, 9th, 11th and 13th day, and in them appeared a serious cardiac damage accompanied with the increase in indicators of heart damage. Multiple exposure of 3 mg/kg for four times of cisplatin increased cardiac cells apoptosis with increased expression of Bax and cleaved-caspase 3, and decreased expression of Bcl-2. Interestingly, supplement of maltol at doses of 50 and 100 mg/kg for 15 days significantly suppressed the cardiac disturbance. In cultured H9C2 cells, maltol enhanced PI3K/Akt expression level during cisplatin treatment, and reduced cisplatin-induced apoptosis. Notably, inhibition of PI3K/Akt by LY294002 and HY-10249A lessened the efficacy of maltol. In mice, maltol apparently induced PI3K/Akt in heart tissues and protected against cisplatin-induced cardiotoxicity. In conclusion, maltol exerted the protective effects against cisplatin-induced cardiotoxicity, at least partially by inhibiting the activation of PI3K/Akt signaling pathways in cardiomyocytes, to ease oxidative stress, and alleviate reactive oxygen species-mediated apoptosis.

Panos-Fermented Extract-Mediated Nanoemulsion: Preparation, Characterization, and In Vitro Anti-Inflammatory Effects on RAW 264.7 Cells.

This study focused on developing Panos nanoemulsion (P-NE) and enhancing the anti-inflammatory efficacy for the treatment of inflammation. The effects of P-NE were evaluated in terms of Nitric oxide (NO production) in Lipopolysaccharide (LPS), induced RAW 264.7 cells, Reactive oxygen species (ROS) generation using Human Keratinocyte cells (HaCaT), and quantitative polymerase chain reaction (qPCR) analysis. Sea buckthorn oil, Tween 80, and span 80 were used and optimize the process. Panos extract (P-Ext) was prepared using the fermentation process. Further high-energy ultra-sonication was used for the preparation of P-NE. The developed nanoemulsion (NE) was characterized using different analytical methods. Field emission transmission electron microscopy (FE-TEM) analyzed the spherical shape and morphology. In addition, stability was analyzed by Dynamic light scattering (DLS) analysis, where particle size was analyzed 83 nm, and Zeta potential -28.20 ± 2 (mV). Furthermore, 90 days of stability was tested using different temperatures conditions where excellent stability was observed. P-NE are non-toxic in (HaCaT), and RAW264.7 cells up to 100 µg/mL further showed effects on ROS and NO production of the cells at 50 µg/mL. The qPCR analysis demonstrated the suppression of pro-inflammatory mediators for (Cox 2, IL-6, IL-1ß, and TNF-α, NF-κB, Ikkα, and iNOS) gene expression. The prepared NE exhibited anti-inflammatory effects, demonstrating its potential as a safe and non-toxic nanomedicine.

Inhibitory Effects of Ginsenoside Ro on the Growth of B16F10 Melanoma via Its Metabolites.

Ginsenoside Ro (Ro), a major saponin derived and isolated from Panax ginseng C.A. Meyer, exerts multiple biological activities. However, the anti-tumour efficacy of Ro remains unclear because of its poor in vitro effects. In this study, we confirmed that Ro has no anti-tumour activity in vitro. We explored the anti-tumour activity of Ro in vivo in B16F10 tumour-bearing mice. The results revealed that Ro considerably suppressed tumour growth with no significant side effects on immune organs and body weight. Zingibroside R1, chikusetsusaponin IVa, and calenduloside E, three metabolites of Ro, were detected in the plasma of Ro-treated tumour-bearing mice and showed excellent anti-tumour effects as well as anti-angiogenic activity. The results suggest that the metabolites play important roles in the anti-tumour efficacy of Ro in vivo. Additionally, the haemolysis test demonstrated that Ro has good biocompatibility. Taken together, the findings of this study demonstrate that Ro markedly suppresses the tumour growth of B16F10-transplanted tumours in vivo, and its anti-tumour effects are based on the biological activity of its metabolites. The anti-tumour efficacy of these metabolites is due, at least in part, to its anti-angiogenic activity.

Synergistic effect of PB2 283M and 526R contributes to enhanced virulence of H5N8 influenza viruses in mice.

Highly pathogenic avian influenza (HPAI) H5N8 virus has caused considerable economic losses to poultry industry and poses a great threat to public health. Our previous study revealed two genetically similar HPAI H5N8 viruses displaying completely different virulence in mice. However, the molecular basis for viral pathogenicity to mammals remains unknown. Herein, we generated a series of reassortants between the two viruses and evaluated their virulence in mice. We demonstrated that 283M in PB2 is a new mammalian virulence marker for H5 viruses and that synergistic effect of amino acid residues 283M and 526R in PB2 is responsible for high virulence of the HPAI H5N8 virus. Analysis of available PB2 sequences showed that PB2 283M is highly conserved among influenza A viruses, while PB2 526R presents in most of human H3N2 and H5N1 isolates. Further study confirmed that the residues 283M and 526R had similar impacts on an HPAI H5N1 virus, suggesting that influenza viruses with both residues may replicate well in mammalian hosts. Together, these results present new insights for synergistic effect of 283M and 526R in PB2 of H5 HPAI virus on virulence to mammalian host, furthering our understanding of the pathogenesis of influenza A virus.

Ameliorative effects of 5-hydroxymethyl-2-furfural (5-HMF) from Schisandra chinensis on alcoholic liver oxidative injury in mice.

The aim of this paper is to evaluate the protective effect of 5-hydroxymethyl-2-furfural (5-HMF) on acute alcohol-induced liver oxidative injury in mice. 5-HMF, a maillard reaction product, was isolated from the fruits of Schisandra chinensis for animal experiments. Experimental ICR mice were pretreated with different doses of 5-HMF (7.5, 15, and 30 mg/kg) for seven days by gavage feeding. Biochemical markers and enzymatic antioxidants from serum and liver tissue were examined. Our results showed that the activities of ALT (alanine aminotransferase), AST (aspartate transaminase), TC (total cholesterol), TG (triglyceride), L-DLC (low density lipoprotein) in serum and the levels of MDA (malondialdehyde) in liver tissue, decreased significantly (p < 0.05) in the 5-HMF-treated group compared with the alcohol group. On the contrary, enzymatic antioxidants CAT (catalase), GSH-Px (glutathione peroxidase), and GSH SOD (superoxide dismutase) were markedly elevated in liver tissue treated with 5-HMF (p < 0.05). Furthermore, the hepatic levels of pro-inflammatory response marker tumor necrosis factor-alpha (TNF-α) and interleukin-1ß (IL-1ß) were significantly suppressed (p < 0.05). Histopathological examination revealed that 5-HMF (30 mg/kg) pretreatment noticeably prevented alcohol-induced hepatocyte apoptosis and fatty degeneration. It is suggested that the hepatoprotective effects exhibited by 5-HMF on alcohol-induced liver oxidative injury may be due to its potent antioxidant properties.

Analytic solution of cut-off wavelength of bend-insensitive fibers based on frustrated total reflection.

An analytic method for solving the cut-off wavelength of single groove-assisted bend-insensitive fiber (BIF) was proposed. Combined with the concepts of frustrated total reflection and numerical aperture, the cut-off wavelength formula for the BIF was deduced. Experimental results showed that, compared to the traditional analytic and numeric methods, this method had higher accuracy in the calculation of cut-off wavelength for BIFs that had a large proportion of depressed inner-cladding layer, which significantly expanded the range of accurately predicting the cut-off wavelength.

Contrasting the distribution kinetics of microplastics and nanoplastics in medaka following exposure and depuration.

Direct ingestion of micro/nanoplastics (MNPs) results in significant accumulation in gastrointestinal (GI) tract of fish. The breathing process of fish makes MNPs easily retained in their gills. However, the uptake of MNPs in other fish organs remains largely unknown, let alone their kinetic processes. Herein, microplastics (MPs) and nanoplastics (NPs) in vivo imaging and precise quantification in various tissues (GI tract, gill, liver, brain, eye, and skin) of seawater (SW)- and freshwater (FW)- acclimated medaka Oryzias melastigma were achieved at an environmentally relevant concentration. Subsequently, the distribution kinetics of MNPs was investigated over a 96-h uptake and 48-h depuration period. MNPs were quickly and mostly captured in GI tract and gill of O. melastigma, and then transferred to liver and brain likely via blood circulation. Such transport was more efficient for NPs as compared to MPs, as evidenced by the consistently higher bioconcentration factors in both SW and FW conditions. The detection of MNPs in eye and skin of O. melastigma was more of an adsorption process, although the specific mechanisms of adsorption and absorption process can hardly be clearly differentiated. This study presented distribution kinetics of MNPs in O. melastigma and highlighted their possible transportation among tissues.

Microplastics and nanoplastics induced differential respiratory damages in tilapia fish Oreochromis niloticus.

With the increasing micro(nano)plastics (MNPs) pollution in aquatic environments, fish respiration is encountering a huge threat. Herein, polystyrene (PS) MNPs with three sizes (80 nm, 2 µm, and 20 µm) were exposed to tilapia Oreochromis niloticus at an environmentally relevant concentration of 100 µg/L for 28 days and their impacts on respiratory function were investigated. Based on the results of oxygen consumption and histological analysis, all the three treatments could induce respiratory damages and such impacts were more severe for the 2 µm and 20 µm treatments than for the 80 nm treatment. These results were explained by the more significant upregulation of egln3 and nadk, and the downregulation of isocitrate. Transcriptomics and metabolomics further revealed that TCA cycle played a key role in respiratory dysfunction induced by micro-sized PS particles, and cytokine and chemokine related functions were simultaneously enriched. Although nano-sized PS particles had the potential to penetrate the respiratory epithelium and reached the internal structure of the O. niloticus gills, they were easily expelled through the blood circulation. Our results highlighted the serious threat of MNPs to fish respiration and provided insights into the differential toxicological mechanisms between micro-sized and nano-sized particles, thus assisting in ecological risk assessments.

Nanoscale dielectric-graphene-dielectric tunable infrared waveguide with ultrahigh refractive indices.

We propose in this paper a dielectric-graphene-dielectric tunable infrared waveguide based on multilayer metamaterials with ultrahigh refractive indices. The waveguide modes with different orders are systematically analyzed with numerical simulations based on both multilayer structures and effective medium approach. The waveguide shows hyperbolic dispersion properties from mid-infrared to far-infrared wavelength, which means the modes with ultrahigh mode indices could be supported in the waveguide. Furthermore, the optical properties of the waveguide modes could be tuned by the biased voltages on graphene layers. The waveguide may have various promising applications in the quantum cascade lasers and bio-sensing.

Resonant coupling in trenched bend-insensitive optical fiber.

We report in this Letter the resonant coupling mechanism in bending trenched bend-insensitive fiber (BIF). It is found that among the trench parameters, the core-trench distance is predominant for optimized BIF design. We reveal that resonant coupling is an intrinsic characteristic of bending trenched BIF, and resonant coupling between the fiber core and the innermost cladding would limit the ultimate bending loss of BIF under tight bend. Resonant coupling is also present in double-trenched BIF, and would impair its bending performance.

Mode-coupling analysis and trench design for large-mode-area low-cross-talk multicore fiber.

The complete analytical solutions of the mode-coupling dynamics for seven-core multicore fibers (MCFs) with identical cores is proposed. All the coupling coefficients C(mn) of adjacent cores and nonadjacent cores as a function of the structural parameters are investigated. It is shown that the coupling coefficients can decrease by adjusting the structural parameters. In addition, the trench-assisted structure could be used for suppressing cross talk in MCF, and the effective area A(eff) can be enlarged without degrading the crosstalk properties. Simulations suggest that low cross talk and/or large A(eff) could be achieved by adjusting the trench parameters. Large mode area could be obtained by utilizing small trench (small trench width c, small refractive index difference Δ(trench)) in the trench-assisted MCF (TA-MCF), whereas low cross talk could be achieved by utilizing larger trench (large trench width c, large Δ(trench)) in the TA-MCF.

Analysis and characterization of Er-Yb codoped-depressed inner cladding fiber.

A 125 µm-diameter erbium-ytterbium-codoped single-mode fiber is reported. The utilization of depressed inner cladding guarantees the improvement of trade-off between the effective area and bending sensitivity compared to step-index profiles. Changes of cutoff wavelength, effective area, and macrobend loss under the influence of various structural parameters, and the balancing selection of core radius and subsidence layer width are investigated systematically. For the laboratory-made depressed inner cladding fiber, a macrobend loss of 0.06 dB/loop for a bending radius as tight as 10 mm was achieved, while maintaining an effective area of 164.22 µm² with intact single-mode properties at 1550 nm. The maximum small signal gain was achieved at 40.9 dB, and the gain fluctuation was less than 1 dB at the C-band. The fiber is suitable for high-power, small, portable, and handy optical fiber devices.

Disturbing ion regulation and excretion in medaka (Oryzias melastigma) gills by microplastics: Insights from the gut-gill axis.

The accumulation of microplastics (MPs) in fish gills has been widely recognized, however, whether such stress could thereby impact the physiological responses of fish gills is still unknown. Here, we investigated the impacts of three sizes (400 nm, 4 µm, 20 µm) of polystyrene (PS) MPs on (Na+, K+, Cl-) ions regulation and ammonia excretion in medaka Oryzias melastigma. Significantly increased net Na+ and K+ flux rates were observed transiently during 0-3 h and 3-9 h, but not during 9-24 h. Such results suggest that the physiological resilience of fish gills regarding Na+ and K+ regulation was unaffected upon the exposure to PS-MPs, probably evidenced by the increased secretion of mucus. However, Cl- regulation and ammonia excretion were significantly impaired, partly in consistent with the damages of ionocytes. The adverse impacts of PS-MPs on Cl- regulation and ammonia excretion were size-dependent, with significant disturbances observed in 4 µm and 20 µm treated group for Cl- regulation, but only in 20 µm treated group for ammonia excretion. The specific enrichment of Shinella and lower abundance of function profiles related to ion transport and metabolism might be responsible for the specific disturbance of Cl- regulation found in the 4 µm treated group. The enrichment of Gemmobacter also accounted for the disturbances of ammonia excretion in 20 µm treated group. Our results highlighted the impacts of PS-MPs on the physiological functions in fish gills.

In utero metabolomic signatures of refined grain intake and risk of gestational diabetes: A metabolome-wide association study.

BACKGROUND: Epidemiologic evidence has linked refined grain intake to a higher risk of gestational diabetes (GDM), but the biological underpinnings remain unclear. OBJECTIVES: We aimed to identify and validate refined grain-related metabolomic biomarkers for GDM risk. METHODS: In a metabolome-wide association study of 91 cases with GDM and 180 matched controls without GDM (discovery set) nested in the prospective Pregnancy Environment and Lifestyle Study (PETALS), refined grain intake during preconception and early pregnancy and serum untargeted metabolomics were assessed at gestational weeks 10-13. We identified refined grain-related metabolites using multivariable linear regression and examined their prospective associations with GDM risk using conditional logistic regression. We further examined the predictivity of refined grain-related metabolites selected by least absolute shrinkage and selection operator regression in the discovery set and validation set (a random PETALS subsample of 38 individuals with and 336 without GDM). RESULTS: Among 821 annotated serum (87.4% fasting) metabolites, 42 were associated with refined grain intake, of which 17 (70.6% in glycerolipids, glycerophospholipids, and sphingolipids clusters) were associated with subsequent GDM risk (all false discovery rate-adjusted P values <0.05). Adding 7 of 17 metabolites to a conventional risk factor-based prediction model increased the C-statistic for GDM risk in the discovery set from 0.71 (95% CI: 0.64, 0.77) to 0.77 (95% CI: 0.71, 0.83) and in the validation set from 0.77 (95% CI: 0.69, 0.86) to 0.81 (95% CI: 0.74, 0.89), both with P-for-difference <0.05. CONCLUSIONS: Clusters of glycerolipids, glycerophospholipids, and sphingolipids may be implicated in the association between refined grain intake and GDM risk, as demonstrated by the significant associations of these metabolites with both refined grains and GDM risk and the incremental predictive value of these metabolites for GDM risk beyond the conventional risk factors. These findings provide evidence on the potential biological underpinnings linking refined grain intake to the risk of GDM and help identify novel disease-related dietary biomarkers to inform diet-related preventive strategies for GDM.

Importance of the average radius of coal particles on determining the methane diffusion coefficient.

The average radius of coal particles is an estimate of the diffusion path in the particle method for determining the diffusion coefficient. It is currently calculated using the arithmetic mean of coal particle sieved intervals. This calculation, however, ignores the coal particle size distribution, resulting in significant deviations when calculating the gas diffusion coefficient. An appropriate average radius calculation method should consider the particle size distribution and the physical essence of diffusion. To accomplish this, a series of methods for calculating the mean particle diameters and their physical significance were reviewed. Next, coal samples were sieved into three intervals, and gas diffusion tests and laser particle size distribution were conducted. Results show that coal particles are within the sieving interval, ranging from 42.01 to 76.18%. By solving the diffusion coefficients using four mean particle diameters based on particle size distribution and diffusive mass transfer, the difference between the arithmetic mean value and these diameters is up to 89.06%. [Formula: see text] and [Formula: see text] are preferred for the calculation of the average radius since they are compatible with coal particle shape and the physical meaning of diffusive mass transfer.

Immunomodulatory Effect of Ginsenoside Rb2 Against Cyclophosphamide-Induced Immunosuppression in Mice.

Ginsenoside Rb2 (Rb2), a fundamental saponin produced and isolated from ginseng (Panax ginseng C.A. Meyer), has a wide range of biological actions. The objective of this investigation was to see if ginsenoside Rb2 has any immunomodulatory properties against cyclophosphamide (CTX)-induced immunosuppression. For the positive control group, levamisole hydrochloride (LD) was used. We discovered that intraperitoneal injection of Rb2 (5, 10, 20 mg/kg) could relieve CTX-induced immunosuppression by enhanced immune organ index, reduced the pathological characteristics of immunosuppression, promoted natural killer (NK) cells viability, improved cell-mediated immune response, boosted the IFN-γ (Interferon-gamma), TNF-α (Tumor necrosis factor-alpha), IL-2 (Interleukin-2), and IgG (Immunoglobulin G), as well as macrophage activity like carbon clearance and phagocytic index. Rb2 significantly elevated the mRNA expression of IL-4 (Interleukin-4), SYK (Tyrosine-protein kinase-SYK), IL-2, TNF-α, and IL-6 (Interleukin-6) in the spleen of CTX-injected animals. Molecular docking results showed that Rb2 had excellent binding properties with IL-4, SYK, IL-2, TNF, and IL-6, indicating the target protein might be strongly correlated with the immunomodulatory effect of Rb2. Taken together, ginsenoside Rb2 can improve the immune function that is declined in CTX-induced immunosuppressed mice, the efficacy maybe due to the regulation of related cytokine and mRNA expression.

How phytoplankton biomass controls metal(loid) bioaccumulation in size-fractionated plankton in anthropogenic-impacted subtropical lakes: A comprehensive study in the Yangtze River Delta, China.

Phytoplankton biomass can significantly affect metal(loid) bioaccumulation in plankton, but the underlying mechanisms are still controversial. We investigated the bioaccumulation of eight metal(loid)s (As, Co, Cu, Hg, Mn, Pb, Se, and Zn) in three size categories of planktonic organisms - seston (0.7-64 µm), mesozooplankton (200-500 µm), and macrozooplankton (>500 µm) - sampled from six freshwater lakes in two seasons in the Yangtze River Delta, China. Our results highlight phytoplankton biomass is the major driver on metal(loid) bioaccumulation in the studied anthropogenic-impacted subtropical lakes, mainly via affecting site-specific water physiochemical characteristics and plankton communities. However, such impact is highly dependent on chlorophyll a (Chl-a) concentration. The bioaccumulation of metal(loid)s in size-fractionated plankton declined significantly with increasing phytoplankton biomass when Chl-a was below ∼50 µg L-1, mainly owing to the reduced metal(loid) bioavailability and subsequent bioaccumulation at more productive sites (with elevated pH and dissolved organic carbon), rather than algal bloom dilution. To a lesser extent, phytoplankton growth dilution and the smaller body-size of zooplankton at more productive sites also contributed to the lower metal(loid) bioaccumulation. The bioaccumulation of metal(loid)s was enhanced under severe algal bloom conditions (when Chl-a concentration was higher than ∼50 µg L-1). Although the underlying mechanisms still require further investigations, the potential risks of metal(loid) bioaccumulation under severe algal bloom conditions deserve special attention.