Advances of microplastics ingestion on the morphological and behavioral conditions of model zebrafish: A review.

Concerns have been conveyed regarding the availability and hazards of microplastics (MPs) in aquatic biota due to their widespread presence in aquatic habitats. Zebrafish (Danio rerio) are widely used as a model organism to study the adverse impacts of MPs due to their several compelling advantages, such as their small size, ease of breeding, inexpensive maintenance, short life cycle, year-round spawning, high fecundity, fewer legal restrictions, and genetic resemblances to humans. Exposure of organisms to MPs produces physical and chemical toxic effects, including abnormal behavior, oxidative stress, neurotoxicity, genotoxicity, immune toxicity, reproductive imbalance, and histopathological effects. But the severity of the effects is size and concentration-dependent. It has been demonstrated that smaller particles could reach the gut and liver, while larger particles are only confined to the gill, the digestive tract of adult zebrafish. This thorough review encapsulates the current body of literature concerning research on MPs in zebrafish and demonstrates an overview of MPs size and concentration effects on the physiological, morphological, and behavioral characteristics of zebrafish. Finding gaps in the literature paves the way for further investigation.

MCF-7 cell - derived exosomes were involved in protecting source cells from the damage caused by tributyltin chloride via transport function.

Tributyltin chloride (TBTC) is a ubiquitous environmental pollutant with various adverse effects on human health. Exosomes are cell - derived signaling and substance transport vesicles. This investigation aimed to explore whether exosomes could impact the toxic effects caused by TBTC via their transport function. Cytotoxicity, DNA and chromosome damage caused by TBTC on MCF-7 cells were analyzed with CCK-8, flow cytometry, comet assay and micronucleus tests, respectively. Exosomal characterization and quantitative analysis were performed with ultracentrifugation, transmission electron microscope (TEM) and bicinchoninic acid (BCA) methods. TBTC content in exosomes was detected with Liquid Chromatography-Mass Spectrometry (LC-MS). The impacts of exosomal secretion on the toxic effects of TBTC were analyzed. Our data indicated that TBTC caused significant cytotoxicity, DNA and chromosome damage effects on MCF-7 cells, and a significantly increased exosomal secretion. Importantly, TBTC could be transported out of MCF-7 cells by exosomes. Further, when exosomal secretion was blocked with GW4869, the toxic effects of TBTC were significantly exacerbated. We concluded that TBTC promoted exosomal secretion, which in turn transported TBTC out of the source cells to alleviate its toxic effects. This investigation provided a novel insight into the role and mechanism of exosomal release under TBTC stress.

Integrated transcriptomic and metabolomic approaches reveal molecular response and potential biomarkers of the deep-sea mussel Gigantidas platifrons to copper exposure.

Metal pollution caused by deep-sea mining activities has potential detrimental effects on deep-sea ecosystems. However, our knowledge of how deep-sea organisms respond to this pollution is limited, given the challenges of remoteness and technology. To address this, we conducted a toxicity experiment by using deep-sea mussel Gigantidas platifrons as model animals and exposing them to different copper (Cu) concentrations (50 and 500 µg/L) for 7 days. Transcriptomics and LC-MS-based metabolomics methods were employed to characterize the profiles of transcription and metabolism in deep-sea mussels exposed to Cu. Transcriptomic results suggested that Cu toxicity significantly affected the immune response, apoptosis, and signaling processes in G. platifrons. Metabolomic results demonstrated that Cu exposure disrupted its carbohydrate metabolism, anaerobic metabolism and amino acid metabolism. By integrating both sets of results, transcriptomic and metabolomic, we find that Cu exposure significantly disrupts the metabolic pathway of protein digestion and absorption in G. platifrons. Furthermore, several key genes (e.g., heat shock protein 70 and baculoviral IAP repeat-containing protein 2/3) and metabolites (e.g., alanine and succinate) were identified as potential molecular biomarkers for deep-sea mussel's responses to Cu toxicity. This study contributes novel insight for assessing the potential effects of deep-sea mining activities on deep-sea organisms.

Congener-specific fate and impact of microcystins in the soil-earthworm system.

Microcystins (MCs) have a significant influence on aquatic ecosystems, but little is known about their terrestrial fate and impact. Here, we investigated the fate of two MCs (MC-LR and MC-RR) in the soil-earthworm system, with consideration of their congener-specific impact on earthworm health, soil bacteria, and soil metabolome. Although MCs had little acute lethal effect on earthworms, they caused obvious growth inhibition and setae rupture. Relative to MC-RR, MC-LR exhibited higher bioaccumulation and the resulting dermal lesions and deformation of longitudinal muscles. While the incorporation of both MCs into soils stimulated pathogenic bacteria and depressed oxidative stress tolerant bacteria, the response among soil nitrification and glutathione metabolism differed between the two congeners. The dissipation kinetics of MCs obeyed the first-order model. Earthworms stimulated soil N-cycling enzyme activities, increased the abundance of MC-degrading bacteria, and promoted bacterial metabolic functions related to glutathione metabolism, xenobiotics biodegradation, and metabolism of amino acids that comprise MCs, which accelerated the dissipation of MC-LR and MC-RR by 227% and 82%, respectively. These results provide evidence of significant congener differences in the terrestrial fate and impact of MCs, which will enable a better understanding of their role in mediating soil functions and ecosystem services.

Illicit Drugs in Surface Waters: How to Get Fish off the Addictive Hook.

The United Nations World Drug Report published in 2022 alarmed that the global market of illicit drugs is steadily expanding in space and scale. Substances of abuse are usually perceived in the light of threats to human health and public security, while the environmental aspects of their use and subsequent emissions usually remain less explored. However, as with other human activities, drug production, trade, and consumption of drugs may leave their environmental mark. Therefore, this paper aims to review the occurrence of illicit drugs in surface waters and their bioaccumulation and toxicity in fish. Illicit drugs of different groups, i.e., psychostimulants (methamphetamines/amphetamines, cocaine, and its metabolite benzoylecgonine) and depressants (opioids: morphine, heroin, methadone, fentanyl), can reach the aquatic environment through wastewater discharge as they are often not entirely removed during wastewater treatment processes, resulting in their subsequent circulation in nanomolar concentrations, potentially affecting aquatic biota, including fish. Exposure to such xenobiotics can induce oxidative stress and dysfunction to mitochondrial and lysosomal function, distort locomotion activity by regulating the dopaminergic and glutamatergic systems, increase the predation risk, instigate neurological disorders, disbalance neurotransmission, and produce histopathological alterations in the brain and liver tissues, similar to those described in mammals. Hence, this drugs-related multidimensional harm to fish should be thoroughly investigated in line with environmental protection policies before it is too late. At the same time, selected fish species (e.g., Danio rerio, zebrafish) can be employed as models to study toxic and binge-like effects of psychoactive, illicit compounds.

miR-584-5p / Ykt6 - mediated autophagy - lysosome - exosome pathway as a critical route affecting the toxic effects of lead on HK-2 cells.

Lead is a widespread environmental pollutant with serious adverse effects on human health, but the mechanism underlying its toxicity remains elusive. This study aimed to investigate the role of miR-584-5p / Ykt6 axis in the toxic effect of lead on HK-2 cells and the related mechanism. Our data suggested that lead exposure caused significant cytotoxicity, DNA and chromosome damage to HK-2 cells. Mechanistically, lead exposure down-regulated miR-584-5p and up-regulated Ykt6 expression, consequently, autophagosomal number and autophagic flux increased, lysosomal number and activity decreased, exosomal secretion increased. Interestingly, when miR-584-5p level was enhanced with mimic, autophagosomal number and autophagic flux decreased, lysosomal number and activity increased, ultimately, exosomal secretion was down-regulated, which resulted in significant aggravated toxic effects of lead. Further, directly blocking exosomal secretion with inhibitor GW4869 also resulted in exacerbated toxic effects of lead. Herein, we conclude that miR-584-5p / Ykt6 - mediated autophagy - lysosome - exosome pathway may be a critical route affecting the toxic effects of lead on HK-2 cells. We provide a novel insight into the mechanism underlying the toxicity of lead on human cells.

Ecotoxicological evaluation of chitosan biopolymer films particles in adult zebrafish (Danio rerio): A comparative study with polystyrene microplastics.

To mitigate the environmental impact of microplastics (MPs), the scientific community has innovated sustainable and biodegradable polymers as viable alternatives to traditional plastics. Chitosan, the deacetylated form of chitin, stands as one of the most thoroughly investigated biopolymers and has garnered significant interest due to its versatile applications in both medical and cosmetic fields. Nevertheless, there is still a knowledge gap regarding the impact that chitosan biopolymer films (CBPF) may generate in aquatic organisms. In light of the foregoing, this study aimed to assess and compare the potential effects of CBPF on the gastrointestinal tract, gills, brain, and liver of Danio rerio against those induced by MPs. The findings revealed that both CBPF and MPs induced changes in the levels of oxidative stress biomarkers across all organs. However, it is essential to note that our star plots illustrate a tendency for CBPF to activate antioxidant enzymes and for MPs to produce oxidative damage. Regarding gene expression, our findings indicate that MPs led to an up-regulation in the expression of genes associated with apoptotic response (p53, casp3, cas9, bax, and bcl2) in all fish organs. Meanwhile, CBPF produced the same effect in genes related to antioxidant response (nrf1 and nrf2). Overall, our histological observations substantiated these effects, revealing the presence of plastic particles and tissue alterations in the gills and gastrointestinal tract of fish subjected to MPs. From these results, it can be concluded that CBPF does not represent a risk to fish after long exposure.

Combined toxic effects of polystyrene nanoplastics and lead on Chlorella vulgaris growth, membrane lipid peroxidation, antioxidant capacity, and morphological alterations.

In recent years, there has been a significant rise in the utilization of amino-functionalized polystyrene nanoplastics (PS-NH2). This surge in usage can be attributed to their exceptional characteristics, including a substantial specific surface area, high energy, and strong reactivity. These properties make them highly suitable for a wide range of industrial and medical applications. Nevertheless, there is a growing apprehension regarding their potential toxicity to aquatic organisms, particularly when considering the potential impact of heavy metals like lead (Pb) on the toxicity of PS-NH2. Herein, we examined the toxic effects of sole PS-NH2 (90 nm) at five concentrations (e.g., 0, 0.125, 0.25, 0.5, and 1 mg/L), as well as the simultaneous exposure of PS-NH2 and Pb2+ (using two environmental concentrations, e.g., 20 µg/L for Pb low (PbL) and 80 µg/L for Pb higher (PbH)) to the microalga Chlorella vulgaris. After a 96-h exposure, significant differences in chlorophyll a content and algal growth (biomass) were observed between the control group and other treatments (ANOVA, p < 0.05). The algae exposed to PS-NH2, PS-NH2 + PbL, and PS-NH2 + PbH treatment groups exhibited dose-dependent toxicity responses to chlorophyll a content and biomass. According to the Abbott toxicity model, the combined toxicity of treatment groups of PS-NH2 and PbL,H showed synergistic effects. The largest morphological changes such as C. vulgaris' size reduction and cellular aggregation were evident in the medium treated with elevated concentrations of both PS-NH2 and Pb2+. The toxicity of the treatment groups followed the sequence PS-NH2 < PS-NH2 + PbL < PS-NH2 + PbH. These results contribute novel insights into co-exposure toxicity to PS-NH2 and Pb2+ in algae communities.

The two faces of antibiotics: an overview of the effects of antibiotic residues in foodstuffs.

Antibiotics, which have been used for many years to treat infections, also play an important role in food contamination with antibiotic residues. There is also unnecessary use of antibiotics, particularly to increase production efficiency. Non-compliance with withdrawal periods and maximum residue limits (MRLs) for antibiotics used in food-producing animals results in undesirable events, such as allergic reactions, teratogenicity, carcinogenicity, changes in the microbiota and, in particular, antibiotic resistance. Therefore, it may be useful to avoid unnecessary use of antibiotics, to limit the use of antibiotics and to turn to alternatives that can be used instead of antibiotics. The aim of this review is to provide information on the undesirable effects of antibiotic residues in food-producing organisms and in the environment, their determination, and the precautions that can be taken.

Nanoplastic toxicity induces metabolic shifts in Populus × euramericana cv. '74/76' revealed by multi-omics analysis.

There is increasing global concern regarding the pervasive issue of plastic pollution. We investigated the response of Populus × euramericana cv. '74/76' to nanoplastic toxicity via phenotypic, microanatomical, physiological, transcriptomic, and metabolomic approaches. Polystyrene nanoplastics (PS-NPs) were distributed throughout the test plants after the application of PS-NPs. Nanoplastics principally accumulated in the roots; minimal fractions were translocated to the leaves. In leaves, however, PS-NPs easily penetrated membranes and became concentrated in chloroplasts, causing thylakoid disintegration and chlorophyll degradation. Finally, oxidant damage from the influx of PS-NPs led to diminished photosynthesis, stunted growth, and etiolation and/or wilting. By integrating dual-omics data, we found that plants could counteract mild PS-NP-induced oxidative stress through the antioxidant enzyme system without initiating secondary metabolic defense mechanisms. In contrast, severe PS-NP treatments promoted a shift in metabolic pattern from primary metabolism to secondary metabolic defense mechanisms, an effect that was particularly pronounced during the upregulation of flavonoid biosynthesis. Our findings provide a useful framework from which to further clarify the roles of key biochemical pathways in plant responses to nanoplastic toxicity. Our work also supports the development of effective strategies to mitigate the environmental risks of nanoplastics by biologically immobilizing them in contaminated lands.

The toxic effects of petroleum pollutants to microalgae in marine environment.

Marine oil pollution is one of the major global environmental pollution problems. Marine microalgae are the foundation of the marine food chain, providing the main primary productivity of the ocean. They not only maintain the energy flow and material cycle of the entire marine ecosystem, but also play an important role in regulating global climate change. Exploring the impact of petroleum pollutants on marine microalgae is extremely important for studying marine environmental pollution. This review first introduced the sources, compositions, and forms of petroleum pollutants and their migration and transformation processes in the ocean. Then, the toxic effects of petroleum pollutants on marine microalgae were summarized. The growth of marine microalgae showed low-concentration promotion and high-concentration inhibition. The population growth and interspecific relationships of marine microalga was changed and the photosynthesis of marine microalgae was influenced. Finally, potential research directions and suggestions for marine microalgae in the future were proposed.

Ecotoxicity and trophic transfer of metallic nanomaterials in aquatic ecosystems.

Metallic nanomaterials (MNMs) possess unique properties that have led to their widespread application in fields such as electronics and medicine. However, concerns about their interactions with environmental factors and potential toxicity to aquatic life have emerged. There is growing evidence suggesting MNMs can have detrimental effects on aquatic ecosystems, and are potential for bioaccumulation and biomagnification in the food chain, posing risks to higher trophic levels and potentially humans. While many studies have focused on the general ecotoxicity of MNMs, fewer have delved into their trophic transfer within aquatic food chains. This review highlights the ecotoxicological effects of MNMs on aquatic systems via waterborne exposure or dietary exposure, emphasizing their accumulation and transformation across the food web. Biomagnification factor (BMF), the ratio of the contaminant concentration in predator to that in prey, was used to evaluate the biomagnification due to the complex nature of aquatic food chains. However, most current studies have BMF values of less than 1 indicating no biomagnification. Factors influencing MNM toxicity in aquatic environments include nanomaterial properties, ion variations, light, dissolved oxygen, and pH. The multifaceted interactions of these variables with MNM toxicity remain to be fully elucidated. We conclude with recommendations for future research directions to mitigate the adverse effects of MNMs in aquatic ecosystems and advocate for a cautious approach to the production and application of MNMs.

Current trends and mismatches on fungicide use and assessment of the ecological effects in freshwater ecosystems.

Despite increasing evidence of off-site ecological impacts of pesticides and policy efforts worldwide, pesticide use is still far from being ecologically sustainable. Fungicides are among the most sold classes of pesticides and are crucial to ensure global food supply and security. This study aimed to identify potential gaps of knowledge and mismatches between research and usage data of fungicides by: (i) systematizing the current trends in global sales of fungicides, focusing on the European context in particular (where they are proportionally important); (ii) reviewing the scientific literature on the impacts of synthetic fungicides on non-target freshwater organisms. Sales data revealed important global and regional asymmetries in the relative importance of fungicides and the preferred active ingredients. The literature review on the ecological effects of fungicides disclosed a mismatch between the most studied and the most sold substances, as well as a bias towards the use of single species assays with standard test organisms. To ensure a proper evaluation, risk scenarios should focus on a regional scale, and research agendas must highlight sensitive aquatic ecorreceptors and improve the crosstalk between analytical and sales data.

Toxic Impact of Sheathmar on Oreochromis mossambicus: A Comprehensive Biomarker Analysis.

This study assessed the toxicity of the Sheathmar pesticide on Oreochromis mossambicus. The 96-h median lethal concentration (LC50) was determined using probit analysis, with survivability assessed using general threshold survival models (GUTS). Over 45 days, fish exposed to sublethal Sheathmar doses were studied for changes in growth, blood profiles, and liver enzymes. According to the findings, the LC50 values of Sheathmar for Oreochromis mossambicus at different time intervals were determined as follows: 3016.64 µg/l at 24 h, 2723.13 µg/l at 48 h, 2415.45 µg/l at 72 h, and 2154.87 µg/l at 96 h. The GUTS-SD model effectively predicted fish survivability after Sheathmar exposure. Sublethal exposure led to significant alterations in growth and hematological, biochemical, and stress enzyme parameters in Oreochromis mossambicus. Moreover, the correlation matrix, integrated biomarker response (IBR), and biomarker response index (BRI) highlighted significant impacts on multiple biomarkers in Oreochromis mossambicus. Thus, the results underscore the harmful effects of Sheathmar on Oreochromis mossambicus, indicating a need for further study and mitigation.

A study on the subchronic toxicity of triclocarban to the early-life development of oryzias melastigma and focused on the analysis of osmoregulatory regulation mechanisms.

Triclocarban (TCC), a novel antimicrobial agent found in personal care products, has been extensively detected in marine environments. However, research on the toxic effects of TCC on marine organisms remains inadequate. This study delved into the subchronic toxic effects of TCC on the early life stages of marine medaka (Oryzias melastigma, O. melastigma), revealing that TCC could reduce embryo heart rate and hatching rate while diminishing the survival rate of larvae. Biomarker assays indicated that TCC could inflict damage on the embryos' antioxidant and nervous systems. Transcriptomic analysis suggested that TCC could impact cell growth, reproduction, and various life processes, activating cancer signaling pathways, increasing the likelihood of cancer, and exerting toxic effects on the immune and osmoregulatory systems. To validate and enhance our understanding of TCC's unique toxic impact on the osmoregulatory system of O. melastigma, we conducted homology modeling and molecular docking analyses on the protein involved in osmoregulation. The study intuitively revealed the potential binding affinity of TCC to sodium/potassium-transporting ATPase subunit alph (ATP1A1), indicating its ability to disrupt osmotic balance in marine fish by affecting this target protein. In summary, the results of this study will further enhance our comprehension of the potential toxic effects and mechanisms of TCC on the early stages of marine fish, with a specific focus on its unique toxic effects in osmoregulation.

[Human Accumulation and Toxic Effects of Microplastics：A Critical Review].

The effect of microplastics on the ecological environment and human health has become a topical issue， and research on the risks and harmful effects of MPs on human health in particular has attracted widespread attention. Due to the characteristics of small size， low degradability， and easy migration， MPs continuously migrate from the environment to the human body， and their main exposure pathways are oral ingestion， inhalation， and dermal contact， with the main exposure media being food， drinking water， dust， personal care products， etc. MPs have been detected in organs， fluids， and excreta of digestive， respiratory， cardiovascular， reproductive systems， etc. The abundance range of MPs in the human body is 0-1 206.94 particles per gram. After entering the human body， MPs can cause cytotoxicity， mitochondrial toxicity， DNA damage， cell membrane damage， and other effects on human cells and organs， leading to serious consequences such as local inflammation， ecological imbalance， metabolic disorders， etc.， in various systems. Owing to their small specific surface area， they can also adsorb pollutants such as heavy metals， organic pollutants， antibiotics， pathogens， and harmful microorganisms， causing combined toxicity and immunotoxicity. In the end， we highlighted general deficiencies in existing studies and provided directions for future research on the influence of MPs on human health.

Adverse effects of fenpropathrin on the intestine of common carp (Cyprinus carpio L.) and the mechanism involved.

Fenpropathrin (FEN), a pyrethroid pesticide, is frequently detected in natural water bodies, unavoidable pose adverse effects to aquatic organisms. However, the harmful effects and potential mechanisms of FEN on aquatic species are poorly understood. In this study, common carp were treatment with FEN at 0.45 and 1.35 µg/L for 14 d, and the toxic effects and underlying mechanisms of FEN on the intestine of carp were revealed. RNA-seq results showed that FEN exposure cause a wide range of transcriptional alterations in the intestine and the differentially expressed genes were mainly enrichment in the pathways related to immune and metabolism. Specifically, FEN exposure induced pathological damage and altered submicroscopic structure of the intestine, elevated the levels of Bacteroides fragilis enterotoxin, altered the contents of claudin-1, occludin, and zonula occluden-1 (ZO-1), and causing injury to the intestinal barrier. In addition, inflammation-related index TNF-α in the serum and IL-6 in the intestinal tissues were generally increased after FEN exposure. Moreover, FEN exposure promoted an increase in reactive oxygen species (ROS), altered the levels of superoxide dismutase (SOD), catalase (CAT), and glutathione (GSH), upregulated the contents of malondialdehyde (MDA) in the intestines. The apoptosis-related parameter cytochrome c, caspase-9, and caspase-3 were significantly altered, indicating that inflammation reaction, oxidative stress, and apoptosis may be involved in the toxic mechanism of FEN on carp. Moreover, FEN treatment also altered the intestinal flora community significantly, which may affect the intestinal normal physiological function and thus affect the growth of fish. Overall, the present study help to clarify the intestinal reaction mechanisms after FEN treatment, and provide a basis for the risk assessment of FEN.

Comprehensive molecular level characterization of protein- and polyphenol-rich tara (Caesalpinia spinosa) seed germ flour suggests novel hypothesis about possible accidental hazards.

Tara (Caesalpinia spinosa, Leguminosae) seed germ (TSG), a by-product of tara gum (E417) extraction, has been used as a protein- and polyphenol-rich food ingredient for human and animal nutrition. Nevertheless, TSG is the alleged culprit for a recent foodborne outbreak of even severe acute illnesses that have affected hundreds of individuals in the USA, perhaps triggered by nonprotein amino acids such as baikiain. Herein, the composition of TSG has been characterized at molecular level, with a focus on proteins, phenolics, lipids, and mineral composition. TSG contains 43.4 % (w/w) proteins, tentatively identified for the first time by proteomics, and 14 % lipids, consisting of 83.6 % unsaturated fatty acids, especially linoleic acid. Ash is surprising high (6.5 %) because of an elevated concentration of P, K, Ca, and Mg. The detection of a rare earth element such as gadolinium (Gd, 1.6 mg kg-1), likely sourced from anthropogenic pollution, suggests alternative hypotheses for the origin of TSG hazards.

Fecal calcium levels of bird nestlings as a potential indicator of species-specific metal sensitivity.

Sensitivity of bird species to environmental metal pollution varies but there is currently no general framework to predict species-specific sensitivity. Such information would be valuable from a conservation point-of-view. Calcium (Ca) has antagonistic effects on metal toxicity and studies with some common model species show that low dietary and circulating calcium (Ca) levels indicate higher sensitivity to harmful effects of toxic metals. Here we measured fecal Ca and five other macroelement (potassium K, magnesium Mg, sodium Na, phosphorus P, sulphur S) concentrations as proxies for dietary levels in 66 bird species to better understand their interspecific variation and potential use as an indicator of metal sensitivity in a wider range of species (the main analyses include 39 species). We found marked interspecific differences in fecal Ca concentration, which correlated positively with Mg and negatively with Na, P and S levels. Lowest Ca concentrations were found in insectivorous species and especially aerial foragers, such as swifts (Apodidae) and swallows (Hirundinidae). Instead, ground foraging species like starlings (Sturnidae), sparrows (Passeridae), cranes (Gruidae) and larks (Alaudidae) showed relatively high fecal Ca levels. Independent of phylogeny, insectivorous diet and aerial foraging seem to indicate low Ca levels and potential sensitivity to toxic metals. Our results, together with information published on fecal Ca levels and toxic metal impacts, suggest that fecal Ca levels are a promising new tool to evaluate potential metal-sensitivity of birds, and we encourage gathering such information in other bird species. Information on the effects of metals on breeding parameters in a wider range of bird species would also help in ranking species by their sensitivity to metal pollution.

In situ monitoring of toxic effects of algal toxin on cells by a novel microfluidic flow cytometry instrument.

Algal toxins produced by microalgae, such as domoic acid (DA)1, have toxic effects on humans. However, toxicity tests using mice only yield lethal doses of algal toxins without providing insights into the mechanism of action on cells. In this study, a fast segmentation of microfluidic flow cytometry cell images based on the bidirectional background subtraction (BBS)2 method was developed to get the visual evidence of apoptosis in both bright-field and fluorescence images. This approach enables mapping of changes in cell morphology and activity under algal toxins, allowing for fast (within 60 s) and automated biological detection. By combining microfluidics with flow cytometry, the intricate cellular-level reaction process can be observed in micro samples of 293 T cells and mouse spleen cells, offering potential for future in vitro experiments.