Zooplankton-based adverse outcome pathways: A tool for assessing endocrine disrupting compounds in aquatic environments.

Endocrine disrupting compounds (EDCs) pose a significant ecological risk, particularly in aquatic ecosystems. EDCs have become a focal point in ecotoxicology, and their identification and regulation have become a priority. Zooplankton have gained global recognition as bioindicators, benefiting from rigorous standardization and regulatory validation processes. This review aims to provide a comprehensive summary of zooplankton-based adverse outcome pathways (AOPs) with a focus on EDCs as toxicants and the utilisation of freshwater zooplankton as bioindicators in ecotoxicological assessments. This review presents case studies in which zooplankton have been used in the development of AOPs, emphasizing the identification of molecular initiating events (MIEs) and key events (KEs) specific to zooplankton exposed to EDCs. Zooplankton-based AOPs may become an important resource for understanding the intricate processes by which EDCs impair the endocrine system. Furthermore, the data sources, experimental approaches, advantages, and challenges associated with zooplankton-based AOPs are discussed. Zooplankton-based AOPs framework can provide vital tools for consolidating toxicological knowledge into a structured toxicity pathway of EDCs, offering a transformative platform for facilitating enhanced risk assessment and chemical regulation.

De Novo Transcriptomic and Life-History Responses of Moina Micrura Under Stress Environment Conditions.

Moina micrura represents a promising model species for ecological and ecotoxicological investigations in tropical freshwater ecosystems. Illumina NovaSeq™ 6000 sequencing was employed in this study to analyze M. micrura across three distinct developmental stages: juvenile, adult, and male. Current study successfully annotated 51,547 unigenes (73.11%) derived from seven (7) different databases. A total of 554 genes were found to be significantly upregulated, while 452 genes showed significant downregulation between juvenile and male. Moreover, 1001 genes were upregulated, whereas 830 genes exhibited downregulation between the adult and male. Analysis of differentially expressed genes revealed upregulation of chitin, cuticle, myosin (MYO), mitogen-activated protein kinases (MAPK), fibrillin (FBN), cytochrome (CYP), glutathione s-transferase (GST), vitellogenin (VTG), acetylcholinesterase (AChE), and transforming growth factor beta (TGFB) under unfavorable environmental conditions (male), as compared to favorable environmental conditions (juveniles and adults). These alterations in gene expression significantly impact the phenological and life-history traits of M. micrura. Furthermore, the upregulation of hemoglobin (HMB), doublesex (DSX), juvenile hormone analogs (JHA), heat shock protein (HSP), and methyltransferase (METT) genes in males initiates the sex-switching effects observed in M. micrura. These findings hold substantial value for researchers interested in determining M. micrura sequences for future investigations of gene expression and comparative reproductive genome analysis within the Moina genus and cladoceran families.

Occurrence, distribution, and ecological risk of bisphenol analogues in marine ecosystem of urbanized coast and estuary.

Bisphenol analogues are prevalent globally because of rampant usage and imprecise processing techniques, prompting alerts about environmental and health hazards. The method employed in this study by solid phase extraction (SPE) and liquid chromatography-tandem quadrupole mass spectrometer (LC-MS/MS) for both quantification and qualitative analysis of the bisphenol compounds in the surface water samples. The coastal and estuarine surface water of Port Dickson and Lukut ranges from 1.32 ng/L to 1890.51 ng/L of bisphenol analogues. BPF mean concentration at 1143.88 ng/L is the highest, followed by BPA and BPS at 59.01 ng/L and 10.96 ng/L, respectively. Based on RQm for bisphenol analogues, the highest for BPF at 2.49 (RQ > 1, high risk), followed by BPS at 0.12 (0.1 < RQ < 1, medium risk) and BPA at 0.09 (0.1 < RQ < 1, medium risk). The presence and current risk of bisphenols analogues should alert the possible water quality degradation soon.

Risk assessment of bisphenol analogues towards mortality, heart rate and stress-mediated gene expression in cladocerans Moina micrura.

Bisphenol A (BPA) is a well-known endocrine-disrupting compound that causes several toxic effects on human and aquatic organisms. The restriction of BPA in several applications has increased the substituted toxic chemicals such as bisphenol F (BPF) and bisphenol S (BPS). A native tropical freshwater cladoceran, Moina micrura, was used as a bioindicator to assess the adverse effects of bisphenol analogues at molecular, organ, individual and population levels. Bisphenol analogues significantly upregulated the expressions of stress-related genes, which are the haemoglobin and glutathione S-transferase genes, but the sex determination genes such as doublesex and juvenile hormone analogue genes were not significantly different. The results show that bisphenol analogues affect the heart rate and mortality rate of M. micrura. The 48-h lethal concentration (LC50) values based on acute toxicity for BPA, BPF and BPS were 611.6 µg L-1, 632.0 µg L-1 and 819.1 µg L-1, respectively. The order of toxicity based on the LC50 and predictive non-effect concentration values were as follows: BPA > BPF > BPS. Furthermore, the incorporated method combining the responses throughout the organisation levels can comprehensively interpret the toxic effects of bisphenol analogues, thus providing further understanding of the toxicity mechanisms. Moreover, the output of this study produces a comprehensive ecotoxicity assessment, which provides insights for the legislators regarding exposure management and mitigation of bisphenol analogues in riverine ecosystems.

Acute toxicity and risk assessment of perfluorooctanoic acid (PFOA) and perfluorooctanesulfonate (PFOS) in tropical cladocerans Moina micrura.

Per- and polyfluoroalkyl substances (PFAS) are gaining worldwide attention because of their toxicity, bioaccumulative and resistance to biological degradation in the environment. PFAS can be categorised into endocrine disrupting chemicals (EDCs) and identified as possible carcinogenic agents for the aquatic ecosystem and humans. Despite this, only a few studies have been conducted on the aquatic toxicity of PFAS, particularly in invertebrate species such as zooplankton. This study evaluated the acute toxicity of two main PFAS, perfluorooctanoic acid (PFOA) and perfluorooctanesulfonate (PFOS), by using freshwater cladocerans (Moina micrura) as bioindicators. This study aimed to assess the adverse effects at different levels of organisations such as organ (heart size and heart rate), individual (individual size and mortality) and population (lethal concentration, LC50). PFOA was shown to be more hazardous than PFOS, with the LC50 values (confidence interval) of 474.7 (350.4-644.5) µg L-1 and 549.6 (407.2-743.9) µg L-1, respectively. As the concentrations of PFOS and PFOA increased, there were declines in individual size and heart rate as compared to the control group. The values of PNECs acquired by using the AF method (PNECAF) for PFOA and PFOS were 0.4747 and 0.5496 µg L-1, respectively. Meanwhile, the PNEC values obtained using the SSD method (PNECSSD) were 1077.0 µg L-1 (PFOA) and 172.5 µg L-1 (PFOS). PNECAF is more protective and conservative compared to PNECSSD. The findings of this study have significant implications for PFOS and PFOA risk assessment in aquatic environments. Thus, it will aid freshwater sustainability and safeguard the human dependency on water resources.

Development of a single-run liquid chromatography-mass spectrometry analysis for the detection of 11 multiclass contaminants of emerging concern using a direct filtration method.

In toxicological analysis, the analytical validation method is important to assess the exact risk of contaminants of emerging concern in the environment. Syringe filters are mainly used to remove impurities from sample solutions. However, the loss of analyte to the syringe filter could be considerable, causing an underestimate of the analyte concentrations. The current study develops and validates simultaneous liquid chromatography-mass spectrometry analysis using a direct filtration method to detect four groups of contaminants of emerging concern. The adsorption of the analyte onto three different matrices and six types of syringe filters is reported. The lowest adsorption of analytes was observed in methanol (16.72%), followed by deionized water (48.19%) and filtered surface lake water (48.94%). Irrespective of the type of the matrices, the lowest average adsorption by the syringe filter was observed in the 0.45 µm polypropylene membrane (15.15%), followed by the 0.20 µm polypropylene membrane (16.10%), the 0.20 µm regenerated cellulose (16.15%), the 0.20 µm polytetrafluoroethylene membrane (47.38%), the 0.45 µm nylon membrane (64.87%) and the 0.20 µm nylon membrane (71.30%). In conclusion, the recommended syringe filter membranes for contaminants of emerging concern analysis are polypropylene membranes and regenerated cellulose, regardless of the matrix used.

Assessment of RNA extraction protocols from cladocerans.

The usage of cladocerans as non-model organisms in ecotoxicological and risk assessment studies has intensified in recent years due to their ecological importance in aquatic ecosystems. The molecular assessment such as gene expression analysis has been introduced in ecotoxicological and risk assessment to link the expression of specific genes to a biological process in the cladocerans. The validity and accuracy of gene expression analysis depends on the quantity, quality and integrity of extracted ribonucleic acid (RNA) of the sample. However, the standard methods of RNA extraction from the cladocerans are still lacking. This study evaluates the extraction of RNA from tropical freshwater cladocerans Moina micrura using two methods: the phenol-chloroform extraction method (QIAzol) and a column-based kit (Qiagen Micro Kit). Glycogen was introduced in both approaches to enhance the recovery of extracted RNA and the extracted RNA was characterised using spectrophotometric analysis (NanoDrop), capillary electrophoresis (Bioanalyzer). Then, the extracted RNA was analysed with reverse transcription polymerase chain reaction (RT-PCR) to validate the RNA extraction method towards downstream gene expression analysis. The results indicate that the column-based kit is most suitable for the extraction of RNA from M. micrura, with the quantity (RNA concentration = 26.90 ± 6.89 ng/µl), quality (A260:230 = 1.95 ± 0.15, A280:230 = 1.85 ± 0.09) and integrity (RNA integrity number, RIN = 7.20 ± 0.16). The RT-PCR analysis shows that the method successfully amplified both alpha tubulin and actin gene at 33-35 cycles (i.e. Ct = 32.64 to 33.48). The results demonstrate that the addition of glycogen is only suitable for the phenol-chloroform extraction method. RNA extraction with high and comprehensive quality control assessment will increase the accuracy and reliability of downstream gene expression, thus providing more ecotoxicological data at the molecular biological level on other freshwater zooplankton species.

Occurrence, abundance, and distribution of microplastics pollution: an evidence in surface tropical water of Klang River estuary, Malaysia.

Microplastics have been considered as contaminants of emerging concern due to ubiquity in the environment; however, the occurrence of microplastics in river estuaries is scarcely investigated. The Klang River estuary is an important ecosystem that receives various contaminants from urbanised, highly populated areas and the busiest maritime centre in Selangor, Malaysia. This study investigates the abundance and characteristics of microplastics in surface water of the Klang River estuary. The abundance of microplastics ranged from 0.5 to 4.5 particles L-1 with a mean abundance of 2.47 particles L-1. There is no correlation between the abundance of microplastics and physicochemical properties, while there is a strong correlation between salinity and conductivity. The microplastics were characterised with a stereomicroscope and attenuated total reflection-Fourier transform infrared spectroscopy to analyse size, shape, colour, and polymer composition. The microplastics in the surface water were predominantly in the 300-1000 µm size class, followed by > 1000 µm and < 300 µm, and were mostly transparent fibres, fragments, and pellets. Polyamide and polyethylene were the main polymer types in the composition of the microplastics, suggesting that the microplastics originated from heavily urbanised and industrial locations such as the port, jetty, and residential areas. The widespread occurrence of microplastics in the environment and subsequent penetration of aquatic food webs may pose a serious threat to organisms. This study provides baseline data and a framework for further investigation of microplastic contamination in estuaries.

Accumulation and risk assessment of heavy metals employing species sensitivity distributions in Linggi River, Negeri Sembilan, Malaysia.

The constant increase of heavy metals into the aqueous environment has become a contemporary global issue of concern to government authorities and the public. The study assesses the concentration, distribution, and risk assessment of heavy metals in freshwater from the Linggi River, Negeri Sembilan, Malaysia. Species sensitivity distribution (SSD) was utilised to calculate the cumulative probability distribution of toxicity from heavy metals. The aquatic organism's toxicity data obtained from the ECOTOXicology knowledgebase (ECOTOX) was used to estimate the predictive non-effects concentration (PNEC). The decreasing sequence of hazardous concentration (HC5) was manganese > aluminium > copper > lead > arsenic > cadmium > nickel > zinc > selenium, respectively. The highest heavy metal concentration was iron with a mean value of 45.77 µg L-1, followed by manganese (14.41 µg L-1) and aluminium (11.72 µg L-1). The mean heavy metal pollution index (HPI) value in this study is 11.52, implying low-level heavy metal pollutions in Linggi River. The risk quotient (RQ) approaches were applied to assess the potential risk of heavy metals. The RQ shows a medium risk of aluminium (RQm = 0.1125) and zinc (RQm = 0.1262); a low risk of arsenic (RQm = 0.0122) and manganese (RQm = 0.0687); and a negligible risk of cadmium (RQm = 0.0085), copper (RQm = 0.0054), nickel (RQm = 0.0054), lead (RQm = 0.0016) and selenium (RQm = 0.0012). The output of this study produces comprehensive pollution risk, thus provides insights for the legislators regarding exposure management and mitigation.

Occurrence and distribution of endocrine-disrupting chemicals in mariculture fish and the human health implications.

The presence and distribution of endocrine-disrupting chemicals (EDCs) in the mariculture fish from Pulau Kukup, Johor of Malaysia have been studied along with the impact on human health. Six different species of mariculture fish were collected, due to their high consumption in the Asian region-especially Malaysia, to assess their levels of EDCs. The highest concentration of EDCs detected in the muscle was dexamethasone (2.37-15.84 ng/g) and (0.77-13.41 ng/g), in the liver was dexamethasone (<2.54-43.56 ng/g) and progesterone (2.23-9.78 ng/g), and in the reproductive organ are dexamethasone (<2.54-37.23 ng/g) and caffeine (0.21-18.92 ng/g). The human health risk assessment in the current study suggested that there is no potential risk to the consumer because the hazard index was below 1 (HI < 1). The present study provides information on the pollution profile of EDCs and the associated human health risk with EDCs in mariculture fish.

Iminodiacetic acid modified kenaf fiber for waste water treatment.

In the present study, iminodiacetic acid (IDA)-modified kenaf fiber, K-IDA formed by the chemical modification of plant kenaf biomass was tested for its efficacy as a sorbent material towards the purification of waste water. The K-IDA fiber was first characterized by the instrumental techniques like Fourier transform infrared (FTIR) analysis, elemental analysis (CHNSO), and scanning electron microscopy (SEM). On testing for the biosorption, we found that the K-IDA has an increment in the adsorption of Cu2+ ions as compared against the untreated fiber. The Cu2+ ions adsorption onto K-IDA fits very well with the Langmuir model and the adsorption maximum achieved to be 91.74mg/g. Further, the adsorption kinetics observed to be pseudo second-order kinetics model and the Cu2+ ions adsorption is a spontaneous endothermic process. The desorption study indicates a highest percentage of Cu2+ of 97.59% from K-IDA under 1M HCl solution against H2SO4 (72.59%) and HNO3 (68.66%). The reusability study indicates that the efficiency did not change much until the 4th cycle and also providing enough evidence for the engagement of our biodegradable K-IDA fiber towards the removal of Cu2+ ions in real-time waste water samples obtained from the electroplating and wood treatment industries.