SWATH-MS-based proteogenomic analysis reveals the involvement of alternative splicing in poplar upon lead stress.

Alternative splicing (AS) regulates gene expression and increases proteomic diversity for the fine tuning of stress responses in plants, but the exact mechanism through which AS functions in plant stress responses is not thoroughly understood. Here, we investigated how AS functions in poplar (Populus trichocarpa), a popular plant for bioremediation, in response to lead (Pb) stress. Using a proteogenomic analysis, we determine that Pb stress induced alterations in AS patterns that are characterized by an increased use of nonconventional splice sites and a higher abundance of Pb-responsive splicing factors (SFs) associated with Pb-responsive transcription factors. A strong Pb(II)-inducible chaperone protein, PtHSP70, that undergoes AS was further characterized. Overexpression of its two spliced isoforms, PtHSP70-AS1 and PtHSP70-AS2, in poplar and Arabidopsis significantly enhances the tolerance to Pb. Further characterization shows that both isoforms can directly bind to Pb(II), and PtHSP70-AS2 exhibits 10-fold higher binding capacities and a greater increase in expression under Pb stress, thereby reducing cellular toxicity through Pb(II) extrusion and conferring Pb tolerance. AS of PtHSP70 is found to be regulated by PtU1-70K, a Pb(II)-inducible core SF involved in 5'-splice site recognition. Because the same splicing pattern is also found in HSP70 orthologs in other plant species, AS of HSP70 may be a common regulatory mechanism to cope with Pb(II) toxicity. Overall, we have revealed a novel post-transcriptional machinery that mediates heavy metal tolerance in diverse plant species. Our findings offer new molecular targets and bioengineering strategies for phytoremediation and provide new insight for future directions in AS research.

Cell Plasticity in a Mouse Model of Benign Prostate Hyperplasia Drives Amplification of Androgen-Independent Epithelial Cell Populations Sensitive to Antioxidant Therapy.

Benign prostate hyperplasia (BPH) is caused by the nonmalignant enlargement of the transition zone of the prostate gland, leading to lower urinary tract symptoms. Although current medical treatments are unsatisfactory in many patients, the limited understanding of the mechanisms driving disease progression prevents the development of alternative therapeutic strategies. The probasin-prolactin (Pb-PRL) transgenic mouse recapitulates many histopathological features of human BPH. Herein, these alterations parallel urodynamic disturbance reminiscent of lower urinary tract symptoms. Single-cell RNA-sequencing analysis of Pb-PRL mouse prostates revealed that their epithelium mainly includes low-androgen signaling cell populations analogous to Club/Hillock cells enriched in the aged human prostate. These intermediate cells are predicted to result from the reprogramming of androgen-dependent luminal cells. Pb-PRL mouse prostates exhibited increased vulnerability to oxidative stress due to reduction of antioxidant enzyme expression. One-month treatment of Pb-PRL mice with anethole trithione (ATT), a specific inhibitor of mitochondrial ROS production, reduced prostate weight and voiding frequency. In human BPH-1 epithelial cells, ATT decreased mitochondrial metabolism, cell proliferation, and stemness features. ATT prevented the growth of organoids generated by sorted Pb-PRL basal and LSCmed cells, the two major BPH-associated, androgen-independent epithelial cell compartments. Taken together, these results support cell plasticity as a driver of BPH progression and therapeutic resistance to androgen signaling inhibition, and identify antioxidant therapy as a promising treatment of BPH.

Offsetting pb induced oxidative stress in Vicia faba plants by foliar spray of chitosan through adjustment of morpho-biochemical and molecular indices.

In the course of their life, plants face a multitude of environmental anomaly that affects their growth and production. In recent decades, lead (Pb) gained an increasing attention as it is among the most significant contaminants in the environment. Therefore, in this study the effects of Pb concentrations (0, 50 and 100 ppm) on Vicia faba plants and attempts to alleviate this stress using chitosan (Chs; 0 and 0.1%) were performed. The results validated that with increasing Pb concentrations, a decline in growth, pigments and protein contents was observed. In the same time, a significant upsurge in the stress markers, both malondialdehyde (MDA) and H2O2, was observed under Pb stress. Nonetheless, foliar spraying with Chs improves the faba bean growth, pigment fractions, protein, carbohydrates, reduces MDA and H2O2 contents and decreases Pb concentrations under Pb stress. Pb mitigation effects by Chs are probably related with the activity of antioxidant enzymes, phenylalanine ammonia lyase (PAL) and proline. The application of Chs enhanced the activities of peroxidase, catalase and PAL by 25.77, 17.71 and 20.07%, respectively at 100 ppm Pb compared to their control. Plant genomic material exhibits significant molecular polymorphism, with an average polymorphism of 91.66% across all primers. To assess the genetic distance created among treatments, the dendrogram was constructed and the results of the similarity index ranged from 0.75 to 0.95, indicating genetic divergence. Our research offers a thorough comprehension of the role of Chs in lessening the oxidative stress, which will encourage the use of Chs in agricultural plant protection.

Triggering antibacterial activity of a common plant by biosorption of selected heavy metals.

The presented study proposes an efficient utilization of a common Thymus serpyllum L. (wild thyme) plant as a highly potent biosorbent of Cu(II) and Pb(II) ions and the efficient interaction of the copper-laden plant with two opportunistic bacteria. Apart from biochars that are commonly used for adsorption, here we report the direct use of native plant, which is potentially interesting also for soil remediation. The highest adsorption capacity for Cu(II) and Pb(II) ions (qe = 12.66 and 53.13 mg g-1, respectively) was achieved after 10 and 30 min of adsorption, respectively. Moreover, the Cu-laden plant was shown to be an efficient antibacterial agent against the bacteria Escherichia coli and Staphylococcus aureus, the results being slightly better in the former case. Such an activity is enabled only via the interaction of the adsorbed ions effectively distributed within the biological matrix of the plant with bacterial cells. Thus, the sustainable resource can be used both for the treatment of wastewater and, after an effective embedment of metal ions, for the fight against microbes.

Simultaneous removal of crude oil and heavy metals by highly adapted bacterial strain Cutibacterium sp. NL2 isolated from Algerian oilfield.

Investigating the ability of bacteria to simultaneously enhance hydrocarbon removal and reduce heavy metals' toxicity is necessary to design more effective bioremediation strategies. A bacterium (NL2 strain) isolated from an Algerian oilfield was cultivated on crude oil as sole carbon and energy sources. Molecular analyses of the 16S rRNA gene sequence placed the strain within the Cutibacterium genera. This isolate was able to tolerate up to 60% of crude oil as sole carbon source. Chemical analyses (GC-MS) evidenced that strain NL2 was able to degrade 92.22% of crude oil (at optimal growing conditions: pH 10, 44 °C, 50 g L-1 NaCl, and 20% of crude oil (v/v) as sole carbon source) in only 7 days. NL2 isolate was also able to produce biosurfactants with reduction of surface tension of growing media (29.4 mN m-1). On the other hand, NL2 strain was able to tolerate high lead (Pb) and copper (Cu) concentrations (up to 60 mM). In fact, NL2 cultivated in the presence of 20% of crude oil, and 0.48 mM of Pb was able to reduce Pb concentration by a 41.36%. In turn, when cultivated on high Pb concentration (15 mM), the strain was able to remove 35.19% of it and 86.25% of crude oil, both in a time frame of 7 days. Our findings suggest that Cutibacterium strain NL2 is able to efficiently use and remove a wide range of crude oil substrates in presence of high Pb concentration. Accordingly, NL2 strain is of extreme interest from a biotechnological standpoint.

Proteome signatures of joint toxicity to arsenic (As) and lead (Pb) in human brain organoids with optic vesicles.

Arsenic (As) and lead (Pb) are toxins found in the natural surroundings, and the harmful health outcomes caused by the co-exposure of such toxins have become a considerable problem. However, the joint neurotoxicity of As and Pb to neurodevelopment and the underlying mechanisms remain unclear. Pluripotent stem cell-derived human brain organoids are emerging animal model alternatives for understanding neurological-related diseases. Therefore, we utilized brain organoids with optic vesicles (OVB-organoids) to systematically analyze the neurotoxicity of As and Pb. After 24 h of As and/or Pb exposure, hematoxylin-eosin staining revealed that As and Pb exposure could cause disorders in the structure of the ventricular zone and general cell disarrangement in OVB-organoids. Immunostaining displayed that OVB-organoids are more susceptible to As and Pb co-exposure than independent exposure in apoptosis, proliferation, and cell differentiation. Meanwhile, even though As and Pb could both hinder cell proliferation, contrary to Pb, As could induce an increasing proportion of mitotic (G2/M) cells. The proteome landscape of OVB-organoids illustrated that Pb synergized with As in G2/M arrest and the common role of As and Pb in carcinogenesis. Besides, proteomics analyses suggested the consequential role of autophagy and Wnt pathway in the neurotoxicity of As and Pb co-exposure. Overall, our findings provide penetrating insights into the cell cycle, carcinogenesis, autophagy, and Wnt pathway underlying the As and Pb binary exposure scenarios, which could enhance our understanding of the mixture neurotoxicity mechanisms.

Cellulolytic potential of mangrove bacteria Bacillus haynesii DS7010 and the effect of anthropogenic and environmental stressors on bacterial survivability and cellulose metabolism.

Cellulose degrading bacterial diversity of Bhitarkanika mangrove ecosystem, India, was uncovered and the cellulose degradation mechanism in Bacillus haynesii DS7010 under the modifiers such as pH (pCO2), salinity and lead (Pb) was elucidated in the present study. The abundance of cellulose degrading heterotrophic bacteria was found to be higher in mangrove sediment than in water. The most potential strain, B. haynesii DS7010 showed the presence of endoglucanase, exoglucanase and ß-glucosidase with the maximum degradation recorded at 48 h of incubation, with 1% substrate concentration at 41 °C incubation temperature. Two glycoside hydrolase genes, celA and celB were confirmed in this bacterium. 3D structure prediction of the translated CelA and CelB proteins showed maximum similarities with glycoside hydrolase 48 (GH48) and glycoside hydrolase 5 (GH5) respectively. Native PAGE followed by zymogram assay unveiled the presence of eight isoforms of cellulase ranged from 78 kDa to 245 kDa. Among the stressors, most adverse effect was observed under Pb stress at 1400 ppm concentration, followed by pH at pH 4. This was indicated by prolonged lag phase growth, higher reactive oxygen species (ROS) production, lower enzyme activity and downregulation of celA and celB gene expressions. Salinity augmented bacterial metabolism up to 3% NaCl concentration. Mangrove leaf litter degradation by B. haynesii DS7010 indicated a substantial reduction in cellulolytic potential of the bacterium in response to the synergistic effect of the stressors. Microcosm set up with the stressors exhibited 0.97% decrease in total carbon (C%) and 0.02% increase in total nitrogen (N%) after 35 d of degradation while under natural conditions, the reduction in C and the increase in N were 4.05% and 0.2%, respectively. The findings of the study suggest the cellulose degradation mechanism of a mangrove bacterium and its resilience to the future consequences of environmental pollution and climate change.

Distribution and bioaccumulation of trace elements in two Cyprinidae fish species in the Indus river, Pakistan, including the impact of hydraulic structure on macroinvertebrates' biodiversity.

The concentration of trace elements (chromium, lead, zinc, copper, manganese, and iron) was determined in water, sediment and tissues of two Cyprinidae fish species - Labeo rohita and Tor putitora - collected from the eight sampling stations of Indus River in 2022 for four successive seasons (autumn, winter, spring, summer), and also study the present condition of macroinvertebrates after the construction of hydraulic structure. The obtained results of trace element concentrations in the Indus River were higher than the acceptable drinking water standards by WHO. The nitrate concentration ranges from 5.2 to 59.6 mg l-1, turbidity ranges from 3.00 to 63.9 NTU, total suspended solids and ammonium ions are below the detection limit (<0.05). In the liver, highest dry wt trace elements (µg/g) such as Cr (4.32), Pb (7.07), Zn (58.26), Cu (8.38), Mn (50.27), and Fe (83.9) for the Labeo rohita; and Tor Putitora has significantly greater accumulated concentration (Cr, Pb, Zn, Cu, Mn, Fe) in muscle and liver than did Labeo rohita species. Additionally, lower number of macroinvertebrates were recorded during the monsoonal season than pre-monsoon and post-monsoon. Local communities surrounded by polluted environments are more probably to consume more fish and expose them to higher concentrations of toxic trace elements (lead and copper). The findings also provide a basis for broader ecological management of the Indus River, which significantly influenced human beings and socioeconomic disasters, particularly in the local community.

Lead exposure induced transgenerational developmental neurotoxicity by altering genome methylation in Drosophila melanogaster.

Heavy metal toxicity is a significant global health concern, with particular attention given to lead (Pb) exposure due to its adverse effects on cognitive development, especially in children exposed to low concentrations. While Pb neurotoxicity has been extensively studied, the analysis and molecular mechanisms underlying the transgenerational effects of Pb exposure-induced neurotoxicity remain poorly understood. In this study, we utilized Drosophila, a powerful developmental animal model, to investigate this phenomenon. Our findings demonstrated that Pb exposure during the developmental stage had a profound effect on the neurodevelopment of F0 fruit flies. Specifically, we observed a loss of correlation between the terminal motor area and muscle fiber area, along with an increased frequency of the ß-lobe midline crossing phenotype in mushroom bodies. Western blot analysis indicated altered expression levels of synaptic vesicle proteins, with a decrease in Synapsin (SYN) and an increase in Bruchpilot (BRP) expression, suggesting changes in synaptic vesicle release sites. These findings were corroborated by electrophysiological data, showing an increase in the amplitude of evoked excitatory junctional potential (EJP) and an increase in the frequency of spontaneous excitatory junctional potential (mEJP) following Pb exposure. Importantly, our results further confirmed that the developmental neurotoxicity resulting from grandparental Pb exposure exhibited a transgenerational effect. The F3 offspring displayed neurodevelopmental defects, synaptic function abnormalities, and repetitive behavior despite lacking direct Pb exposure. Our MeDIP-seq analysis further revealed significant alterations in DNA methylation levels in several neurodevelopmental associated genes (eagle, happyhour, neuroglian, bazooka, and spinophilin) in the F3 offspring exposed to Pb. These findings suggest that DNA methylation modifications may underlie the inheritance of acquired phenotypic traits resulting from environmental Pb exposure.

Root-associated bacterial microbiome shaped by root selective effects benefits phytostabilization by Athyrium wardii (Hook.).

The root-associated microbiome assembly substantially promotes (hyper)accumulator plant growth and metal accumulation and is influenced by multiple factors, especially host species and environmental stress. Athyrium wardii (Hook.) is a phytostabilizer that grows in lead (Pb)-zinc (Zn) mine tailings and shows high root Pb accumulation. However, there remains little information on the assembly of the root-associated microbiome of A. wardii and its role in phytostabilization. A field study investigated the structural and functional variation in the root-associated bacterial microbiome of Athyrium wardii (Hook.) exposed to different levels of contamination in Pb-Zn mine tailings. The root compartment dominated the variation in the root-associated bacterial microbiome but the levels of contaminants showed less impact. Bacterial co-occurrence was enhanced in the rhizosphere soil and rhizoplane but tended to be much simpler in the endosphere in terms of network complexity and connectivity. This indicates that the microbial community assembly of A. wardii was non-random and shaped by root selective effects. Proteobacteria, Chloroflexi, Actinobacteria, Cyanobacteria, and Acidobacteriota were generally the dominant bacterial phyla. The genera Crossiella and Bradyrhizobium were enriched in the rhizosphere and cyanobacterial genera were enriched in the endosphere, demonstrating substantial advantages to plant survival and adaptation in the harsh mine environment. Functional categories involved in amino acid and carbohydrate metabolism were abundant in the rhizosphere soil, thus contributing to metal solubility and bioavailability in the rhizosphere. Membrane transporters, especially ATP-binding cassette transporters, were enriched in the endosphere, indicating a potential role in metal tolerance and transportation in A. wardii. The study shows substantial variation in the structure and function of microbiomes colonizing different compartments, with the rhizosphere and endophytic microbiota potentially involved in plant metal tolerance and accumulation during phytostabilization.

Synergistic application of Pseudomonas strains and compost mitigates lead (Pb) stress in sunflower (Helianthus annuus L.) via improved nutrient uptake, antioxidant defense and physiology.

Lead (Pb) is one of the most dreadful non-essential elements whose toxicity has been well reported worldwide due to its interference with the major plant functions and its overall yield. Bioremediation techniques comprising the application of beneficial microorganisms have gained attention in recent times owing to their ecofriendly nature. Addition of organic matter to soil has been reported to stimulate microbial activities. Compost application improves soil structure and binds toxic contaminants due to its larger surface area and presence of functional groups. Furthermore, it stimulates soil microbial activities by acting as C-source. So, in current study, we investigated the individual and synergistic potential of two lead (Pb)-tolerant Pseudomonas strains alongwith compost (1% w/w) in sustaining sunflower growth under Pb contaminated soil conditions. Lead chloride (PbCl2) salt was used for raising desired Pb concentration (500 mg kg-1). Results revealed that Pb stress drastically affected all the measured attributes of sunflower plant, however joint application of rhizobacteria and compost counteracted these adverse effects. Among them, co-application of str-1 and compost proved to be significantly better than str-2, as its inoculation significantly improved shoot and root lengths (64 and 76%), leaf area and leaves plant-1 (95 and 166%), 100-achene weight (200%), no. of flowers plant-1 (138%), chl 'a', 'b' and carotenoid (86, 159 and 33%) contents in sunflower as compared to control treatments. Furthermore, inoculation of Pseudomonas fluorescens along with compost increased the NPK in achene (139, 200 and 165%), flavonoid and phenolic contents (258 and 185%) along with transpiration and photosynthetic rates (54 and 72%) in leaves as compared to control treatment under Pb contamination. In addition, Pb entry to roots, shoots and achene were significantly suppressed under by 87, 90 and 91% respectively due to integrated application of compost and str-1 as evident by maximum Pb-immobilization efficiency (97%) obtained in this treatment. Similarly, bioconcentration factors for roots, shoots and achene were found to be 0.58, 0.18 and 0.0055 with associated translocation factor (0.30), which also revealed phytostabilization of Pb under combined application of PGPR and compost. Since, phytoremediation of heavy metals under current scenario of increasing global population is inevitable, results of the current study concluded that tolerant PGPR species along with organic amendments such as compost can inhibit Pb uptake by sunflower and confer Pb tolerance via improved nutrient uptake, physiology, antioxidative defense and gas exchange.

Interaction between Haematococcus pluvialis microalgae and lead nitrate: lead adsorption from water.

Our study aims to investigate the response of the unicellular alga, Haematococcus pluvialis, to the toxicity of lead and propose a low-cost, highly efficient biological adsorbent for the purification of wastewater and lead-contaminated water. The first part examines the effects of lead toxicity on certain physiological indicators of this alga. In the second part, the potential of this alga in lead removal and its adsorption capacity was assessed. The alga was cultivated in a BG11 medium and treated with lead nitrate concentrations of 10, 50, and 200 mg/L during its exponential growth. The results showed that with an increase in lead concentration up to 200 mg/L, the growth rate, chlorophyll a, chlorophyll b, carotenoid and total protein content decreased, while malondialdehyde (MDA) content increased. The astaxanthin content slightly increased at the 10 mg/L but decreased at the 200 mg/L treatment. Maximum lead adsorption was observed at 98.69% under optimal conditions, including a pH of 6, an adsorbent dose of 1 g/L, a lead concentration of 25 mg/L, a temperature of 25 °C, and an exposure time of 120 min. The results of this study demonstrate that Haematococcus pluvialis has the potential for effective lead removal from aquatic environments.

While the influence of heavy metals on certain algae species has been explored, research on the impact of lead on Haematococcus pluvialis­a microalga of significant interest for astaxanthin production­remains uncharted territory. Therefore, understanding the impact of this heavy metal and the alga's metal absorption capabilities has profound implications for biotechnology and bioremediation applications. This study promotes H. pluvialis as an economically viable lead absorbent suitable for both industrial and domestic purposes.

Metal toxicity in Bryum coronatum Schwaegrichen: impact on chlorophyll content, lamina cell structure, and metal accumulation.

This research examined the impact of heavy metals, including Cd, Pb, and Zn, on chlorophyll content and lamina cell structure in Bryum coronatum. After exposure to varying metal concentrations (0.015, 0.065, 0.250, 1, and 4 mg/L), chlorophyll content, chloroplast numbers, lamina cell change, and metal accumulation were investigated. Chlorophyll content was assessed using spectrophotometry, whereas chloroplast numbers and lamina cell changes were examined under a light microscope. Metal accumulation was quantified through ICP-MS. The findings revealed that Cd notably reduced chlorophyll a content, while Pb and Zn showed minimal influence. Cd and Pb exposure decreased the number of chloroplasts in lamina cells, with no impact from Zn. The moss's capacity to absorb metals increased with higher exposure levels, indicating its potential as a biomonitor for heavy metal pollution. Cell mortality occurred in response to Cd and Pb, primarily in the median and apical lamina regions, while Zn had no effect. This study sheds light on heavy metal toxicity in B. coronatum, underscoring its significance for environmental monitoring. Further research on the mechanisms and consequences of heavy metal toxicity in bryophytes is essential for a comprehensive understanding of this critical issue.

The capacity of moss B. coronatum to absorb metals increased with higher exposure levels, providing quantitative data on heavy metal pollution around it.

Integrated Transcriptomic and Metabolomic Analysis Reveals the Molecular Regulatory Mechanism of Flavonoid Biosynthesis in Maize Roots under Lead Stress.

Flavonoids are secondary metabolites that play important roles in the resistance of plants to abiotic stress. Despite the widely reported adverse effects of lead (Pb) contamination on maize, the effects of Pb on the biosynthetic processes of flavonoids in maize roots are still unknown. In the present work, we employed a combination of multi-omics and conventional assay methods to investigate the effects of two concentrations of Pb (40 and 250 mg/kg) on flavonoid biosynthesis in maize roots and the associated molecular regulatory mechanisms. Analysis using conventional assays revealed that 40 and 250 mg/kg Pb exposure increased the lead content of maize root to 0.67 ± 0.18 mg/kg and 3.09 ± 0.02 mg/kg, respectively, but they did not result in significant changes in maize root length. The multi-omics results suggested that exposure to 40 mg/kg of Pb caused differential expression of 33 genes and 34 metabolites related to flavonoids in the maize root system, while 250 mg/kg of Pb caused differential expression of 34 genes and 31 metabolites. Not only did these differentially expressed genes and metabolites participate in transferase activity, anthocyanin-containing compound biosynthetic processes, metal ion binding, hydroxyl group binding, cinnamoyl transferase activity, hydroxycinnamoyl transferase activity, and flavanone 4-reductase activity but they were also significantly enriched in the flavonoid, isoflavonoid, flavone, and flavonol biosynthesis pathways. These results show that Pb is involved in the regulation of maize root growth by interfering with the biosynthesis of flavonoids in the maize root system. The results of this study will enable the elucidation of the mechanisms of the effects of lead on maize root systems.

Responses of plankton community to threshold metal concentrations of cadmium and lead in a mesocosm experiment at Bay of Bengal.

Metals are essential at trace levels to aquatic organisms for the function of many physiological and biological processes. But their elevated levels are toxic to the ecosystem and even brings about shifts in the plankton population. Threshold limits such as Predicted No Effect Concentration (PNEC - 0.6 µg/l of Cd; 2.7 µg/l of Pb), Criterion Continuous Concentration (CCC - 3.0 µg/l of Cd; 4.5 µg/l of Pb) and Criterion Maximum Concentration (CMC - 23 µg/l of Cd; 130 µg/l of Pb) prescribed for Indian coastal waters were used for the study. Short-term mesocosm experiments (96 h) were conducted in coastal waters of Visakhapatnam to evaluate responses of the planktonic community on exposure to threshold concentrations of cadmium and lead for the first time. Four individual experimental bags of 2500 L capacity (Control, PNEC, CCC & CMC) were used for the deployment and ambient water samples were analysed simultaneously to evaluate the impacts of the threshold levels in the natural waters. Chaetoceros sp. were dominant group in the control system whereas, Prorocentrum sp. Ceratium sp. Tintinopsis sp. Chaetoceros sp. and Skeletonema sp. were major groups in the test bags. Throughout the experiment the phytoplankton community did not show any significant differences with increased nutrients and plankton biomass (Chl-a <8.64 mg/m3). Positive response of plankton community was observed in the experimental bags. High abundance of diatoms were observed in PNEC, CCC & CMC bags at 48 h and the abundance decreased with shift in the species at 72-96 h. The catalase activity in phytoplankton (5.99 nmol/min/ml) and the zooplankton (4.77 nmol/min/ml) showed induction after exposure to PNEC. The present mesocosm study is confirmed that short-term exposure to threshold metal concentration did not affects the phytoplankton community structure in PNEC, but CCC and CMC affects the community structure beyond 24 h. The insights from this study will serve as a baseline information and help develop environmental management tools. We believe that long-term mesocosm experiments would unravel metal detoxification mechanisms at the cellular level and metal transfer rate at higher trophic levels in real-world environment.

Protective role of selenium and selenium-nanoparticles against multiple stresses in Pangasianodon hypophthalmus.

Pollution and climate change pose significant threats to aquatic ecosystems, with adverse impacts on aquatic animals, including fish. Climate change increases the toxicity of metal in aquatic ecosystems. To understand the severity of metal pollution and climate change, an experiment was conducted to delineate the mitigation potential of selenium (Se) and selenium nanoparticles (Se-NPs) against lead (Pb) and high temperature stress in Pangasianodon hypophthalmus. For the experiment, five isonitrogenous and isocaloric diets were prepared, varying in selenium supplementation as Se at 0, 1, and 2 mg kg-1 diet, and Se-NPs at 1 and 2 mg kg-1 diet. The fish in stressor groups were exposed to Pb (1/20th of LC50 concentration, 4 ppm) and high temperature (34 °C) throughout the experiment. The results demonstrated that dietary supplementation of Se at 1 and 2 mg kg-1 diet, as well as Se-NPs at 1 mg kg-1 diet, significantly reduced (p < 0.01) the levels of lactate dehydrogenase and malate dehydrogenase in both liver and muscle tissues. Additionally, the levels of alanine aminotransferase and aspartate aminotransferase in both gill and liver tissues were significantly decreased (p < 0.01) with the inclusion of Se and Se-NPs in the diets. Furthermore, the enzymes glucose-6-phosphate dehydrogenase in gill and liver tissues, fructose 1,6-bisphosphatase in liver and muscle tissues, and acid phosphatase in liver tissue were remarkably reduced (p < 0.01) due to the supplementation of Se and Se-NPs. Moreover, dietary supplementation of Se and Se-NPs significantly enhanced (p < 0.01) the activity of pyruvate kinase, glucokinase, hexokinase, alkaline phosphatase, ATPase, protease, amylase, lipase, and RNA/DNA ratio in the fish. Histopathological examination of gill and liver tissues also indicated that Se and Se-NPs protected against structural damage caused by lead and high-temperature stress. Moreover, the study examined the bioaccumulation of selenium and lead in muscle, water, and diets. The aim of the study revealed that Se and Se-NPs effectively protected the fish from lead toxicity and high-temperature stress, while also improving the function of cellular metabolic enzymes in P. hypophthalmus.

Comparative transcriptomics revealed the mechanism of Stenotrophomonas rhizophila JC1 response and biosorption to Pb2.

Nowadays, there is limited research focusing on the biosorption of Pb2+ through microbial process, particularly at the level of gene expression. To overcome this knowledge gap, we studied the adsorption capacity of Stenotrophomonas rhizophila JC1 to Pb2+, and investigated the physiological mechanism by means of SEM, EDS, FTIR, membrane permeability detection, and investigated the molecular mechanism through comparative transcriptomics. The results showed that after 16 h of cultivation, the biosorption capacity of JC1 for 100 mg/L of Pb2+ reached at 79.8%. The main mechanism of JC1 adsorb Pb2+ is via intracellular accumulation, accounting for more than 90% of the total adsorption. At the physiological level, Pb2+ can precipitate with anion functional groups (e.g., -OH, -NH) on the bacterial cell wall or undergo replacement reaction with cell component elements (e.g., Si, Ca) to adsorb Pb2+ outside of the cell wall, thus accomplishing extracellular adsorption of Pb2+ by strains. Furthermore, the cell membrane acts as a "switch" that inhibits the entry of metal ions into the cell from the plasma membrane. At the molecular level, the gene pbt specificity is responsible for the adsorption of Pb2+ by JC1. In addition, phosphate permease is a major member of the ABC transporter family involved in Pb2+, and czcA/cusA or Co2+/Mg2+ efflux protein plays an important role in the efflux of Pb2+ in JC1. Further, cellular macromolecule biosynthesis, inorganic cation transmembrane transport, citrate cycle (TCA) and carbon metabolism pathways all play crucial roles in the response of strain JC1 to Pb2+ stress.

Pollution pressure drives microbial assemblages that improve the phytoremediation potential of heavy metals by Ricinus communis.

Due to the rapid expansion of industrial activity, soil pollution has intensified. Plants growing in these polluted areas have developed a rhizobiome uniquely and specially adapted to thrive in such environments. However, it remains uncertain whether pollution acts as a sufficiently selective force to shape the rhizobiome, and whether these adaptations endure over time, potentially aiding in long-term phytoremediation. Therefore, in the present study, we aimed to compare whether the microbiome associated with roots from plants germinated in polluted riverbanks will improve the phytoremediation of Cd and Pb under mesocosm experiments compared with plants germinating in a greenhouse. The experimental design was a factorial 2 × 2, i.e., the origin of the plant and the presence or absence of 100 mg/L of Cd and 1000 mg/L of Pb. Our results showed that plants germinated in polluted riverbanks have the capacity to accumulate twice the amount of Pb and Cd during mesocosm experiments. The metagenomic analysis showed that plants from the river exposed to heavy metals at the end of mesocosm experiments were rich in Rhizobium sp. AC44/96 and Enterobacter sp. EA-1, Enterobacter soli, Pantoea rwandensis, Pantoea endophytica. In addition, those plants were uniquely associated with Rhizobium grahamii, which likely contributed to the differences in the levels of phytoremediation achieved. Furthermore, the functional analysis revealed an augmented functional potential related to hormones, metallothioneins, dismutases, and reductases; meanwhile, the plants germinated in the greenhouse showed an unspecific strategy to exceed heavy metal stress. In conclusion, pollution pressure drives stable microbial assemblages, which could be used in future phytostabilization and phytoremediation experiments.

Accumulation of Cd and Pb in sediments and Asian swamp eels (Monopterus albus) from downstream area of Cisadane River, Indonesia.

This study examined the presence of two heavy metals (Cd and Pb) in the sediments and Asian swamp eels (Monopterus albus) in the downstream area of Cisadane River. The average concentrations of Cd and Pb in the sediments from all sampling locations were 0.594 ± 0.230 mg/kg and 34.677 ± 24.406 mg/kg, respectively. These concentrations were above the natural background concentration and the recommended value of interim sediment quality guidelines (ISQG), suggesting an enrichment process and potential ecological risk of studied metals to the ecosystem of Cisadane River. The increase in contamination within this region may be attributed to point sources such as landfill areas, as well as the industrial and agricultural land activities in surrounding area, and experienced an increasing level leading towards the estuary of Cisadane River. Meanwhile, the average concentrations of Cd and Pb in the eels from all sampling locations were 0.775 ± 0.528 µg/g and 28.940 ± 12.921 µg/g, respectively. This study also discovered that gill tissues contained higher levels of Cd and Pb than the digestive organ and flesh of Asian swamp eels. These concentrations were higher than Indonesian and international standards, suggesting a potential human health risk and therefore the needs of limitations in the consumption of the eels. Based on the human health risk assessment, the eels from the downstream of Cisadane River are still considered safe to be consumed as long as they comply with the specified maximum consumption limits.

Validation and calibration of a novel GEM biosensor for specific detection of Cd2+, Zn2+, and Pb2.

BACKGROUND: In this study, we designed a novel genetic circuit sensitive to Cd2+, Zn2+ and Pb2+ by mimicking the CadA/CadR operon system mediated heavy metal homeostasis mechanism of Pseudomonas aeruginosa. The regular DNA motifs on natural operon were reconfigured and coupled with the enhanced Green Fluorescent Protein (eGFP) reporter to develop a novel basic NOT type logic gate CadA/CadR-eGFP to respond metal ions mentioned above. A Genetically Engineered Microbial (GEM)-based biosensor (E.coli-BL21:pJET1.2-CadA/CadR-eGFP) was developed by cloning the chemically synthesised CadA/CadR-eGFP gene circuit into pJET1.2-plasmid and transforming into Escherichia coli (E. coli)-BL21 bacterial cells. RESULTS: The GEM-based biosensor cells indicated the reporter gene expression in the presence of Cd2+, Zn2+ and Pb2+ either singly or in combination. Further, the same biosensor cells calibrated for fluorescent intensity against heavy metal concentration generated linear graphs for Cd2+, Zn2+ and Pb2+ with the R2 values of 0.9809, 0.9761 and 0.9758, respectively as compared to non-specific metals, Fe3+ (0.0373), AsO43- (0.3825) and Ni2+ (0.8498) making our biosensor suitable for the detection of low concentration of the former metal ions in the range of 1-6 ppb. Furthermore, the GEM based biosensor cells were growing naturally within the concentration range of heavy metals, at 37 °C and optimum pH = 7.0 in the medium, resembling the characteristics of wildtype E.coli. CONCLUSION: Finally, the novel GEM based biosensor cells developed in this study can be applied for detection of targeted heavy metals in low concentration ranges (1-6 ppb) at normal bacterial physiological conditions.