Search Results (6)

In this study, we developed a label-free biosensor based on aptamer-modified multi-walled carbon nanotube extended gate field-effect transistor (MWCNT-EG-FET) for easy and selective detection of microcystin-LR (MC-LR), a prominent cyanotoxin associated with liver damage, bleeding, and necrosis. EG-FET had two parts: a MOSFET and an extended-gate Au/SiO₂ electrode, which serves as the sensitive membrane. A custom-designed DNA oligonucleotide (5-NH₂-C₆-AN6) was used as MC-LR-targeting aptamer (MCTA). MWCNTs were functionalized with MCTA and then stably fixed on the sensitive membrane. The detection of MC-LR in freshwater was effectively achieved within 5 min by assessing the variations in electrical resistance that occur due to the selective interactions between MC-LR and MCTA. The detection limit and analytical sensitivity of the biosensor for MC-LR were found to be 0.134 ng/mL and 0.024 ng/mL, respectively. The sensitive membrane could be readily discarded if damaged, eliminating the need to replace the main transducer MOSFET. The developed sensor exhibits features such as straightforward preparation, swift response, potential for miniaturization, and ease of use, making it an attractive candidate for future integrated lab-on-chip systems for MC-LR detection in freshwater environments. Full article

Microcystin-LR (MC-LR) are biologically active cycloheptapeptide compounds that are released by cyanobacteria during water blooms and are extensively found in aquatic ecosystems. The Penaeus vannamei is a significant species in global aquaculture. However, the high level of eutrophication in aquaculture water frequently leads to outbreaks of cyanobacterial blooms, posing a significant threat to its sustainable cultivation. Astaxanthin (AX) is commonly utilized in aquaculture for its physiological benefits, including promoting growth and enhancing immune function in cultured organisms. This study aimed to examine the protective effect of astaxanthin on P. vannamei exposed to microcystin-induced stress. The experiment consisted of three groups: one group was fed formulated feed containing MC (100 μg/kg), another group was fed formulated feed containing MC (100 μg/kg) + AX (100 mg/kg), and the third group was fed basic feed (control group). After 15 days of feeding, the specific growth rate (SGR) was significantly higher in the MCAX group (2.21% day⁻¹) compared to the MC group (0.77% day⁻¹), and there was no significant difference between the MCAX group (2.21% day⁻¹) and the control group (2.24% day⁻¹). Similarly, the percent of weight gain (PWG) was also significantly higher in the MCAX group (14.61%) compared to the MC group (13.44%) and the control group (16.64%). Compared to the control group, the epithelial cells in the MC group suffered severe damage and detachment from the basement membrane. However, in the MCAX group, although there was still a gap between the intestinal epithelial cells and the basement membrane, the overall intestinal morphology was slightly less impaired than it was in the MC group. The analysis of the intestinal microbiota revealed a significant disparity in the community composition (chao 1 and ACE) between the MC and MCAX groups. When comparing the various bacterial genera, the MC group exhibited an increase in Vibrio abundance, whereas the MCAX group showed a decrease in both Shewanella and Vibrio abundance. The results indicate that AX has a positive impact on the growth performance and resistance of P. vannamei against MC by regulating the composition of the intestinal microbiota. AX can be utilized to mitigate the detrimental effects of MC in aquaculture practices. This function could be attributed to the role of AX in preserving the structural integrity of the intestinal mucosa and regulating the composition of the intestinal microbiota. Full article

The objective of the present study was to investigate the effects of different doses of dietary Eucommia ulmoides leaf extract (ELE) on juvenile red claw crayfish (Cherax quadricarinatus). A total number of 720 red claw crayfish (initial body weight of 0.24 ± 0.01 g) were randomly assigned to six groups and fed diets containing 0 (Diet 1), 0.5 (Diet 2), 1 (Diet 3), 2 (Diet 4), 4 (Diet 5) and 10 (Diet 6) g dry weight (dw) ELE kg (dw)⁻¹ diets for eight weeks and challenged with microcystin-LR stress. The results indicated that dietary supplementation with 1–2 g dw ELE kg (dw)⁻¹ diet could significantly improve the weight gain rate (WGR) and specific growth rate (SGR) of crayfish. Muscle crude protein contents of crayfish fed Diet 2, Diet 3, and Diet 4 were significantly higher than those of the control group. Compared with the control group, dietary ELE could increase total antioxidant capacity (T-AOC), superoxide dismutase (SOD), glutathione peroxidase (GPx), acid phosphatase (ACP), alkaline phosphatase (AKP), and phenoloxidase (PO) activities and decrease malondialdehyde (MDA) level of crayfish. Dietary ELE significantly increased the relative expression levels of SOD, thioredoxin 1 (TRX1), GPx, selenium-dependent glutathione peroxidase (Se-GPx), cytochrome P450 (CYP450), anti-lipopolysaccharide factor (ALF) and C-type lysozyme (C-LZM) mRNA of crayfish compared with the control group during the feeding experiment. When subjected to MC-LR stress for 48 h, the mRNA expression levels of SOD, GPx, Se-GPx, glutathione-s-transferase 1 (GST1), ALF, hemocyanin (HEM), and C-LZM in the hepatopancreas could be improved to varying degrees compared with the Diet 1. Supplementation of 1–2 g dw ELE kg (dw)⁻¹ diet could improve the survival rate (SR) of crayfish under MC-LR stress. These results indicated that dietary ELE (1–2 g dw ELE kg (dw)⁻¹ diet) could improve the growth performance, muscle protein, and non-specific immune response and increase the SR of crayfish under MC-LR stress by regulating the mRNA expression levels of the immune- and antioxidant-related genes. Full article

Microalgae blooms are natural processes that occur in eutrophic aquatic ecosystems. Microalgae blooms, namely those constituted by cyanobacteria, are undergoing a significant expansion as a result of anthropogenic pollution and climate change. Many of these blooms cause environmental and public health concerns due to the production and accumulation of toxic substances by some cyanobacterial species. Despite the burdens that cyanobacteria may cause in the environment and human health, cyanobacterial biomasses are interesting sources of compounds in biotechnology. Cyanobacteria also have interesting plant growth properties, and their biomass is an excellent soil amendment. In order to promote safe use of this type of material in biotechnology and agriculture, a research work was outlined, which consisted in seeking inexpensive and environmentally sustainable methods of treatment of Microcystis aeruginosa biomass and to reduce the content of the toxin microcystin (MC) in the biomass. Lyophilized or hydrated biomass from laboratory cultures of M. aeruginosa were subjected to treatments by heat (50 °C), ultraviolet radiation, ozone, and solar radiation for periods ranging from 2 to 12 h. The results demonstrate a significant reduction in the amount of MC in the biomass exposed to natural radiation for 12 h, from 0.0042 to 0.0028 mg of MC-LR/mg of dry biomass, equivalent to a reduction of about 33% of the total toxin. Efforts are currently being made to characterize the chemical transformation of the toxin catalyzed by natural radiation. No other treatment allowed us to reduce the amount of toxin present in the biomass, which suggests a strong chemical resistance of MC. This method of treatment of cyanobacterial biomass is quite interesting, and its use on a large scale depends on a confirmation of the preservation of the biotechnological properties of biomass after the applied treatment. Full article

Cyanobacteria, which form water blooms all over the world, can produce a wide range of cyanotoxins such as hepatotoxic microcystins (MCs) and other biologically active metabolites harmful to living organisms, including humans. Microcystin biodegradation, particularly caused by bacteria, has been broadly documented; however, studies in this field focus mainly on strains isolated from natural aquatic environments. In this paper, the biodegradation of microcystin-RR (MC-RR), microcystin-LR (MC-LR), and microcystin-LF (MC-LF) after incubation with Spirodela polyrhiza and the associated microorganisms (which were cultured under laboratory conditions) is shown. The strongest MC biodegradation rate after nine days of incubation was observed for MC-RR, followed by MC-LR. No statistically significant decrease in the concentration of MC-LF was noted. Products of MC decomposition were detected via the HPLC method, and their highest number was found for MC-RR (six products with the retention time between 5.6 and 16.2 min), followed by MC-LR (two products with the retention time between 19.3 and 20.2 min). Although the decrease in MC-LF concentration was not significant, four MC-LF degradation products were detected with the retention time between 28.9 and 33.0 min. The results showed that MC-LF was the most stable and resistant MC variant under experimental conditions. No accumulation of MCs or their biodegradation products in S. polyrhiza was found. The findings suggest that the microorganisms (bacteria and algae) associated with S. polyrhiza could be responsible for the MC biodegradation observed. Therefore, there is a need to broaden the research on the biodegradation products detected and potential MC-degraders associated with plants. Full article

Cyanobacteria harmful algal blooms (CHABs) are primarily caused by man-made eutrophication and increasing climate-change conditions. The presence of heavy metal runoff in affected water systems may result in CHABs alteration to their ecological interactions. Certain CHABs produce by-products, such as microcystin (MC) cyanotoxins, that have detrimentally affected humans through contact via recreation activities within implicated water bodies, directly drinking contaminated water, ingesting biomagnified cyanotoxins in seafood, and/or contact through miscellaneous water treatment. Metallothionein (MT) is a small, metal-sequestration cysteine rich protein often upregulated within the stress response mechanism. This study focused on zinc metal resistance and stress response in a toxigenic cyanobacterium, Microcystis aeruginosa UTEX LB 2385, by monitoring cells with (0, 0.1, 0.25, and 0.5 mg/L) ZnCl₂ treatment. Flow cytometry and phase contrast microscopy were used to evaluate physiological responses in cultures. Molecular assays and an immunosorbent assay were used to characterize the expression of MT and MC under zinc stress. The results showed that the half maximal inhibitory concentration (IC₅₀) was 0.25 mg/L ZnCl₂. Flow cytometry and phase contrast microscopy showed morphological changes occurred in cultures exposed to 0.25 and 0.5 mg/L ZnCl₂. Quantitative PCR (qPCR) analysis of selected cDNA samples showed significant upregulation of Mmt through all time points, significant upregulation of mcyC at a later time point. ELISA MC-LR analysis showed extracellular MC-LR (µg/L) and intracellular MC-LR (µg/cell) quota measurements persisted through 15 days, although 0.25 mg/L ZnCl₂ treatment produced half the normal cell biomass and 0.5 mg/L treatment largely inhibited growth. The 0.25 and 0.5 mg/L ZnCl₂ treated cells demonstrated a ~40% and 33% increase of extracellular MC-LR(µg/L) equivalents, respectively, as early as Day 5 compared to control cells. The 0.5 mg/L ZnCl₂ treated cells showed higher total MC-LR (µg/cell) quota yield by Day 8 than both 0 mg/L ZnCl₂ control cells and 0.1 mg/L ZnCl₂ treated cells, indicating release of MCs upon cell lysis. This study showed this Microcystis aeruginosa strain is able to survive in 0.25 mg/L ZnCl₂ concentration. Certain morphological zinc stress responses and the upregulation of mt and mcy genes, as well as periodical increased extracellular MC-LR concentration with ZnCl₂ treatment were observed. Full article