Search Results (8)

This study assessed the effects of tetracycline (TC) on growth of Lemna aoukikusa and Spirodela polyrhiza under batch conditions. The duckweeds were exposed to a range of 0.0–5.0 mg L⁻¹ of TC for 7 days in a medium containing 10 mg L⁻¹ total nitrogen (TN) and 1 mg L⁻¹ total phosphorus (TP). The relative growth rate (RGR) of each species was determined from the frond area measurement using image analysis. The EC₅₀ values as the TC concentrations causing a 50% reduction in RGR, were 4.4 mg L⁻¹ for L. aoukikusa and 0.65 mg L⁻¹ for S. polyrhiza. At 5.0 mg L⁻¹ TC, TP removal decreased to 60% in the L. aoukikusa culture and 77% in the S. polyrhiza culture, compared to 85–91% and 96%, respectively, under lower TC exposure. Nevertheless, TN and TP removals were not significantly impaired at TC concentrations found in swine wastewater. The TC removals were 76–94% for the L. aoukikusa culture and 68–91% for the S. polyrhiza culture, which were attributed to adsorption and plant uptake. These findings highlight the feasibility of duckweed-based stabilization ponds for simultaneous antibiotic attenuation and nutrient removal. Full article

The identification of natural and environmentally friendly pesticides is a key area of interest for the agrochemical industry, with many potentially active compounds being sourced from numerous plant species. In this study, we report the bioassay-guided isolation and identification of phytotoxic and antifungal compounds from the ethyl acetate extract of Helietta parvifolia stems. We identified eight compounds, consisting of two coumarins and six alkaloids. Among these, a new alkaloid, 2-hydroxy-3,6,7-trimethoxyquinoline-4-carbaldehyde (6), was elucidated, along with seven known compounds. The phytotoxicity of purified compounds was evaluated, and chalepin (4) was active against Agrostis stolonifera at 1 mM with 50% inhibition of seed germination and it reduced Lemna pausicotata (duckweed) growth by 50% (IC₅₀) at 168 μM. Additionally, we evaluated the antifungal activity against the fungal plant pathogen Colletotrichum fragariae using a thin-layer chromatography bioautography assay, which revealed that three isolated furoquinoline alkaloids (flindersiamine (3), kokusagenine (7), and maculine (8)) among the isolated compounds had the strongest inhibitory effects on the growth of C. fragariae at all tested concentrations. Our results indicate that these active natural compounds, i.e., (3), (4), (7), and (8), could be scaffolds for the production of more active pesticides with better physicochemical properties. Full article

Aquatic environment are often contaminated with heavy metals from various industrial sources. However, physicochemical techniques for pollutant detection are limited, thus prompting the need for additional bioassays. We investigated the use of greater duckweed (Spirodela polyrhiza) as a bioindicator of metal pollution. We exposed S. polyrhiza to four pollutants (namely, silver, cadmium, copper, and chromium) and assessed metal toxicity by measuring its frond area and the length of its regrown roots. The plant displayed significant differences in both frond size and root growth in response to the four metals. Silver was the most toxic (EC₅₀ = 23 µg L⁻¹) while copper the least (EC₅₀ = 365–607 µg L⁻¹). Direct comparisons of metal sensitivity and the reliability of the two endpoint assays showed that root growth was more sensitive (lower in terms of 50% effective concentration) to chromium, cadmium, and copper, and was more reliable (lower in terms of coefficient of variation) than those for frond area. Compared to conventional Lemna-based tests, the S. polyrhiza test is easier to perform (requiring only one 24-well plate, 3 mL of medium and a 72-h exposure). Moreover, it does not require livestock cultivation/maintenance, making it more suitable for repeated measurements. Measurements of S. polyrhiza root length may be suitable for assessment when copper and chromium in municipal and industrial wastewater exceed the environmentally permissible levels. Full article

Toxic cyanobacterial blooms are a well-known problem in eutrophic water bodies, but cyanobacterial toxicity in ambient springs is unexplored. Therefore, we studied the toxin content and effects of two extracts obtained from epilithic and benthic microbial mats containing different algae and filamentous cyanobacteria, Phormidium breve and Oscillatoria limosa, respectively, on fresh biomass, number of roots, and pigment content in duckweed Spirodela polyrhiza and on survivorship of Daphnia magna (Cladocera). Mat samples, used to prepare extracts for bioassays, were collected in the anthropogenically transformed limnocrenic Górecko spring, located (50°31′08″ N and 22°57′10″ E) in the Roztocze region (Eastern Poland). It drains an abundant aquifer built with Cretaceous sediments. Neither anatoxin-a (ANTX) nor microcystins (MCs) were detected in the extracts using HPLC techniques; however, negative effects of the extracts on tested organisms were observed. The Phormidium extract contained concentrations of cyanobacterial metabolites a few times higher than the Oscillatoria extract. In general, both extracts affected plants in a dose-dependent manner; however, the Oscillatoria extract influenced pigment production without a clear trend. The highest concentrations of Phormidium extract (p < 0.05) significantly decreased the number of roots and the content of chlorophylls and carotenoids in S. polyrhiza. The Oscillatoria extract caused a statistically significant (p < 0.05) decrease in biomass and number of roots; however, it generally positively influenced the production of pigments. The extract containing O. limosa metabolites was more toxic to D. magna than the extract containing higher amounts of metabolites of P. breve. Cyanobacteria inhabiting temperate springs may negatively affect hydrobionts by producing secondary metabolites other than ANTX and MCs; however, the contribution of algae cannot be excluded. Extensive research on cyanobacteria in springs, their metabolites, and their effects on living organisms should be conducted. Full article

European Pharmacopoeia monograph 2371 describes the production of homeopathic preparations. A specific efficacy of these preparations in high dilution levels is questionable in view of basic scientific principles. There is empirical evidence for such effects, for example in a Lemna-intoxication bioassay published 2010. To test the replicability and robustness of this bioassay, we conducted two experimental series (five independent blinded and randomised experiments each). The specimen of Lemna gibba L., clone-number 9352, were stressed in arsenic solution for 48 h (158 mg/L AsNa₂HO₄ (250 mg/L in series 2)), then grew in either As₂O₃ preparations produced according to Eu. Pharm. Monogr. 2371 or control solution. Comparing the area-related relative growth rate of day 3–9 (rgr 3–9) between treatment and control groups for each series showed differences that were not significant in series 1 (p = 0.10), significant in series 2 (p = 0.04) and significant in the pooled data of both series (p < 0.01). The effect direction (rgr 3–9 increase) was comparable to experiments of 2010, but the effect size was smaller, likely due to a changed light cycle. These results are not compatible with the hypothesis that the application of European Pharmacopoeia monograph 2371 results in pharmaceutical preparations without specific effects. Further studies are needed to investigate a potential mode of action explaining these effects. Full article

The common duckweed (Lemna minor), a freshwater monocot that floats on the surfaces of slow-moving streams and ponds, is commonly used in toxicity testing. The novel Lemna root- regrowth test is a toxicity test performed in replicate test vessels (24-well plates), each containing 3 mL test solution and a 2–3 frond colony. Prior to exposure, roots are excised from the plant, and newly developed roots are measured after 3 days of regrowth. Compared to the three internationally standardized methods, this bioassay is faster (72 h), simpler, more convenient (requiring only a 3-mL) and cheaper. The sensitivity of root regrowth to 3,5-dichlorophenol was statistically the same as using the conventional ISO test method. The results of interlaboratory comparison tests conducted by 10 international institutes showed 21.3% repeatability and 27.2% reproducibility for CuSO₄ and 21.28% repeatability and 18.6% reproducibility for wastewater. These validity criteria are well within the generally accepted levels of <30% to 40%, confirming that this test method is acceptable as a standardized biological test and can be used as a regulatory tool. The Lemna root regrowth test complements the lengthier conventional protocols and is suitable for rapid screening of wastewater and priority substances spikes in natural waters. Full article

The application of layered double hydroxide (LDH) nanomaterials as catalysts has attracted great interest due to their unique structural features. It also triggered the need to study their fate and behavior in the aquatic environment. In the present study, Zn-Fe nanolayered double hydroxides (Zn-Fe LDHs) were synthesized using a co-precipitation method and characterized by X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FT-IR), scanning electron microscopy (SEM), and nitrogen adsorption-desorption analyses. The toxicity of the home-made Zn-Fe LDHs catalyst was examined by employing a variety of aquatic organisms from different trophic levels, namely the marine photobacterium Vibrio fischeri, the freshwater microalga Pseudokirchneriella subcapitata, the freshwater crustacean Daphnia magna, and the duckweed Spirodela polyrhiza. From the experimental results, it was evident that the acute toxicity of the catalyst depended on the exposure time and type of selected test organism. Zn-Fe LDHs toxicity was also affected by its physical state in suspension, chemical composition, as well as interaction with the bioassay test medium. Full article

The discovery of potent natural and ecofriendly pesticides is one of the focuses of the agrochemical industry, and plant species are a source of many potentially active compounds. We describe the bioassay-guided isolation of antifungal and phytotoxic compounds from the ethyl acetate extract of Ambrosia salsola twigs and leaves. With this methodology, we isolated and identified twelve compounds (four chalcones, six flavonols and two pseudoguaianolide sesquiterpene lactones). Three new chalcones were elucidated as (S)-β-Hydroxy-2′,3,4,6′-tetrahydroxy-5-methoxydihydrochalcone (salsolol A), (S)-β-Hydroxy-2′,4,4′,6′-tetrahydroxy-3-methoxydihydrochalcone (salsolol B), and (R)-α, (R)-β-Dihydroxy-2′,3,4,4′,6′-pentahydroxydihydrochalcone (salsolol C) together with nine known compounds: balanochalcone, six quercetin derivatives, confertin, and neoambrosin. Chemical structures were determined based on comprehensive direct analysis in real time-high resolution mass spectrometry (HR-DART-MS), as well as 1D and 2D NMR experiments: Cosy Double Quantum Filter (DQFCOSY), Heteronuclear Multiple Quantum Coherence (HMQC) and Heteronuclear Multiple Bond Coherence (HMBC), and the absolute configurations of the chalcones were confirmed by CD spectra analysis. Crystal structure of confertin was determined by X-ray diffraction. The phytotoxicity of purified compounds was evaluated, and neoambrosim was active against Agrostis stolonifera at 1 mM, while confertin was active against both, Lactuca sativa and A. stolonifera at 1 mM and 100 µM, respectively. Confertin and salsolol A and B had IC₅₀ values of 261, 275, and 251 µM, respectively, against Lemna pausicotata (duckweed). The antifungal activity was also tested against Colletotrichum fragariae Brooks using a thin layer chromatography bioautography assay. Both confertin and neoambrosin were antifungal at 100 µM, with a higher confertin activity than that of neoambrosin at this concentration. Full article