Immobilization Assay Research Articles

Size-dependent ability of AtaA to immobilize cells in Acinetobacter sp. Tol 5

Microbial cells serve as efficient and environmentally friendly biocatalysts, but their stability and reusability in practical applications must often be improved through immobilization. Acinetobacter sp. Tol 5 shows high adhesiveness to materials due to its large cell surface protein AtaA, which consists of 3630 amino acids (aa). Previously, we developed a method for immobilizing bacteria using AtaA. Herein, we investigated the cell immobilization ability of in-frame deletion (IFD) mutants of AtaA with different sizes in Tol 5. Mini-AtaA, which consists of 775 aa and is functional in Escherichia coli, was produced and present on the cell surface; however, mini-AtaA showed no immobilization ability in Tol 5. A cell immobilization assay was performed with cells expressing 16 IFD mutants of AtaA with different sizes, revealing that a length of at least 1417 aa was required for the sufficient immobilization of Tol 5 cells; thus, the minimum length needed to achieve the adhesive function of AtaA varies among bacterial species. The constructed mutant library of AtaA ranging from 3630 to 775 aa will allow researchers to quickly and easily explore the optimal size of AtaA, even for bacteria newly introduced to AtaA.

Pepsin immobilization on activated carbon and functionalized with glutaraldehyde and genipin for the synthesis of antioxidant peptides of goat casein

In this article, the synthesis of antioxidant peptides in the enzymatic hydrolysis of caprine casein was analyzed at three different time points (60 min, 90 min, and 120 min) using immobilized pepsin on activated and modified carbon (AC, ACF, ACG 50, ACG 100). The immobilization assays revealed a reduction in the biocatalysts' activity compared to the free enzyme. Among the modified ones, ACG 50 exhibited greater activity and better efficiency for reuse cycles, with superior values after 60 min and 90 min. Peptide synthesis was observed under all studied conditions. Analyses (DPPH, β-carotene/linoleic acid, FRAP) confirmed the antioxidant potential of the peptides generated by the immobilized enzyme. However, the immobilized enzyme in ACG 50 and ACG 100, combined with longer hydrolysis times, allowed the formation of peptides with an antioxidant capacity greater than or equivalent to those generated by the free enzyme, despite reduced enzymatic activity.

Preliminary assessment of environmental safety (ecosafety) of dextrin-based nanosponges for environmental applications

The ability to employ waste products, such as vegetable scraps, as raw materials for the synthesis of new promising adsorbing materials is at the base of the circular economy and end of waste concepts. Dextrin-based nanosponges (D_NS), both cyclodextrin (CD) and maltodextrin (MD), have shown remarkable adsorption abilities in the removal of toxic compounds from water and wastewater, thus representing a bio-based low-cost solution which is establishing itself in the market. Nevertheless, their environmental safety for either aquatic or terrestrial organisms has been overlooked, raising concern in terms of potential hazards to natural ecosystems. Here, the environmental safety (ecosafety) of six newly synthesized batches of D_NS was determined along with their full characterization by means of dynamic light scattering (DLS), thermogravimetric analysis (TGA), Fourier transformed infrared spectroscopy with attenuated total reflection (FTIR-ATR) and transmission electron microscopy (SEM). Ecotoxicity evaluation was performed using a battery of model organisms and ecotoxicity assays, such as the microalgae growth inhibition test using the freshwater Raphidocelis subcapitata and the marine diatom Dunaliella tertiolecta, regeneration assay using the freshwater cnidarian Hydra vulgaris and immobilization assay with the marine brine shrimp Artemia franciscana. Impact on seedling germination of a terrestrial plant of commercial interest, Cucurbita pepo was also investigated. Ecotoxicity data showed mild to low toxicity of the six batches, up to 1 mg/mL, in the following order: R. subcapitata >H. vulgaris >D. tertiolecta >A. franciscana > C. pepo. The only exception was represented by one batch (NS-Q+_BDE_(GLU2) which resulted highly toxic for both freshwater species, R. subcapitata and H. vulgaris. Those criticalities were solved with the synthesis of a fresh new batch and were hence attributed to the single synthesis and not to the specific D_NS formulation. No effect on germination of pumpkin but rather more a stimulative effect was observed. To our knowledge this is the first evaluation of the environmental safety of D_ NS. As such we emphasize that current formulations and exposure levels in the range of mg/mL do not harm aquatic and terrestrial species thus representing an ecosafe solution also for environmental applications.

Influence of Silver Fiber Morphology on the Dose-Response Relationship and Enrichment in Daphnia magna Studied by Elemental Imaging with LA-ICP-TOF-MS.

This study aims to enhance the understanding of the environmental risks associated with nanomaterials, particularly nanofibers. Previous research suggested that silver fibers exhibit higher toxicity (EC50/48h 1.6-8.5 μg/L) compared to spherical silver particles (EC50/48h 43 μg/L). To investigate the hypothesis that toxicity is influenced by the morphology and size of nanomaterials, various silver nanofibers with different dimensions (length and diameter) were selected. The study assessed their toxicity toward Daphnia magna using the 48 h immobilization assay. The EC50 values for the different fibers ranged from 122 to 614 μg/L. Subsequently, the study quantified the uptake and distribution of two representative nanofibers in D. magna neonates by employing digestion and imaging mass spectrometry in the form of laser-ablation-ICP-MS. A novel sample preparation method was utilized, allowing the analysis of whole, intact daphnids, which facilitated the localization of silver material and prevented artifacts. The results revealed that, despite the similar ecotoxicity of the silver fibers, the amount of silver associated with the neonates differed by a factor of 2-3. However, both types of nanofibers were primarily found in the gut of the organisms. In conclusion, the findings of this study do not support the expectation that the morphology or size of silver materials affect their toxicity to D. magna.

Polymers of low concern? Assessment of microplastic particles used in 3D printing regarding their toxicity on Raphidocelis subcapitata and Daphnia magna

The potential effects of microplastic particle exposure on environmental organisms has sparked intense research activities. Various studies have been conducted, however on a limited set of mostly pristine polymer materials. In parallel to the ongoing research activities, it is discussed to include non-natural polymers into the registration process under REACH. Currently, non-natural polymers are exempted from registration, based on the general assumption of being non-hazardous due to their high molecular weight. In addition, the extensive number of polymers exceeding the registration capacities was mentioned as a reason for exemption. Hence, relevant polymers requiring a registration shall be selected according to specific criteria that help to identify those with a concern for hazardous effects. In the line of these developments we here present the results of a systematic ecotoxicity testing of 16 microplastic particles of different polymer composition, part of which are micronized polymer powders used in 3D printing (intentionally produced primary microplastic, losing particle shape by the 3D printing). All polymer materials were systematically varied and extensively characterised with regard to their properties (e.g. particle size, cross-linking, molar mass distribution, end groups, reactivity). Most of the polymers exerted toxicity in 48 h acute Daphnia magna immobilisation assay and 72 h chronic Raphidocelis subcapitata growth inhibition assay, except one PA and one HDPE material showing toxicity in D. magna, and one HDPE material showing toxicity in green algae. From these results we conclude that none of the microplastic particles studied here, independent of their polymer properties, give raise to concern for hazardous effects.

Cyclic Disulfide Liposomes for Membrane Functionalization and Cellular Delivery

Liposomes are effective therapeutic delivery nanocarriers due to their ability to encapsulate and enhance the pharmacokinetic properties of a wide range of therapeutics. Two primary areas in which improvement is needed for liposomal drug delivery is to enhance the ability to infiltrate cells and to facilitate derivatization of the liposome surface. Herein, we report a liposome platform incorporating a cyclic disulfide lipid (CDL) for the dual purpose of enhancing cell entry and functionalizing the liposome membrane through thiol-disulfide exchange. In order to accomplish this, CDL-1 and CDL-2, composed of lipoic acid (LA) or asparagusic acid (AA) appended to a lipid scaffold, were designed and synthesized. A fluorescence-based microplate immobilization assay was implemented to show that these compounds enable convenient membrane decoration through reaction with thiol-functionalized small molecules. Additionally, fluorescence microscopy experiments indicated dramatic enhancements in cellular delivery when CDLs were incorporated within liposomes. These results demonstrate that multifunctional CDLs serve as an exciting liposome system for surface decoration and enhanced cellular delivery.

Production and characterization of a human antisperm monoclonal antibody against CD52g for topical contraception in women.

BackgroundApproximately 40% of human pregnancies are unintended, indicating a need for more acceptable effective contraception methods. New antibody production systems make it possible to manufacture reagent-grade human monoclonal antibodies (mAbs) for clinical use. We used the Nicotiana platform to produce a human antisperm mAb and tested its efficacy for on-demand topical contraception.MethodsHeavy and light chain variable region DNA sequences of a human IgM antisperm antibody derived from an infertile woman were inserted with human IgG1 constant region sequences into an agrobacterium and transfected into Nicotiana benthamiana. The product, an IgG1 mAb [“Human Contraception Antibody” (HCA)], was purified on Protein A columns, and QC was performed using the LabChip GXII Touch protein characterization system and SEC-HPLC. HCA was tested for antigen specificity by immunofluorescence and western blot assays, antisperm activity by sperm agglutination and complement dependent sperm immobilization assays, and safety in a human vaginal tissue (EpiVaginal™) model.FindingsHCA was obtained at concentrations ranging from 0.4 to 4 mg/ml and consisted of > 90% IgG monomers. The mAb specifically reacted with a glycan epitope on CD52g, a glycoprotein produced in the male reproductive tract and found in abundance on sperm. HCA potently agglutinated sperm under a variety of relevant physiological conditions at concentrations ≥ 6.25 µg/ml, and mediated complement-dependent sperm immobilization at concentrations ≥ 1 µg/ml. HCA and its immune complexes did not induce inflammation in EpiVaginal™ tissue.InterpretationHCA, an IgG1 mAb with potent sperm agglutination and immobilization activity and a good safety profile, is a promising candidate for female contraception.FundingThis research was supported by grants R01 HD095630 and P50HD096957 from the National Institutes of Health.

Synthesis of Biogenic High-Magnesium Calcite and its Experimental Immobilization Effect on Cd2+

Biogenic carbonates usually differ significantly both in physical and chemical characteristics from those of the same chemical origin, and heavy metal immobilization is prominent among these differences. In this research, a biosynthetic method of high-magnesium calcite (HMC) generated by Bacillus velezensis (B. velezensis) in the presence of humic acid (HA) was developed to ascertain the immobilization characteristics of biogenic high-magnesium calcite (BHM) for Cd2+. The study showed that the presence of HA was crucial to the formation of crystalline BHM. The Cd2+ immobilization assay results indicated that significant amounts of Cd2+ could be adsorbed and the obtained data could fit the Langmuir equation (R 2 = 0.98, Q max = 65.36 mg/g), and the adsorption process conformed to the pseudo-second order kinetic model (R 2 = 0.99). In particular, BHM can maintain the adsorption capacity at above 100 mg/g even under acidic conditions (pH ≥ 3). Further analysis showed that the adsorption was mainly monolayer physical adsorption, and a small amount of chemisorption also occurred. Thermodynamic parameters (ΔG > 0, ΔH > 0, and ΔS < 0) indicated that the adsorption process was a non-spontaneous endothermic reaction. XRD and SEM-EDS analyses demonstrated that the culture precipitate obtained was mainly HMC, with numerous 10-nm pores based on Barrett-Joyner-Halenda theory of surface area. The results of TG-DTG/DTA analysis showed that the BHM contained about 31.31 ± 3.77% organic matter. Amorphous CdCO3 and the complex or chelate bearing Cd2+ formed in adsorption process could ensure low desorption, implying a novel prospect for remediation of heavy metal pollution using BHM.

A Novel Isoallergen Dau c 1.0401 in Carrot: Stability, Allergenicity, and Comparison with Other Isoallergens

Around 25% of food allergic persons in Central Europe suffer from carrot allergy caused by the major carrot allergen Dau c 1. Three different isoallergens, Dau c 1.01, Dau c 1.02 and Dau c 1.03 are identified. However, information about the qualitative and quantitative composition of natural (n)Dau c 1 is scarce. The new carrot allergen Dau c 1.0401 is identified on the mRNA and protein level by RT-PCR and mass spectrometry. It displays only around 60% sequence identity to the other known Dau c 1 isoallergens. NMR and CD-spectra are typical for a well-folded protein containing both α-helices and β-strands. It showed a poor refolding capacity after incubation at 95 °C. IgE-binding is impaired in immunoblots, whereas in inhibition assays IgE binding to soluble Dau c 1.0401 is detected and it clearly provoked a response in mediator release assays. Dau c 1.0401 is a new isoallergen which contributes to the allergenicity of carrots. The absence of immunoreactivity in immobilized assays indicates that IgE binding is impaired when the protein is blotted on a solid phase. Altogether, the results point out that its allergenicity can be reduced upon carrot processing.

Greening effect of slag cement-based concrete: Environmental and ecotoxicological impact

Materials containing ground granulated blast furnace slag (GGBFS) display a transient green–bluecolor after demolding. This greening effect has been investigated for leaching behavior and ecotoxicological impact. Color of concretes and pure pastes containing GGBFS was assessed with a portable spectrophotometer, and samples were then submitted to a tank monolith leaching test. Ecotoxicological tests were conducted on reference sample and a green concrete sample at both natural and adjusted pH of 8.1. Main results support that the temporary greening effect of GGBFS-containing materials has no particular impact neither on the chemistry of leachates, nor ecotoxicity. Additionally, alkaline leachates are the main issue of leached cement or GGBFS based materials with pH around 11.5–12.5. Alkaline pH is a preponderant factor of ecotoxicity to sensitive organisms such as Daphnia magna , immobilization assay (48 h) resulting in 5.10 Toxic Units (TU) for reference sample at pH 12.50 against 1.38 TU at pH 8.10. Furthermore, sulfides are a specific issue of GGBFS materials concentrated up to 0.94 mmol m −2 in leachates, having an ecotoxic impact on living organisms at all trophic levels. At pH 8.10, green concrete leachates have 4.85 TU for Raphidocelis subcapitata growth assay (sulfides concentration of 0.63 mmol m −2 ) against 3.0 TU for green concrete sample. However, sulfides are easily removed from natural solution by oxidation or evaporation. • Greening effect is not associated with hazardous leachates or ecotoxic effects. • Alkaline pH (> 11) is the main issue with cement or slag-based materials. • The algae R. subcapitata is not affected by high pH unlike the crustacean D. magna . • GBFS contain sulfides up to 1 wt.% release in eluates of concretes and pastes. • Sulfides are quickly neutralized in natural environment.

Application of effect-based methods (EBMs) in a river basin: a preliminary study in Central Italy.

Effect-based methods (EBMs), i.e. in vitro and in vivo bioassays, represent innovative tools for the effect detection of environmental chemical pollutants on living organisms. The aim of this study was to evaluate the water quality of a river ecosystem implementing two in vivo bioassays on target freshwater animal species: the crustacean Daphnia magna and the small fish Danio rerio, also known as zebrafish. The methods applied in this study, i.e. the Daphnia sp. Acute Immobilisation assay and the Fish Embryo Acute Toxicity (FET) test, are commonly used in water quality research and their application in short-term ecotoxicity detection is suggested by recent European projects. Two sampling sites were chosen in the urban part of the Tiber River in Rome, while a third one was chosen as a reference site in the Farfa River, a tributary upstream of the city. The sites in the Tiber River are potentially affected by different pollution sources, including urban and industrial wastewater discharges, the pesticide release, livestock waste products, and waste dumps. The results of the study showed wide differences between the two applied bioassays. The FET test was generally more sensitive in detecting even low effects in all the water samples, but the strongest statistically results were observed with the D. magna Acute Immobilisation test. The results of this research confirm the effectiveness of EBMs in investigating and monitoring water chemical pollution, and stress the need for performing further studies, e.g. chemical analyses and other bioassays, to improve the knowledge of the health status of the Tiber River basin. Further results will aim to support the local authorities in adopting measures to reduce and to eliminate the sources of chemical pollution in the study area.

Evaluation of acute toxicity of vinasse by means of Daphnia magna and Aliivibrio fischeri: a comparative study.

In the bioethanol industry, per liter of the produced alcohol 9 to 14 liters of vinasse are obtained as a byproduct. If the vinasse is directly shed into bodies of water without an adequate treatment, it may have negative effects on the existing biota and human health due to its high turbidity and color, low pH and high content of organic material. The purpose of this study was to assess the acute toxicity of vinasse by means of a rapid test with Aliivibrio fischeri and compare it with a standard immobilization assay with Daphnia magna. The standard assay of D. magna by means of its EC50 of 4.7% showed that organism was more sensitive to the contaminant, in comparison with the 69.6% obtained with the A. fischeri which suggests that it should be continuesly used as one of the organisms of first choice for the evaluation of the acute toxicity of this effluent.

Rapid screening for ecotoxicity of plating and semiconductor wastewater employing the heartbeat of Daphnia magna

Industrial wastewater discharge is one of major threats to the sustainability of aquatic environment. Rapid and sensitive detection of toxic wastewater discharge and appropriate control if necessary are therefore crucial. In the present study, a 1 h Daphnia magna exposure with heartbeat as an observation endpoint was developed and assessed for its utility as a rapid toxicity screening measure. Two types of metal-rich industrial wastewater, i.e., metal plating and the semiconductor industry were chosen as target samples, and the 1 h heartbeat assay was applied. Based on a literature search, four metals, i.e., Cu, Cr, Ni and Zn were identified as major chemicals of ecotoxicological concerns in these wastewaters. The effective concentrations determined for each metal from the D. magna 1 h heartbeat test were comparable to those derived from the conventional D. magna 48 h immobilization test. Copper sulfate (CuSO4) was determined as the most toxic, followed by potassium dichromate (K2Cr2O7), nickel sulfate (NiSO4) and zinc sulfate (ZnSO4). For ternary mixtures, the 1 h heartbeat test showed also comparable responses to those of the 48 h immobilization test, suggesting its utility for screening the toxicity of simple metal mixtures. For the site-sampled metal plating water, the rapid heartbeat assay showed similar responses to those of the 48 h immobilization assay. However, for the semiconductor industry wastewater, clearly different responses were observed in both the heartbeat and immobilization assays, probably due to the influence of other contaminants with different modes of action that are present in the wastewater. Our observations showed that the D. magna 1 h heartbeat test can be considered as a rapid ecotoxicity screening measure for certain wastewaters with simple metal mixtures.

A Rapid Response Electrochemical Biosensor for Detecting Thc In Saliva

Marijuana is listed as a Schedule I substance under the American Controlled Substances Act of 1970. As more U.S. states and countries beyond the U.S. seek legalization, demands grow for identifying individuals driving under the influence (DUI) of marijuana. Currently no roadside DUI test exists for determining marijuana impairment, thus the merit lies in detecting the primary and the most sought psychoactive compound tetrahydrocannabinol (THC) in marijuana. Salivary THC levels are correlated to blood THC levels making it a non-invasive medium for rapid THC testing. Affinity biosensing is leveraged for THC biomarker detection through the chemical reaction between target THC and THC specific antibody to a measure signal output related to the concentration of the targeted biomarker. Here, we propose a novel, rapid, electrochemical biosensor for the detection of THC in saliva as a marijuana roadside DUI test with a lower detection limit of 100 pg/ml and a dynamic range of 100 pg/ml – 100 ng/ml in human saliva. The developed biosensor is the first of its kind to utilize affinity-based detection through impedimetric measurements with a rapid detection time of less than a minute. Fourier transform infrared spectroscopy analysis confirmed the successful immobilization of the THC immobilization assay on the biosensing platform. Zeta potential studies provided information regarding the stability and the electrochemical behavior of THC immunoassay in varying salivary pH buffers. We have demonstrated stable, dose dependent biosensing in varying salivary pH’s. A binary classification system demonstrating a high general performance (AUC = 0.95) was employed to predict the presence of THC in human saliva. The biosensor on integration with low-power electronics and a portable saliva swab serves as a roadside DUI hand-held platform for rapid identification of THC in saliva samples obtained from human subjects.

Ecotoxicity of Nitrogen, Sulfur, or Oxygen Heterocycles and Short-Chained Alkyl Phenols Commonly Detected in Contaminated Groundwater.

Nitrogen, sulfur, or oxygen heterocyclic aromatic hydrocarbons (NSO-HETs) and short-chained alkyl phenols (SCAPs) are commonly detected in groundwater at contaminated sites and in the surrounding environment. It is now scientific consensus that these chemicals pose a risk to human and ecosystem health. However, toxicity data are comparably fragmentary, and only few studies have addressed the ecotoxicity of NSO-HETs and SCAPs in a systematic and comparative fashion. To overcome this shortcoming, we tested 18 SCAPs, 16 NSO-HETs, as well as the homocyclic hydrocarbons indane and indene in the Microtox® assay with Aliivibrio fischeri, the growth inhibition test with Desmodesmus subspicatus, the acute immobilization assay with Daphnia magna, as well as the fish embryo toxicity test with embryos of the zebrafish (Danio rerio). Because of the physicochemical properties of the tested chemicals (limited water solubility, volatility, and sorption to test vessels), actual exposure concentrations in test media and their dissipation over time were analytically quantified by means of gas chromatography with mass spectrometry. Analytically corrected effect levels (median effect and lethal concentrations) ranged from 0.017 to 180 mg L-1 , underlining the environmental relevance of some NSO-HETs and SCAPs. Para-substituted phenols showed the overall greatest toxicities in all 4 toxicity tests. We provide, for the first time, a complete high-quality data set in support of better environmental risk assessments of these chemicals. Environ Toxicol Chem 2019;38:1343-1355. © 2019 SETAC.

Biofilm formation by ESBL-producing strains of Escherichia coli and Klebsiella pneumoniae

ObjectivesBiofilm production in extended spectrum β-lactamase (ESBL)-producing Enterobacteriaceae provides a favourable environment for the exchange of antibiotic-resistance genes and could facilitate widespread dissemination. We aimed to assess biofilm development in ESBL-producing E. coli and K. pneumoniae isolates and determine how development relates to microbiological characteristics and clinical outcomes. Methods147 ESBL-producing E. coli and 82 K. pneumoniae were genetically characterized. Biofilm formation was measured at 1.5, 4, 6, and 24 h during culture in blood heart infusion using a microbead immobilization assay (BioFilm Ring test®). Results were given as biofilm formation index (BFI) with lower values indicating increased presence of biofilm (range = 0–21). ResultsIn total, 57.1% of strains were strong producers of biofilm (BFI < 2), whereas 13.4% lacked biofilm production (BFI > 18). Standard biofilm production (BFI < 7) was common in E. coli isolates (61.9%). For E. coli, biofilm production was less frequently observed in ST131 clones (p = 0.03) but more frequently in strains harbouring toxin (p = 0.008) or adhesin (p = 0.008) virulence factor genes. Despite almost all K. pneumoniae having standard biofilm production (90.2%), there was a 2.4-times higher odds of observing biofilm in ST29/147/323 versus other ST-types (p = 0.13). Patients with standard biofilm producing isolates were not at increased risk of transfer to intensive-care (odds-ratio=2.80, 95%CI=0.59–13.21) or death within 12-months (odds-ratio=1.61, 95%CI=0.75–3.43). ConclusionIn these ESBL-producing strains, biofilm production is linked to certain virulence factors in E. coli and is common in K. pneumoniae. Further exploration of whether biofilm production increases dissemination and risk of severe clinical outcomes is needed in larger collections of isolates.

Aquatic toxicity and loss of linear alkylbenzenesulfonates alone and in a neonicotinoid insecticide formulation in surface water

Substance losses of linear alkylbenzene sulfonates (LASs) in a neat surfactant mixture, in an insecticide formulation Mospilan 20 SG, and in solutions with different neonicotinoid active ingredients (AIs) was studied in distilled water and in surface water samples originated from River Danube. Analytical measurements were performed both by HPLC-UV and commercial ELISA methods. Loss rates of LASs were found different in these aqueous matrices, with decomposition rates higher for the neat surfactant mixture than for Mospilan 20 SG (nearly 2- and 9-fold in distilled water and in surface water from River Danube, respectively). Half-lives determined in surface water from River Danube were shown to be affected by the presence of neonicotinoid AIs thiacloprid > imidacloprid > acetamiprid (ACE), while clothianidin and thiamethoxam did not affect LAS decomposition. Aquatic toxicity of Mospilan 20 SG, along with that of its AI ACE and co-formulant LAS, as well as the mixture of ACE and LAS was also investigated in the 48-h acute immobilisation assay on the water flea (Daphnia magna) aquatic indicator organism. LAS appeared to be significantly (8-fold) more toxic in the D. magna test than ACE, and the toxicity of the formulated insecticide was found to be 1.3 and 19.6 times higher than explained by its AI and LAS content, respectively, indicating synergistic toxicity. The strongest synergy between ACE and LASs was observed, when the neat forms of the two substances were applied in combination at concentrations equivalent to those in Mospilan 20 SG.

Aquatic toxicity of biofuel candidates on Daphnia magna

The increasing need for carbon-neutral, low-emission transportation sector has led to the development of advanced biofuels with tailor-made production and combustion processes. Even though the large-scale deployment of these advanced biofuels also increases the risk for their release into the environment, their toxic potency remains largely unknown. To identify hazardous biofuel candidates as early as possible, the fuel development process can be expanded by “Green Toxicology”. To demonstrate such early Green Toxicology testing, this study investigates the aquatic toxicity for the two biofuel candidates 2-methyltetrahydrofuran (2-MTHF) and 2-methylfuran (2-MF) on Daphnia magna. We performed the prolonged acute immobilisation assay (96 h) and the D. magna reproduction test. 2-MF induced acute effects on D. magna that were two orders of magnitude stronger than those of 2-MTHF. Furthermore, both substances affected the growth and reproductive output of D. magna in a 21 d reproduction test, with 2-MF already inducing effects with concentrations one to two orders of magnitude lower than those of 2-MTHF. Thus, our assessment of the aquatic toxicity suggests that further biofuel development should focus on 2-MTHF. Furthermore, the acute immobilisation test with D. magna was identified as a promising tool for a rapid and sensitive “Green Toxicology” screening of further biofuel candidates.

Carbon fibers with a nano-hydroxyapatite coating as an excellent biofilm support for bioreactors

A biofilm support with high biocompatibility is needed for bioreactors. A nano-hydroxyapatite (HA) coating on carbon fibers (CFs) was prepared by electrochemical deposition (ECD). The sludge immobilization assays, bacterial cells adhesion assays and Derjaguin–Landau–Verwey–Overbeek (DLVO) theory were used to evaluate the capacity of CF supports to immobilize activated sludge and bacterial cells. The sludge immobilization and bacterial cells adhesion assays illustrated that HA coating could enhance the capacity of CFs to immobilize microorganisms. SEM images showed that HA and bacterial cells formed a dense film on CFs surface. In addition, HA, acting as a glue, could combine CFs with bacterial cells or between cells, which helped CFs capture more bacterial cells. DLVO theory illustrated that CFs with HA coating had a lower total interaction energy than CFs without handling, explaining the higher capacity of CFs with HA coating to immobilize bacterial cells. This result was owning to the less negative zeta potential and higher hydrophilicity of CFs with HA coating, and the hydrophilicity made a greater contribution to the lower total interaction energy. Experiments and theory reveal that HA coating could enhance the biocompatibility of CFs, and CFs with HA coating could be used as an excellent biofilm support for bioreactors.

The significance of nanomaterial post-exposure responses in Daphnia magna standard acute immobilisation assay: Example with testing TiO2 nanoparticles

One of the most widely used aquatic standarized tests for the toxicity screening of chemicals is the acute toxicity test with the freshwater crustacean Daphnia magna, which has also been applied in the toxicity screening of manufactured nanoparticles (NPs). However, in the case of non-soluble NPs most of the results of this test have showed no effect. The aim of the work presented here was to modify the standardized test by the least possible extent to make it more sensitive for non-soluble particles. The standard acute immobilisation assay with daphnids was modified by prolonging the exposure period and by measuring additional endpoints. Daphnids were exposed to TiO2 NPs in a standard acute test (48h of exposure), a standard acute test (48h of exposure) followed by 24h recovery period in clean medium or a prolonged exposure in the NPs solutions totaling 72h. Together with immobility, the adsorption of NPs to body surfaces was also observed as an alternative measure of the NPs effects. Our results showed almost no effect of TiO2 NPs on D. magna after the 48h standard acute test, while immobility was increased when the exposure period to TiO2 NPs was prolonged from 48h to 72h. Even when daphnids were transferred to clean medium for additional 24h after 48h of exposure to TiO2 NPs the immobility increased. We conclude that by transferring the daphnids to clean medium at the end of the 48h exposure to TiO2 NPs, the delayed effects of the tested material can be seen. This methodological step could improve the sensitivity of D. magna test as a model in nanomaterial environmental risk assessment.