Validation of Salamander Dermal Mucus Swabs as a Novel, Nonlethal Approach for Amphibian Metabolomics and Glutathione Analysis Following Pesticide Exposure.

Evaluating biomarkers of stress in amphibians is critical to conservation, yet current techniques are often destructive and/or time-consuming, which limits ease of use. In the present study, we validate the use of dermal swabs in spotted salamanders (Ambystoma maculatum) for biochemical profiling, as well as glutathione (GSH) stress response following pesticide exposure. Thirty-three purchased spotted salamanders were acclimated to laboratory conditions at Washington College (Chestertown, MD, USA) for 4 weeks. Following acclimation, salamanders were randomly sorted into three groups for an 8-h pesticide exposure on soil: control with no pesticide, 2,4-dichlorophenoxyacetic acid (2,4-D), or chlorpyrifos. Before and after exposure, mucus samples were obtained by gently rubbing a polyester-tipped swab 50 times across the ventral and dorsal surfaces. Salamanders were humanely euthanized and dissected to remove the brain for acetylcholinesterase and liver for GSH and hepatic metabolome analyses, and a whole-body tissue homogenate was used for pesticide quantification. Levels of GSH were present in lower quantities on dermal swabs relative to liver tissues for chlorpyrifos, 2,4-D, and control treatments. However, 2,4-D exposures demonstrated a large effect size increase for GSH levels in livers (Cohen's d = 0.925, p = 0.036). Other GSH increases were statistically insignificant, and effect sizes were characterized as small for 2,4-D mucosal swabs (d = 0.36), medium for chlorpyrifos mucosal swabs (d = 0.713), and negligible for chlorpyrifos liver levels (d = 0.012). The metabolomics analyses indicated that the urea cycle, alanine, and glutamate metabolism biological pathways were perturbed by both sets of pesticide exposures. Obtaining mucus samples through dermal swabbing in amphibians is a viable technique for evaluating health in these imperiled taxa. Environ Toxicol Chem 2024;43:1126-1137. © 2024 SETAC.

Magnetic graphene derivates for efficient herbicide removal from aqueous solution through adsorption.

2,4-Dichlorophenoxyacetic acid (2,4-D) is an herbicide and is among the most widely distributed pollutant in the environment and wastewater. Herein is presented a complete comparison of adsorption performance between two different magnetic carbon nanomaterials: graphene oxide (GO) and its reduced form (rGO). Magnetic functionalization was performed employing a coprecipitation method, using only one source of Fe2+, requiring low energy, and potentially allowing the control of the amount of incorporated magnetite. For the first time in literature, a green reduction approach for GO with and without Fe3O4, maintaining the magnetic behavior after the reaction, and an adsorption performance comparison between both carbon nanomaterials are demonstrated. The nanoadsorbents were characterized by FTIR, XRD, Raman, VSM, XPS, and SEM analyses, which demonstrates the successful synthesis of graphene derivate, with different amounts of incorporate magnetite, resulting in distinct magnetization values. The reduction was confirmed by XPS and FTIR techniques. The type of adsorbent reveals that the amount of magnetite on nanomaterial surfaces has significant influence on adsorption capacity and removal efficiency. The procedure demonstrated that the best performance, for magnetic nanocomposites, was obtained by GO∙Fe3O4 1:1 and rGO∙Fe3O4 1:1, presenting values of removal percentage of 70.49 and 91.19%, respectively. The highest adsorption capacity was reached at pH 2.0 for GO∙Fe3O4 1:1 (69.98 mg g-1) and rGO∙Fe3O4 1:1 (89.27 mg g-1), through different interactions: π-π, cation-π, and hydrogen bonds. The adsorption phenomenon exhibited a high dependence on pH, initial concentration of adsorbate, and coexisting ions. Sips and PSO models demonstrate the best adjustment for experimental data, suggesting a heterogeneous surface and different energy sites, respectively. The thermodynamic parameters showed that the process was spontaneous and exothermic. Finally, the nanoadsorbents demonstrated a high efficiency in 2,4-D adsorption even after five adsorption/desorption cycles.

Meta-analytical review of antioxidant mechanisms responses in animals exposed to herbicide 2,4-D herbicide.

The 2,4-Dichlorophenoxyacetic acid (2,4-D) is a low-cost herbicide to eradicate broadleaf weeds. Since the development of 2,4-D resistant transgenic crops, it has been described as one of the most widely distributed pollutants in the world, increasing concern about its environmental impacts. This study aimed to elucidate the antioxidant system response in animals exposed to 2,4-D by different routes of exposure. It focused on determining if tissue, phylogenetic group, and herbicide formulation would influence the antioxidant mechanisms. A careful literature search of Scopus, WoS, and Science Direct retrieved 6983, 24,098, and 20,616 articles, respectively. The dataset comprised 390 control-treatment comparisons and included three routes of exposure: transgenerational, oral, and topical. The data set for transgenerational and oral exposure revealed oxidative stress through a decrease in enzymatic activities and the level of molecules of the antioxidant system. In contrast, topical exposure increased the oxidative stress. Tissue-specific analyses revealed that the transgenerational effects reduced hepatic catalase (CAT) activity. Oral exposure caused a variety of effects, including increased CAT activity in the prostate and decreased activity in various tissues. Mammals predominate in the transgenerational and oral groups, showing a significantly reduced activity of the antioxidant system. In contrast, in the topical exposure, an increased activity of oxidative stress biomarkers was observed in fish, earthworms, and mollusks. The effects of the 2,4-D formulation on oxidative stress responses showed significant differences between pure and commercial formulations, with oral exposure resulting in decreased activity and topical exposure increasing responses. In summary, orally exposed animals exhibited a clear decrease in enzyme activities, transgenerational exposure elicited tissue-specific prompted biochemical reductions, and topical exposure induced increased responses, emphasizing the need for unbiased exploration of the effects of 2,4-D on biomarkers of oxidative stress while addressing publication bias in oral and topical datasets.

Taguchi optimisation of the synthesis of vine-pruning-waste hydrochar as potential adsorbent for pesticides in water.

This research aimed to synthesise an effective hydrochar adsorbent from vineyard pruning wastes to remove emerging contaminants as a potential valorisation product. The adsorption capacity of the hydrochar was optimised using the Taguchi method. Four synthesis variables were evaluated: hydrothermal reaction temperature, use of H3PO4 as a catalyst, number of acetone washes, and type of chemical cold activation. The simultaneous adsorption of five model pesticides (clothianidin (CTD), acetamiprid (ACE), 2,4-D, metalaxyl (MET), and atrazine (ATZ)) at an initial pH of 7 was studied. At optimum conditions, the hydrochar presented a total adsorption capacity of 22.7 µmol/g, representing a 2.7-fold improvement with respect to pristine hydrochar performance. High percentage removals were achieved for all pollutants (85 % CTD, 94 % ACE, 86 % MET, and 95 % ATZ) except for 2,4-D (4 %). This research provides a valuable reference for developing hydrochar adsorbents for pollution control and the valorisation of biomass wastes.

Pesticide-induced alterations in zebrafish (Danio rerio) behavior, histology, DNA damage and mRNA expression: An integrated approach.

To assess the impact of glyphosate and 2,4-D herbicides, as well as the insecticide imidacloprid, both individually and in combination, the gills of adult zebrafish were used due to their intimate interaction with chemicals diluted in water. Bioassays were performed exposing the animals to the different pesticides and their mixture for 96 h. The behavior of the fish was analyzed, a histological examination of the gills was carried out, and the genotoxic effects were also analyzed by means of the comet assay (CA) and the change in the expression profiles of genes involved in the pathways of the oxidative stress and cellular apoptosis. The length traveled and the average speed of the control fish, compared to those exposed to the pesticides and mainly those exposed to the mixture, were significantly greater. All the groups exposed individually exhibited a decrease in thigmotaxis time, indicating a reduction in the behavior of protecting themselves from predators. Histological analysis revealed significant differences in the structures of the gill tissues. The quantification of the histological lesions showed mild lesions in the fish exposed to imidacloprid, moderate to severe lesions for glyphosate, and severe lesions in the case of 2,4-D and the mixture of pesticides. The CA revealed the sensitivity of gill cells to DNA damage following exposure to glyphosate, 2,4-D, imidacloprid and the mixture. Finally, both genes involved in the oxidative stress pathway and those related to the cell apoptosis pathway were overexpressed, while the ogg1 gene, involved in DNA repair, was downregulated.

Synthetic auxin herbicide 2,4-D and its influence on a model BY-2 suspension.

2,4-D is a broadly used auxin herbicide. The presence of the 2,4-D synthetic auxin in the medium is imperative for long-term BY-2 tobacco suspension viability. The precise mechanism of this symbiosis of the suspension and the synthetic auxin remains unclear. Our goal was to study the hormonal regulation of the growth of the cell suspension; and to describe the experiments clarifying the interaction between the chosen growth regulators and phytohormones on the cellular level, specifically between the 2,4-D synthetic auxin and the native stress phytohormone - ethylene. This study examined the influence of low 2,4-D concentrations stimulating cell growth in vitro as well as the influence of high herbicide concentrations on the model tobacco BY-2 suspension. The culture took 6 days. Different parameters were evaluated, including the influence of different 2,4-D concentrations on the production of the phytohormone ethylene and its precursor 1-Aminocyclopropane-1-carboxylic acid (ACC) in the tobacco cells. The content of 2,4-D in the cells and the medium was established. The observations of the morphological changes showed that a heavy impregnation of the cell walls taking place depending on the concentration of 2,4-D. A dramatic increase in protective polysaccharides and a remodulation of the cell walls by the formation of a pectin shield in artificial conditions were expected and observed. At the same time, massive production of the stress phytohormone ethylene took place, and, because of that, plant mutagenicity, anomalous tumour-type proliferation growth, and the production of supercells were observed. The hypothesis of the protective shield is discussed.

Effects of pasture intensification and sugarcane cultivation on non-target species: A realistic evaluation in pesticide-contaminated mesocosms.

Conventional soil management in agricultural areas may expose non-target organisms living nearby to several types of contaminants. In this study, the effects of soil management in extensive pasture (EP), intensive pasture (IP), and sugarcane crops (C) were evaluated in a realistic-field-scale study. Thirteen aquatic mesocosms embedded in EP, IP, and C treatments were monitored over 392 days. The recommended management for each of the areas was simulated, such as tillage, fertilizer, pesticides (i.e. 2,4-D, fipronil) and vinasse application, and cattle pasture. To access the potential toxic effects that the different steps of soil management in these areas may cause, the cladoceran Ceriophania silvestrii was used as aquatic bioindicator, the dicot Eruca sativa as phytotoxicity bioindicator in water, and the dipteran Chironomus sancticaroli as sediment bioindicator. Generalized linear mixed models were used to identify differences between the treatments. Low concentrations of 2,4-D (<97 µg L-1) and fipronil (<0.21 µg L-1) in water were able to alter fecundity, female survival, and the intrinsic rate of population increase of C. silvestrii in IP and C treatments. Similarly, the dicot E. sativa had germination, shoot and root growth affected mainly by 2,4-D concentrations in the water. For C. sancticarolli, larval development was affected by the presence of fipronil (<402.6 ng g-1). The acidic pH (below 5) reduced the fecundity and female survival of C. silvestrii and affected the germination and growth of E. sativa. Fecundity and female survival of C. silvestrii decrease in the presence of phosphorus-containing elements. The outcomes of this study may improve our understanding of the consequences of exposure of freshwater biota to complex stressors in an environment that is rapidly and constantly changing.

Adverse effects of 2,4-D dimethylammonium based-herbicide on Acetylcholinesterase expression in Nile tilapia (Oreochromis niloticus).

In this study, the evaluation of a 2,4-D dimethylammonium based-herbicide impacted on Nile tilapia was done. The effects focus on Acetylcholinesterase (AChE) expression in the brain, gill, muscle, and plasma using antibody techniques. Our findings revealed a decrease in AChE expression with prolonged exposure. For these, AChE was purified using hydroxyapatite column chromatography. Moreover, the isolated protein was characterized as AChE by Polyclonal Ab specific to AChE through the Western blot. For interpretation at the cellular and molecular level, we employed two analytical techniques, histology, and optical coherence tomography (OCT). Alterations in the gill, liver, and muscle were observed to increase with increased exposure time. Field study concludes that AChE could serve as a biomarker to detect herbicide contamination in water and its accumulation in aquatic animals. This study may aid in surveillance and strategy formulation for managing contamination from such substances in various water sources.

Application of mid-infrared spectroscopy to the prediction and specification of pesticide sorption: A promising and cost-effective tool.

The cocktail of pesticides sprayed to protect crops generates a miscellaneous and generalized contamination of water bodies. Sorption, especially on soils, regulates the spreading and persistence of these contaminants. Fine resolution sorption data and knowledge of its drivers are needed to manage this contamination. The aim of this study is to investigate the potential of Mid-Infrared spectroscopy (MIR) to predict and specify the adsorption and desorption of a diversity of pesticides. We constituted a set of 37 soils from French mainland and West Indies covering large ranges of texture, organic carbon, minerals and pH. We measured the adsorption and desorption coefficients of glyphosate, 2,4-dichlorophenoxyacetic acid (2,4-D) and difenoconazole and acquired MIR Lab spectra for these soils. We developed Partial Least Square Regression (PLSR) models for the prediction of the sorption coefficients from the MIR spectra. We further identified the most influencing spectral bands and related these to putative organic and mineral functional groups. The prediction performance of the PLSR models was generally high for the adsorption coefficients Kdads (0.4 < R2 < 0.9 & RPIQ >1.8). It was contrasted for the desorption coefficients and related to the magnitude of the desorption hysteresis. The most significant spectral bands in the PLSR differ according to the pesticides indicating contrasted interactions with mineral and organic functional groups. Glyphosate interacts primarily with polar mineral groups (OH) and difenoconazole with hydrophobic organic groups (CH2, CC, COO-, C-O, C-O-C). 2,4-D has both positive and negative interactions with these groups. Finally, this work suggests that MIR combined with PLSR is a promising and cost-effective tool. It allows both the prediction of adsorption and desorption parameters and the specification of these mechanisms for a diversity of pesticides including polar active ingredients.

Transient efflux inhibition improves plant regeneration by natural auxins.

Plant genome editing and propagation are important tools in crop breeding and production. Both rely heavily on the development of efficient in vitro plant regeneration systems. Two prominent regeneration systems that are widely employed in crop production are somatic embryogenesis (SE) and de novo shoot regeneration. In many of the protocols for SE or shoot regeneration, explants are treated with the synthetic auxin analog 2,4-dichlorophenoxyacetic acid (2,4-D), since natural auxins, such as indole-3-acetic acid (IAA) or 4-chloroindole-3-acetic acid (4-Cl-IAA), are less effective or even fail to induce regeneration. Based on previous reports that 2,4-D, compared to endogenous auxins, is not effectively exported from plant cells, we investigated whether efflux inhibition of endogenous auxins could convert these auxins into efficient inducers of SE in Arabidopsis immature zygotic embryos (IZEs). We show that natural auxins and synthetic analogs thereof become efficient inducers of SE when their efflux is transiently inhibited by co-application of the auxin transport inhibitor naphthylphthalamic acid (NPA). Moreover, IZEs of auxin efflux mutants pin2 or abcb1 abcb19 show enhanced SE efficiency when treated with IAA or efflux-inhibited IAA, confirming that auxin efflux reduces the efficiency of Arabidopsis SE. Importantly, in contrast to the 2,4-D system, where only 50-60% of the embryos converted to seedlings, all SEs induced by transport-inhibited natural auxins converted to seedlings. Efflux-inhibited IAA, like 2,4-D, also efficiently induced SE from carrot suspension cells, whereas IAA alone could not, and efflux-inhibited 4-Cl-IAA significantly improved de novo shoot regeneration in Brassica napus. Our data provides new insights into the action of 2,4-D as an efficient inducer of plant regeneration but also shows that replacing this synthetic auxin for efflux-inhibited natural auxin significantly improves different types of plant regeneration, leading to a more synchronized and homogenous development of the regenerated plants.

Seed germination and vegetative and in vitro propagation of Hieracium lucidum subsp. lucidum (Asteraceae), a critically endangered endemic taxon of the Sicilian flora.

Hieracium lucidum subsp. lucidum is a critically endangered endemic taxa of the Sicilian flora. It is a relict of the Tertiary period surviving on the cliffs of Monte Gallo (NW-Sicily). This research focused on finding the best protocols for seed germination and vegetative and in vitro propagation to contribute to ex situ conservation. Seed germination tests were carried out using constant temperatures of 15 °C, 20 °C and 25 °C in continuous darkness and an alternating temperature of 30/15 °C (16 h/8 h, light/dark). The seeds had no dormancy, and a high germination capacity (70-95%) was obtained at all tested thermoperiods. The possibility of vegetative propagation of the taxon was evaluated through the rooting capacity of stem cuttings treated or not treated with indole-3-butyric acid (IBA). All cuttings were treated with IBA rooted within 2 months, while only 50% of the untreated cuttings were rooted within a longer time. An efficient protocol for rapid in vitro propagation from leaf portions was developed. The response of explants was tested on hormone-free Murashige and Skoog (MS) basal medium and MS enriched with different types of cytokinins: 6-Benzylaminopurine (BAP) and meta-Topolin (mT) in combination with naphthaleneacetic acid (NAA) and 2,4-Dichlorophenoxyacetic acid (2,4-D) at the same concentration. The combination of mT (2 mg L-1) and 2,4-D (1 mg L-1) in the medium was the most effective and showed the highest percentage of callus induction and the mean number of regenerated shoots. The maximum rate of root regeneration and the maximum number and length of roots were obtained on hormone-free MS and MS enriched with IBA at concentrations of 1 mg L-1. From the results obtained, it can be concluded that H. lucidum subsp. lucidum can be successfully propagated using one of the tested techniques, subject to the availability of the material for reproduction.

Association of UGT1A1 gene variants, expression levels, and enzyme concentrations with 2,3,7,8-TCDD exposure in individuals exposed to Agent Orange/Dioxin.

Among the congener of dioxin, 2,3,7,8-TCDD is the most toxic, having a serious long-term impact on the environment and human health. UDP-glucuronosyltransferase 1A1 (UGT1A1) plays a crucial role in the detoxification and excretion of endogenous and exogenous lipophilic compounds, primarily in the liver and gastrointestinal tract. This study aimed to investigate the association of UGT1A1 gene polymorphisms, expression levels, and enzyme concentration with Agent Orange/Dioxin exposure. The study included 100 individuals exposed to Agent Orange/Dioxin nearby Da Nang and Bien Hoa airports in Vietnam and 100 healthy controls. UGT1A1 SNP rs10929303, rs1042640 and rs8330 were determined by Sanger sequencing, mRNA expression was quantified by RT-qPCR and plasma UGT1A1 concentrations were measured by ELISA. The results showed that UGT1A1 polymorphisms at SNPs rs10929303, rs1042640 and rs8330 were associated with Agent Orange/Dioxin exposure (OR = 0.55, P = 0.018; OR = 0.55, P = 0.018 and OR = 0.57, P = 0.026, respectively). UGT1A1 mRNA expression levels and enzyme concentration were significantly elevated in individuals exposed to Agent Orange/Dioxin compared to controls (P < 0.0001). Benchmark dose (BMD) analyses showed that chronic exposure to 2,3,7,8-TCDD contamination affects the UGT1A1 mRNA and protein levels. Furthermore, UGT1A1 polymorphisms affected gene expression and enzyme concentrations in individuals exposed to Agent Orange/Dioxin. In conclusion, UGT1A1 gene polymorphisms, UGT1A gene expression levels and UGT1A1 enzyme concentrations were associated with Agent Orange/Dioxin exposure. The metabolism of 2,3,7,8-TCDD may influence UGT1A gene expression and enzyme concentrations.

A Ratiometric Biosensor Containing Manganese Dioxide Nanosheets and Nitrogen-Doped Quantum Dots for 2,4-Dichlorophenoxyacetic Acid Monitoring.

Nanomaterials are desirable for sensing applications. Therefore, MnO2 nanosheets and nitrogen-doped carbon dots (NCDs) were used to construct a ratiometric biosensor for quantification of 2,4-dichlorophenoxyacetic acid. The MnO2 nanosheets drove the oxidation of colorless o-phenylenediamine to OPDox, which exhibits fluorescence emission peaks at 556 nm. The fluorescence of OPDox was efficiently quenched and the NCDs were recovered as the ascorbic acid produced by the hydrolyzed alkaline phosphatase (ALP) substrate increased. Owing to the selective inhibition of ALP activity by 2,4-D and the inner filter effect, the fluorescence intensity of the NCDs at 430 nm was suppressed, whereas that at 556 nm was maintained. The fluorescence intensity ratio was used for quantitative detection. The linear equation was F = 0.138 + 3.863·C 2,4-D (correlation coefficient R2 = 0.9904), whereas the limits of detection (LOD) and quantification (LOQ) were 0.013 and 0.040 µg/mL. The method was successfully employed for the determination of 2,4-D in different vegetables with recoveries of 79%~105%. The fluorescent color change in the 2,4-D sensing system can also be captured by a smartphone to achieve colorimetric detection by homemade portable test kit.

Degradation of atrazine, glyphosate, and 2,4-D in soils collected from two contrasting crop rotations in Southwest Chaco, Argentina.

Argentina stands as one of the leading consumers of herbicides. In a laboratory incubation experiment, the persistence and production of degradation metabolites of Atrazine, 2,4-D, and Glyphosate were investigated in a loamy clay soil under two contrasting agricultural practices: continuous soybean cultivation (T1) and intensified rotations with grasses and legumes (T2). The soils were collected from a long-term no-till trial replicating the influence of the meteorological conditions in the productive region. The soil was enriched with diluted concentrations of 6.71, 9.95, and 24 mg a.i./kg-1 of soil for the respective herbicides, equivalent to annual doses commonly used in the productive region. Samples were taken at intervals of 0, 0.5, 1, 2, 4, 6, 8, 16, 32, and 64 days, and analysis was conducted using high-resolution liquid chromatography UPLC MS/MS. An optimal fit to the first-order kinetic model was observed for each herbicide in both rotations, resulting in relatively short half-lives. Intensified crop sequences favored the production of biotic degradation metabolites. The impact of the high frequency of soybean cultivation revealed a trend of soil acidification and a reduced biological contribution to attenuation processes in soil contamination.

Controlled release herbicide formulation for effective weed control efficacy.

Controlled release formulation (CRF) of herbicide is an effective weed management technique with less eco-toxicity than other available commercial formulations. To maximise the effectiveness of CRFs however, it is crucial to understand the herbicide-releasing behaviour at play, which predominately depends on the interaction mechanisms between active ingredients and carrier materials during adsorption. In this study, we investigated and modelled the adsorption characteristics of model herbicide 2,4-D onto two organo-montmorillonites (octadecylamine- and aminopropyltriethoxysilane-modified) to synthesise polymer-based CRFs. Herbicide-releasing behaviour of the synthesised CRF microbeads was then analysed under various experimental conditions, and weed control efficacy determined under glasshouse conditions. Results revealed that adsorption of 2,4-D onto both organo-montmorillonites follows the pseudo-second-order kinetics model and is predominately controlled by the chemisorption process. However, multi-step mechanisms were detected in the adsorption on both organoclays, hence intra-particle diffusion is not the sole rate-limiting step for the adsorption process. Both organoclays followed the Elovich model, suggesting they have energetically heterogeneous surfaces. Herbicide-releasing behaviours of synthesised beads were investigated at various pH temperatures and ionic strengths under laboratory and glasshouse conditions. Furthermore, weed control efficacy of synthesised beads were investigated using pot studies under glasshouse condition. Desorption studies revealed that both synthesised microbeads have slow releasing behaviour at a wide range of pHs (5-9), temperatures (25-45 °C), and ionic strengths. The results also revealed that synthesised microbeads have excellent weed control efficacy on different broad-leaf weed species under glasshouse conditions.

2,4-D-based herbicide underdoses cause mortality, malformations, and nuclear abnormalities in Physalaemus cuvieri tadpoles.

Amphibians are considered bioindicators of the environment due to their high sensitivity and involvement in terrestrial and aquatic ecosystems. In the last two decades, 2,4-D has been one of the most widely used herbicides in Brazil and around the world, as its use has been authorized for genetically modified crops and therefore has been detected in surface and groundwater. Against this background, the aim of this work was to investigate the effects of environmentally relevant concentrations of 2,4-D-based herbicides on survival, malformations, swimming activity, presence of micronuclei and erythrocyte nuclear abnormalities in Physalaemus cuvieri tadpoles. The amphibians were exposed to six concentrations of 2,4-D-based herbicides: 0.0, 4.0, 30.0, 52.5, 75.0, and 100 µg L-1, for 168 h. At concentrations higher than 52.5 µg L-1, significantly increased mortality was observed from 24 h after exposure. At the highest concentration (100 µg L-1), the occurrence of mouth and intestinal malformations was also observed. The occurrence of erythrocyte nuclear abnormalities at concentrations of 30.0, 52.5, 75.0 and 100 µg L-1 and the presence of micronuclei at concentrations of 52.5, 75.0, and 100 µg L-1 were also recorded. These effects of 2,4-D in P. cuvieri indicate that the ecological risk observed at concentrations above 10.35 µg L-1 2,4-D may represent a threat to the health and survival of this species, i.e., exposure to 2,4-D at concentrations already detected in surface waters in the species' range is toxic to P. cuvieri.

Adsorption of 2,4-dichlorophenoxyacetic acid on activated carbons from macadamia nut shells.

This study reports on the application of activated carbons from macadamia nut shells as adsorbents for the removal of 2,4-dichlorophenoxyacetic acid, a commonly used pesticide, from water. Different activating agents (FeCl3, ZnCl2, KOH and H3PO4) were used to obtain adsorbents within a wide range of porous texture and surface properties. The characterization of the resulting activated carbons was performed by N2 adsorption-desorption, elemental analysis, TG and pHPZC. The adsorption experiments were conducted in batch at 25, 45 and 65 °C. The adsorption kinetics on activated carbons obtained with FeCl3 H3PO4 or KOH was well described by the pseudo-second order model, whereas for the resulting from ZnCl2 activation the experimental data fit better the pseudo-first order model. The equilibrium studies were performed with the KOH- and ZnCl2-activated carbons, the two showing higher surface area values. In both cases, high adsorption capacities were obtained (c.a. 600 mg g-1) and the experimental data were better described by the Langmuir and Toth models. The thermodynamic study allows concluding the spontaneous and endothermic character of the adsorption process, as well as an increase of randomness at the solid/liquid interface. Breakthrough curves were also obtained and fitted to the logistic model.

Machine learning-mediated Passiflora caerulea callogenesis optimization.

Callogenesis is one of the most powerful biotechnological approaches for in vitro secondary metabolite production and indirect organogenesis in Passiflora caerulea. Comprehensive knowledge of callogenesis and optimized protocol can be obtained by the application of a combination of machine learning (ML) and optimization algorithms. In the present investigation, the callogenesis responses (i.e., callogenesis rate and callus fresh weight) of P. caerulea were predicted based on different types and concentrations of plant growth regulators (PGRs) (i.e., 2,4-dichlorophenoxyacetic acid (2,4-D), 6-benzylaminopurine (BAP), 1-naphthaleneacetic acid (NAA), and indole-3-Butyric Acid (IBA)) as well as explant types (i.e., leaf, node, and internode) using multilayer perceptron (MLP). Moreover, the developed models were integrated into the genetic algorithm (GA) to optimize the concentration of PGRs and explant types for maximizing callogenesis responses. Furthermore, sensitivity analysis was conducted to assess the importance of each input variable on the callogenesis responses. The results showed that MLP had high predictive accuracy (R2 > 0.81) in both training and testing sets for modeling all studied parameters. Based on the results of the optimization process, the highest callogenesis rate (100%) would be obtained from the leaf explant cultured in the medium supplemented with 0.52 mg/L IBA plus 0.43 mg/L NAA plus 1.4 mg/L 2,4-D plus 0.2 mg/L BAP. The results of the sensitivity analysis showed the explant-dependent impact of the exogenous application of PGRs on callogenesis. Generally, the results showed that a combination of MLP and GA can display a forward-thinking aid to optimize and predict in vitro culture systems and consequentially cope with several challenges faced currently in Passiflora tissue culture.

Bioinspired Herbicides-Ionic Liquids or Liquid Cocrystals?

This research provides information about combinations of several amino acids, including l-proline (Pro), l-arginine (Arg), and l-histidine (His), with phenoxyacetic acid herbicides (MCPA and 2,4-D). Five amino acid ionic liquids (AAILs), one amino acid higher-melting salt (AAHMS), and two amino acid liquid cocrystals (AALCs) were obtained in high yields (>90%). The ionization of the six new structures was confirmed by NMR, IR, and molecular modeling. X-ray crystallography was used to definitively confirm the binding location of the mobile hydrogen. Furthermore, we propose a computational method for estimating the energy of specific hydrogen bond(s) in AAIL crystals based on the NBO and QTAIM hydrogen bond parameters obtained by model calculations. An in-depth analysis of the structures allowed to answer the question posed in the title, ionic liquids or liquid cocrystals? AAILs based on arginine and histidine were obtained. In contrast, combining proline with MCPA and 2,4-D led to AALCs. Finally, the compounds were analyzed to measure their herbicidal activity. These studies proved that the novel form of MCPA or 2,4-D improved its ability to control weeds compared to commercial formulations containing the same active ingredients.

Machine learning, conventional and statistical physics modeling of 2,4-Dichlorophenoxyacetic acid (2,4-D) herbicide removal using biochar prepared from Vateria indica fruit biomass.

The adsorption properties of 2,4-Dichlorophenoxyacetic acid (2,4-D) onto biochar, obtained through HCl-assisted hydrothermal carbonization process of Vateria indica fruits (VI-BC), were extensively studied using traditional and statistical physics approaches. The traditional adsorption investigations encompassed kinetics, equilibrium, and thermodynamics studies. Subsequently, the Hill statistical physics model was employed to interpret the mechanism. Also, artificial neural network (ANN) and adaptive neuro-fuzzy inference system (ANFIS) machine learning tools were successfully employed to model the adsorption data wherein both models had high prediction potential (R2 > 0.99). The outcomes demonstrated that the produced VI-BC exhibited remarkable adsorptive traits, having a considerable specific surface area (111.54 m2/g), pore size (5.89 nm), a variety of functional groups, and appropriate attributes for efficiently adsorbing 2,4-D. For 10 mg/L 2,4-D, at pH 2.0 and with 0.3 g/L dose, an impressive 91.67% adsorption efficiency was achieved within a 120-min. Pseudo-second-order model aptly depicted the kinetic behavior of 2,4-D adsorption, while the Freundlich model provided a more accurate representation of the isotherms. 2,4-D maximum adsorption capacity stood at 131.39 mg/g at 303 K. The Hill statistical physics model elucidated that the adsorption primarily occurred via physisorption mechanisms, involving electrostatic attractions, π-π conjugation, and pore filling. This conclusion was further substantiated by post-adsorption characterization of the VI-BC. Thermodynamic analysis indicated that the interactions between VI-BC and 2,4-D were favorable, spontaneous, and exothermic. The calculated low energy of adsorption (1.255 kJ/mol) and ΔH° value (-20.49 kJ/mol) further supported physisorption as the dominant mechanism. In summary, this study underscores the significant potential of the newly developed biochar as a promising alternative material for efficiently removing the 2,4-D herbicide from polluted environments.