Bioassay-guided isolation and structure elucidation of anti-mycobacterium tuberculosis compounds from Galatella grimmii (Regel & Schmalh.) Sennikov.

BACKGROUND: Galatella is a genus in the family Asteraceae, represented by 35-45 species. Considering the high effectiveness of the ethyl acetate (EtOAc) fraction of G. grimmii against Mycobacterium tuberculosis (MIC = 0.5 µg/mL), a bioassay-directed fractionation of this extract was carried out. METHODS: The methanolic extract of the aerial parts of G. grimmii was obtained using maceration, then it was suspended in water and partitioned with petroleum ether, dichloromethane (CH2Cl2), EtOAc, and n-butanol (n-BuOH), successively. The most potent fraction (EtOAc), was selected for further isolation by Sephadex LH-20 and semi-preparative HPLC to obtain active compounds. RESULTS: Fractionation of the EtOAc solvent fraction resulted in the characterization of five compounds, among them, compounds 1 and 2 showed the highest anti-mycobacterial effects with MICs of 0.062 and 1.00 µg/mL against H37Rv M. tuberculosis, respectively, which were higher than those of rifampin (MIC of 1.25 µg/mL) and isoniazid (MIC of 0.31 µg/mL), as positive controls. Also, compound 1 ​​inhibited all tested strains of drug-resistant Mycobacterium (MDR and XDR). Notably, the isolated compounds have been reported for the first time from G. grimmii. CONCLUSION: Due to the potent anti-mycobacterial effect of isolated compounds from G. grimmii, this study could pave the way for developing a novel class of natural anti-tuberculosis compounds.

The antidepressant, sertraline, impacts growth and reproduction in the benthic deposit feeder, Tubifex tubifex.

Among emerging contaminants, pharmaceuticals are considered one of the most pertinent substances that may threaten aquatic ecosystems. Pharmaceuticals are designed to be directed at specific metabolic- and molecular pathways. Thus, they are assumed to be still biologically active when entering the ecosystem and may result in unpremeditated impacts on non-target organisms. One of the most widely used selective serotonin reuptake inhibitors, sertraline (an antidepressant), is regularly found in aquatic environments. However, knowledge about the effects, and in particular, of sediment-associated sertraline in benthic invertebrates is limited. We examined the impacts of chronic exposure (28 d) to sediment-associated sertraline (3.3, 33, 330 µg/g dw sed.) on survival, growth and reproduction in the deposit-feeding oligochaete, Tubifex tubifex. Sertraline significantly decreased T. tubifex survival and growth. Worms exposed to high sertraline concentrations (330 µg/g) had a lower growth rate and reproduction, as indicated by a significantly lower number of cumulated cocoons. Worms exposed to an environmentally relevant concentration (3.3 µg/g) decreased growth but maintained a reproduction rate similar to that of the control. The implications are that adult worms exposed to high sertraline concentrations presumably required more energy for maintenance and detoxification, thereby reducing available energy for reproduction and growth. This represents a trade-off between survival, reproduction and growth. In contrast, T. tubifex exposed to environmentally relevant concentrations allocated more energy to reproduction by slightly increasing the number of cocoons produced and reducing growth. However, the quantity and quality of offspring may be impacted as we observed fewer juveniles in the environmentally relevant treatment than in the control. Overall, the results indicate that sediment-associated sertraline is bioavailable and negatively impacts T. tubifex survival, growth, and reproduction even at environmentally relevant concentrations.

Niche construction and niche choice by aphids infesting wheat ears.

The niche of aphids is largely defined by their consumption of plant phloem sap and its composition, including nutrients and specialized metabolites. Niche construction is the change of the environment by organisms, which may influence the fitness of these organisms and their offspring. To better understand interactions between plants and aphids, it is necessary to investigate whether aphids modify the chemical composition of the phloem sap of their host plants and whether conspecifics are affected by previous infestation. In the current study, ears of wheat (Triticum aestivum) plants were infested with clonal lineages of the English grain aphid (Sitobion avenae) or were left uninfested. The metabolic composition of ear phloem sap exudates was analyzed through amino acid profiling and metabolic fingerprinting. Aphids of the clonal lineages were either put on previously aphid-infested or on uninfested ears and their colony sizes followed over time. Furthermore, it was investigated whether aphids choose one treatment group over another. Sitobion avenae infestation affected the relative concentrations of some metabolites in the phloem exudates of the ears. Compared to uninfested plants, the relative concentration of asparagine was higher after aphid infestation. Colonies grew significantly larger on previously aphid-infested ears, which the aphids also clearly chose in the choice experiment. The pronounced positive effect of previous infestation on aphid colonies indicates niche construction, while the choice of these constructed niches reveals niche choice by S. avenae on wheat. The interplay between these different niche realization processes highlights the complexity of interactions between aphids and their hosts.

Ecotoxicological effects of suspended sediments on marine microalgae using flow cytometry and pulse-amplitude modulation (PAM) fluorometry.

Microalgal bioassays were conducted to evaluate the ecotoxicological effects of suspended sediments (SS) collected from coastal environments. Growth inhibition was assessed for six microalgal species, and multiple endpoints were measured using flow cytometry (FCM) and pulse-amplitude modulation (PAM) fluorometry for three species (Dunaliella tertiolecta, Isochrysis galbana, and Phaeodactylum tricornutum). Among these, the EC50 for growth inhibition of D. tertiolecta (6700 mg L-1) was notably lower compared to the other species, and among several endpoints, esterase activity was the most inhibited. Species-specific responses to SS exposure were identified, with D. tertiolecta exhibiting greater susceptibility across most endpoints. Meanwhile, measurements of Fo', Fm', and Y(NPQ) in P. tricornutum using PAM fluorometry revealed greater sensitivity. Based on the results of this study and review, the tentative predicted no-effect concentration was calculated as 12.1 mg L-1. Overall, this study provides novel insights into SS ecotoxicity, establishing a crucial baseline for future investigations.

Application of Microbiological Screening Tests in Assessment of Environmental Exposure to Antibiotics: Preliminary Studies.

Contact of aquatic microbiocenoses with antibiotics present in the environment can cause the former to develop resistance to antimicrobial drugs. Therefore, the search for methods to detect antibiotics and drug-resistant microorganisms in the environment is important. The presented paper proposes a simple procedure to assess environmental exposure to antibiotics and the presence of non-susceptible microorganisms. Medium solutions with selected antibiotics and a microbial growth indicator were applied to test plates, and were inoculated with water samples from various ecosystems. After incubation, the susceptibility of the microorganisms to antibiotics was determined and presented in chronic microbial toxic concentration (MTC) values. It was confirmed that the presented procedure enables the assessment of the antibiotic susceptibility and adaptation potential of unselected microorganisms from different aquatic ecosystems. However, the MTC values depend on the inoculum volume, the density and seasonal activity of the microorganisms, the method of inoculum preparation, and the incubation time of the test plate. The described procedure may be practically applied as a screening test to identify the presence of drug-resistant microorganisms. Additionally, it may also be suitable as a method to assess environmental exposure to antibiotics. However, prior standardisation is required before implementing this procedure in quantitative studies.

Chemosensory protein 22 in Riptortus pedestris is involved in the recognition of three soybean volatiles.

Riptortus pedestris (Hemiptera: Alydidae), a common agricultural pest, is the major causative agent of "soybean staygreen." However, the interactions between chemosensory proteins (CSPs) in R. pedestris and host plant volatiles have yet to be comprehensively studied. In this study, we performed real-time fluorescence quantitative polymerase chain reaction (PCR) to analyze the antennal expression of RpedCSP22 and subsequently analyzed the interactions between 21 soybean volatiles, five aggregation pheromones, and RpedCSP22 protein in vitro using a protein expression system, molecular docking, site-directed mutagenesis, and fluorescence competitive binding experiments. The RpedCSP22 protein showed binding affinity to three soybean volatiles (benzaldehyde, 4-ethylbenzaldehyde, and 1-octene-3-ol), with optimal binding observed under neutral pH conditions, and lost binding ability after site-directed mutagenesis. In subsequent RNA interference (RNAi) studies, gene silencing was more than 90 %, and in silenced insects, electroantennographic responses were reduced by more than 75 % compared to non-silenced insects. Moreover, Y-tube olfactory behavioral assessments revealed that the attraction of R. pedestris to the three soybean volatiles was significantly attenuated. These findings suggest that RpedCSP22 plays an important role in the recognition of host plant volatiles by R. pedestris andprovides a theoretical basis for the development of novel inhibitors targeting pest behavior.

Methodological advances in formulation and assay of herbal resources-based topical drug delivery systems.

Medicinal plants have been utilized for centuries as a source of healing compounds, which consist of thousands of known bioactive molecules with therapeutic potentials. This article aims to explore and emphasize the significance of medicinal plants and bioactive compounds in the development of topical pharmaceutical formulations. The journey from the extraction of phytochemicals to the development of topical pharmaceutical formulations is described with the aid of scientific evidence selected from PubMed, Google Scholar, ScienceDirect, and Web of Science. Articles published in English during 2018-2023 period were considered and selected randomly. The review discusses the extraction process of medicinal plants, solvent selection, and green synthesis of metal nanoparticles. Subsequently, various biological activities of plant extracts are elaborated especially focusing on antimicrobial, antioxidant, anti-inflammatory, and sun protection activities, along with the corresponding in vitro assays commonly employed for the evaluation. The article presents the process of compound isolation through bioactivity-guided fractionation and also the toxicity evaluation of isolated fractions. Finally, the formulation of medicinal plant extracts into topical pharmaceuticals is addressed, emphasizing the stability evaluation procedures necessary for ensuring product quality and efficacy.

Bioactivity Profiling and Phytochemical Analysis of Carissa carandas Extracts: Antioxidant, Anti-Inflammatory, and Anti-Urinary Tract Infection Properties.

Carissa carandas L. (Apocynaceae) is widely distributed in tropical and subtropical regions of Asia including Pakistan, India, Afghanistan, and Sri Lanka. C. carandas is considered as an integral component of traditional medicinal systems to combat several health ailments. The present study aimed to assess this plant's phytochemical contents and biological potential by performing sequential extraction, adopting a bioassay-guided approach. C. carandas powder was extracted with n-hexane to remove fatty substances and then residues were sequentially extracted with dichloromethane, methanol, and 50% methanol. All the sequential crude extracts were evaluated for phytochemical contents (total phenolics, flavonoids, and anthocyanins), in vitro antioxidant activity (FRAP, DPPH), in vitro anti-inflammatory activity (serum and egg albumin denaturation), in vivo anti-inflammatory activity (carrageenan- and formaldehyde-induced paw edema), and in vitro antimicrobial activity. Active crude extract was then partitioned using the liquid-liquid separation method followed by further separation of the active fraction by RP-HPLC. The active fraction was then subjected to LC-ESI-MS/MS analysis for tentative identification of bioactive metabolites responsible for its bioactive properties, followed by HPLC quantification. The analysis revealed methanol extract to have more phytochemical contents, radical scavenging properties, reduced inflammation in both models (in vitro and in vivo), and antimicrobial properties against urinary tract infection-causing agents as compared to dichloromethane and 50% methanol extracts. The ethyl acetate fraction obtained after liquid-liquid partitioning (LLP) of the active methanol extract exhibited more activity as compared to C. carandas methanol extract. RP-HPLC sub-fractionation yielded seven sub-fractions, but a slight decrease in biological potential was recorded. Therefore, LLP fraction B was subjected to further analysis. LC-ESI-MS/MS analysis led to the tentative identification of phenolic acids (chlorogenic acid, quinic acid), flavonoids (quercetin), and anthocyanins (peonidin-3-arabinoside, delphinidin-3-galactoside, delphinidin-3-rutinoside) in the active LLP ethyl acetate fraction. Chlorogenic acid, ellagic acid, and quinic acid were quantified as 17.6 µg/mg, 5.90 µg/mg, and 3.30 µg/mg, respectively, on a dry weight basis by HPLC. C. carandas may be considered a promising therapeutic plant, and the results of the current study provide more evidence to support the assertions made in ancient medical traditions. These findings highlight its promising applications in health, medicine, cosmetics, preservatives, and as a natural coloring agent.

Repellent Effects of Coconut Fatty Acid Methyl Esters and Their Blends with Bioactive Volatiles on Winged Myzus persicae (Sulzer) Aphids (Hemiptera: Aphididae).

Myzus persicae (Sulzer) (Hemiptera: Aphididae) is one of the most important aphid crop pests, due to its direct damage and its ability to transmit viral diseases in crops. The objective is to test whether spraying nanoemulsions of botanical products repels winged individuals of M. persicae in a bioassay in culture chambers. The bioactive volatiles were applied on pepper plants at a dose of 0.2% alone or at 0.1% of each component in blends. A treated plant and a control plant were placed at each side of an entomological cage inside a growth chamber. The winged individuals were released between the plants, in a black-painted Petri dish suspended by wires in the upper half of the cage. The most repellent products were farnesol (repellency index, RI = 40.24%), (E)-anethole (RI = 30.85%) and coconut fatty acid methyl ester (coconut FAME) (RI = 28.93%), alone or in the following blends: farnesol + (E)-anethole + distilled lemon oil (RI = 36.55%) or (E)-anethole + distilled lemon oil + coconut FAME (RI = 30.63%). The observed effect of coconut FAME on aphids is the first report of this product having a repellent effect on a crop pest. Repellent substances for viral disease vectors should be further investigated to develop new strategies for plant protection.

Fungal Attachment-Resistant Polymers for the Additive Manufacture of Medical Devices.

This study reports the development of the first copolymer material that (i) is resistant to fungal attachment and hence biofilm formation, (ii) operates via a nonkilling mechanism, i.e., avoids the use of antifungal actives and the emergence of fungal resistance, (iii) exhibits sufficient elasticity for use in flexible medical devices, and (iv) is suitable for 3D printing (3DP), enabling the production of safer, personalized medical devices. Candida albicans (C. albicans) can form biofilms on in-dwelling medical devices, leading to potentially fatal fungal infections in the human host. Poly(dimethylsiloxane) (PDMS) is a common material used for the manufacture of medical devices, such as voice prostheses, but it is prone to microbial attachment. Therefore, to deliver a fungal-resistant polymer with key physical properties similar to PDMS (e.g., flexibility), eight homopolymers and 30 subsequent copolymers with varying glass transition temperatures (Tg) and fungal antiattachment properties were synthesized and their materials/processing properties studied. Of the copolymers produced, triethylene glycol methyl ether methacrylate (TEGMA) copolymerized with (r)-α-acryloyloxy-ß,ß-dimethyl-γ-butyrolactone (AODMBA) at a 40:60 copolymer ratio was found to be the most promising candidate by meeting all of the above criteria. This included demonstrating the capability to successfully undergo 3DP by material jetting, via the printing of a voice prosthesis valve-flap using the selected copolymer.

Aligning Post-Column ESI-MS, MALDI-MS, and Coagulation Bioassay Data of Naja spp., Ophiophagus hannah, and Pseudonaja textillis Venoms Chromatographically to Assess MALDI-MS and ESI-MS Complementarity with Correlation of Bioactive Toxins to Mass Spectrometric Data.

Snakebite is a serious health issue in tropical and subtropical areas of the world and results in various pathologies, such as hemotoxicity, neurotoxicity, and local swelling, blistering, and tissue necrosis around the bite site. These pathologies may ultimately lead to permanent morbidity and may even be fatal. Understanding the chemical and biological properties of individual snake venom toxins is of great importance when developing a newer generation of safer and more effective snakebite treatments. Two main approaches to ionizing toxins prior to mass spectrometry (MS) analysis are electrospray ionization (ESI) and matrix-assisted laser desorption ionization (MALDI). In the present study, we investigated the use of both ESI-MS and MALDI-MS as complementary techniques for toxin characterization in venom research. We applied nanofractionation analytics to separate crude elapid venoms using reversed-phase liquid chromatography (RPLC) and high-resolution fractionation of the eluting toxins into 384-well plates, followed by online LC-ESI-MS measurements. To acquire clear comparisons between the two ionization approaches, offline MALDI-MS measurements were performed on the nanofractionated toxins. For comparison to the LC-ESI-MS data, we created so-called MALDI-MS chromatograms of each toxin. We also applied plasma coagulation assaying on 384-well plates with nanofractionated toxins to demonstrate parallel biochemical profiling within the workflow. The plotting of post-column acquired MALDI-MS data as so-called plotted MALDI-MS chromatograms to directly align the MALDI-MS data with ESI-MS extracted ion chromatograms allows the efficient correlation of intact mass toxin results from the two MS-based soft ionization approaches with coagulation bioassay chromatograms. This facilitates the efficient correlation of chromatographic bioassay peaks with the MS data. The correlated toxin masses from ESI-MS and/or MALDI-MS were all around 6-8 or 13-14 kDa, with one mass around 20 kDa. Between 24 and 67% of the toxins were observed with good intensity from both ionization methods, depending on the venom analyzed. All Naja venoms analyzed presented anticoagulation activity, whereas pro-coagulation was only observed for the Pseudonaja textillis venom. The data of MALDI-MS can provide complementary identification and characterization power for toxin research on elapid venoms next to ESI-MS.

Utility of Cry1Ja for Transgenic Insect Control.

Insect control traits are a key component of improving the efficacy of insect pest management and maximizing crop yields for growers. Insect traits based on proteins expressed by the bacteria Bacillus thuringiensis (Bt) have proven to be very effective tools in achieving this goal. Unfortunately, the adaptability of insects has led to resistance to certain proteins in current commercial products. Therefore, new insecticidal traits representing a different mode of action (MoA) than those currently in use are needed. Cry1Ja has good insecticidal activity against various lepidopteran species, and it provides robust protection against insect feeding with in planta expression. For Bt proteins, different MoAs are determined by their binding sites in the insect midgut. In this study, competitive binding assays are performed using brush border membrane vesicles (BBMVs) from Helicoverpa zea, Spodoptera frugiperda, and Chrysodeixis includens to evaluate the MoA of Cry1Ja relative to representatives of the various Bt proteins that are expressed in current commercial products for lepidopteran insect protection. This study highlights differences in the shared Cry protein binding sites in three insect species, Cry1Ja bioactivity against Cry1Fa resistant FAW, and in planta efficacy against target pests. These data illustrate the potential of Cry1Ja for new insect trait development.

Insights into the Detoxification of Spruce Monoterpenes by the Eurasian Spruce Bark Beetle.

Plant defence mechanisms, including physical barriers like toughened bark and chemical defences like allelochemicals, are essential for protecting them against pests. Trees allocate non-structural carbohydrates (NSCs) to produce secondary metabolites like monoterpenes, which increase during biotic stress to fend off pests like the Eurasian spruce bark beetle, ESBB (Ips typographus). Despite these defences, the ESBB infests Norway spruce, causing significant ecological damage by exploiting weakened trees and using pheromones for aggregation. However, the mechanism of sensing and resistance towards host allelochemicals in ESBB is poorly understood. We hypothesised that the exposure of ESBB to spruce allelochemicals, especially monoterpenes, leads to an upsurge in the important detoxification genes like P450s, GSTs, UGTs, and transporters, and at the same time, genes responsible for development must be compromised. The current study demonstrates that exposure to monoterpenes like R-limonene and sabiene effectively elevated detoxification enzyme activities. The differential gene expression (DGE) analysis revealed 294 differentially expressed (DE) detoxification genes in response to R-limonene and 426 DE detoxification genes in response to sabiene treatments, with 209 common genes between the treatments. Amongst these, genes from the cytochrome P450 family 4 and 6 genes (CP4 and CP6), esterases, glutathione S-transferases family 1 (GSTT1), UDP-glucuronosyltransferase 2B genes (UDB), and glucose synthesis-related dehydrogenases were highly upregulated. We further validated 19 genes using RT-qPCR. Additionally, we observed similar high expression levels of detoxification genes across different monoterpene treatments, including myrcene and α-pinene, suggesting a conserved detoxification mechanism in ESBB, which demands further investigation. These findings highlight the potential for molecular target-based beetle management strategies targeting these key detoxification genes.

Evaluation of the Pharmaceutical Activities of Chuanxiong, a Key Medicinal Material in Traditional Chinese Medicine.

Szechwan lovage rhizome (SLR, the rhizome of Ligusticum chuanxiong Hort., Chuanxiong in Chinese transliteration) is one Chinese materia medica (CMM) commonly used to activate blood circulation and remove blood stasis. SLR is applicable to most blood stasis syndromes. It has significant clinical efficacy in relation to human diseases of the cardiocerebrovascular system, nervous system, respiratory system, digestive system, urinary system, etc. Apart from China, SLR is also used in Singapore, Malaysia, the European Union, and the United States of America. However, the current chemical markers in pharmacopeia or monography for the quality assessment of SLR are not well characterized or specifically characterized, nor do they fully reflect the medicinal efficacy of SLR, resulting in the quality of SLR not being effectively controlled. CMM can only have medicinal efficacy when they are applied in vivo to an organism. The intensity of their pharmaceutical activities can more directly represent the quality of CMM. Therefore, the chemical constituents and pharmacological actions of SLR are reviewed in this paper. In order to demonstrate the medicinal efficacy of SLR in promoting blood circulation and removing blood stasis, bioassay methods are put forward to evaluate the pharmaceutical activities of SLR to improve hemorheology, hemodynamics, and vascular microcirculation, as well as its anti-platelet aggregation and anticoagulation properties. Through comprehensive analyses of these pharmaceutical properties, the quality and therapeutic value of SLR are ascertained.

Bioassay-Guided Isolation and Identification of Xanthine Oxidase Inhibitory Constituents from the Fruits of Chaenomeles speciosa (Sweet) Nakai.

Chaenomeles speciosa (Sweet) Nakai (C. speciosa) is a traditional Chinese herbal medicine that possesses not only abundant nutritional value but also significant medicinal properties. The extracts of C. speciosa fruits effectively reduce urate levels, but the specific chemical constituents responsible for this effect in C. speciosa fruits are still unknown. Therefore, this study aims to investigate and analyze the structure-activity relationships of these constituents to better understand their ability to lower uric acid. Activity-guided fractionation and purification processes were used to isolate compounds with xanthine oxidase (XO) inhibitory activity from C. speciosa fruits, resulting in three extracts: petroleum ether, ethyl acetate, and n-butanol. The ethyl acetate and n-butanol fractions showed strong activity and underwent further separation and purification using chromatographic techniques. Twenty-four compounds were isolated and identified, with nine showing potent activity, including chlorogenic acid, methyl chlorogenate, butyl chlorogenate, ethyl chlorogenate, cryptochlorogenic acid methyl ester, caffeic acid, p-coumaric acid, benzoic acid and protocatechuic acid. The docking analysis showed that these compounds interacted with amino acid residues in the active site of XO through hydrogen bonding and hydrophobic interactions. These findings suggest that these compounds help reduce uric acid in C. speciosa, supporting further investigation into their mechanism of action.

Toxicity assessment of powdered laundry detergents: an in vivo approach with a plant-based bioassay.

Powdered laundry detergents, encompassing a diverse blend of organic and inorganic compounds, are crucial in efficiently removing dirt in household cleaning. This study investigated the cytotoxic and genotoxic potential of commonly used powdered laundry detergents in Sri Lanka using the Allium cepa bioassay. Five detergents (four branded A, B, C, and D, and one non-branded E) were selected for assessment. Toxicity evaluations were conducted across a range of predetermined aqueous detergent concentrations (0-2500 mg/L) using the A. cepa bioassay, with all experiments being triplicated and following standard protocols. Exposure to detergent concentrations up to 500 mg/L resulted in mitosis suppression, nuclear aberrations, and chromosomal abnormalities in A. cepa, indicating concentration-dependent cytotoxicity and genotoxicity. Condensed nuclei were notably prevalent among nuclear abnormalities, while vagrant chromosomes and chromosomal adherence were the most frequent chromosomal aberrations observed. At higher concentrations (> 500 mg/L), the selected detergents induced necrotic cell death in A. cepa root meristematic cells. This study warns to avoid the unnecessary use of detergents as they cause significant ecological risks and advocates for further research to comprehensively assess detergent toxicity across diverse organisms within ecosystems to safeguard ecosystem health effectively.

Faster ciguatoxin extraction methods for toxicity screening.

Ciguatera poisoning (CP) is a severe global public health problem caused by the consumption of seafood products contaminated with ciguatoxins (CTXs). The growing demand for seafood products requires high-throughput testing for CTX-susceptible seafood, however complex extraction and slow cleanup methods inhibit this goal. Herein, several methods for extracting CTXs from fish tissue were established and compared; these methods are sensitive, specific, and valid while achieving higher sample extraction throughput than currently established protocols. The trial fish material was generated from multiple species, with different physical conditions (wet and freeze-dried tissue), and naturally contaminated with various CTXs (i.e., CTX-1B, CTX-3C, and C-CTX-1), thus ensuring these methods are robust and broadly applicable. The extraction methods used were based on mechanical maceration with acetone or methanol or enzymatic digestion followed by acetone and ethyl acetate extraction. Crude extracts were investigated for CTX-like toxicity using an in vitro mouse neuroblastoma (N2a) cell-based assay (CBA). Among the three methods, there was no significant difference in toxin estimates (p = 0.219, two-way ANOVA), indicating their interchangeability. For speed (> 16 samples/day), accuracy (100%), and CTX analog retention confirmation by liquid chromatography-tandem mass spectrometry (LC‒MS/MS), the preferred extraction methods were both methanol and enzyme-based. All extraction methods post hoc confirmation of CTX analogs successfully met international seafood market-based CTX contaminant guidance. These methods can drastically increase global CTX screening capabilities and subsequently relieve sample processing bottlenecks, inhibiting environmental and human health-based CTX analysis.

Assessment of receptor-mediated activity (AhR and ERα), mutagenicity, and teratogenicity of metal shredder wastes in Wallonia, Belgium.

In this study, hazardous wastes including fluff, dust, and scrubbing sludge were sampled in 2019 from two metal shredding facilities located in Wallonia, Belgium. To assess the extent of the contamination, a global approach combining chemical and biological techniques was used, to better reflect the risks to health and the environment. The samples investigated induced significant in vitro aryl hydrocarbon receptor (AhR) agonistic bioactivities and estrogenic receptor (ERα) (ant)agonistic bioactivities in the respective CALUX (chemical activated luciferase gene expression) bioassays. The mutagenicity of the samples was investigated with the bacterial reverse gene mutation test using the Salmonella typhimurium TA98 and TA100 strains. Except for the sludge sample (site 3), all samples induced a mutagenic response in the TA98 strain (± S9 metabolic fraction) whereas in the TA100 strain (+ S9 metabolic fraction), only the sludge sample (site 2) showed a clear mutagenic effect. The in vivo toxicity/teratogenicity of the shredder wastes was further evaluated with zebrafish embryos. Except for the dust sample (site 2), all samples were found to be teratogenic as they returned teratogenic indexes (TIs) > 1. The high levels of contamination, the mutagenicity, and the teratogenicity of these shredder wastes raise significant concerns about their potential negative impacts on both human health and environment.

Isolation and genetic characterization of Toxoplasma gondii from chickens from public markets in Pernambuco State, Brazil.

This study aimed to evaluate the presence and viability of Toxoplasma gondii in chickens intended for human consumption in the Pernambuco State, Brazil. Blood and tissue samples were collected from 25 chickens sold in markets in Recife, Pernambuco. Samples were evaluated by indirect immunofluorescence assay (IFA) to detect antibodies to T. gondii. Pools of brain and heart of seropositive chickens were subjected to bioassay in two Swiss Webster mice, which were evaluated for 45 days then tested by IFA to detect seroconversion. The mice were euthanized, and their brains were evaluated for cysts. Peritoneal lavage was also conducted in mice that exhibited clinical signs. Brains containing cysts or peritoneal lavage with tachyzoites were inoculated into MA-104 cells. Brains of mice inoculated with the same tissue were pooled and analysed by ITS1-PCR. We obtained a frequency of antibodies to T. gondii of 68.00% (17/25) in chickens, and a seroconversion rate of 70.58% (24/34) in mice. Detection of Toxoplasma ITS1 DNA confirmed an isolation rate of 41.1% (7/17). Three isolates were characterized by mnPCR-RFLP as genotypes ToxoDB#36 and ToxoDB#114. We highlight the occurrence of ToxoDB#36 in chickens in Pernambuco State and the parasites' viability in chickens intended for human consumption.

Quantitative analysis and characterization of floral volatiles, and the role of active compounds on the behavior of Heortia vitessoides.

This study explores the role of floral volatile organic compounds (FVOCs) in insect behavior, focusing on Aquilaria sinensis (AS), a valuable tropical plant threatened by Heortia vitessoides Moore. Despite H. vitessoides' attraction to AS and non-host plants like Elaeocarpus decipiens (ED) and Dalbergia odorifera (DO), little is known about their chemical interactions. FVOCs from these plants were analyzed at 9:00 and 18:00 using GC×GC-QTOF-MS and HS-SPME. The results showed that ED exhibiting the highest concentration (92.340 ng/mg), followed by DO (75.167 ng/mg) and AS (64.450 ng/mg). Through GC-EAD and EAG, a total of 11 FVOC compounds with electrophysiological activates were identified. These compounds, except linalool, showed dose-dependent responses. Y-Tube bioassays confirmed phenylethyl alcohol or the mixture of EAD-active compounds produced positive chemotactic responses in both males and females. FVOCs have the potential to be used as a natural and sustainable alternative to chemical insecticides in pest control.