Introduction: Scorpion venoms are complex mixtures of bioactive molecules, including neurotoxins, cytolytic peptides, enzymes, and other components that target ion channels, membrane receptors, and various physiological pathways. Although many medically important species have been extensively studied, little is known about the molecular composition of venoms in rare or endemic taxa. The scorpion genus Teuthraustes Simon, 1878 (family Chactidae) comprises 27 described species, with 15 recorded from Ecuador. These species exhibit a remarkable concentration in the Andean highlands and Amazonian regions, although the genus also occurs in Colombia, Venezuela, Peru, and Brazil. Specifically, Teuthraustes atramentarius, the type species, was originally described from Ecuador and remains endemic to the country’s central highlands. Despite its early description, no omics-based studies have investigated its venom composition. Here, we present the first exploration of the venom gland transcriptome of T. atramentarius.

Methods: RNA from dissected venom glands of three specimens, following venom induction by electrostimulation, was sequenced using Illumina RNA-Seq. The assembled transcriptome was analyzed with the DeTox pipeline to identify toxin candidates. Top candidates with signal peptides, cysteine patterns, and similarity to known toxins were manually inspected based on best-hit alignments and preliminary phylogenetic analysis.

Results: Among 40,083 predicted ORFs, 288 matched ToxProt entries, with seven retained as strong toxin candidates. These included one homologous to Phi-liotoxin-Lw1a, a ryanodine receptor modulator; one Cathepsin D-like aspartic peptidase; two invertebrate defensins; two CAP superfamily cysteine-rich venom proteins (CRVPs); and one Hge-scorpine, a multifunctional peptide with antimicrobial and ion channel-blocking properties.

Remarks: To our knowledge, this study constitutes the first omics-based approach to investigating the venom composition of T. atramentarius, revealing seven strong toxin candidates related to known scorpion and animal toxins. However, further research is needed, particularly proteomic validation in venom samples, broader specimen sampling, and integration with proteotranscriptomic and evolutionary approaches.

Cannabis plants are susceptible to microbial contamination, including fungi capable of producing harmful mycotoxins. The presence of these toxins in cannabis products poses serious health risks, particularly to immunocompromised people. This study evaluated the safety of dried cannabis buds intended for medicinal use by examining microbial contamination and residual mycotoxins through culture-based techniques, PCR/qPCR, and ELISA. Irradiation significantly reduced viable fungal and bacterial colony-forming units (CFUs) and eliminated culturable bacteria but did not achieve complete sterilization. Viable spores of toxigenic genera such as Aspergillus, Penicillium, and Fusarium persisted. Sanger sequencing of the PCR product of the sample's DNA generated using Internal Transcribed Spacer (ITS) primers identified dominant mycotoxigenic fungi such as Aspergillus, Cladosporium, Fusarium, and Penicillium in non-irradiated (NR) samples, while next-generation sequencing (NGS) revealed additional non-culturable species. PCR and qPCR studies detected biosynthetic genes for aflatoxins (Nor1), trichothecenes (Tri5), ochratoxins (PKS), and deoxynivalenol (DON) across all samples. While band intensity decreased post-irradiation, gene copy numbers remained comparable, suggesting DNA damage without full degradation. ELISA confirmed the presence of aflatoxin, ochratoxin, DON, and T2 toxins in both irradiated and licensed producer (LP) marketplace samples, with variable concentrations. LP samples showed lower microbial counts and mycotoxin gene abundance but still contained detectable residual DNA and toxins. These findings indicate that while irradiation lowers microbial loads, it does not eliminate mycotoxigenic fungi or their metabolites. Therefore, culture-based assays alone are insufficient for comprehensive safety assessments and must be complemented by molecular and immunological techniques. Given the persistence of toxigenic fungi and their toxins in irradiated and licensed products, we recommend stricter microbial safety standards, particularly for medicinal cannabis, which may put immunocompromised patients at higher risk. For high-risk pathogens like Aspergillus and Fusarium, which pose significant health concerns, a zero-tolerance threshold (<10 CFU/g) is advised, supported by stringent decontamination and testing.

Mycotoxins are naturally occurring toxic metabolites produced by fungi that contaminate animal feed, posing significant risks to livestock health and food safety. The molecular interactions of these toxins with key proteins are critical for elucidating toxicity mechanisms and developing preventive strategies. This study explores the application of computational methods, particularly molecular docking and molecular dynamics (MD) simulations, to predicting and validating the interactions between major mycotoxins and biologically relevant target proteins in food-producing animals.

Molecular docking was first employed to estimate the binding affinity of representative mycotoxins, including aflatoxin B1 and ochratoxin A, with proteins directly implicated in toxin transport, metabolism, and bioavailability and the interaction profiles. These included bovine serum albumin (BSA), a primary carrier protein mediating systemic distribution; porcine organic anion transporter 1 (OAT1), which facilitates renal elimination; and cytochrome P450 enzymes, responsible for metabolic activation. The resulting complexes were subsequently evaluated using MD simulations to assess the stability, flexibility, and dynamic behaviour of the ligand–protein interactions under physiological conditions. Parameters including the root-mean-square deviation (RMSD), root-mean-square fluctuation (RMSF), hydrogen bond occupancy, and molecular mechanics Poisson–Boltzmann surface area (MM-PBSA) energy calculations were analysed to validate the binding stability and interaction persistence over time.

The findings underscore the potential of in silico approaches in veterinary toxicology for identifying high-risk feed contaminants and elucidating their molecular mechanisms of action through interactions with biologically relevant proteins. This framework not only advances our understanding of mycotoxin toxicity but also supports evidence-based risk assessments, regulatory monitoring, and the rational design of protective feed additives. Importantly, characterising these protein–mycotoxin interactions provides mechanistic insights into toxicokinetics; informs the development of targeted mitigation strategies to reduce mycotoxin exposure in livestock; and contributes to safeguarding public health by limiting the transfer of residues into the human food chain. The integration of computational toxicology into veterinary research aligns with One Health principles, offering scalable tools for protecting animal health and ensuring food safety.

Introduction: Deoxynivalenol (DON), a trichothecene mycotoxin produced by Fusarium graminearum, is a potent virulence factor and a critical determinant of Fusarium Head Blight (FHB) in small grain cereals like wheat and barley. Previous research, utilizing a pooled chemical–genetic screen of Chlamydomonas reinhardtii mutants, identified that DON activates the chloroplast unfolded protein response (cpUPR), a chloroplast retrograde signaling pathway. This demonstrated that trichothecenes induce chloroplast stress. The present research leveraged trichothecenes as chemical probes to elucidate the chloroplast-to-nucleus communication during stress. Translating insights from algal systems to higher plants offers crucial clues to ways of mitigating crop diseases like FHB.

Methods: We investigated the impact of trichothecenes on chloroplasts in Chlamydomonas, Arabidopsis, wheat, and barley. The chloroplast damage was assessed via changes in the autofluorescence and general chloroplast morphology. Using a miniaturized Fluorescence Induction and Relaxation (FIRe) system developed by Max Gorbunov at Rutgers University, we measured the photosynthetic rates and photooxidation in Arabidopsis leaf tissues exposed to vacuum-infused trichothecenes.

Results: Trichothecenes consistently caused significant chloroplast damage in higher plants. The exposure of Arabidopsis leaves to 50 μM DON for 1 hour reduced the quantum efficiency of PSII (Fv/Fm) by approximately 20%. DON exposure also significantly reduced the chloroplasts' autofluorescence. In Chlamydomonas, the disruption of Mars1, a key cpUPR kinase, inhibited the induction of proteotoxic stress proteins, including heat shock protein CrHSP22F. Testing orthologous cpUPR-related nuclear genes in wheat showed the robust induction of TaHSP22E. The screen identified additional mycotoxin-sensitive Chlamydomonas mutants exhibiting defective cpUPR activation, providing further insights for higher plant research.

Conclusions: Our findings underscore that trichothecenes are valuable tools for investigating the chloroplast retrograde signaling pathway and identifying novel players, including heat shock proteins, in the cpUPR. These comprehensive analyses, from detailed chloroplast damage assessments to genetic screens, have deepened our understanding of plant stress responses and the mechanisms by which these toxins contribute to causing cereal crop diseases.

Introduction

In Argentina, accidents caused by venomous animals are a public health problem. Snake venom toxicity tests are usually performed in mice and rats, which causes pain and suffering. These tests are not only expensive but also highly regulated. To overcome these limitations, we evaluated the potential use of alternative invertebrate models for snake venom testing. Artemia salina is a small crustacean that lives in high-salinity environments and is commonly used by aquarists as fish food.

Methodology

A. salina cysts were incubated in a thermostatic bath at 28 °C in a 3.2% w/v NaCl solution until hatching. Ten nauplii larvae per well were pipetted into a 96-well microplate. Venoms from four snake species—Naja kaouthia, Micrurus pyrrhocryptus, Bothrops neuwiedi, and Bothrops diporus—were added to each well. Larval motility was quantified after 24 hours of incubation at 28 °C using the Wmicrotracker device in the 3.2% w/v NaCl solution. Each well was also video recorded to monitor potential morphological and/or motility changes caused by the venoms.

Results

The observed EC₅₀s values on motility inhibition suggest that the highest toxicity was observed in N. kaouthia, followed by B. neuwiedi, M. pyrrhocryptus, and B. diporus. Nauplii incubated with N. kaouthia and M. pyrrhocryptus venoms began to show signs of damage at 1.0 µg/mL. B. diporus and B. neuwiedi induced damage at higher concentrations. Structural damage was observed, including appendage destruction and cuticle detachment.

Conclusions

Venoms from four different species were tested for their effects on A. salina motility and morphology. All venoms caused structural damage and complete cessation of larval motility at the highest concentrations tested. These results suggest that A. salina is sensitive to snake venoms in a dose-dependent manner. Further studies will be necessary to test their usefulness in correlating the action of venom and its neutralization by antivenoms in the murine model.

Ricin, one of the most potent toxins known, and the Shiga toxins (Stxs) produced by Escherichia coli (STEC) bind to the C-termini of ribosomal P-stalk proteins to depurinate the sarcin/ricin loop at a different location on the large subunit of the ribosome. The ribosome binding sites of ricin and Stxs have not yet been targeted using small molecules. We established a fluorescence anisotropy-based competition assay to evaluate small molecule inhibitors against RTA. A unique feature of this assay is its ability to identify small molecule inhibitors that bind to the ribosome binding site of RTA and allosterically inhibit its catalytic activity. The FA assay was used to measure the binding constants of small molecule inhibitors that blocked the interaction between a BODIPY-TMR labeled peptide probe mimicking the conserved C-terminal end of the P-stalk proteins (P11) and the A subunit of ricin toxin (RTA). Initially, we used the Biacore SPR to screen the Maybridge Ro3 Core library of compounds and identified CC10501 as the lead molecule. Then, we improved CC10501 using a structure-based design, and using the fluorescence anisotropy-based competition assay, we identified RU-NT-206 and RU-NT-192 as the lead molecules, which showed over 50- and 30-fold improved affinity. The K_i values measured using the FA assay showed a positive correlation with the results of a Vero cell protection assay. These results validated the P-stalk pocket as a target for inhibition and identified RU-NT-206 and RU-NT-192 as lead molecules for further optimization.

Introduction: Parkinson’s disease (PD) is characterized by the progressive degeneration of dopaminergic neurons in the nigrostriatal pathway, primarily due to misfolded α-synuclein. Lysosomes play a crucial role in degrading these proteins through cathepsins, a mechanism impaired in PD. Trigonelline (TGN), an alkaloid found in plants and identified in jellyfish by our research group, has been shown to inhibit caspase-1. Its analog, nicotinic acid, has demonstrated beneficial effects in PD models. This study investigates the modulation of cathepsin D (CTSD) and cathepsin B (CTSB) activity using TGN in the substantia nigra (SN) and caudate putamen (CPu) of PD-induced rats. Methods: Male Wistar rats were divided into three groups: Control (striatal saline injection and oral vehicle), PD (striatal 6-OHDA and oral vehicle), and PD and TGN (striatal 6-OHDA and oral TGN treatment). Fourteen days after striatal injections, animals were treated with TGN (50 mg/Kg; once daily for five consecutive days). One hour after the final treatment, the animals were euthanized, and the SN and CPu were collected for enzymatic activity analysis using specific substrates. Fluorescence was measured every minutes over a 50-minute period. Results: Preliminary findings indicate that in the PD model, CTSD activity in the right SN was reduced (Control = 1.65 AUF/min; PD = 0.14 AUF/min), as was CTSB activity (Control = 2.37 AUF/min; PD = 1.37 AUF/min). TGN treatment enhanced the activity of both enzymes (CTSD = 0.79 AUF/min; CTSB = 3.07 AUF/min). In the CPu, CTSD activity remained similar among groups, whereas CTSB activity was reduced in PD but was not restored by TGN. Conclusion: TGN enhances the activity of key lysosomal enzymes in the SN, which is associated with the removal of protein aggregates. These findings suggest a potential role for TGN in maintaining lysosomal functionality in this critical brain region affected by PD.

Introduction: Alzheimer's disease (AD) is characterized by the accumulation of the amyloid-β (Aβ) peptide in its oligomeric and fibrillar conformations, which disrupts several neuronal functions, including the autophagy–lysosomal pathway, essential for eliminating misfolded, aggregated, or mutated proteins and maintaining proteostasis. Cathepsin D, the most abundant lysosomal peptidase, is dysfunctional in AD, and in this study, we evaluated the effects of molecules from marine invertebrates on restoring its activity. Methods: Differentiated SH-SY5Y cells were pre-treated with oAβ42 (5 μM) for 48 hours, followed by treatment with venom from six Brazilian marine invertebrate species (1 mg/mL) for 24 hours. After the treatment, cell lysates were obtained, and the cathepsin D activity was measured using a specific substrate (GKPILFFRLK(Dnp)-D-R-NH2-MCA) and the fluorescence intensity monitored every five minutes (λex =: 405 nm, λem: 495 nm). The secretions were fractionated through solid-phase extraction in a C18 cartridge, eluted using acetonitrile/0.1% trifluoroacetic acid, and tested again. Results: oAβ42 decreased the velocity of cathepsin D's enzymatic reactions by 40% compared to those in the control (cells without treatment). Venom from the corals Renilla reniformis and Tubastraea tagusensis the sea anemone Anthopleura cascaia; and the sea urchin Echinometra lucunter restored the cathepsin D activity, bringing it back to the control levels or even exceeding them. The venom from A. cascaia, which exhibited the most intense activity, was fractionated further and analyzed, and an active fraction rich in peptides and small molecules was identified. Conclusion: Marine-derived molecules could restore the activity of the main lysosomal enzyme, cathepsin D, important for the elimination of the amyloid peptide and reductions in neuron death, which could contribute significantly to new therapies for the treatment of AD.

The aim of this paper is to investigate the protective effect of curcumin on zearalenone-induced liver injury in piglets and to explore its molecular mechanism to provide a theoretical basis for the alleviation of ZEN-induced liver injury in piglets by nutritional means in production. Twenty-four female weaned piglets (Duroc×Long White×Large White, 7.43±0.88 kg) were randomly divided into three groups: control (CON), ZEN (2 mg/kg), and ZEN+CUR (2 mg/kg ZEN + 300 mg/kg CUR). The trial lasted 28 days following a 3-day pre-feeding period. The results showed that ZEN significantly reduced piglets' body weight on day 28 compared to the CON group, but CUR supplementation restored body weight. ZEN also decreased average daily gain (ADG) and feed intake (ADFI), while CUR improved these parameters in the ZEN+CUR group. ZEN increased the liver index, causing hepatomegaly, but CUR treatment reduced the liver index. Histologically, ZEN caused liver cell swelling, hemorrhage, and vacuolar degeneration, while CUR improved these pathological changes. Serum analysis revealed that ZEN increased alanine aminotransferase (ALT), alkaline phosphatase (ALP), total bile acids (TBA), total bilirubin (TBIL), and γ-glutamyltransferase (GGT) levels, but CUR effectively restored these levels. ZEN had no significant effect on serum aspartate aminotransferase (AST) or albumin (ALB) levels. Overall, CUR demonstrated a protective effect against ZEN-induced liver injury in piglets.

Cytotoxin (CTX) is a three-finger toxin found predominantly in cobra venoms, responsible for dermonecrosis. CTX is generally low in immunogenicity, resulting in the ineffectiveness of existing antibody-based antivenoms in attenuating the local symptoms of dermonecrosis. Aptamers emerge as promising new antivenoms for affordability, manufacturing feasibility and low immunogenicity. We found four epitopes within its functional loops: 'KLVPLFYK', 'AGKNL', 'MFMVSTPKVPV', and 'DVCPKNSLL'. This study aimed to discover aptamers that can specifically recognise these functional epitopes. The binding affinity, neutralisation efficacy, and mechanisms were determined for the aptamer candidates. Our results show that four different aptamer candidates exhibited different degrees of binding to functional epitopes 1, 2, 3, and 4. These aptamers were Apt15, Apt33, Apt26, and Apt31, respectively. Both functional epitopes 1 and 4 demonstrated the highest synergistic cytotoxicity in human skin keratinocytes, with elevated levels of receptor-interacting protein kinase-3 (RIPK3), implying that necroptosis associated with the cytotoxicity. Based on the enzyme-linked aptamer assay (ELAA), Apt31 had the strongest binding affinity towards functional epitope 4, with a K_D value of 187.7 nM. In the in vitro neutralisation assay, Apt31 conferred neutralisation potency with an ED₅₀ of 0.03 nM. However, Apt31 did not rescue human skin keratinocytes in experimental post-envenomed conditions, although increasing concentrations of Apt31 reduced the RIPK3 levels in the post-envenomed keratinocytes. The results conclude that Apt31 potently neutralised the in vitro cytotoxicity caused by functional epitopes but not the post-envenomed conditions.