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.