Mycotoxins represent secondary metabolic products generated by fungal genera, including Aspergillus, Penicillium, and Fusarium, and are known to trigger hepatotoxic, nephrotoxic, carcinogenic, and immunotoxic effects in humans. While well-defined regulatory thresholds exist to manage dietary mycotoxin intake for major mycotoxins such as aflatoxins, ochratoxins, fumonisins, deoxynivalenol, and zearalenone, the health consequences stemming from inhalation among the agricultural and food-processing workforce are still poorly understood. Importantly, there are no health-based occupational exposure limits specifically for airborne mycotoxins, which creates a substantial knowledge gap within occupational health practice. Data indicate that local cytochrome-P450-dependent bioactivation may put exposed individuals at risk of airway inflammation, impaired immune responses, alongside plausible mechanistic risk for lung-carcinogenic effects following mycotoxin inhalation. Even so, robust human epidemiological data supporting such adverse respiratory outcomes remain scarce. This narrative review compiles available toxicological findings, field-measured exposure levels, and health-related observations, centering on occupational inhalation scenarios. By sorting evidence according to its inferential strength, the present work aims to unpack inhalation-linked hazards and lay the groundwork for subsequent risk assessment work and targeted protective measures for susceptible working groups.
The connecting peptide (C-peptide) of human insulin has been implicated in native disulfide bond formation and diverse biological functions; however, its structural significance and potential functional roles in venom insulins remain poorly understood. The present study compares the structural significance of the C-peptide in human insulin (hum-ins) and Conus geographus venom insulin G1 (Vcon-ins G1) using computational methods. The proinsulin of both species is identical in length (86 residues); however, mature Vcon-ins G1 is seven residues shorter, while its C-peptide is correspondingly seven residues longer. VCon-ins G1 also exhibits distinctive features, including posttranslational modifications such as γ-carboxyglutamic acid (Gla), 4-trans-hydroxyproline, and C-terminal amidation, as well as a monomeric solution state and rapid activation of the human insulin receptor. Using AlphaFold 3-predicted proinsulin structures, this study investigates hydrophobicity profiles, disulfide conformations, and oxidative folding pathways. A contrasting aggregation propensity is observed: in hum-ins, the C-peptide is less aggregation-prone than the mature chain, whereas the reverse is seen in Vcon-ins G1. Disulfide conformations also differ markedly: hum-ins displays relatively confined disulfide conformations, whereas Vcon-ins G1 exhibits more diffuse and strained disulfides, with the C-peptide influencing both systems. The predicted sequential order of disulfide formation differed markedly between hum-ins and Vcon-ins G1, indicating distinct oxidative folding pathways influenced by the C-peptide. A putative γ-glutamyl carboxylase recognition motif and differential accessibility of glutamic acid residues with and without disulfide bonds suggest that the C-peptide of Vcon-ins G1 may serve as a potential recognition site for γ-glutamyl carboxylase. Comparative phylogenetic analysis of proinsulin and C-peptide sequences revealed region-specific diversification of C-peptides, potentially reflecting evolution of features that facilitate recognition by modifying enzymes such as γ-glutamyl carboxylase. These findings suggest that the C-peptide of Vcon-ins G1 has distinct structural and evolutionary roles that may contribute to its oxidative folding, posttranslational modification, and functional specialization.
Aflatoxin B1(AFB1), zearalenone(ZEN), and alternariol(AOH) are major mycotoxins that threaten food safety, yet enzymes capable of detoxifying these structurally diverse toxins remain limited. In this study, two enzymes from Bacillus velezensis HNGD-JQ06, laccase BVLac and superoxide dismutase BV2SOD, were identified as broad-spectrum detoxification candidates. BVLac degraded 93.33% of AFB1 at 60 °C and pH 7.0, 94.10% of ZEN at 60 °C and pH 8.0, and 96.87% of AOH at 50 °C and pH 8.0. BV2SOD degraded 92.59% of AFB1 and 91.99% of ZEN at 50 °C and pH 7.0, and 98.94% of AOH at 60 °C and pH 8.0. UHPLC-MS/MS analysis suggested that AFB1, ZEN, and AOH were transformed into products tentatively assigned as AFQ1/epi-AFQ1, 15-OH-ZEN, and a hydroxylated AOH derivative, respectively. Zebrafish assays confirmed reduced toxicity of the products, while molecular docking supported favorable enzyme-toxin binding. Both enzymes also retained activity in peanut, corn, and wheat flour matrices, highlighting their potential for multi-mycotoxin biodetoxification.
Snake venoms are complex biochemical phenotypes whose composition can diverge dramatically among taxa, yet the functional and biomedical consequences of such variation remain understudied for many lineages. Here, we characterized the functional activities of venom from multiple rattlesnake species distributed across insular and mainland localities in the Baja California Peninsula and Gulf of California. We quantified phospholipase A2 (PLA2), proteolytic, and fibrinogenolytic activities as well as median lethal doses (LD50) for 55 individuals across Crotalus enyo, C. mitchellii, C. pyrrhus sensu lato, and C. ruber. We also evaluated the neutralization efficacy of the commercial polyvalent antivenom Antivipmyn®. Venom functional activities differed significantly among species. Crotalus mitchellii expressed a neurotoxic phenotype characterized by the highest PLA2 activity, the lowest LD50 values, and minimal proteolysis. Crotalus pyrrhus and C. ruber expressed proteolytic phenotypes with high enzymatic activity and ubiquitous dual-chain fibrinogenolysis. Crotalus enyo exhibited intermediate functional activities with moderate enzymatic output and the most efficient antivenom neutralization among species. Although Antivipmyn® neutralized the lethal activity of all venoms, efficacy differed significantly among species in both mass-normalized (mgAV/mgV) and potency-scaled (LD50-equivalents per vial) metrics. Notably, insular and mainland populations within species showed qualitative variation in both functional activities and antivenom responses. These results establish that the focal species possess functionally distinct venom phenotypes with species-specific consequences for antivenom neutralization, underscoring the relevance of venom functional characterization for clinical management of envenomation across the Baja California region.
Snake venom vascular endothelial growth factors (svVEGFs) are homodimeric proteins comprising two equal subunits bound by two intermolecular disulfide bonds. The main biological effects of svVEGFs are local vasodilatation, increasing of vascular permeability, and hypotension. Barietin, a svVEGF purified from the venom of puff adder Bitis arietans, differs from other representatives of this toxin family by its shortened C-terminal amino acid sequence. No data about barietin effects on hemodynamics are available. Studying those in rats we found that barietin at a dose of 1 mg/kg significantly increased heart rate, exerting only a minor influence on blood pressure. In this respect, it differs from other svVEGFs having a significant effect on blood pressure. We aimed to find the barietin active fragment which may be responsible for its tachycardic activity. Because in literature the fragments corresponding to the so-called loop 1 of VEGF-A (a mammalian homologue of barietin) manifested some affinity to VEGF receptors, we synthesized a peptide homologous to this loop comprising barietin fragment 24-36, with two Cys residues added at the N- and C-termini to form a disulfide. At intravenous injection at a dose of 5 mg/kg, it showed a tendency to increase heart rate without any significant effect on blood pressure, thus demonstrated a trend to reproduce the effect of barietin. So, barietin is the first svVEGF which increases the heart rate without substantial effect on the blood pressure, with its fragment demonstrating a trend for a similar activity.
Deoxynivalenol (DON) is a mycotoxin that severely contaminates agri-food products and causes liver injury. Quercetin (Que) has anti-ferroptosis and hepatoprotective effects. This study employed a network pharmacology approach to investigate ferroptosis-associated targets underlying Que's mitigation of DON-induced liver injury, with experimental validation of its hepatoprotective mechanisms. We identified four ferroptosis regulatory targets of Que associated with liver injury: GPX4, HMOX1, SLC7A11, and TFRC. Enrichment analysis revealed key signaling pathways modulated by Que in liver injury: ferroptosis. The results showed that DON significantly decreased the levels of SLC7A11 and GPX4 and increased the levels of TFRC in liver tissue. DON decreased GSH level and GSH-Px activity, and increased MDA and iron levels, resulting in structural and functional damage to hepatocyte mitochondria and liver tissue. All of these DON-induced adverse liver changes were reversed by Que treatment, thereby improving the growth performance of mice. In vitro, DON decreased GPX4, SLC7A11, GCLC, NQO1, and Nrf2 expression; promoted TFRC expression and MDA, 4-HNE, and total ROS production; accelerated GSH depletion; and enhanced lipid ROS accumulation and Fe(II) overload, leading to ferroptosis. Que pretreatment reversed these changes and alleviated DON-induced ferroptosis, thereby increasing cell viability and cell growth. In addition, Que decreased erastin and RSL3-induced lipid ROS level. Altogether, Que attenuated DON-induced liver injury by activating the SLC7A11/GSH/GPX4 signalling pathway, and thereby inhibiting hepatic ferroptosis. This study indicated that Que has a positive impact on liver health in toxin pollution.
Deficiency of feed and fodder year-round is the biggest challenge for livestock rearing in the context of climate change and global warming. Among the unconventional forage sources, Lantana camara has recently gained attention because of its widespread availability. The plant leaves contain a significant amount of protein and nutrients, which makes them a potential forage source. However, the occurrence of lantadenes is also responsible for hepatotoxicity in animals. Therefore, this study investigated the comparative evaluation of different detoxification treatments on L. camara leaves and their possible effects on nutritional composition, Lantadene A concentration, and in vitro cytotoxicity. The L. camara leaves were given eight different treatments, viz. shade-drying, sun-drying, soaking in water, chopping, activated-charcoal-treatment, aqueous Berberis lycium extract-treatment, aqueous Picrorhiza kurroa extract-treatment, and untreated leaves. Later, lantadenes were isolated from all the treated lantana leaves. Thin-layer chromatography was carried out for qualitative estimation of lantadenes, followed by quantification of lantadene A by UHPLC-PDA method. The quantitative analysis revealed a decrease in Lantadene A content in aqueous P. kurroa-treated leaves (453.2 mg/g) and those soaked in water (460.5 mg/g) as compared to untreated (490.9 mg/g). In vitro cytotoxicity evaluation on Caco-2 cell lines was done by MTT assay to determine the IC50 values. The analysis of the results revealed that soaking in water for 18 h is expected to decrease the toxicity of L. camara leaves. The soaked leaves had higher crude protein (23.9%), ether extract (8.7%), and crude fibre (13.7%) contents. Therefore, this treatment needs to be further investigated in an in vivo model to find the safe level of inclusion in animal feed.
Centratherum punctatum Cass. (Asteraceae) is a widely distributed plant in northeastern Brazil and has been associated with natural poisoning in cattle and goats, as well as experimental intoxication in sheep. However, naturally occurring poisoning in sheep has not previously been documented. This study describes the epidemiological, clinical, pathological, and histopathological findings of two outbreaks of C. punctatum poisoning in sheep from the semiarid region of Paraíba State, northeastern Brazil. In Outbreak 1, 20 sheep grazing in a forage cactus (Opuntia spp.) field heavily invaded by C. punctatum developed soft feces, diarrhea, dehydration, and rapid loss of body condition. All animals recovered spontaneously after access to the plant was prevented, while forage cactus consumption continued. In Outbreak 2, four sheep received freshly harvested forage predominantly composed of C. punctatum. The adult ewe and ram developed only soft to pasty feces and recovered within five days after plant withdrawal, whereas two lambs developed severe gastrointestinal disease and died. Fecal egg counts were low, hematological findings were unremarkable, and hepatic and renal biochemical parameters evaluated in Outbreak 2 remained within their respective reference intervals. Necropsy of the lambs revealed lesions predominantly affecting the gastrointestinal tract, including ruminal reddening, ruminal tympany and dry, compacted gastrointestinal contents in one lamb, and abundant fluid gastrointestinal contents in the other, abomasal edema with multifocal superficial erosions, and intestinal hyperemia. Histologically, the forestomachs showed epithelial degeneration, cytoplasmic vacuolation, multifocal erosions, adherent necrotic cellular debris, mild mixed inflammation, and vascular congestion. The abomasum showed multifocal superficial erosions, while the small intestine exhibited moderate diffuse lymphoplasmacytic enteritis without identifiable parasitic structures. The convergence of epidemiological, clinical, laboratory, and pathological findings, together with recovery after plant withdrawal, supports C. punctatum as the cause of disease. This study provides the first detailed characterization of naturally occurring C. punctatum poisoning in sheep.
Structural motifs can be conserved among proteins with low primary-sequence identity, potentially supporting distinct yet related molecular functions. Endogenous phospholipase A2 inhibitors from snake plasma (sbPLIs) include the γ-type inhibitor from Crotalus durissus terrificus, known as Crotalus neutralizing factor (CNF), which protects the snake by selectively inhibiting the lethal and PLA2 activities of crotoxin (CTx). CD59, a complement regulator and single-LU-domain member of the Ly6/uPAR superfamily, shares the three-finger fold with CNF, raising the possibility of structural and functional convergence. Here, CD59 linear peptide arrays produced by SPOT synthesis were probed with CB, the PLA2 subunit of CTx, to identify CD59 regions capable of binding CB. Two reactive clusters were mapped, and derived peptides were synthesized and tested for their ability to inhibit the PLA2 activity of CTx and CB in vitro. Four CD59-derived peptides interacted with CB on the arrays; however, none inhibited PLA2 activity, in contrast to native CNF used as a positive control. These findings indicate that, although CD59 contains linear segments that recognize CB, this interaction is not sufficient to reproduce the PLA2-inhibitory function of CNF, suggesting that any functional interplay between these Ly6/uPAR proteins is likely to depend on more complex structural determinants.
Snakebite envenoming (SBE) remains a significant public health issue in Brazil, where venom diversity, barriers to timely healthcare, and widespread ethnomedical practices intersect. Venom variability across medically relevant snake species poses important challenges for antivenom design and antigenic coverage in the country. In this context, this integrative review examines how venom diversity, antivenom efficacy and protective coverage, and traditional first aid and pre-hospital practices influence the management of snakebite envenoming. Preclinical studies revealed substantial inter- and intraspecific variation in the venoms of Bothrops, Crotalus, Lachesis, and Micrurus species, resulting in heterogeneous venom compositions and biological activities, including lethality that may affect antigenic recognition and neutralization. Although Brazilian antivenoms generally neutralize major toxic activities and reverse life-threatening systemic manifestations of envenoming, there are important limitations in their protective capacity against well-established severe complications such as local tissue damage and neurological syndromes. These limitations are highlighted by the incomplete representation of regional venom diversity in immunization pools and differential toxin immunorecognition. Delayed access to antivenom further worsens clinical outcomes, particularly in remote areas. Concurrently, medicinal plants and first aid and pre-hospital practices were widely used despite limited scientific validation and, in some cases, potential iatrogenic risks to human health. Overall, the available evidence indicates that effective snakebite management in Brazil requires addressing the combined challenges posed by venom variability and current antivenom programs, optimizing antivenom design and coverage, expanding equitable access to healthcare, promoting evidence-based community education, encouraging rigorous scientific evaluation of safe complementary practices.
Thrombocytopenia is a common manifestation in viperid snakebite envenomings, contributing to systemic bleeding and hemodynamic alterations. Despite its relevance, this effect is not evaluated when assessing the neutralizing ability of antivenoms. In this study we present an in vivo method in mice for the quantification of thrombocytopenic effect. Representative venoms of Costa Rican viperid snake genera (Atropoides, Bothrops, Bothriechis, Cerrophidion, Crotalus, Lachesis, Metlapilcoatlus, and Porthidium) were administered by the intramuscular route in mice and platelet counts were carried out 3 h after envenoming. Only the venoms of Bothrops asper and Bothriechis lateralis induced a significant drop in platelet numbers as compared to controls receiving saline solution. The polyvalent antivenom used in Central America was effective in the neutralization of the effect, even in the case of B. lateralis venom, which is not included in the immunizing mixture. In the case of B. asper venom, a C-type lectin-like (SNACLEC), which indues platelet aggregation in the presence of plasma, is responsible for the thrombocytopenic effect. Thus, an in vitro test was designed to assess the efficacy of the antivenom, based on the neutralization of the in vitro aggregating effect of the SNACLEC and of crude venoms of B. asper and B. lateralis in which proteinase activity was inhibited to avoid the coagulation of plasma. This in vitro assay could be useful for assessing antivenom efficacy against thrombocytopenic components of venoms. It is suggested that the neutralization of thrombocytopenic effect of venoms by antivenoms be added to the list of supplementary tests used in the preclinical assessment of antivenom efficacy.
Ant toxinology in Brazil is a small but growing field that has revealed wide biochemical diversity and clear potential for bioprospecting therapeutic molecules. This review compiles the Brazilian contribution, focusing on advances in the characterization of venoms from medically and ecologically important species of Dinoponera, Solenopsis, Paraponera, Pachycondyla, Neoponera, and Ectatomma. Brazilian groups applied omics approaches, including transcriptomics and proteomics, to resolve the composition of these venoms and identified peptide-rich arsenals with antimicrobial, antiparasitic, antitumor, and neuroactive activity. Studies of venom phenotypic plasticity showed ecological factors such as diet and seasonality shape venom composition. Two challenges persist: assigning function to still unidentified components and obtaining venom in the quantities that broad analysis requires. The near-term prospects are the rational design of peptide analogues with improved activity and continued bioprospecting of new species, which together position Brazil as a central contributor to ant toxinology.
INTRODUCTION:Ziyuglycoside I, a hemostatic saponin isolated from Sanguisorba officinalis (Di-Yu), has reported anti-hemorrhagic properties. Deinagkistrodon acutus venom (DAV) is associated with venom-induced consumptive coagulopathy, but its effects on whole-blood coagulation and potential adjunctive interventions remain insufficiently characterized. This study evaluated whether Ziyuglycoside I attenuates DAV-induced coagulation abnormalities using thromboelastography (TEG) and molecular docking. METHODS:Citrated whole blood from healthy donors was exposed to 10, 20, or 30 μg DAV to characterize concentration-related coagulation changes and select a challenge dose. Based on these results, 20 μg DAV was used for subsequent intervention experiments with Ziyuglycoside I, antivenom serum (ASV), or their combination. TEG assessed reaction time (R), clot formation time (K), α-angle, maximum amplitude (MA), and lysis at 30 min (LY30). Molecular docking evaluated the potential interaction between Ziyuglycoside I and a representative snake venom metalloproteinase (SVMP). RESULTS:DAV induced progressively more pronounced TEG abnormalities with increasing concentrations, and 20 μg was selected as a marked but quantifiable challenge dose. At this concentration, DAV prolonged R and K, decreased α-angle and MA, and increased LY30. Ziyuglycoside I significantly improved clot formation kinetics and clot strength, whereas ASV markedly suppressed LY30 elevation. Compared with ASV alone, combination treatment further increased MA, indicating additional improvement in clot strength. Molecular docking predicted a favorable binding pose of Ziyuglycoside I near the active region of the representative SVMP (-8.2 kcal/mol). CONCLUSION:Ziyuglycoside I attenuated DAV-induced coagulation abnormalities mainly by improving clot formation and strength, whereas ASV more strongly inhibited hyperfibrinolysis. Their combination further improved clot strength, supporting Ziyuglycoside I as a potential adjunctive compound for DAV-induced hemostatic disturbances.
