Poster preparation and presentation information

Thank you for considering presenting your work as a poster at this conference. Below are the key details and submission guidelines.

Deadlines

• Poster abstracts (as MS Word) must be submitted by 15th August.
• Digital posters (as PDF) and flash-talk videos must be submitted by 20th August.

Poster preparation & format

Poster size

Digital-only presenters

• Save your poster in A1 or A0, landscape or portrait format.
• Page size is flexible if presenting only digitally.

In-person presenters

• Print your poster in A1 portrait format only
• Larger or landscape posters may not be displayed due to space constraints

File naming

For submission, name your digital poster files as follows:

• <your surname>-Ven26-Poster.
• Example: for David Jones, name your file as Jones-Ven26-Poster.
• Do not use generic names such as: Oxford-poster, Venoms2026, Oxford-venoms-poster.

Digital poster submission

All poster presenters, whether attending virtually or in person, are required to submit a digital version ( as PDF) of their poster via the designated upload link. Posters will be made accessible to all conference participants via the secure ‘Download VEN26 Documents’ page, ensuring visibility across both virtual and in-person audiences.

• Submit your final PDF poster (<5MB) via the designated upload link
• Do not send posters or abstracts by email
• Ensure your submission is final—once published, it cannot be replaced
• Late submissions may not be included in the programme

Poster presentation

Presentation timing

Presenting digitally only

• Currently, there is no scheduled time for presenting digital posters. Instead, virtual attendees will be able to interact with poster presenters via the Zoom chatbox during the conference.
• A specific poster presentation session may be announced later.

Presenting in-person

• Bring a printed A1 portrait poster for display
• Larger or landscape posters may not be displayed due to space constraints
• You are responsible for printing and transporting your poster
• You may be assigned a specific day and time for display

Presentation via a flash-talk video

To maximise visibility, exposure and engagement, we encourage all poster presenters, whether presenting digitally or in-person, to submit a short video (max 5 minutes) introducing their work. Videos will be featured on the LPMHealthcare YouTube channel.

Instructions

• Download the official opening slide (VenOx26 first slide) and use it as the first slide of your presentation (see example: https://youtu.be/XatqenCd_IU?si=Yu1PooCD4JmSLAiz).
• Record your presentation using Zoom or any preferred platform. Keep it under 5 minutes.
• Save your video in a YouTube-compatible format (e.g., MP4)
• Send your video via a file transfer service such as MailBigFile or WeTransfer to: VenomsOxford@gmail.com

Any further information about the poster presentations will be available in the future on this page.

Before uploading your poster, you must make sure that you follow ALL of the instructions above!

Upload your poster

Accepted posters (unedited)

(Presenters in Bold; virtual only posters are marked V)

If your abstract has been accepted for presentation but it does not appear in the list below, please let us know as soon as possible by emailing VenomsOxford@gmail.com.

A Sweet Approach to Venom Identification: Glycan Probes for Detection of Snake Venom Lectins

Princy Anna Moncy Abraham¹, Alexander N Baker

University of Warwick, Department of Chemistry, Coventry, CV4 7AL, UK

Snakebite envenomation (SBE) is a neglected tropical disease (NTD) caused by a complex concoction of biologically active molecules injected into the bloodstream. The WHO estimates around 80,000 to 138,000 deaths occur yearly due to snakebites, disproportionately affecting low-income communities with limited access to adequate testing facilities and antivenoms. To reduce mortality rates, the correct monovalent antivenom must be administered within an appropriate timeframe as such identification of the species responsible is critical. Although many antibody-based detection methods, such as ELISA, have been developed, they are costly, require trained staff and cold storage, limiting their use in tropical communities where SBE is most prevalent. Glycans offer a promising alternative as sensing units to antibodies as they are more thermally stable and cheaper to produce. Snake venom can contain around 20 to over 100 toxins; venom from vipers contain 4-7% C-type lectins that predominantly bind to terminal galactose and N-acetylgalactosamine residues. Previous work in the Baker group established the first prototype of a glycan-based UV-Vis assay to detect Western Diamondback Rattlesnake (C. atrox) venom. This is a protein-free platform, utilising various glycan-terminated poly(hydroxyethyl acrylamide) (pHEA) gold nanoparticles to detect Rattle Snake Venom Lectin (RSVL) in crude venom. By varying the glycan presented, the length of the pHEA and the size of the gold nanoparticle, the selectivity of the system could be adjusted, achieving a limit of detection (LoD) of 20 μg mL^-1 for the best particle system. This current work focuses on synthesising more complex glycans that have a higher binding affinity to RSVL according to literature. This should further improve the selectivity of the system for C. atrox and lower the LoD (nm mL^-1).

Recombinant Expression, Structural Characterization and Ligand Binding of the Cysteine-Rich Domain of a P-III Snake Venom Metalloproteinase from Bothrops pictus

Juan J Cabrejos¹, Fanny Lazo¹; Armando Yarlequé¹, Diego A Leonardo², Eloy Condori², Victoriano Pérez², Dan E Vivas-Ruiz¹

¹Animal Toxins and Antivenoms Research Group (TOXIVEN) – Molecular Biology Laboratory, Faculty of Biological Sciences, National University of San Marcos, Lima, Peru

²Institute of Physics of São Carlos (IFSC), University of São Paulo (USP), São Carlos, São Paulo, Brazil

Bothrops pictus, a pit viper endemic to the Peruvian coastal desert, produces P-III snake venom metalloproteinases (SVMPs) whose hemorrhagic activity depends on a cysteine-rich domain (CRD) that engages basement membrane components. The CRD of a P-III SVMP from B. pictus was cloned and expressed in Escherichia coli BL21 using the pET SUMO system; induction at 18 °C gave the highest soluble yield. The SUMO-CRD fusion migrated at ~22 kDa and, after SUMO protease cleavage, the free domain at ~11 kDa, with ~5 mg/mL of purified protein recovered. Far-UV circular dichroism (0.2 mg/mL) showed a single minimum at 202–203 nm and a negative shoulder at 222 nm (θ₂₂₂/θ₂₀₈ = 0.39), corresponding to ~24% β-sheet and ~6% α-helix over a predominantly disordered background. Thermal ramping from 4 to 60 °C produced no cooperative unfolding transition but an isodichroic point at ~210 nm, consistent with a polyproline II ⇌ unordered coil equilibrium, with aggregation above 55 °C. Free-thiol quantification detected [N] unpaired cysteine(s) per molecule, indicating incomplete disulfide pairing and providing a mechanistic basis for the limited conformational order attained in the reducing cytoplasm of E. coli. Despite this plasticity, indirect ELISA demonstrated specific binding of the recombinant CRD to collagen IV and laminin, and molecular docking predicted favorable contacts supporting the formation of both complexes. Together, these data indicate that the isolated CRD retains its β-scaffold while remaining conformationally flexible, and that recognition of basement membrane ligands does not require a fully ordered fold, consistent with a coupled folding-upon-binding mechanism. This work provides the first structural and functional characterization of a recombinant CRD from an endemic Peruvian pit viper, and identifies disulfide completion as the key determinant to address in future refolding and functional studies. This work was funded by PROCIENCIA (contract PE501088550-2024).

Small Molecule Scaffold Development for Binding Analysis of Snake Venom CDRs

Alexander J Cantor, Alexander N Baker, Sébastien Perrier

Department of Chemistry, University of Warwick, Gibbet Hill, Coventry, CV4 7AL, UK

Rapid diagnostics play a crucial role in global health by enabling timely detection and treatment of diseases. In low-resource settings, where access to advanced medical infrastructure is limited, affordable and fast diagnostic tools are essential for effective healthcare delivery.  This need is particularly urgent for cases of snakebite, a WHO designated neglected tropical disease affecting over 5.4 million people annually, with the highest burden in less economically developed countries. Accurate detection of envenomation could reduce dependence on polyvalent antivenoms, which are costly and pose a higher risk of adverse effects.  What if rapid, polymer-based diagnostics using peptide chemistry could offer a fast, affordable solution to help tackle this global health challenge? This poster will demonstrate the potential of antibody derived complementarity-determining region (CDR) peptide sequences for use in low-cost diagnostics. While the CDRs used have a high binding affinity for α-cobratoxin – a key neurotoxin in N. kaouthia snake venom, peptide-based diagnostics developed in this fashion could also help tackle some traditional disadvantages of antibody-based diagnostics, such as temperature instability and the molecular weight/absorption relationship. This work introduces a promising new platform for low-cost diagnostics aiming to address critical gaps in current global health responses.

Delayed access to care and severity of scorpion envenomation: a dose-response analysis from Central Brazil

Isadora M L R Castro¹, Ana Clara A Dias¹, Carine R M G Amoroso¹, Laís M L R Castro¹, Laura R Jacomini¹, Layane B F Pinto¹, Maria Eduarda R Hallal¹, Marco Aurélio M Elias², Nelson J Silva Jr¹, Juliany G G Silva¹

¹Pontifical Catholic University of Goiás, Goiânia, Brazil

²Federal University of Goiás, Goiânia, Brazil

Scorpion envenomation is an important public health problem in Brazil, particularly in regions where access to timely medical care remains uneven. This study evaluated factors associated with moderate or severe scorpion envenomation in Goiás, Central Brazil. A retrospective analytical study was conducted using secondary data from notifications of venomous animal exposures recorded by the Goiás Toxicological Information and Assistance Center (CIATox-GO) and the Hospital Estadual de Doenças Tropicais Dr. Anuar Auad (HDT) between 2003 and 2018. Of 28,984 notifications initially identified, 8,328 scorpion envenomation cases met the eligibility, consistency, and completeness criteria. Clinical severity was classified as mild or moderate/severe. Adjusted prevalence ratios (aPRs) were estimated using Poisson regression with robust variance, including sex, age group, zone of occurrence, time from sting to care, and local and systemic manifestations. Longer time to care was independently associated with greater severity; each additional day between the recorded dates was associated with a 3.1% increase in the prevalence of moderate/severe cases (aPR=1.031; 95% CI 1.013–1.049; p=0.001). Zone of occurrence was also associated with severity (aPR=1.124; 95% CI 1.055–1.198; p<0.001). Children were the most vulnerable group: adolescents (aPR=0.483), adults (aPR=0.471), and older adults (aPR=0.616) had significantly lower prevalence than children (all p<0.001). These findings indicate that clinical severity is associated with both biological vulnerability and the timeliness and territorial context of access to care. Reducing delays, strengthening regional emergency and referral networks, and prioritizing pediatric assessment may help reduce avoidable morbidity in endemic areas.

Coral Snake Envenomations in the Metropolitan Region of Goiânia: Epidemiological Rarity and Clinical Importance

Lais M L R de Castro¹, Isadora M L R de Castro², Juliany G G Silva², Nelson J Silva Jr²

¹Evangelical University of Goiás, Anápolis, Goiás, Brazil

²Pontifical Catholic University of Goiás, Goiânia, Brazil

Coralsnake (Micrurus spp.) envenomations are among the least frequently reported snakebite accidents in Brazil but remain medically relevant because of their potential severity and the need for prompt diagnosis and specific antivenom therapy. However, epidemiological data describing these uncommon events, particularly in highly urbanized regions, remain scarce. This study aimed to characterize the epidemiological profile of Micrurus envenomations in the Metropolitan Region of Goiânia (MRG), Central Brazil. A retrospective descriptive study was conducted using data from the Brazilian Notifiable Diseases Information System (SINAN) from 2007 to 2026 in contrast to data derived directly from medical records within the Goiás state health system (SUPES-GO). Data were initially analysed to contextualize the burden of venomous animal accidents, followed by state-level analysis in Goiás and detailed evaluation of Micrurus envenomations occurring in the 21 municipalities of the MRG. During the study period, Brazil reported 4,237,991 venomous animal accidents, including 589,810 snakebites, of which only 0.95% were attributed to Micrurus. In Goiás, 183 (0,53%) Micrurus envenomations were identified, with 18 cases occurring in the MRG. Most patients were male (83.3%), and one-third of the cases occurred in Goiânia. Bites predominantly involved the lower limbs (50.0%), 88.9% of patients received medical care within six hours, and 55.6% of cases were classified as severe. Despite this, 88.9% of patients evolved to cure and no deaths were recorded. These findings demonstrate that, although coralsnake envenomations represent a very small proportion of snakebite notifications, they frequently present with severe clinical classification linked to proper taxonomical identification. Their occurrence in a highly urbanized metropolitan region reinforces the need for continuous epidemiological surveillance, maintenance of clinical suspicion, and timely availability of specific antivenom.

Fear Without Persecution: Knowledge and Myths, Rather Than Risk Perception, Shape Conservation Attitudes Toward Venomous Animals

Ana Clara A Dias², Laura R Jacomini¹, Carine R M G Amoroso¹, Isadora M L R Castro¹, Layane B F Pinto¹, Maria Eduarda R Hallal¹, Bruna, G de Aguiar¹, Ivone F Sousa², Nelson J S Júnior¹, Juliany G G Silva^{1 e 2}

¹Pontifical Catholic University of Goiás, School of Medical and Life Sciences, Graduate Program in Environmental and Health Sciences, Goiânia, Goiás, Brazil

²Pontifical Catholic Universityof Goias, School of Social Sciences and Health, Psychology Course, Goiânia, Goiás, Brazil

Venomous animals often evoke fear and negative perceptions that may affect public support for their conservation. Previous studies suggest that misconceptions, folklore, and cultural beliefs can promote persecutory attitudes, whereas knowledge and education may favor coexistence and conservation. To investigate the relationships among risk perception, knowledge/experience, social influence, myths and beliefs, and conservation attitudes toward venomous animals, while exploring the concept of Fear Without Persecution, whereby fear does not necessarily translate into negative conservation attitudes. A cross-sectional study was conducted with 108 participants using a structured questionnaire composed of five domains: risk perception, knowledge/experience, social influence, conservation attitudes, and myths/beliefs. Descriptive statistics, reliability analyses, Spearman correlations, and multiple linear regression were performed. Snakes were the animals most frequently recalled (81.5%; 88/108), and fear was the predominant emotion reported (59.3%), followed by references to previous accidents (29.6%). Mean scores were 3.68 for risk perception, 3.53 for knowledge/experience, 3.34 for social influence, 4.17 for conservation attitudes, and 2.08 for myths/beliefs. Reliability was satisfactory across domains (Cronbach’s α = 0.679-0.872). Knowledge was positively associated with conservation attitudes (ρ = 0.406, p < 0.001), whereas myths/beliefs showed a strong negative association (ρ = −0.571, p < 0.001). Social influence was positively associated with risk perception (ρ = 0.384, p < 0.001), while knowledge was not significantly associated with risk perception. Previous experience with venomous animal accidents was associated with greater knowledge (p = 0.002), but not with higher risk perception. Multiple regression explained 38.6% of the variance in conservation attitudes (R² = 0.386, p < 0.001). Myths/beliefs emerged as the strongest negative predictor (β = −0.504, p < 0.001), whereas knowledge was a significant positive predictor (β = 0.280, p < 0.001). Risk perception was not an independent predictor of conservation attitudes (p = 0.483). Although fear of venomous animals was highly prevalent, conservation attitudes were influenced primarily by knowledge and myths rather than by risk perception itself. These findings support the concept of Fear Without Persecution, suggesting that fear alone does not necessarily result in persecution or opposition to conservation. Instead, misinformation and misconceptions appear to be the main barriers to coexistence, whereas knowledge promotes more positive conservation attitudes, reinforcing previous evidence that cultural beliefs are stronger drivers of anti-conservation behavior than fear alone.

Can in silico predicted repurposed drugs prevent dermonecrosis caused by Echis romani and Bitis arietans venom?

Anna Groves¹, Stephanie French^1,2, Rachael Da-Silva¹, Rohit Patel¹, Martijn ten Have¹, Cassandra M Modahl¹ 

¹CSRI, Liverpool School of Tropical Medicine, Pembroke Place, Liverpool, L3 5QA, UK

²Keele University, Keele, Staffordshire, ST5 5GB, UK

Snakebite envenoming is a neglected tropical disease responsible for >100,000 deaths annually and long-term morbidity in an additional >400,000 individuals each year. Currently the only treatment for snakebite envenoming is antivenom; however, its use is associated with several limitations, including a high risk of adverse reactions such as anaphylactic shock, low specific antibody titres, and high production costs. Furthermore, antivenoms are ineffective at preventing local tissue damage, which contributes to the high morbidity. Repurposed small molecule drugs have emerged as a promising snakebite envenoming therapy due to several advantages, including reduced costs and the absence of cold-chain storage requirements. Current research has focused on small molecule drugs that inhibit specific snake venom toxins, such as the compounds marimastat and varespladib. However, host-directed small molecule therapeutics represent an alternative approach. Here, we aim to identify candidate drugs capable of targeting the downstream effects of venom by reversing the gene expression signatures of cytotoxic snake envenoming. Using a human keratinocyte cell model and RNA-sequencing, we determined which genes are up- and down-regulated following treatment with cytotoxic snake venoms. From these results, we applied an in silico prediction algorithm to identify small molecule drugs likely to reverse venom-induced gene expression changes. Candidate compounds were then evaluated using resazurin-based cytotoxicity assays in the presence and absence of venom treatments. These findings demonstrate the utility of in silico approaches for identifying novel small molecule drugs that target host response pathways as potential adjunct therapeutics to mitigate dermonecrosis and local tissue damage caused by snakebite envenoming.

Eco-epidemiology, taxonomic precision, and antivenom adequacy in snakebite envenoming in Central Brazil: novel indicators for surveillance quality and public health decision-making

Maria Eduarda R Hallal¹, Yasmin J Mendes², Ana Clara A Dias¹, Isadora M L R Castro¹, Laura R Jacomini¹, Layane B F Pinto¹, Darlan Feitosa Tavares¹, Nelson J S Júnior¹, Juliany G G Silva¹.

¹Pontifical Catholic University of Goiás, School of Medical and Life Sciences, Goiânia, Brazil

²School of Medical Sciences of Minas Gerais, Belo Horizonte, Brazil

Snakebite envenoming remains a neglected tropical disease and a major public health challenge in tropical and subtropical regions. Recognized by the World Health Organization as a global priority, reducing snakebite mortality by 50% by 2030 requires improvements in surveillance and antivenom management. This study evaluated snakebite notifications from a referral center in Central Brazil and proposed indicators to assess surveillance quality and therapeutic adequacy. A retrospective observational study analyzed 11,015 notifications recorded between 2003 and 2018 at a specialized tropical diseases hospital. Demographic, environmental, clinical, and therapeutic variables were analyzed. Two indicators were developed: the Taxonomic Precision Indicator (TPI/IPT), measuring valid species identification, and the Ecoepidemiological Surveillance Index (EESI/IVE), based on species identification, environment of occurrence, time to medical care, clinical severity, and patient outcome. An Antivenom Adequacy Audit compared the recorded snake species with the antivenom administered. Bothrops spp. accounted for 61.6% of cases, Crotalus spp. for 16.3%, and 22.1% lacked taxonomic identification. Most patients received care within three hours, and antivenom administration exceeded 96% of envenoming cases. The TPI/IPT reached 77.9%, indicating moderate taxonomic surveillance performance. The audit identified therapeutic inconsistencies, including antivenom use in non-venomous snakebites and unexpected antivenom combinations. Missing environmental and clinical data reduced surveillance quality. The proposed indicators identified weaknesses in surveillance and treatment while providing a practical framework to evaluate notification quality and optimize antivenom management. Their application may strengthen surveillance systems and support the WHO 2030 targets for reducing snakebite mortality.

Closed loop design of glycans for snakebite envenomation diagnostics

Anthony DQ Hoang^1,2, Gabriele C Sosso¹, Alexander N Baker¹

¹Department of Chemistry, University of Warwick, Coventry CV4 7AL, UK

²Warwick Medical School, University of Warwick, Coventry CV4 7AL, UK

Despite its impact, snakebite envenoming (SBE) remains a global health crisis that causes over 100,000 deaths and leaves a further ~400,000 permanently disabled annually. Traditional animal-derived antivenom is categorised into two main groups: monovalent (effective against one species) or polyvalent (effective against several). Where the biting species is known, monovalent antivenom is preferred as its greater specificity allows for a lower dosage, which reduces the risk of severe adverse immune reactions. Ideal treatment therefore depends on species-level diagnosis, which can be difficult to achieve due to the lack of effective diagnostic approaches. Lateral flow devices are a promising diagnostic format as they are inexpensive, portable, simple to use, and can return results within 5–30 minutes, making them well suited to low- and middle-income countries. Because many medically important snakes have lectins in their venom, glycans can serve as the recognition element, and their low cost and thermal stability makes them ideal for use in these countries. Our group has shown that glycopolymer-functionalised gold nanoparticles can detect Crotalus atrox venom via UV-vis within 20 minutes, with no response to plant lectin controls or to lectin-free Naja naja (Elapidae) venom. Sensitivity remains a barrier due to the current limit of detection (~20 µg/mL) sitting well above clinically relevant levels. Because the glycan is among the strongest determinants of sensitivity, we are building a closed-loop in silico/in vitro pipeline to identify better binders. Machine learning models trained on published glycan microarray data will predict binders, which molecular dynamics simulations then filter for experimental testing, and the resulting data fed back into the models to close the loop.

Development of Novel Glycan Functionalised Architectures for Rapid Detection of Snake Venom

Garrett D Jackson, Alexander N Baker

Chemistry Department, University of Warwick, Coventry, CV4 7SH, UK

Envenomation from snake bites is a WHO designated neglected tropical disease (NTD) which affects between 1-5 million people in tropical and subtropical countries annually. Without adequate treatment, around 20% of these bites are fatal and around 3-4 times that will result in permanent disabilities. With many of these incidences occurring in rural communities, where fast access to appropriate healthcare is minimal, the need for a simple, affordable point-of-care test (POCT) is of utmost importance. The Baker group has previously demonstrated the use of synthetic glyco-polymers functionalised nanoparticles with gold nanoparticle aggregation assays, to detect SARS-COV-2 in both patient and viral samples. This work will explore ideas centred around the synthesis and use of novel glycan architectures, to further the development of an LFD for detecting snake venom (C. atrox, western diamondback rattlesnake) with relatively low detection limits (μgml^-1 range).

A One Health Perspective on Spider Envenomation in Central Brazil: Eco-Epidemiology, Surveillance Quality, Professional Training, and Antivenom Stewardship

Laura R Jacomini, Ana Clara A Dias, Isadora M L R Castro, Maria Eduarda R Hallal, Layane B F Pinto, Ivone F Sousa, Darlan Feitosa Tavares, Nelson J Silva Jr, Juliany G G Silva

Pontifical Catholic University of Goiás, Goiânia, Brazil

Spider envenomation remains an underrecognized challenge for translational toxinology in tropical regions, where species-specific care depends on taxonomic identification, adequate therapy and robust surveillance. This retrospective study analysed 1,482 spider-bite notifications recorded over 16 years at Hospital Estadual de Doenças Tropicais Dr. Anuar Auad, a reference centre in Central Brazil. Epidemiological, ecological, clinical, therapeutic and surveillance-quality variables were assessed. Taxonomic identification was unavailable in 1,314 cases (88.7%), representing the main surveillance limitation. Among identified cases, Loxosceles spp. accounted for 5.9%, Phoneutria spp. for 4.5%, Theraphosidae for 0.8% and Latrodectus spp. for 0.1%. Distinct eco-epidemiological patterns were observed: Loxosceles spp. and Theraphosidae predominated in urban settings, whereas Phoneutria spp. was more represented in rural areas. Adult males were most affected, suggesting occupational, domestic and environmental exposure. Cases clustered during warmer and wetter months, supporting climatic influence on human-arachnid encounters. Most patients received care within three hours and evolved with mild manifestations; however, severe outcomes occurred more often after Phoneutria spp. (6.0%) and Loxosceles spp. (4.6%) accidents, consistent with their neurotoxic and dermonecrotic potential. Renal injury, haemorrhagic complications and compartment syndrome indicated morbidity despite low lethality. Antivenom use concentrated in Phoneutria spp. and Loxosceles spp., but anti-Bothrops, anticrotalic and antiscorpion records revealed therapeutic inconsistencies. Integrating taxonomic resolution, eco-epidemiological patterns, clinical severity and antivenom-use consistency provides a framework to audit surveillance quality and support evidence-based antivenom stewardship. Strengthening species identification, notifier training and treatment audit may improve toxinology surveillance.

Sugar-Coated Sensors: The Synthesis of Glycans for Snake Venom Detection

Aleena J Kombanal, Alexander N Baker

Department of Chemistry, University of Warwick, Coventry, CV4 7AL, UK Snakebite envenomation (SBE) claims an estimated 81,000-138,000 lives annually and roughly 400,000 of the surviving victims suffer permanent physical or psychological disabilities. Antivenom remains the primary treatment for envenomation; however, effective treatment relies on administration of the appropriate antivenom. Accurate and rapid identification of the offending snake is therefore critical. Current research in the Baker Humanitarian Chemistry Group aims to develop a glycan-based lateral flow assay (LFA) for the detection of SBE. In 2025, the group developed the first proof-of-concept glycopolymer-based UV-visible assay for detecting Western Diamondback Rattlesnake (Crotalus atrox) venom. A library of glycan-terminated poly(hydroxyethyl acrylamide) functionalised gold nanoparticles with a range of polymer tether lengths, glycans and AuNP sizes was investigated with the optimal particle system exhibiting a LoD of ~20 ug mL^-1. This work will build on these findings by synthesising a range of complex glycans with reference to previously reported glycan-array data against rattlesnake venom lectin (RSVL). These complex glycans will then be evaluated to identify structures that exhibit stronger and more selective RSVL recognition, informing the development of an antibody-free diagnostic platform for SBE.

Discovery of V_HHs Targeting Three-Finger Toxins from Micrurus alleni and Micrurus mosquitensis for development of a genus-wide recombinant antivenom

Maissa Mathlouthi, Melisa Bénard-Valle, Matilde Cotrim, Kasper H Björnsson, Pelle D Knudsen, Andreas H Laustsen

Department of Biotechnology and Biomedicine, Technical University of Denmark, Lyngby, Denmark

Snakebite envenomation remains a major global health concern. Coral snakes, distributed across North and South America, are estimated to cause approximately 2,400 envenomation cases annually. Coral snake venom exerts a potent neurotoxic effect which can be lethal if untreated. The major toxin families in coral snake venom are phospholipases A₂ (PLA₂s) and three-finger toxins (3FTxs). Current treatment relies on polyclonal antibodies obtained from the plasma of hyperimmunized animals; however, they often suffer from batch-to-batch variation and low neutralization of poorly immunogenic toxins. Venom variability among coral snake species also limits cross-neutralization. Our group has previously shown that a mix of only two single-domain antibodies (V_HHs) neutralize the lethality of whole coral snake venoms from Mexico and the United States. This project aims to discover V_HHs that can extend the neutralization capacity of this mixture to coral snake species from central and south America. To this end, we performed the discovery and initial characterization of V_HHs against 3FTxs from Micrurus alleni  and  Micrurus  mosquitensis. This poster presents the purification and isolation of specific 3FTx toxins from crude venom, the discovery of toxin-specific V_HHs through screening of phage display outputs and assessment of V_HH binding. To date, high affinity binders have been identified and expressed and are currently under further characterization. This work may ultimately contribute to the development of a broad-spectrum antivenom effective against coral snake species across the Americas.

Galleria Envenoming Model (G.E.M.): an alternative in vivo non-mammalian model using Galleria mellonella (wax moth) larvae

Rachel Bridge¹, Stefanie K Menzies², James Wakefield³, James Pearce³, Aarón Gómez Argüello⁴, Guillermo León Montero⁴, Stuart Ainsworth⁵ and Rachel H Clare¹

¹Biology Department, Edge Hill University, St Helens Road, Ormskirk, UK

²Biomedical & Life Sciences, Lancaster University, Bailrigg, Lancaster, UK

³Living Systems Institute, University of Exeter, Stocker Road, Exeter, UK

⁴Instituto Clodomiro Picado, Universidad de Costa Rica, Dulce Nombre of Coronado, San José, Costa Rica

⁵Institute of Infection, Veterinary & Ecological Sciences, University of Liverpool, Brownlow Hill, Liverpool, UK

Snakebite envenoming (SBE) annually kills ~100,000 people worldwide, with 4-fold more suffering life-altering morbidity. Antivenom, the only approved treatment, requires preclinical models to assess efficacy and safety as an essential part of the regulated manufacturing  process. The only validated assays to determine the potential clinical effectiveness of an antivenom are preclinical rodent models (typically mice). These procedures are rated ‘severe’ in the UK and EU, are resource-intensive and require high animal numbers. Galleria mellonella have gained momentum as a replacement for vertebrate models in the last decade across many scientific fields. Developing a G. mellonella model for SBE would provide a low-cost, low-resource intensive, high-throughput non-vertebrate alternative in vivo model. Our pilot data demonstrates that G. mellonella envenoming pathology is comparable to mammalian envenoming, resulting in discrete pathology representing clotting abnormalities, paralysis and cytotoxicity. Through our NC3Rs project grant titled the Galleria Envenoming Model (G.E.M) we are i) developing the model to allow for quantitative non-bias analysis of clotting, paralysis and cytotoxicity, ii) validating this model through testing 14 diverse venoms (LD₅₀s) and 3 antivenoms (ED₅₀s) including correlation to the directly matched mouse data, before iii) validation of the model in 2 academic groups and within the antivenom manufacturer Instituto Clodomiro Picado. In parallel we will host workshops (online and in person) to train additional laboratories to establish the same standardised model. To date we have tested 6 of the 12 venoms and one antivenom (EchiTAb plus-ICP) against three of these venoms.

Potential use of G-CSF therapy in skeletal muscle alterations induced by Bothrops leucurus venom

Clara M Mimoso¹, Marcos Lázaro S Guerreiro², Girlaine C Santos³, Simone G Macambira^3,4, Luciana L Casais-e-Silva¹

¹Laboratory of Neuroimmunoendocrinology and Toxinology (LABNIET), Department of Bioregulation, Institute of Health Sciences (ICS), Federal University of Bahia (UFBA), Salvador, BA, Brazil

²Department of Life Sciences, State University of Bahia (UNEB), Salvador, BA, Brazil

³Laboratory of Tissue Engineering and Immunopharmacology, Gonçalo Moniz Institute, FIOCRUZ/Bahia, Salvador, BA, Brazil

⁴Department of Biochemistry and Biophysics, Institute of Health Sciences (ICS), Federal University of Bahia (UFBA), Salvador, BA, Brazil

Snakebite is a major public health issue in tropical regions. In Bahia, Brazil, around 70% of these incidents are caused by Bothrops leucurus snake (BL). The local effects of envenomation are not effectively neutralized by antivenom therapy, often resulting in irreversible tissue damage. Although alternative treatments have been proposed, none are clinically implemented. Experimental Granulocyte Colony-Stimulating Factor (G-CSF) treatments in skeletal muscle (SM) showed an increase in the diameter of regenerative fibers, a rise in myocyte numbers, and enhanced angiogenic effects, improving tissue recovery. This study aimed to characterize the temporal progression of SM injury and repair induced by BL venom in an experimental model, including structural damage, inflammatory cell infiltration, and cytokine expression. The effect of G-CSF treatment (Filgrastim®) on these parameters was also evaluated. Swiss mice (n=5) were inoculated with BL venom (50 μg/paw) and euthanized at 3 h, 6 h, 24 h, 7 d, 14 d, and 28 d (CEUA-ICS 3470020420). G-CSF (0,3 μg/μL) was administered to distinct groups at 7 d and 28 d post-inoculation. Local damage was assessed through plasma creatine kinase (CK) quantification, histological analysis, inflammatory cell counts, and cytokine gene expression (RT-qPCR). Venom-induced myotoxicity was confirmed by elevated CK at 6 h, along with edema, hemorrhage, myonecrosis, myofibrillar hypercontraction, and delta lesions. Inflammatory infiltrates were observed at all time points, with the highest levels at 7 d and 14 d. Gene expression revealed increased levels of TNF-α, IL-1β, IFN-γ, IL-6, IL-10, ARG, and VEGF at different time points. G-CSF treatment reduced pro-inflammatory cytokine expression, improved vascular growth, and lowered collagen deposition, suggesting enhanced muscle recovery. These findings indicate that G-CSF may be an effective therapeutic strategy to reduce musculoskeletal damage following BL envenomation.

Comparative Biochemical and Biological Characterization of the Venoms of Echis coloratus and Echis carinatus sochureki (V)

Nafiseh Nasri Nasrabadi¹, Majid Tebianian², Morteza Taghizadeh², Hossein Vatanpour³, Mehdi Kheirollahpour^4,5

¹Quality Control Department, Razi Vaccine & Serum Research Institute, Agricultural Research, Education and Extension Organization (AREEO), Karaj, Iran

²Razi Vaccine & Serum Research Institute, Agricultural Research, Education and Extension Organization (AREEO), Karaj, Iran

³Department of Toxicology, School of Pharmacy, Shahid Beheshti University of Medical Sciences, Tehran, Iran

⁴Department of Analytical Chemistry, Chemistry & Chemical Engineering Research Center of Iran (CCERCI), Tehran, Iran

⁵Department of Venomous Animals and Anti-venom, Razi Vaccine and Serum Research Institute, Agricultural Research, Education and Extension Organization, Karaj, Iran

Echis coloratus (saw-scaled viper, Middle Eastern) and Echis carinatus sochureki (Sochurek’s saw-scaled viper, Asiatic) are venomous snakes with clinical significance responsible for significant morbidity and mortality. They are members of the genus Echis and their differing venom composition could be the cause for divergent clinical course and success of antivenom therapy. This study provides a comparative analysis of the biochemical composition and biological activities of venoms from E. coloratus and E. carinatus sochureki, focusing on enzymatic profiles, toxicity parameters, and hematological effects. Venom proteins were analyzed using SDS-PAGE and RP-HPLC. Bioactivities were assessed through parameters, including LD₅₀, hemorrhagic, procoagulant, defibrinogenating, and necrotic activities, as well as by measuring different blood coagulation times (PT, APTT, TCT, and FCT). Blood-related perturbations, induced by venom, were measured in in vitro and in vivo models. In addition, neutralizing capability of a commercially polyvalent antivenom against the venom’s procoagulant and defibrinogenating activities was assayed. Both venoms share a predominance of snake venom metalloproteinases (SVMPs) and serine proteases, but significant quantitative differences were observed. E. carinatus sochureki venom exhibited higher PLA₂ activity and greater lethal potency (lower LD₅₀), whereas E. coloratus venom showed stronger hemorrhagic activity and higher SVMP content. Similar SDS-PAGE and RP-HPLC analyses confirmed there were qualitative and quantitative difference between the protein constituents of the two venoms in accordance with the previously demonstrated difference in their biologic activities Both venoms demonstrated potent procoagulant effects via factor X and prothrombin activation, albeit with differential kinetics. Antivenom neutralization studies indicated variable cross-reactivity. This report confirms that envenomation by Echis carinatus sochureki can lead to severe hematological disorders that become evident within the first hours after envenoming, necessitating prompt medical and clinical attention to reduce the severity of symptoms. To the best of our knowledge, this is the first study to report a comparative analysis of the biochemical and biological profiles of the venoms of E. coloratus and E. carinatus sochureki in Iran. The venoms of these two species exhibits both overlapping and species-specific biochemical and toxicological profiles. These differences underscore the importance of species-specific antivenom development and have direct implications for the clinical management of envenomation in endemic regions.

Evolution of alkaloid sequestration in poison frogs: a phylogenomic study

Julia A de Pinna^1,2, Daniel C Pimenta², Kim Roelants¹

¹Ecology, Evolution and Genetics group, Vrije Universiteit Brussel, Brussels, Belgium

²Ecology and Evolution Lab, Instituto Butantan, São Paulo, Brazil

The majority of frogs use a proteinaceous skin secretion to fend off predators. However, in Dendrobatidae and several other frog families, these gene-encoded toxins have been evolutionarily replaced by lipophilic alkaloids sequestered from a specialized diet. As these replacements imply the evolution of mechanisms that protect the frogs from the alkaloids’ neurotoxic effects and promote the accumulation of the alkaloids in the frogs’ skin glands, they represent a remarkable case of evolutionary innovation. To elucidate the origin of toxin sequestration, we investigated patterns of adaptative evolution in (1) Saxiphilin and Alkaloid-Binding Globulin (ABG), two secretory proteins previously proposed to be involved in detoxication, transport and storage of alkaloids and (2) voltage-gated sodium channels (Nav) known as the most common target of neurotoxic alkaloids. Genomic screening and phylogenetic analyses show that the Saxiphilin and ABG genes of alkaloid-sequestering frogs have orthologues in a wide range of non-sequestering frogs, suggesting repurposing of preexisting genes at the origin of toxin sequestration. Gene duplication events and signatures of positive selection within Dendrobatidae indicate that protein diversification and adaptive evolution may have further optimized alkaloid sequestration in this clade. Predicted structures for the secretory proteins and sodium channels demonstrate aminoacid substitutions in positions that confirm strong selective pressure for toxin processing and resistance. This is the first comprehensive phylogenetic study that targets evolutionary trends in alkaloid sequestration and brings new insights on how this special mechanism evolved in poison frogs.

In Silico Development of Antibody Biomimetics for Snakebite Diagnostics

Neil Prabhakar, Gabriele C Sosso, Alexander N Baker

Chemistry Department, University of Warwick, Coventry, CV4 7SH, UK

Every year, snakebite is responsible for an estimated 1.8-5.5 million envenomations, causing over 100,000 deaths and over four times as many disabilities. Current treatments are antibody-based therapies that are either monoclonal or polyclonal and are traditionally made from immunising large animals such as horses. As such, patients are at risk of anaphylaxis and serum sickness from the only approved therapy. Monoclonal treatments are species-specific, require less treatment than broader polyvalent antivenoms, and therefore reduce the likelihood of an adverse reaction. This necessitates accurate, timely identification of the offending snake, as most bites occur in rural and remote communities. Developing rapid, low-cost, and accessible diagnostic methods is essential. The Baker Humanitarian Chemistry Group has previously shown proof-of-concept glycan aggregation assays against snake venom lectins as potential point-of-care (POC) alternatives. However, glycan sensors may not achieve the low nanomolar and, in some cases, picomolar binding affinities that specific nanobodies can. The CDR-H3 domain of these nanobodies is widely considered the most important domain for target binding. Peptide nanoprobes built from cyclised CDR-H3 domains have previously shown low-nanomolar binding affinities while being more tuneable than whole nanobodies and antibodies. This work will explore machine learning-guided affinity development of potential CDR-H3 sequences and ensemble-based evaluation of structural similarity between cyclised CDR-H3 domains on small organic linkers compared to the native nanobody framework. This aims to develop a workflow for synthetically accessible nanoprobes with high target specificity and broad functionality against snake venom toxins and other target proteins.

Development and Evaluation of RasFab: A Region-Specific Heptavalent F(ab’)₂ Antivenom Against Medically Important Iraqi Snake Venoms (V)

Seyed Fakhroddin Rahmani¹, Seyed Arshia Rahmani², Seyed Bardia Rahmani²

¹Head of production Department, Kimia Wargah Sazan, Tehran, Iran

²Quality Control Department, Kimia Wargah Sazan., Tehran, Iran

Snakebite envenoming remains a major neglected tropical disease in Iraq. The diversity of medically important venomous snakes in Iraq represents a significant therapeutic challenge. The efficacy of antivenoms is greatly influenced by the taxonomic composition of the immunizing venoms, therefore, it is expected that region-specific antivenoms provide a better clinical coverage. The present study was conducted to develop and pre-clinically evaluate a heptavalent equine F(ab’)₂ antivenom against the major venomous snake species in Iraq. Based on a comprehensive review of the medically important snake fauna of Iraq, venoms from seven species (Macrovipera lebetina, Walterinnesia morgani, Montivipera raddei, Echis carinatus, Pseudocerastes persicus, Pseudocerastes fieldi, and Cerastes gasperettii) were selected for immunization. Horses were hyperimmunized with a mixture of venoms and plasma-derived immunoglobulins were enzymatically digested and purified to produce an equine F(ab’)2 antivenom. The antivenom was tested for its capacity to neutralize toxins in standard pre-clinical neutralization assays. RasFAb passed all quality control criteria and demonstrated potent neutralisation efficacy against all seven venoms tested. The antivenom neutralized a minimum of 50 LD₅₀ of each venom in pre-clinical neutralization assays, indicating extensive paraspecific efficacy against the medically important venomous snakes of Iraq. The RasFAb is a new, region-specific, heptavalent equine F(ab’)2-derived antivenom that has been developed for the treatment of snakebite envenomation in Iraq. Its favourable preclinical neutralization profile supports its potential as a therapeutic alternative for the treatment of envenomation caused by the country’s most prevalent venomous snake species. Additional toxicological, stability and clinical studies are required to establish its safety and therapeutic efficacy in human subjects.

In Vivo Myotoxic Activity, Morphological Analysis and Transcriptional Profiling of Native and Recombinant Bothrops atrox Myotoxin (V)

Angie Regalado¹, Jacquelyne Zarria-Romero², Alex Proleón¹, Raul Rivas¹, Daniel Torrejón¹, Fanny Lazo¹ Armando Yarlequé¹, Dan Vivas-Ruiz¹

¹Laboratory of Molecular Biology, Faculty of Biological Sciences, Universidad Nacional Mayor de San Marcos, Av. Venezuela cuadra 34 s/n, Ciudad Universitaria, Lima 15081, Perú

²Laboratory of Reproduction and Developmental Biology, Faculty of Biological Sciences, Universidad Nacional Mayor de San Marcos, Av. Venezuela cuadra 34 s/n, Ciudad Universitaria, Lima 15081, Perú

The present study evaluates the in vivo myotoxic and transcriptional effects of native (BaMtx) and recombinant (rBaMtx) myotoxin from Bothrops atrox venom. Intramuscular administration in mice induced a dose-dependent increase in plasma creatine kinase (CK) activity at all evaluated doses (25, 50, and 75 µg), demonstrating acute skeletal muscle injury. Notably, rBaMtx produced significantly higher CK levels than native BaMtx at each dose. Consistent with this finding, the minimum myotoxic dose (MMD) was lower for rBaMtx (9.90 µg; 95% CI: 7.39–12.80) than for native BaMtx (15.53 µg; 95% CI: 12.90–18.49). Scanning electron microscopy (SEM), performed one hour after toxin administration, provided complementary morphological evidence of acute muscle damage. Control tissues exhibited well-organized, parallel myofibers with continuous sarcolemmal surfaces. In contrast, muscles exposed to either native BaMtx or rBaMtx displayed marked sarcolemmal disorganization, membrane irregularities, and focal surface protrusions. Cross-sectional SEM images further revealed fiber swelling, fragmentation, and severe disruption of the normal parallel muscle architecture in toxin-treated tissues. We evaluated the gene expression profiling of muscle tissue at 3, 6 and 24 h post inoculation to analyze the regenerative phase. Expression kinetics of CD68, CD163 and Ang-1 displayed no significant differences between treatments indicating a comparable magnitude of immune and vascular response. However, rBaMtx induced a significantly higher expression of NGF at 24 h. Together, these findings demonstrate that rBaMtx retains the characteristic in vivo myotoxic activity of native BaMtx, exhibiting greater quantitative myotoxic potency, and initial quantitative differences in toxin induced tissue destruction determine the magnitude of subsequent localized neuro-regenerative signalling. Financial support: PROCIENCIA-Peru (PE501087970-2024); VRIP-UNMSM-Peru (B25100011).

Functional characterization of recombinant Pictolysin-III metalloprotease domains produced in prokaryotic and eukaryotic expression systems (V)

Jorge Alejandro Roque¹, Daniel Torrejón¹, Felix Urra, Fanny Lazo¹, Edith Rodriguez¹, Armando Yarlequé¹, Dan Vivas-Ruiz¹

¹Animal Toxins and Antivenoms Research Group (TOXIVEN) – Molecular Biology Laboratory, Faculty of Biological Sciences, National University of San Marcos, Lima, Peru

²Laboratorio de Plasticidad Metabólica y Bioenergética, Núcleo Interdisciplinario de Farmacología e Inmunología, Instituto de Ciencias Biomédicas (ICBM), Universidad de Chile, Santiago, Chile

Pictolysin-III (Pic-III) is a P-III snake venom metalloproteinase from Bothrops pictus with proteolytic, fibrino(geno)lytic, hemorrhagic, platelet aggregation-inhibitory, and reported cellular effects. However, the contribution of its isolated metalloprotease domain remains incompletely defined. Following recombinant production in Escherichia coli (rMD-Ec) and Pichia pastoris (rMD-Pp), both domains were comparatively evaluated against native Pic-III. Both proteins showed azocaseinolytic activity, with rMD-Pp and rMD-Ec retaining 59.4% and 23.5% of native activity, respectively. Their catalytic responses differed with temperature, pH, incubation time, antivenom, inhibitors, and divalent cations, while EDTA and DTT abolished detectable activity. Both domains preferentially degraded the Aα chain of human fibrinogen and showed time-dependent fibrinolysis. At 60 min, rMD-Pp reached 87.9% of Pic-III fibrinolytic activity, whereas rMD-Ec reached 36.7%. In vivo, rMD-Pp produced dose-dependent hemorrhage, with an estimated minimum hemorrhagic dose of 1.02 µg, whereas rMD-Ec caused no visible hemorrhage up to 4 µg. Exploratory molecular dynamics of theoretical glycosylated and non-glycosylated models showed similar global compactness but numerical differences in solvent exposure and localized flexibility, particularly in the omega-loop. Impedance-based real-time cell analysis also revealed treatment- and cell line-dependent response profiles. Overall, the isolated metalloprotease domain retained several functional properties of native Pic-III, although their magnitude depended on the recombinant preparation. rMD-Pp more closely reproduced the proteolytic, fibrinolytic, and hemorrhagic behavior of native Pic-III than rMD-Ec, supporting an important contribution of this domain to the functional profile of the full-length toxin. Financial support: CONTRATO N.° PE501088550-2024-PROCIENCIA; VRIP B25100061

Cytotoxic Effects of Aflatoxins Produced by Aspergillus flavus and Aspergillus parasiticus Isolated from Feed on Murine Lymphocytes (V)

Nooshin Sohrabi

Department of Biology, Payame Noor University, Tehran, Iran

Aflatoxins are powerful toxins produced by Aspergillus flavus and Aspergillus parasiticus. Ingestion of these mycotoxins is linked to immunotoxicity and adverse health outcomes. This study investigated the cytotoxic effects of aflatoxins on murine T lymphocytes from standard and feed-derived Aspergillus isolates. Two standard isolates and two feed isolates of A. flavus and A. parasiticus were studied. The synthesis of aflatoxins B1 (AFB1) and aflatoxins G1 (AFG1) were evaluated by thin layer chromatography (TLC) and high performance liquid chromatography (HPLC). The cytotoxicity was determined in the murine EL4 T-lymphocyte cell line after incubation with different aflatoxin concentrations for different incubation times. The MTT assay and flow cytometry (FCM) were used to determine cell viability and apoptosis. The standard isolates of A. parasiticus and A. flavus from the feed showed the highest production levels of AFG1 and AFB1 respectively. A concentration of 1:10 dilution of aflatoxin extract from A. parasiticus caused the most significant decrease in EL4 cell viability and the highest incidence of apoptosis. Cytotoxicity increased in a concentration-dependent manner, corresponding to the aflatoxin production profile of the isolates. Feed-derived Aspergillus isolates could produce high levels of AFB1 and AFG1 with remarkable cytotoxic effects on murine T lymphocytes. The findings demonstrate the toxicological significance of feed contamination by aflatoxigenic Aspergillus species and emphasize the need for regular monitoring of animal feed to mitigate exposure to immunotoxic mycotoxins.

How close are Bothrops venom proteins to anticancer translation? A systematic audit of 58 studies

Ana G Amado-Argüelles¹, Armando Yarlequé², Dan E Vivas-Ruiz², Félix A Urra³˒

¹Programa Doctoral en Ciencias de la Salud, Facultad de Medicina Humana, Universidad Nacional Mayor de San Marcos (UNMSM), Av. Grau 755, Lima 15001, Perú

²Laboratorio de Biología Molecular, Facultad de Ciencias Biológicas, Universidad Nacional Mayor de San Marcos (UNMSM), Av. Venezuela Cdra 34 S/N, Ciudad Universitaria, Lima 15081, Perú

³Laboratory of Metabolic Plasticity and Bioenergetics, Interdisciplinary Nucleus of Pharmacology & Immunology (ICBM), Medical Faculty, University of Chile, Av. Independencia 1027, Santiago, Chile

Snake venom proteins are increasingly investigated as molecular scaffolds for anticancer drug discovery, yet the experimental maturity of this evidence remains unclear. We systematically evaluated 58 primary studies addressing the in vitro effects of Bothrops venoms and venom-derived molecules on tumor cells. Evidence was examined across major enzymatic and non-enzymatic protein families, including L-amino acid oxidases, phospholipases A₂, snake venom metalloproteinases, serine proteases, C-type lectins, and disintegrins. L-amino acid oxidases and selected phospholipase A₂ isoforms showed particularly prominent cytotoxic effects, whereas other protein families exhibited more heterogeneous responses involving oxidative stress, mitochondrial dysfunction, apoptosis, membrane perturbation, extracellular matrix remodeling, and altered cell adhesion and migration. However, methodological auditing revealed substantial heterogeneity in toxin characterization, experimental models, concentration reporting, selectivity assessment, and mechanistic validation. Using a three-level framework proposed in this review, 13/58 studies (22.4%) were classified as Level I (baseline cytotoxicity/viability), 31/58 (53.4%) as Level II (functional and cellular validation), and only 14/58 (24.1%) as Level III (molecular mechanistic validation). Thus, although Bothrops venom proteins constitute a diverse source of biologically active anticancer-associated scaffolds, only a limited fraction of current evidence reaches advanced mechanistic characterization. Progress in oncovenomics will require standardized toxin identification, orthogonal biological validation, tumor-versus-normal selectivity assessment, and more physiologically relevant experimental models to distinguish promising macromolecules from preliminary cytotoxic candidates. Financial Support:  PE501088550-2024-PROCIENCIA.

Using a lung transwell model for the in vitro assessment of inhaled defence- and national security-relevant compounds, using ricin as an example

Tiffany Morcom, Matthew Sheavyn, Lucy Roberts, Lucy Cork, Dominic Jenner, Alex Stoll, Millie Fuller, Charlotte Whitmore

Dstl, Porton Down, UK

As scientific research seeks to reduce the reliance on animal models, alternative, validated approaches to accurately model human respiratory biology are necessary to study the effects of compounds in vitro. As an example, inhalation of ricin causes severe lung injury and respiratory failure and is often studied in vivo. Pathological changes in the lung epithelia include necrosis and the apoptotic deletion of epithelial cells at all levels of the respiratory tract, and marked, neutrophilic inflammation. This study examined ricin’s cytotoxic interactions with transwell models of the human lower respiratory tract to identify if in vitro results correlate with these in vivo observations. SmallAir™ transwell models (Epithelix) were exposed to aerosolised ricin, delivered by the VITROCELL® Cloud Alpha MOVE, at deposition densities of ~0.004, 0.4, and 40 ng/cm² . At various timepoints after exposure, assays were conducted to assess cytotoxicity, cytokine release, histological profiles, and ricin permeation. Cytotoxicity was determined at 0-, 4-, 24-, and 48-hours with a LDH assay. The highest deposition density of ricin (40 ng/cm²) increased LDH release in a time-dependent manner, indicating increased cytotoxicity. At the lower densities LDH release did not increase, indicating a minimal cytotoxic effect. This observation was consistent with measures of the barrier integrity of the SmallAir™ models, quantified using lucifer yellow permeability assays and transepithelial resistance measurements. Here, the highest deposition density of ricin caused the most disruption to barrier integrity whilst little effect was observed at the lower densities. These preliminary results are consistent with observations reported in vivo, supporting the use of the SmallAir™ lung transwell model as an in vitro platform for assessing the respiratory effects of ricin. © Crown Copyright 2026, Dstl. This material is licensed under the terms of the Open Government Licence.