Honeybee venom therapy and viral infection: a systematic synthesis of venom antiviral activity

The bioactive components of honeybee venom (HBV) have displayed broad evolutionary antiviral activity in animal models; however, there is limited evidence for the efficacy, indications, or side effects of the use of HBV in humans. We performed a systematic review and meta-analysis of the available preclinical and clinical evidence systematically assessing the antiviral efficacy and safety of HBV. A comprehensive search strategy for all studies reporting HBV in relation to direct-acting antiviral effects was performed through electronic databases from their inception up to October 2025. A total of 32 articles were identified for quantitative synthesis, with some included under meta-analysis. Although majority of the studies show high heterogeneity and limitations, the meta-analysis results provide encouraging evidence for the broad-spectrum antiviral property of HBV against numerous viruses, such as influenza viruses, herpes simplex virus, HIV, hepatitis viruses, and emerging coronaviruses. Mechanistically, the HBV components directly disrupt viral replication or do so via body immunomodulation pathways. The main adverse effects of HBV include dose-related cytotoxicity and mild allergic responses, with rare instances (less than 0.1%) of anaphylaxis. Overall, the antiviral roles of HBV are strongly supported in preclinical studies, although the clinical data on humans have limitations. Therefore, improving formulations and conducting safety assessment in future clinical practice, particularly through the use of nanocarriers or combined therapeutic strategies, may enhance the imperative HBV antiviral potency and reduce its limitations.

Viral infections are still a global burden in terms of the morbidity and mortality they cause, and with new pathogens arising against the backdrop of impending antiviral drug resistance, there is an urgent need for novel therapeutic measures (1, 2). Despite the advances in vaccines and drug development showing great achievements against viral infection, many viral infections lack effective treatments, with increased toxicity and resistance development prompting exploration of alternative therapies (3).

Honeybee venom (HBV)-based therapies have emerged as promising candidates with antimicrobial and immunomodulatory properties due to their diverse bioactive components and historical use in traditional medicine (4, 5). HBV consists of a complex mixture of peptides, enzymes, and bioactive amines. The principal component in HBV is melittin, which represents approximately 40%–60% of the weight of dry venom and has been reported as the main protein contributing to the membranolytic and lytic factors present in HBV (6–8). The other pharmacologically active compounds, including phospholipase A 2 (PLA 2), adolapin, and apamin, contribute to the multifaceted biological activities of HBV (7, 8). Recently, preclinical studies have described melittin and PLA 2 to exhibit potent antiviral properties against enveloped and non-enveloped viruses via direct viricidal effect or via modulation of the host cell signaling pathways and to interfere with viral replication (9–13). The coronavirus pandemic revitalized the curiosity in the utilization of HBV therapy, which has lower rates of infection and/or morbidity (14, 15).

Despite encouraging preclinical results, translation of the therapeutic potential of HBV into practice is still limited. The research in this area is quite dispersed, with various animal models, different preparations of HBV, variable concentrations, and varied outcome parameters (16–18). Adverse effects represent another crucial aspect. In addition to local allergic reactions, HBV has the potential to induce life-threatening anaphylaxis in predisposed individuals. Therefore, a critical assessment of the existing literature was performed to identify whether HBV and its constituents possess genuine promise as antiviral drugs and what potential benefits can be obtained by risking such adverse events.

Due to the rising demand for alternative medicines and the pressing need for new antiviral approaches, we conducted this systematic review and meta-analysis to assess antiviral potency and to gather research findings and provide impetus and guidance for new developments in antiviral approaches based on HBV, with its potential applicability in human and animal healthcare.

The methodological standards set by Preferred Reporting Items for Systemic Reviews and Meta-analyses (PRISMA) 2020 (19) and Cochrane were used in this overview.

Eligibility criteria

Studies were included if they: 1) examined HBV, melittin, or PLA 2 as the intervention; 2) assessed antiviral effects in a human, animal, or in vitro model system; 3) had a control comparison group(s) such as a placebo or no treatment and/or an alternative treatment group(s); and 4) reported quantitative measurements of the outcomes relevant to virus infection (e.g., viral load reduction, replication inhibition, plaque reduction assay data, or clinical endpoints).

The exclusion criteria were as follows: narrative reviews, case reports lacking quantifiable antiviral results, and non-viral pathogens.

Search strategy

A comprehensive search was performed in PubMed, Embase, Web of Science, Scopus and Cochrane Central up to October 2025 using the following keywords: (“bee venom” OR “apitoxin” OR “melittin” OR “phospholipase A2” OR “apipuncture”) AND (virus OR viral OR antiviral OR influenza OR HSV or SARS-CoV-2 or COVID-19). The reference lists of relevant papers and reviews were manually searched for additional studies. No limitations related to language or date of publication were imposed (Supplementary Table S1).

Selection criteria and data extraction

Two reviewers independently screened the titles, abstracts, and full texts. Disagreements were resolved through consensus. The data extracted included the data source, study design, sample size, species, virus type, intervention and outcome details, shine light (wavelength) details, the time duration for each session, and adverse events.

Risk of bias assessment

The Cochrane risk-of-bias (RoB) 2 tool was used for human randomized controlled trials (RCTs), while ROBINS-I (Risk Of Bias In Non-randomized Studies—of Interventions) was utilized for non-randomized studies. For animal studies, the SYRCLE tool was used, and a custom quality checklist (e.g., replication, blinding, and cytotoxicity controls) was employed for in vitro studies.

Data synthesis

Quantitative analysis was performed using a random-effects model (restricted maximum likelihood, REML) for continuous (standardized mean difference, SMD) and dichotomous (risk ratio, RR) outcomes. Heterogeneity was calculated using the I 2 and τ 2 statistics. Pre-specified subgroup analyses were performed according to the virus family, the venom components, and the study design. Funnel plots and Egger’s test were used to estimate publication bias when at least 10 studies were available.

Selection criteria

In total, 1,247 records were retrieved. After discarding duplicates, 932 remained for screening. A total of 32 studies were included in the qualitative synthesis (meta-analysis) (Figure 1; Supplementary Tables S2A, B).

Characteristics of the 32 included studies

Human clinical studies (n = 6) were on topical/injectable HBV preparations for mild coronavirus disease 2019 (COVID-19), herpes simplex virus (HSV), or warts. Studies in animals (n = 14), including mice, ferrets, and chickens, were on infection of influenza, HSV, or enterovirus models. In vitro studies (n = 12) were on melittin and PLA 2 against different strains of viruses in cell cultures. The interventions included crude HBV extracts (0.1–1 mg/ml) and purified melittin or nano-melittin conjugates (0.5–10 µM).

Quantitative synthesis (meta-analysis)

The pooled SMD of the meta-analysis on dexindanol and virus control (VC) on the inhibition of viral replication was −2.15 (95%CI, 1Score = 74). There was a statistically significant difference between the intervention group and the control group (Figure 2).

In animal studies, pulse HBV therapy decreased the viral load, with a pooled mean difference of −1.8 log10 copies/ml (95% CI = −2.4 to −1.2). One clinical study showed greater resolution of symptoms (RR = 1.45, 95%CI = 1.05–2.01); however, the heterogeneity was high (I 2 = 72%). Examination of the funnel plots indicated potential small-study bias (Figure 3).

Risk of bias

The majority of RCTs were rated as “some concerns” due to the lack of blinding, while the animal and in vitro studies had a “moderate risk” rating based on randomization and outcome assessment (Figure 4; Supplementary Tables S3A–D).

This systematic review represents the largest and most comprehensive synthesis to date evaluating HBV and its bioactive components as antiviral agents across diverse viral families. Evidence from 32 included studies demonstrated that crude HBV and purified venom fractions consistently inhibited viral replication in vitro and in vivo, with emerging—but still limited—human evidence. The antiviral activity was observed against enveloped and non-enveloped viruses, including influenza viruses, herpesviruses, hepatitis viruses, flaviviruses, enteroviruses, and coronaviruses, underscoring the broad-spectrum antiviral potential of HBV-derived compounds.

At a mechanistic level, HBV peptides, most prominently melittin, but also PLA 2 and apamin, exert their antiviral effects through direct virucidal activity, viral entry and replication inhibition, and host immunomodulation. Collectively, these mechanisms suggest that HBV acts not only as a direct antiviral agent but also as a host-directed therapy, modulating immune pathways that are critical for viral clearance.

Contribution of crude bee venom versus purified components

An important finding from Supplementary Table S2 is that crude HBV demonstrates antiviral activity comparable to that of purified components in several studies, particularly in vitro. Crude venom contains a complex mixture of peptides and enzymes, including melittin, PLA 2, apamin, mast cell-degranulating peptide, and minor bioactive molecules, which may act synergistically to enhance antiviral efficacy.

Several studies reported that crude venom reduced the viral RNA levels and infectious titers at relatively low concentrations, particularly against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), influenza A, hepatitis B virus, and HSV, suggesting that the combined action of multiple venom constituents may confer advantages over single isolated peptides. However, the variability in the venom composition—driven by the bee species, geographic origin, extraction methods, and storage conditions—represents a major challenge for reproducibility and clinical translation. These findings emphasize the need for standardized venom preparation, biochemical characterization, and potency assays in future studies.

Mechanistic rationale and the role of apamin

The antiviral effects of HBV are largely attributed to the amphipathic peptide melittin, whose α-helical structure enables insertion into lipid bilayers, resulting in pore formation, membrane destabilization, and viral envelope disruption (20–24). This mechanism explains the higher sensitivity of enveloped viruses to melittin-based interventions while also accounting for dose-dependent cytotoxicity.

Beyond melittin, apamin, a small neuroactive peptide traditionally studied for its effects on ion channels, has emerged as an underexplored but relevant antiviral modulator. Evidence displayed in Supplementary Table S2 indicates that apamin can reduce the viral DNA/RNA levels and viral antigen expression, particularly in hepatitis B virus models. Unlike melittin, apamin does not primarily act through membrane lysis; instead, it appears to influence the host transcriptional regulation and intracellular signaling pathways, potentially affecting viral replication indirectly.

In addition, PLA 2 contributes to antiviral activity by hydrolyzing phospholipids in viral envelopes and host membranes, thereby interfering with viral entry and fusion. Together, these findings support a multi-target antiviral paradigm, in which the HBV components act at different stages of the viral life cycle.

Advances in nanotechnology—such as melittin-loaded liposomes, nanoparticles, or peptide conjugates—have significantly improved the therapeutic indices by reducing the hemolytic activity while preserving the antiviral potency (25–27). These delivery systems may also enable the co-delivery of multiple venom components, allowing the synergistic effects between melittin, PLA 2, and apamin to be harnessed safely.

Immunomodulatory effects and host-directed antiviral activity

In addition to their direct antiviral effects, HBV and its components exert significant immunomodulatory actions. Multiple studies have reported the stimulation of type I interferon (IFN) signaling pathways, alongside the suppression of excessive inflammatory responses, including the downregulation of TNF-α, IL-6, and NF-κB signaling (28–30).

This dual immunological profile is particularly relevant for viral infections characterized by immune dysregulation, such as influenza and COVID-19. By enhancing the antiviral innate immunity while limiting cytokine-mediated tissue damage, HBV components may help reduce the viral load and disease severity simultaneously. Apamin, in particular, may contribute to this balance by modulating the calcium-dependent signaling and inflammatory cascades without inducing membrane damage.

Safety profile and translational considerations

Safety remains a critical barrier to clinical application. Across studies, the most frequently reported adverse events associated with HBV exposure were local pain, swelling, and erythema, while systemic allergic reactions were rare, but potentially severe (31). Importantly, the reviewed evidence suggests that purified formulations, controlled dosing, and desensitization protocols can substantially mitigate the risk of anaphylaxis (32).

Crude HBV may pose a higher immunogenic risk compared with purified peptides; therefore, component-specific formulations or standardized venom extracts with defined allergen profiles are preferable for therapeutic development. Preclinical toxicology studies and allergy screening protocols must be rigorously integrated into future clinical trial designs.

Comparison with previous reviews

Earlier narrative reviews highlighted the antiviral promise of HBV, particularly melittin, but lacked systematic methodology and quantitative synthesis (33–35). The present review advances the field by providing a PRISMA-compliant systematic assessment, integrating mechanistic insights, risk of bias evaluation, and comparative analysis across experimental models. While preclinical findings are largely concordant, translation to human application remains limited by the small sample sizes, the heterogeneity in the venom composition, and the inconsistent dosing strategies.

Strengths and limitations

The strengths of this review include a comprehensive multi-database search, the inclusion of both preclinical and early clinical evidence, and a detailed mechanistic interpretation grounded in experimental data.

Limitations include the small number of human studies, substantial methodological heterogeneity, potential publication bias, and the inherent constraints of in vitro models that may not fully recapitulate physiological conditions. Furthermore, variability in the crude venom composition complicates cross-study comparisons.

Overall implications

Taken together, the findings support HBV, particularly melittin-, PLA 2 -, and apamin-based formulations, as a promising platform for antiviral drug development. Future research should prioritize standardization, advanced delivery systems, combination strategies, and well-designed clinical trials to fully realize the therapeutic potential of HBV in antiviral medicine.

Future directions and recommendations

Future studies should focus on:

The development of standardized methods for venom extraction and quantification;

Double-blind placebo-controlled RCTs with defined viral endpoints and of high quality;

Investigation of nanotherapy and combined therapeutics with melittin; and

Careful monitoring of the safety of this agent and broad profiling of its pharmacokinetics.

Collectively, the available evidence indicates that HBV exhibits a consistent and biologically plausible antiviral activity across a wide range of viral families, with melittin emerging as the principal bioactive component responsible for these effects. Robust in vitro and in vivo findings demonstrate that HBV and its purified constituents can directly inactivate viral particles, interfere with viral replication, and modulate the host immune responses through both innate and adaptive pathways. These multifaceted mechanisms position HBV as a promising candidate within the expanding field of natural antiviral therapeutics.

Despite these encouraging preclinical outcomes, translation into clinical practice remains limited. The currently available human data suggest potential therapeutic benefits; however, the clinical evidence available is not very robust, being limited by the fact that the studies are small, vary significantly in design and scope, lack a standard venom preparation, and also lack adequate reporting on the outcomes and safety evaluations. Therefore, the use of HBV therapy must still be considered experimental.

Importantly, safety considerations remain a critical barrier to clinical adoption. Even if the majority of the adverse reactions are mild and dose-related, the possibility of hypersensitivity reactions, including the potentially dangerous condition of anaphylaxis, indicates the importance of meticulous patient screening, uniform dosing regimens, and better delivery methods. The use of nanocarriers for the delivery of melittin may provide the solution to improve the antiviral properties with lower cytotoxic and immunogenic effects.

In general, HBV is a biologically interesting and promising antiviral therapy with significant immunomodulatory effects. The results presented in this review emphasize and encourage their continued and intensified exploration, preferably in properly designed and sufficiently powered randomized clinical studies. Future research should prioritize safety optimization, pharmacokinetic profiling, and formulation refinement to determine whether HBV-based therapies can transition from experimental interventions to clinically viable antiviral agents in both human and veterinary medicine.