Bee Venom

Bee venom, also known as apitoxin, is a biologically active substance produced in the venom glands located in the abdomen of honeybees (Apis mellifera) and stored in the venom sac. This substance is one of the key defense and attack mechanisms of bees.

Production and Secretion

In worker bees, the egg-laying organ (ovipositor) has undergone structural modification to become a stinger. Both queen and worker bees possess the ability to sting, while male bees lack a stinger and therefore cannot sting. The venom is produced in the bee’s acid and alkaline secretion glands and stored in the venom sac. When a bee stings, it injects this secretion into the target organism through its stinger. The venom glands become active when a worker bee transitions from the pupal stage to the adult stage and can begin secreting venom within approximately three days. Venom quantity peaks during spring and summer months. An average worker bee contains about 3–4 microliters (µl) of venom, equivalent to approximately 0.3 mg. The amount of dry venom obtainable from a single bee is 0.1 microgram (µg) or less.

Physical Properties

Bee venom is normally a liquid, pale yellow in color, with a sharp odor and bitter taste. It is aromatic due to the presence of alarm pheromones. It has a clear, acidic structure with a pH between 5.0 and 5.5. Upon exposure to air, it dries and crystallizes within about 20 minutes, losing 65–70% of its weight during this process. Approximately 88% of bee venom is water. Dry venom is light yellow in color. The brownish hue of some commercial preparations is due to oxidation of venom proteins.

Chemical Composition

Bee venom has a highly complex chemical structure, with its main components being various enzymes, proteins, and peptides. Some of these components exhibit anti-inflammatory and analgesic effects, while others are toxic. The venom contains major constituents including melittin, apamin, MCD-peptide (Mast Cell Degranulation Peptide), histamine, hyaluronidase, and phospholipase-A2. The primary components by dry weight and their approximate proportions are as follows:

Peptides

• Melittin: 40–50% (also referred to as Melittin F and its derivatives)
• Apamin: 2–3% or 1–3%
• MCD-peptide: 1–2%
• Secapin: 0.5–2%
• Adolapin: 1%
• Tertiapin: 0.1%
• Protease inhibitors: <0.8% or 0.8%
• Prokamin A and B: 1.4% or 1–2%
• Minimin and Cardiopeptin: 13–15% (also classified as other small peptides (<5 amino acids))

Enzymes

• Phospholipase A2: 10–12%
• Hyaluronidase: 1–3% or 1–2% (listed as 1–3% in table but mentioned as 1–2% in text)
• Acid phosphomonoesterase: 1%
• α-Glucosidase: 0.6%
• Lysophospholipase: 1%

Active Amines (Biogenic Amines)

• Histamine: 0.6–1.6% or 0.5–2.0%
• Dopamine: 0.13–1% or 0.2–1.0%
• Norepinephrine (Noradrenaline): 0.1–0.7%

Amino Acids

• α-Amino acids: 1%
• Aminobutyric acid: <0.5% or 0.5%

One study reported the presence of 18 distinct bioactive molecules in bee venom.

Effects of Major Components

• Melittin (50–55%): The primary component of bee venom. It reduces surface tension of cell membranes and stabilizes them. At low doses, it has anti-inflammatory effects; it stimulates smooth muscle, increases capillary permeability and blood circulation, lowers blood pressure, and reduces blood clotting. It exhibits both immunostimulatory and immunosuppressive properties and has radioprotective effects. It affects the central nervous system (CNS); it has anticancer, antibacterial, antifungal, and antiviral properties. At high doses, it is inflammatory and hemolytic. It is a small, linear, basic peptide with strong hemolytic activity. It consists of a 26-amino acid sequence with the chemical formula C₁₃₁H₂₂₈N₃₈O₃₂ and a molecular weight of 2847.5 Da.

• Phospholipase A2 (PLA2) (10–12%): Breaks down phospholipids and lyses cell membranes of blood cells. It reduces blood clotting and blood pressure. It is one of the most lethal peptides in bee venom and constitutes 12–15% of the dry weight of venom. It triggers inflammation, is a potent allergen, and is the most harmful component of bee venom. Melittin enhances the effects of PLA2.

• Apamin (2–3%): Has anti-inflammatory effects by triggering cortisol release and exhibits anti-serotonin activity. It enhances defense capacity and shows immunosuppressive effects. At low doses, it stimulates the CNS; at high doses, it is neurotoxic.

• MCD-peptide (1–2%): A peptide that degranulates mast cells, thereby increasing the release of histamine, heparin, and serotonin. It increases capillary permeability, enhances anti-inflammatory effects, and stimulates the CNS. It exhibits immunosuppressive and nociceptive (pain-sensitizing) effects.

• Hyaluronidase (1–2%): Catalyzes the hydrolysis of hyaluronic acid, the fundamental structural component of tissues. By catalyzing protein hydrolysis, it facilitates the penetration of venom into tissues. It dilates blood vessels, increases permeability, and enhances blood circulation. It has allergenic properties.

• Adolapin (1%): Inhibits specific brain enzymes such as cyclooxygenase and lipoxygenase, reducing inflammation. It has anti-rheumatic effects, alleviates pain, and exhibits antipyretic (fever-reducing) activity. It prevents erythrocyte aggregation and has low toxicity.

• Protease Inhibitors (3–4%): Reduce inflammation and exhibit anti-rheumatic effects by inhibiting the activity of various proteases such as trypsin, chymotrypsin, plasmin, and thrombin. They have low toxicity.

• Histamine (0.7–1.5%): A neurotransmitter that dilates blood vessels, increases capillary permeability and blood flow. It stimulates smooth muscle and has allergenic properties.

• Dopamine and Noradrenaline (0.2–1.5%): Neurotransmitter substances. They have no physiological effect in mammals at low concentrations but are active in invertebrates.

• Alarm Pheromones (4–8%): Complex ethers that trigger alarm responses in bee colonies and activate defensive behaviors.

Bee venom affects the immune system, central and peripheral nervous systems, and the cardiovascular system. It also exhibits numerous biological effects including antibacterial, antifungal, antiviral, anti-inflammatory, antiarthritis, anticancer, and wound-healing properties.

Production of Bee Venom

In traditional medicine, venom was collected either by surgically removing the venom gland or by squeezing the bee until it emptied its venom. A modern method involves applying an electric shock to the bees. In this method, a wire grid is placed inside the hive and subjected to periodic electric pulses (typically every 30 minutes). The bees perceive the electric current as a threat and, upon contacting the grid, sting it and inject their venom onto a permeable surface beneath the grid (usually a sterile cloth). The venom collected by this method, when dried, forms a white powder.

After a 30-minute session, only about 1 gram of dry venom can be collected from approximately 10,000 bees in a hive. Another method involves drying the entire bee, but this can contaminate the product with pollen, feces, dust, nectar, and honey. Bee venom can be stored without degradation for up to five years as long as it is protected from moisture and humidity; the optimal storage method is freezing in a deep freezer.

Analytical Methods

Various chromatographic methods have been developed for the characterization of bee venom, including capillary electrophoresis (CE), capillary zone electrophoresis-diode array detector (CZE-DAD), high-performance capillary electrophoresis (HPCE), thin-layer chromatography (TLC), ultra-performance liquid chromatography (UPLC), high-performance liquid chromatography (HPLC), HPLC-DAD-MS/MS, and MALDI-TOF. SDS-PAGE electrophoresis has also been investigated for characterization purposes.

Applications and Apitherapy

Apitherapy refers to the therapeutic use of bee products, including venom. Its origins date back 6,000 years to ancient Egypt. The Romans and Greeks also used bee products for medical purposes. The first publications on the therapeutic use of bee venom began in 1864. Today, apitherapy centers are widespread. In medical literature, bee venom therapy is referred to as Bee Venom Therapy (BVT) (via intradermal or subcutaneous injections) or Bee Sting Therapy (BST) (via live bee stings).

In traditional medicine, bee venom has been used to treat arthritis, rheumatism, pain, tumors, and skin diseases. Its use in modern medicine is also under investigation. Major applications and researched effects include:

• Rheumatic and Joint Diseases: Used in conditions such as osteoarthritis and rheumatoid arthritis. In one study, 544 out of 660 patients with osteoarthritis showed complete recovery. Improvements have also been observed in patients with rheumatoid arthritis.

• Nervous System Disorders: Investigated for use in multiple sclerosis (MS), back pain, sciatica, polyneuritis, facial nerve inflammation, trigeminal neuralgia, Parkinson’s disease, and post-stroke paralysis. In a study by Ludyanski on MS, 175 out of 210 patients (83%) showed improvement. Bee venom pharmacopuncture has been used to control symptoms of chemotherapy-induced peripheral neuropathy.

• Analgesic and Anti-inflammatory: Used as an anti-inflammatory and pain-relieving agent. The component adolapin alleviates pain.

• Cancer: Investigated for effects such as apoptosis induction, necrosis, cytotoxicity, and inhibition of cancer cell proliferation in various cancers (ovarian, hepatoma, prostate, bladder, melanoma, kidney, breast, lung, and liver). Melittin, and therefore bee venom, is considered a potential animal-derived drug source for cancer therapy. Bee venom has been reported to counteract the HIV virus’s ability to compromise the immune system and to strengthen the patient’s immune response, slowing disease progression and halting its development.

• Skin Diseases: Reported use in eczema, dermatitis, psoriasis, scar tissue healing, alopecia, acne, cutaneous tuberculosis, and skin hardening in the elderly. Positive effects of bee venom serum on facial wrinkles were observed from the eighth week onward. It has been suggested to be beneficial in preventing inflammatory skin conditions caused by Propionibacterium acnes.

• Cardiovascular System: Investigated for use in hypertension, arteriosclerosis, and arrhythmia.

• Other Applications: Reported use in eye diseases, gastroenterology (colitis, ulcers), pulmonary diseases (asthma, bronchitis), ENT disorders (pharyngitis, tonsillitis), urology, endocrinology, gynecology, migraine, epilepsy, cholesterol, sinusitis, and chronic fatigue syndrome. Effects against Lyme disease (melittin inhibits Borrelia burgdorferi) and as an anti-diabetic agent (lowers blood glucose and increases insulin secretion) have been studied.

• cosmetics Industry: Bee venom, available in forms such as creams, ointments, and solutions, is used especially in products targeting facial wrinkles.

• Veterinary Medicine: Studies have been conducted on its use in cats, dogs, and horses, with successful results reported. It has been shown to reduce somatic cell counts in mastitis infections in dairy cows and accelerate wound healing in rats. It has been used in spray formulations as an immunoprophylactic agent against Salmonella gallinarum in broiler chicks.

Bee venom therapy is practiced in many countries worldwide, including the United States, China, Korea, Russia, Bulgaria, Japan, Hungary, the Czech Republic, Slovakia, Romania, Poland, Germany, Austria, Switzerland, and France. In Europe, 22 different products containing bee venom are used in nine countries.

Anticancer Properties of Melittin Peptide

Melittin (MEL), the main component of bee venom (BV), constitutes approximately 40–50% of its dry weight. MEL has demonstrated anticancer properties in various cell culture and animal model studies, including cytotoxicity, hemolytic activity, and growth inhibition. The antitumor effect of BV is largely attributed to MEL. Early studies showed that MEL inhibits the growth of human leukemia cells and astrocytoma cells by acting as a calmodulin inhibitor. Leukemia cells were found to be more sensitive to MEL than normal mouse spleen and bone marrow cells. This may be due to the high number of carbohydrate-binding sites on the membranes of bone marrow cells, a reduction in these sites in adult spleen cells, and their near-complete disappearance in neoplastic cells, making tumor cells more vulnerable.

MEL is particularly effective against cells expressing high levels of ras oncogenes, selectively eliminating them by enhancing PLA2 activation. MEL’s cytotoxicity is linked to both necrotic and apoptotic cell death. Although MEL’s potential as an anticancer agent is well recognized, its nonspecific cytolytic activity and rapid degradation in the bloodstream pose challenges for human use. To overcome these issues, optimization studies are underway using appropriate delivery systems such as nanoparticles. Recombinant viruses carrying the MEL gene have been reported to exhibit inhibitory effects on hepatocellular carcinoma.

Precautions and Contraindications

Before initiating bee venom therapy, an allergy test must be performed, and treatment must be administered under the supervision of a qualified physician. Individuals with tuberculosis, cold abscesses, endocarditis, and pregnant women should not use bee venom. The dosage, route of administration, and duration of treatment in apitherapy vary depending on the individual. The average lethal dose (LD50) for adults is estimated at 2.8 mg/kg, equivalent to approximately 560 bee stings (assuming each sting delivers 0.3 mg of venom).