Earthworm belongs to class clitellata of phylum Annelida. Earthworms are a large source of proteins, peptides, enzymes, and the most active substances. Earthworms have several types of leukocytes which synthesize immune-protective molecules. The immune-protective system involves agglutination, opsonization, encapsulation, phagocytosis, clotting, and lysis of foreign substances. In this review paper, we are discussing the therapeutic values of earthworms. Eisenia fetida and Lumbricus rubellas have fibrinolytic enzymes which are potent and safe to use. Earthworm extracts have antioxidant, anti-cancer, sperm killing, wound healing, antibacterial, anti-inflammatory, nerve regeneration, etc. properties.

Earthworm (also known as earth dragon) has been used for many decades as fever reducer. Crude extract from different species of earthworms after performing fibrin assay revealed its protease property. Earthworm extracted protease performs a variety of functions including as an antithrombotic agent and minimizes platelets aggregation and shows high specific nature for substrates. Enzymes have been extracted from various species of earthworm and these enzymes are collectively called lumbrokinase. Lumbrokinase (LK) activates the plasminogen and dissolves the blood clot that entangles in blood vessels. The experimental and computational studies of lumbrokinase enzymes showed serine protease nature and the amino acid sequence of lumbrokinase is similar with the serine proteases. The domains of lumbrokinase revealed its serine protease nature that belongs to PA clan and trypsin family. To target thrombolysis different scientists introduced different delivery systems for the accurate aiming of lumbrokinase towards the target, LKs loaded small nano micelles have been targeted towards thrombus. Thus, through different analyses it was recorded that extracts of earthworm exhibit different intermediates. The protease provides good hemorheological efficacy and is significant for the treatment of thrombosis.

Earthworm fibrinolytic enzyme from Chinese earthworm Eisenia foetida was isolated to investigate its antitumor activity in breast cancer cells. The protein isolated was characterized as an earthworm fibrinolytic enzyme using the fibrin plate method. The molecular weight of the earthworm fibrinolytic enzyme component was determined to be 25 kDa by sodium dodecyl sulfate polyacrylamide gel electrophoresis assay using a standard protein ladder. Earthworm fibrinolytic enzyme can markedly inhibit the growth and migration of MCF-7 cells in a dose and time-dependent manner. In addition, MCF-7 cells treated with earthworm fibrinolytic enzyme (40 μg/ml) began to undergo apoptosis after 24 h. Expression of focal adhesion kinase and CD44v6 measured using reverse transcription polymerase chain reaction and Western blot was down-regulated in a concentration-dependent manner (20-80 μg/ml), resulting in the suppression of MCF-7 cells adhesion. The obtained earthworm fibrinolytic enzyme displayed an antitumor effect on MCF-7 cells in vitro, revealing the therapeutic potential of E. foetida.

Earthworms have long been used as traditional medicine. The purposes of this research were to create bioactive peptides from the unique Amynthas arenulus earthworm (PAAEs) and test their potentials on liver cancer bioprophylactic activity, antioxidant, oxidative stress protection, and immune cell activation. This earthworm had a high protein content ratio, at 55.39%. Besides, PM 35 is one out of 58 bacteria isolated from the earthworm carcasses that exhibited the highest protease and yield protein production which was chosen as the protease-producing bacteria to hydrolyze the protein. The genera were identified by 16S rRNA and 16S–23S rRNA comparison and confirmed as Bacillus velezensis PM 35. The response surface methodology was applied to optimize these hydrolysis parameters, i.e., the enzyme/substrate (E/S) concentration ratio [1%–3% (v/v)] and time (1–3 h) of the hydrolyzing earthworm’s proteins. The optimal hydrolyzing conditions were 3% (v/v) of E/S concentration ratio and 3 h of hydrolysis time, which found protein-hydrolysate yield (24.62%) and degree of hydrolysis (85.45%) as the highest. After being challenged in the gastrointestinal tract-resistant model, these PAAEs (MW <3 and 3–5 kDa) induced liver cancer cell (HepG2) death via apoptotic action modes (cell morphological change and DNA fragmentation). The PAAEs (MW <3 kDa) exhibited significant antioxidant activity via DPPH, ABTS, and FRAP with IC 50 values of 0.94, 0.44, and 6.34 mg/ml, respectively. The PAAEs (MW < 3 kDa) were non-cytotoxic and protected the mouse fibroblast cells (L929) against oxidative stress. These PAAEs (MW < 3 kDa, 0.2 mg/ml) stimulated the B lymphocytes (122.3%), and T lymphocytes (126.7%) proliferation. This research suggests that PAAEs can be used in a variety of applications, especially in the food and pharmaceutical industries.

Most of the radiosensitizers employed are not satisfying because of their untoward site reactions. An extract from an earthworm named earthworm capsule-912 was developed by the author. It has many virtues as follows: 1. Anti-cancer effect both in vivo and in vitro. In vivo, the investigation revealed inhibition on various transplanted tumors including S180, S794, H22, and EMT-6 in mice (BALB/c, LACA, and TA2). In vitro evidences demonstrated inhibition of HEp-2 cell. 2. Radioenhancement effects as evidenced by a ER of 1.54 on transplanted tumors of mice was found. 3. Chemoenhancement action was observed in combination with 5-Fu at the ER of 1.18. 4. Hyperthermic enhancement role was noted on HEp-2 cell and transplanted tumor (EMT-6) of mice.

Earthworm extracts have become an interesting research concept in cancer studies because they have various cancer-stopping and preventive substances. Determining the anticancer effect of the extract on cancer cells may lead to new treatment strategies and alternative medicine against the toxic effects of chemotherapeutic agents. Eisenia Foetida was determined as the most studied type of earthworm, while Colon cancer was the most frequently studied cancer type. Coleomic Fluid was found to be the commonly used extract to test for anticancer activity with positive results. The study contributes to the literature by providing information about earthworm extracts' antiproliferative and antitumor effects.

In conclusion, this study demonstrated that coelomic fluid of the earthworm D. veneta showed anticancer activity against A549 lung cancer cells but did not exhibit cytotoxicity toward the bronchial epithelial cell line BEAS-2B. It was demonstrated that the heated coelomic fluid caused the death of cancer cells via apoptosis. The coelomic fluid from earthworm Dendrobaena veneta for use in the treatment of lung cancer A549’ has been submitted for patent protection.

Conclusion The results of the present study contribute additional evidence for the antitumor activity of earthworm extracts. Therefore, further research should concentrate on isolating and identifying the earthworm’s active biomolecules that have antitumor activity by investigating the molecular mechanism, genetics, and pathways responsible for the antitumor activity of these biomolecules. Keywords Earthworms Breast cancer cells (MCF-7) Prostate cancer cells (PC-3) Antitumor.

Earthworm coelomic fluid has been acknowledged as a rich source of several bioactive substances that may exhibit promising anticancer activity. Therefore, the objective of the present review is to evaluate the findings of the reported studies exploring the antitumor effects of coelomic fluid in the context of its possible utilization as a natural therapeutic agent to cure different types of cancer. The possible mechanisms underlying the coelomic fluid’s anticancerous potential as well as the possibility for future development of cutting-edge therapeutic agents utilizing coelomic fluid-derived natural bioactive compounds to treat cancer disorders have been discussed along with future challenges. In addition, the feasibility of encapsulation of bioactive compounds derived from coelomic fluid with nanomaterials that could be further explored to attain more effective anticancer competence is discussed.

Research on pharmaceuticals derived from earthworms, known as green biomedicine, has been increasing. As a result, earthworms have become an international medicine, even though their original utilization in traditional medicine has been known for thousands of years. With the development of biomedicine, scientists have rediscovered the medicinal value of earthworms related to many chemical components, including (1) earthworm proteases (lumbrokinase, collagenase, superoxide dismutase, cholinesterase, catalases, glycosidases); (2) metal-binding protein (metallothionein, calmodulin-binding protein); (3) other active proteins including those with proliferative improving activity like lysenin, eiseniapore, antitumor proteins, and glycoprotein; (4) active peptides (gut mobility regulation peptide, antibacterial peptide); (5) earthworm metabolites (carbamidine, lumbrinin, lumbrofobrin, terrestrolumbrolysin); (6) special organic acids (succinic acid, lauric acid, and unsaturated fatty acid) and (7) other components such as purin, vitamin B, tyrosine and Se. In this paper, we mainly describe earthworm fibrinolytic enzymes and antibacterial peptides in particular.

Results revealed that the earthworm, Eisenia fetida extract exhibit anti-diabetic activity by inhibiting alpha-amylase activity and also by increasing the glucose uptake in the cells. Thus, earthworm Eisenia fetida extract may serve as an alternative anti-diabetic drug to prevent diabetic complications in near future.

The transfer of antibiotics and antibiotic resistance (AR) to the soil systems poses ecological hazards to various organisms, including earthworms. Apart from this toxicity, earthworms can also mitigate against antibiotics, antibiotic-resistant bacteria and antibiotic-resistance genes by reducing bacterial diversity and abundance. Overall, earthworms mitigate antibiotics and AR via various proximal and distal mechanisms, while dual but contradictory functions (i.e., mitigatory and facilitatory) were reported for AR. 

In recent times, naturally occurring substances such as earthworm extracts have been used successfully as an antimicrobial and anti-inflammatory agent in wound healing. It has also shown promising antitumor activity in cervical and gastric cancer.

This literature review examines the potential use of earthworm extract (Pheretima javanica K) as an alternative treatment for typhus disease. The study analyzes multiple research findings regarding the antibacterial properties of earthworm extracts against Salmonella typhi, the causative agent of typhus. Through a comprehensive review of studies from Web of Science, PubMed, and Google Scholar databases, this research evaluates the effectiveness of different earthworm species and extraction methods. The findings indicate that Pheretima javanica shows promising antimicrobial properties, particularly when extracted with ethanol, while other species such as Lumbricus rubellus demonstrate limited effectiveness. Pheretima javanica shows promise, other species like Lumbricus rubellus demonstrate limited or no significant antibacterial activity against Salmonella typhi.

Introduction Some studies have found that probiotics have the potential to treat PD, and earthworm protein is a traditional Chinese medicine used for the treatment of PD. The purpose of this study was to evaluate the safety and efficacy of Bacteroides fragilis 839 (BF839) + earthworm protein supplement as an adjunctive therapy for PD and to observe changes in the gut microbiota. Methods Forty-six patients with PD were recruited for a 12-week 1:1 randomized, double-blind, placebo-controlled clinical trial to evaluate changes in motor and some non-motor symptom scores and detect metagenomic changes in the gut microbiota. Results From baseline to 12 weeks, compared with placebo, the trial group showed significant reductions in the United Parkinson’s Disease Rate Scale (UPDRS) total score (−7.74 ± 5.92 vs. –1.83 ± 4.14, p < 0.001), UPDRS part I (−0.72 ± 0.81 vs. –0.20 ± 0.72, p = 0.026), UPDRS part II (−2.50 ± 2.24 vs. –0.22 ± 1.98, p = 0.001), UPDRS part III (−3.43 ± 3.42 vs. –1.33 ± 2.65, p = 0.024), and UPDRS part IV (−1.13 ± 1.19 vs. –0.15 ± 0.57, p = 0.001). Significant reductions in the Hamilton Depression Scale-24 score (−3.91 ± 3.99 vs. +1.15 ± 3.42, p < 0.001), Self-Rating Anxiety Scale scores (−7.04 ± 5.71 vs. –1.23 ± 2.34, p < 0.001), and Constipation scoring system scores (−8.59 ± 4.75 vs. 0.27 ± 1.24, p < 0.001), were also noted. In the trial group, one patient experienced mild eczema and one suffered low blood pressure, which could not be conclusively attributed to supplementation. Compared to the placebo group, the trial group showed a marked increase in Enterococcus faecium and a decrease in Klebsiella. Conclusion This study is the first to report that probiotics plus earthworm protein can remarkably improve the motor and some non-motor symptoms of PD without serious adverse effects. However, further clinical trials and exploration of the underlying mechanisms are required. Clinical trial registration Clinical trial registry http://www.chictr.org.cn/, Identification No: ChiCTR2000035122.

Diabetic chronic wound is a worldwide medical burden related to overdosed methylglyoxal (MGO) synthesis, which is the major precursor of glycation of proteins and DNA and is related to the dysfunction of dermal cells thus leading to chronic refractory wounds. Previous studies proved that earthworm extract accelerates diabetic wound healing and possesses cell proliferation and antioxidative effects. However, the effects of earthworm extract on MGO-damaged fibroblasts, the inner mechanisms of MGO-induced cell damage and the functional components in earthworm extract are still poorly understood. Firstly, we evaluated the bioactivities of the earthworm extract PvE-3 on the diabetic wound model and the diabetic related cell damage model. Then the mechanisms were investigated through transcriptomics, flow cytometry and fluorescence probe. The results revealed that PvE-3 promoted diabetic wound healing and protected fibroblast function in cell-damaged conditions. Meanwhile, the high-throughput screening implied the inner mechanisms of diabetic wound healing and PvE-3 cytoprotection effect were involved in the muscle cell function, the cell cycle regulation and the mitochondrial transmembrane potential depolarization. The functional glycoprotein isolated from PvE-3 possessed EGF-like domain which had a strong binding affinity with EGFR. The findings provided references to explore the potential treatments of diabetic wound healing.

The purpose of this study was optimal hydrolysis of Amynthas arenulus earthworm powder with protease-producing bacteria isolated from a soaked dried earthworm. The bacteria were isolated as 10 isolates. The bacteria were screened to find protease production and examined protein contents of each isolate. The A. arenulus earthworm powder hydrolysis was performed with pH 5, temperature 37 °C for 0-60 min. After that, pH 7.0, temperature 37 °C and hydrolysis period from 0-60 min were carried out. The optimal temperature was performed using pH 7.0, temperature from 30-80 °C and hydrolysis period from 0-60 min. The hydrolyzed earthworm examined the protein contents using the Lowry method. The results were found that 10 bacteria were isolated from soaked dried earthworm for 24 h, which can be protease-producing bacteria. The EDNP 9 isolate produced protein contents the highest as 824.14 µg/mL. The EDNP 9 isolate were protease activity at 0.63 U/mL. Meanwhile, hydrolyzed earthworm with pH 7.0, at 37 °C was found hydrolysis period of 60 min, which presented 394.83 µg/mL of protein contents (P < 0.05). Moreover, the optimal temperature that pH of 7.0, a temperature of 30-80 °C for 0-60 min was found the highest protein content (725.29 µg/mL) (P > 0.05). Therefore, the optimal hydrolysis of the A. arenulus earthworm powder was pH 7.0, temperature 60 °C and hydrolysis period 30 min. The earthworm that was hydrolyzed was applied in medical applications further.

Earthworm, Eudrilus eugeniae cannot survive and regenerate without clitellum segments. In regenerating worms, the clitellum’s epithelial and circular muscular layers are reduced to one-third, and longitudinal cell layers to half. In C2C12 cells, Clitellum Factors (CF − 5, 25 and 50%) and Regenerative Clitellum Factors (RCF − 5, 25, 50, 75%) ameliorate the cell viability up to 20–28% and 30–38% respectively than the control. In contrast, extracts from body segments negatively influence cell viability up to 80%. In a scratch-wound assay, 25% RCF and 5% CF achieved 99.86% and 81.54% wound closure in 24 h, respectively, compared to 40% in controls. RCF and CF also possess enhanced anti-microbial activity against gram + ve bacteria. Western Blotting reveals that Wnt3a, HoxD3 and VEGF were remarkably upregulated in RCF and CF treated samples and their upregulated stemness property is effectively regulated by p53, TCTP, H2AX, Cleaved Caspase-3 proteins. Immunofluorescence data clearly states that Wnt3a and Caspase-3 signals are more profoundly observed in nuclear over cytoplasm in RCF treated samples and H2AX shows less nuclear signals than CF. In in-vivo earthworm model conditions, RCF remarkably promotes the survivability and wound healing ability by promoting the Wnt3a and VEGF expression together with downregulation of Cox2.

Although vertebrates are indispensable to biomedical research, studies are often limited by factors such as cost, lengthy internal review, and ethical considerations. We present the earthworm as an alternative, low-cost, invertebrate applicable to certain preliminary vasculature studies. Due to the surgical availability of the earthworm's dorsal vessels, ventral vessels, and five pairs of pseudo hearts, earthworms are readily accessible, offer low-cost maintenance, and require administration of only small doses of a given compound. The earthworm model provides a simple closed vascular circulatory system with a hemoglobin structure similar to human blood. A protocol is provided for anaesthetizing the earthworms and performing surgical incisions to expose relevant blood vessels. Micropipettes for compound administration are formed by heating and pulling glass with a pipette puller and using a beveling system to create a micron-scale fine needle tip. The tips are then used with a micropositioner and microinjector to inject arbitrary compounds into the vascular system of an earthworm, repeatably, with the availability of large sample sizes and small compound volumes. Details on the intricacies of injection procedure are provided. The small vessel size of the earthworm is challenging, particularly in the case of the ventral vessel; however, mastery of the techniques presented offers high repeatability as a low-cost solution, making studies of very large sample size practical.

The medical value of earthworms has been known for centuries. This is evident from the history of the ancient Southeastern Asian medicine (China, Japan, Vietnam). The earthworms are the source of proteins, peptides, enzymes and physiologically active substances. Thus, the extracts prepared from the earthworm tissue have been used for the treatment of numerous diseases. Earthworms, like other complex invertebrates, possess several types of leukocytes which synthesize and secrete a variety of immunoprotective molecules. The immunoprotective system is involved in phagocytosis, encapsulation, agglutination, opsonization, clotting and lysis of foreign components. The lytic reactions against several targets are mediated by two major leukocytes, small and large coelomocytes. In the last 10 years a number of earthworms' clot-dissolving, lytic and immune-boosting compounds have been isolated and tested in laboratory and clinical studies. In particular, research has been focused on clot-dissolving molecules. Fibrinolytic enzymes, which are regarded as potent and safe, have been purified and studied from several species of earthworms, including Lumbricus rubellas and Eisenia fetida. Its therapeutic and preventive effects on thrombosis-related disease have been clinically confirmed. However, several studies have shown that earthworm extracts contain different macromolecules, which exhibit a variety of activities, such as antioxidative, antibacterial, antiinflammatory, antitumor, etc. Some of these activities are involved in wound healing using an earthworm preparation.

Abstract: As a traditional Chinese medicine (TCM), earthworm (Dilong) was applied in China for ages. Some active proteins and compounds in earthworm were characteristic composition as a pharmaceutical, including fibrinolytic enzymes, lumbrokinase, collagenase, superoxide dismutase, cholinesterase, catalases, glycosidases, metallothionein, calmodulin-binding protein, proteins with proliferation improving activity, lysenin, eiseniapore, antitumor protein, glycoprotein extract, gut mobility regulation peptide, antibacterial peptide, carbamidine, lumbritin, lumbrofobrim, terrestrolumbrolysin, purin, vitamin B, tyrosine, succinic acid, lauric acid, unsaturated fatty acid and so on. 

G-90 is a glycolipoprotein mixture obtained from the tissue homogenate of earthworm Eisenia foetida

Earthworm plays a major role in the proper functioning of the soil ecosystem. It acts as scavenger and

Earthworms have provided ancient cultures with food and sources of medicinal cures. Ayurveda, traditional Chinese medicine