Leech Saliva Research Articles

Effect of leech therapy in dermatophytosis: A randomized standard controlled trial

Background and purposeIn Unani medicine leech therapy is used as analgesic and resolvent purposes. The saliva of leech contains various biological substances like Hirudin, hyaluronidase, vasodilators, anesthetics, antibacterial, fibrinases, collagenase etc. To compare the efficacy of Leech therapy with local antifungal clotrimazole (1 %) cream in dermatophytosis on long term basis. Material and methodsA randomized, standard controlled, open labeled, clinical study was conducted on 40 patients of dermatophytosis, aged 18–60 years, and were randomly assigned into two groups; 20 patients in test group and 20 in control group. Patients were diagnosed clinically and confirmed through KOH examination. Complete haemogram, bleeding and clotting times, random blood sugar, HIV I & II, and HBsAg tests were carried out before starting the therapy, while haemogram was repeated after completion of treatment too. In test group, five sittings of leech therapy were applied, with intervals of one week. The control group was given clotrimazole (1 %) cream for topical application twice daily, continuously for a period of 28 days. Post-treatment follow-ups were done on 35th day in person, and then over the phone on 45th, 60th, and 90th days. Data were collected using Signs and Symptoms Severity Score for study, and Visual Analogue Scale (VAS). After completion of trial, appropriate tests were applied for statistical analysis of data. ResultsThe study shows, in most of the parameters, statically significant therapeutic effect in both the groups (p<0.001). No patients of either group reported any adverse event throughout the trial. Conclusionit appeared that leech therapy can contribute significantly in treating dermatophytosis (Qūbā) without any adverse effect, and with prolonged efficacy compared to standard treatment.

Enhancing Quality of Life in Venous Ulcer Patient with Irsal-e-Alaq (Hirudotherapy): A Case Report

Background: In general practice, venous ulcers (stasis ulcers) are the most common and recurrent leg ulcers. Ignorance of it can have a severe detrimental effect on one's quality of life, cause gangrene, and even the limb amputation. Venous ulcers are the most dangerous and disabling side effect of chronic venous insufficiency. Leech therapy seems to be effective for the treatment of varicose ulcers due to its salivary secretions which it injects during bloodsucking. The saliva of leeches contains certain biochemical with vaso-dilating, anticoagulant, anesthetic, thrombolytic, antibiotic, analgesic, and anti-inflammatory properties. As a result, Irsal-e-Alaq (Hirudotherapy), one of the Unani regimens, can effectively control it. Objective: The purpose of this case report is to provide insight into the effects of Irsal-e-Alaq (Hirudotherapy) on venous ulcers. Intervention: A 60-year-old male has a complaint of ulceration in his left lower leg near the medial malleolus with swelling, pain, discharge, and darkening of the skin of the left lower limb. Four leeches were applied once every week for 2 months. Efficacy of leech therapy was assessed on every 15th day by pain /leg discomfort on 4 point scale ranging from 0-3, limb girth at calf, ankle &amp; foot, pigmentation color on 4 point scale, pigmentation area, pain in ulcer on 4 point scale from 0-3, ulcer depth, PUSH Score. Hb% was assessed every 15th day to check anemia, colour flow Doppler was done to assess any change in vein anatomy. Result: In this case report, the ulcers were completely healed, and also showed remarkable improvement in other variables. Conclusion: Irsal-e-Alaq has been proven beneficial in treating venous ulcers and enhancing patient’s quality of life. Keywords: Venous ulcer, Hirudotherapy, Unani Medicine, Pain, Pigmentation.

Leeches and Their Use in Medicine (Annellida: Hirudinea: Rhynchobdelliformes)

Leech therapy relieves pain and inflammation, and there are explanations for its success. In leech saliva, in addition to substances that prevent blood clotting, there are also analgesic and anesthetic compounds that reduce pain. This article aims to study the characteristics of the leech (Annellida: Hirudinea: Rhynchobdelliformes) and its use in Medicine. This is a narrative review of the literature, which is indicated to explain and discuss a certain subject from a theoretical or contextual perspective, to allow the reader to ask questions and update knowledge on a specific topic. The search for scientific articles that made up this review was carried out on Google Scholar, Biological Abstract, HAL, Qeios, ResearchGate, Scielo, and SSRN. The following descriptors in Health Sciences (DeCS) were used: biological therapy, larva, wound, debridement, and healing. The following inclusion criteria were considered: original articles and reviews, published nationally and internationally in full, available electronically, and published in Portuguese, English, and Spanish. The exclusion criteria were dissertations, theses, monographs, and conclusion work, as well as duplicates and those that require payment to access the content during integration.

Cytotoxic effect of silver nanoparticles biosynthesized from Hirudo medicinalis saliva on HepG2 cells

Primary hepatocellular carcinoma is a devastating type of liver cancer. Silver nanoparticles (AgNPs) have been assessed for a variety of purposes, including being tested as an anticancer agent. The aim of this study was to assess the cytotoxicity of AgNPs that were biosynthesized from leech saliva on HepG2 cells, through the undertaking of a simple MTT assay. HepG2 cells were obtained from the cell bank of the Pasteur Institute of Iran. In this study, AgNP-treated HepG2 cells were cultured at a density of 104 cells per well, and 100 μL of MTT at a concentration of 0.5 mg/mL were added to each well; the treated cells were then let to incubate for 4 h. Subsequently, a plate reader device operating at a wavelength of 570 nm was used in order to determine the concentration of the chemical dissolved in isopropanol. Representative images of the cells show remarkable changes in their morphology at AgNP concentrations of 25 and 50 μg/mL. At 48 h, the nanoparticle’s IC50 value was 50 μg/mL. Our study shows that leech salivary extract-derived AgNPs are cytotoxic to HepG2 cells.

The Therapeutic Action of the Leech Hirudo medicinalis

The article under discussion examines the mechanism of the therapeutic effect of the medicinal leech Hirudo medicinalis. Today, studying the composition of the saliva of medicinal leeches is of the greatest relevance. The secretion of leech saliva contains a whole complex of biologically active substances — it contains more than a hundred medicinal components, enzymes, hormones, vitamins and amino acids, as well as a special substance hirudin, which makes the blood more fluid, which prevents the formation of unnecessary platelets. This is an indispensable way to prevent heart attacks, strokes and thrombosis. The author of the article believes that proof of the medicinal properties of specific components of saliva can lead to the development of new drugs or therapeutic methods.

IMPLICATION OF LEECH THERAPY AND ASBAB-E-SITTA ZAROORIYAH IN THE PREVENTION AND TREATMENT OF VARICOSE VEINS: A COMPREHENSIVE REVIEW

Varicose veins are characterized by abnormally twisted, dilated superficial vessels that are caused by the inefficient or defective valves within the vein. It is currently a prevalent disease with the incidence varying globally between 10% and 60%. The Unani Medicine (also called Greco-Arab medicine) is the ancient system of medicine practiced all around the world. According to the Unani classical text, Leech therapy (Irsal-e-Alaq) is one of the most important regimenal therapy for the successful treatment and management of varicose vein and a unique combination of Asbab-e-Sitta Zarooriyah (Six essential factors) is involved in the prevention of this disease. The research studies have proved that medicinal leech therapy displays anticoagulant, antimicrobial, anti-inflammatory and analgesic effects. Leech saliva contains medicinally useful bioactive molecules like hirudin, histamine, etc. that shows anti-coagulant effect and hence improve microcirculation in diseases like varicose vein. Leech therapy is reliable, well tolerated, cost effective treatment that has the potential of preventing varicose vein complications. This review seeks to describe medicinal leech therapy, present therapeutic leech species, and highlight the processes by which leech saliva affects wound healing. Additionally, the review paper highlights the role of Asbab-e-Sitta Zarooriyah in prevention and control of varicose veins. Irregularity in diet and lifestyle may cause disturbance in normal body functioning, which may lead to lifestyle disorder that is a major risk factor thought to influence the susceptibility of varicose vein. The Unani System exerts a major influence on preventing the development of varicose vein by making modification in Asbab-e-Sitta Zarooriyah.

Treatment of Cutaneous Leishmaniasis lesion caused by Leishmania major Using Leech Therapy in BALB/c Mice.

Amphotericin B, Allopurinol, Glucantime and Beta-Glucan are the main drugs currently used for the treatment of cutaneous leishmaniasis. In present study, we evaluated the effect of medical leeches on BALB/c mice models infected with Leishmania parasites. To create animal models of leishmaniasis, BALB/c mice were inoculated with the standard Leishmania major strain MRHO / IR / 75 / ER. Treatment was performed through blood sampling with leeches. We used medical leech (Hirudo medicinalis) species. This leech is a European medicinal leech, one of several species known used as medicinal leech. 22 mice infected with Leishmania major were used for the tests .The studied mice were divided into two groups which were named with the letters L and D. The duration of blood sampling was 25 min for initial treatment, 20 min for the 2nd blood sampling and 15 min for the third times and onwards. Meglumine antimoniate (Glucantime) is a pentavalent antimony (SbV) that recommended by the World Health Organization as a golden standard treatment for all kind of leishmaniasis, but many side effects are still reported. Criteria for anti-leshmania assessment of Leech were including: Leech saliva has an anti-leishmania effect by acting on the purin pathway. Leech saliva prevents the growth and multiplication of Leishmania by a mechanism of action similar to Glucantim (breaking ATP). The duration of leech treatment in BALB/c mice infected with L.major parasites was about 6-8 weeks. Out of the 22 leishmaniasis mice models that underwent treatment, 13 survived until the end of the treatment duration. The recovery rate was about 56%; lesions improved in five out of the 13 surviving mice models. Paired t-test showed a significant difference between the mean wound size at the beginning of treatment and the 8th week of treatment (PV <0.0001). Considering the fact that the standard drugs for the treatment of leishmaniasis have many side effects, alternative therapies such as leech therapy can be effectively used to treat these chronic wounds. There are about 20 active substances in leech saliva. These substances include antistasin, eglins, guamerin, hirudin, saratin, bdellins, complement, and carboxypeptidase inhibitors. Leech saliva is an anticoagulant, anti-inflammation and prevents the platelet. Leech therapy was effective in treating leishmaniasis wounds in BALB/c mice. In this study, there was complete wound healing in 56% of the mice models.

New anticoagulant protein from medicinal leech

The saliva of the medicinal leech contains various anticoagulants. Some of them, such as hirudin, are well known. However, it is reasonable to believe that not all anticoagulant proteins from medicinal leech saliva have been identified. We previously performed a comprehensive study of the transcriptome, genome, and proteome of leech salivary gland cells, which led to the discovery of several previously unknown hypothetical proteins that may have anticoagulant properties. Subsequently, we obtained a series of recombinant proteins and investigated their impact on coagulation in in vitro assays. We identified a previously undescribed protein that exhibited a high ability to suppress coagulation. The His-tagged recombinant protein was expressed in Escherichia coli and purified using metal chelate chromatography. To determine its activity, commonly used coagulation methods were used: activated partial thromboplastin time, prothrombin time, and thrombin inhibition clotting assay. Clotting and chromogenic assays for factor Xa inhibition were performed to evaluate anti-Xa activity. We used recombinant hirudin as a control anticoagulant protein in all experiments. The new protein showed significantly greater inhibition of coagulation than hirudin at the same molar concentrations in the activated partial thrombin time assay. However, hirudin demonstrated better results in the direct thrombin inhibition test, although the tested protein also exhibited the ability to inhibit thrombin. The chromogenic analysis of factor Xa inhibition revealed no activity, whereas the clotting test for factor Xa showed the opposite result. Thus, a new powerful anticoagulant protein has been discovered in the medicinal leech. This protein is homologous to antistatin, with 28 % identical amino acid residues. The recombinant protein was expressed in E. coli. This protein is capable of directly inhibiting thrombin, and based on indirect evidence, other proteases of the blood coagulation cascade have been identified.

Transcriptomic profiling and the first spatial expression analysis of candidate genes in the salivary gland of the East Asian medicinal leech, Hirudo nipponia

Hirudo nipponia, a blood-sucking leech native to East Asia, possesses a rich repertoire of active ingredients in its saliva, showcasing significant medical potential due to its anticoagulant, anti-inflammatory, and antibacterial effects against human diseases. Despite previous studies on the transcriptomic and proteomic characteristics of leech saliva, which have identified medicinal compounds, our knowledge of tissue-specific transcriptomes and their spatial expression patterns remains incomplete. In this study, we conducted an extensive transcriptomic profiling of the salivary gland tissue in H. nipponia based on de novo assemblies of tissue-specific transcriptomes from the salivary gland, teeth, and general head region. Through gene ontology (GO) analysis and hierarchical clustering, we discovered a novel set of anti-coagulant factors—i.e., Hni-Antistasin, Hni-Ghilanten, Hni-Bdellin, Hni-Hirudin—as well as a previously unrecognized immune-related gene, Hni-GLIPR1 and uncharacterized salivary gland specific transcripts. By employing in situ hybridization, we provided the first visualization of gene expression sites within the salivary gland of H. nipponia. Our findings expand on our understanding of transcripts specifically expressed in the salivary gland of blood-sucking leeches, offering valuable resources for the exploration of previously unidentified substances with medicinal applications.

The Actions of Leech Saliva Components and Their Mechanisms in Antitumor Activity

Malignant tumor (hereinafter referred to as tumor) has become a major category of chronic diseases that curb human life and seriously threaten human health.For advanced tumors, chemotherapy, radiotherapy, targetedtherapy and immunotherapy effects are still not satisfactory, and they are particularly urgent to find new therapeutic drugs.The medical leech therapy(MLT) can be traced back to the period of ancient Greece and ancient India for disease treatment.The discovery and appraisal of the leech saliva of hirudin aroused the research interest on the saliva of the lequale.Yet of the more than 100 compounds of different molecular weights detected in leech saliva, only a small fraction have been identified as having therapeutic potential.Studies have shown that the saliva activity ingredients have certain advantages in anti-tumor metastasis, which has attracted the attention of scholars.This article reviews the role of the active ingredients of the saliva and its anti-tumor mechanism in recent years, and provides a reference for the application of the leech saliva.

Ayurvedic Management of Shwitra : The Case Study

Kushtha is one among the Ashtamahagad explained in Ayurveda as there is structural changes in appearance of skin[i]. In Ayurveda Vitiligo is known as "Shwitra”. It is one of the varieties of Kushta in Ayurvedic classics, caused due to vitiation of Tridosha and Dhatus like Rasa, Rakta, Mamsa and Medas. Unbalanced diet (Virrudhahara) is also an important cause. It signifies production of toxic metabolites as an important cause for the development of the disease. Vitiligo is a specific type of a acquired leukoderma and this is idiopathic, patterned, circumscribed hypomelanosis of the skin and hair in which other cause of leukoderma have been excluded. The disease is important principally because of the social stigma associated with vitiligo. Diet restriction and psychological counselling was also a necessary part of the treatment. Leech saliva is reported to have many therapeutic contents like hirudin, bdellins, Hyaluronidase, etc.; among them, eglins and bdellins have anti-inflammatory and antifungal property which gives relief in symptoms. The Shamanaushadhis are also had a crucial part in easing the symptoms by reducing the size of lesion, and restoring normal skin colour.&#x0D; &#x0D; Daga, H. , Toshikhane,H. , Thakur K. , Raole V.,(2020) “Ayurvedic Management of Vicharchika – Single Case Study, International Journal Of Pharmaceutical Research, Vol 12 | Issue 4. org/10.31838/ijpr/2021.13.02.075.&#x0D;

Clinical efficacy of Jaloukavacharana (Leech Therapy) in Thrombosed External Haemorrhoids – A Case Study

Jalaukavacharana or Leech therapy is medicinal leech therapy which is used in a variety of inflammatory conditions. It is indicated in Raktaja and Pittaja Vikaras. Its mode of action depends on the injection of leech saliva into patient's tissues during the process of blood sucking. Leech’s saliva contains numerous bioactive constituent which possesses anti-inflammatory, analgesic, thrombolytic, anti-coagulant and blood circulation enhancing properties. Thrombosed hemorrhoid is an emergency condition that occurs due to high venous pressure associated with severe pain in the anus. In the present study a thrombosed external haemorrhoids is managed with Jaloukavacharana and its efficacy was observed.

Protein Profiling of Hirudo orientalis During Different Seasons for Obtaining Accurate Results in Leech Therapy

Objective: Leech therapy has been used for centuries as a recommended approach to cure several diseases, such as; psoriasis, arthrosclerosis, urinary tract diseases, and wound healing. The present study aimed to analyze the number, quantity, and distribution differences of medicinal leech (Hirudo orientalis) proteins throughout various seasons and in laboratory conditions as well. Methods: Protein profiling of salivary gland secretion from leech was studied by SDS-PAGE and 2D Electrophoresis on the proteins with the molecular weight range of 5 - 250 KDa in the lyophilized salivary gland secretion (SGS) during the seasons of summer and winter, and also in the laboratory conditions. Results: Our results indicated differences in the number and quality of leech saliva proteins in different seasons. We observed a higher number of proteins in summer than in winter. These results demonstrated the presence of Calin and Manillase in summer and Hyaluronidase and Collagenase in winter. Conclusion: This study could help us in choosing the best and most favorable conditions for using H. orientalis proteins for the treatment of different diseases.

Limitation of Jalauka (medicinal leech) application in the prevention of recurrence of keloid

Keloid can be defined as a benign dermal fibroproliferative disorder with no malignant potential. The keloids tend to occur following trivial physical trauma in genetically predisposed individuals. Jalauka (medicinal leech, Hirudo medicinalis) is famed to use in all skin disorders caused by vitiation of Rakta (~blood) and Pitta Dosha (a functional regulatory factor of the body). Many studies also have claimed that the saliva of medicinal leeches potentiates wound healing. It helps in healthy scar formation by inhibiting the conversion of fibrinogen to fibrin. Therefore, a hypothesis was made that medicinal leech application can be helpful in the healing of the wound without keloid formation in patients with keloid formation tendencies. In the present study, case of a 38-year-old male patient infected sebaceous cyst at the right upper back (scapular region) with a previous history of keloid formation after surgical scar was treated by weekly medicinal leech application five times after excision of the cyst under local anesthesia. Initiation wound healing was normal in the studied case until complete epithelization. After 2 months of complete healing, gradual hardening and shiny appearance with irregular margin became very prominent and seemed to convert in keloid, although other associated symptoms were absent. The findings of this case report indicate the limited effect of the medicinal leech application and the requirement of a multimodel therapeutic approach for healthy wound healing in keloid tendency patients.

Keratitis following leech therapy for periocular eczematous dermatitis: a case report

BackgroundThe medicinal leech therapy (MLT) is a kind of complementary treatment method used for various diseases. The leeches (Hirudo medicinalis) have been used for more than 2500 years by surgeons. The substances presenting in the saliva of leeches have anti-inflammatory, anticoagulant, platelet inhibitory, thrombin regulatory, analgesic, extracellular matrix degradative and antimicrobial effects. The method is cheap, easy to apply, effective and its mechanisms of action have been clarified for specific diseases. Infection particularly Aeromonas infection is the most common complication of MLT.Case presentationIn this case report, a keratitis case developing after leech therapy applied for the periocular and facial eczematous dermatitis lesions will be presented. The patient referred to our hospital with decreased vision, ocular pain, stinging, redness and lacrimation complaints. A large corneal epithelial defect with irregular margins, dying by fluorescein, involving more than inferior half of cornea and conjunctival hyperemia were seen in the right eye. No agent was determined in microbiological investigation, as the patient had used topical moxifloxacin eye drop which was commenced in another clinic before applying to us. The patient was treated with fortified vancomycin and ceftazidime, before using besifloxacin with the diagnosis of bacterial keratitis. Three weeks later epithelial defect improved completely leaving an opacity and neovascularization.ConclusionsMLT should be performed by certified physicians with sterile medicinal leeches and precautious antibiotics should be used before MLT for prevention against potential infections.

The Investigation of Effects of Medicinal Leech Saliva Extract on the Breast Fibroblast Cell Line In Vitro: An Experimental Study

The Investigation of Effects of Medicinal Leech Saliva Extract on the Breast Fibroblast Cell Line In Vitro: An Experimental Study

Case Reports and Experts Opinions about Current Use of Leech Therapy in Dermatology and Cosmetology

This article aimed to determine the experts’ opinion on the use of hirudotherapy in cosmetology, to supplement knowledge, and to identify factors that, in the opinion of experts, affect the effectiveness and frequency of use of this method in eliminating skin problems. The study was conducted using the expert interview method. The research tool was a categorized expert interview questionnaire using the CAWI (Computer Assisted Web Interview) technique. Hirudotherapy is one of the oldest healing methods. The mechanism of action is based on the therapeutic properties of bioactive substances isolated from leech saliva. It has been shown to have a wide range of applications in the treatment of numerous diseases in various fields of medicine, including dermatology and cosmetology. Despite its therapeutic properties and effectiveness, hirudotherapy is a rarely used tool in cosmetology. It has been found effective in the treatment of many dermatoses. The most spectacular effects have been observed in the treatment of rosacea, local inflammations, lipomas, skin scars, hard to heal wounds and contusions.

A Taxonomic Study comparing the two types of Medicinal Leeches available in Iraq

Medical leeches are ectoparasites invertebrates which are widely used in medical and surgical treatments because it was found since the ancient time that many diseases can be cured by medicinal leeches, then it was discovered that leech saliva contains anti-inflammatory, analgesic, antibacterial, anticoagulant, and even anticancer enzymes, also they found that medicinal leeches have a great role in treating diabetic ulcers. Tow samples of leeches were collected from two regions in Iraq from September to the end of December 2020. These samples were examined a live by dissecting microscope and then they were examined after fixing them in formaldehyde 5%. The samples of leeches that were used to analyze their genetic DNA were fixed and preserved in absolute ethanol. The classification process was done through a molecular diagnosis of samples of medical leeches by using polymerase chain reaction PCR and technique tracking nitrogenous bases in the nucleic acid chain, and the molecular diagnosis depending on Cytochrome oxidase subunit 1 (CO1), and then the result of PCR of medical leech samples was sent with the primers of the resulting bands and the gene sequences were read depending on the Genetic Analyser Device 3130 supplied by Japanese Hitachi company. Then the gene sequences were matched with gene sequences documented in the National Center Biotechnology Information NCBI. The results were analyzed depending on the BLAST program through which two types of medical leeches were detected Hirudo medicinalis and Hirudo verbena.

Leech Saliva and its Potential Use in Animal Health and Production: A Review

Leech Saliva and its Potential Use in Animal Health and Production: A Review

Effect Of Leech Therapy On TEP (Thrombosed External Pile): A Case Study

The Thrombosed external pile is the one which is located distal to the dentate line and develop as a result of tortuosity and distension of External Hemorrhoidal venous system. The engorgement of a hemorrhoidal vessel with acute swelling may allow blood to pool and subsequently clot; this leads to the Acute Thrombosed External Pile mass, a bluish purplish discolored swelling with severe pain and tenderness. The present case is of TEP (Thrombosed external pile) and the effect of Leech application is evaluated on this. The main objective is to assess the effective role of Leech on essential parameters of the patient,that are troublesome sign and symptoms - Pain, edema, tenderness and discoloration. The case is successfully managed with Leech therapy. Hirudin, calin, Factor Xa present in the saliva of Leech helps in oppressing the process of blood clot in an External pile mass and provides a symptomatic relief to the patient.