Talks, Abstracts

APPLICATION OF LEECHES IN TREATMENT OF MICROCIRCULATION DISTURBANCES IN RECONSTRUCTIVE AND HAND SURGERY

 Paruzel Maciej, Jabłecki J., Domanasiewicz A., Elsaftawy A.

Subdepartment of Replantation of Limbs St. Hedwig District Hospital in Trzebnica, Poland

Purpose: Possibilities of application of leeches in cases of disturbances of venal out-flaw in replanted digits and insufficiency circulation in reconstructive surgery cases.

Material and methods. The indication for treatment were venal out-flaw disturbances of different etiology and their prophylaxis. Leeches were usually applied every 6-8 hours (one-two leech on one finger), the duration of treatment was from 2 to 6 days (4 days on average). After an effective application the animals were utilized. The antibiotics of ciprofloxacin group were used as a prophylaxis of Aeromonas hydrophilia’s infection.

Results. All of the fingers and skin flaps treated with leeches survived. A dramatic improvement of local state of the fingers could be seen soon after the application of leeches. All of the patients did not complain of any discomfort associated with the therapy.

Conclusion. The application of leeches in cases of disturbances venal out – flaw in replanted digits and insufficiency circulation skin flaps seems to be a valuable alternative of other methods commonly used.

HIRUDOTHERAPY AND MAGGOT DEBRIDEMENT THERAPY AS ALTERNATIVE MANAGEMENT OF A CHRONIC NONHEALING ISCHEMIC WOUNDS

OUR EXPERIENCE IN HIRUDOTHERAPY IN SLOVAKIA

Peter Takáč^1,2,  Mária Habrmanová², Henrieta Galátová¹, Viliam Slezák^1,3

¹Scientica, s.r.o, Bratislava, Slovakia; ²Institute of Zoology, Slovak Academy of Sciences, Bratislava, Slovakia; ³University Hospital of the Merciful Brothers, Bratislava, Slovakia

Non healing chronic ischemic wounds are a major clinical challenge very frequently encountered in the clinical practice. Generally is known that in all kinds of non healing wounds micro vascular angiopathy is one of the major factors for hypoxic condition, leading to non-healing. Recently many new devices and techniques are evolved for this purpose, but the efficacy in rectifying tiny micro vascular thrombus is still a matter of discussion. Alternative treatment of wounds with leeches and maggots , has approved very good results for healing different forms of wounds, with great perspective in the standard use in practice.

It was found that leech application improves the hypoxic condition, providing a moist environment for wound healing, and that it helps in slough debridement in non healing ulcers and most importantly leeches possess properties that make them uniquely able to assist with venous compromised tissues.

At the same time the maggot debridement therapy is a modern biotherapeutic method based on the use of necrophagous flies to treat chronic non-healing wounds. In last two decades, maggot debridement therapy became routinely used therapeutic method and thousands of patients over the world had benefit of its healing power. Maggot debridement therapy is a safe, reliable, cost-saving and very effective method in management of chronic non-healing wounds. In our presentation we present the results of treatment with Hirudotherapy in combination with Maggot debridement therapy. Alternative treatment of wounds with leeches and maggots has approved very good results for healing different forms of wounds, with great perspective in the standard use in practice.

This work had been funded by the Operational Program of Research and Development and co-financed with the European Fund for Regional Development (EFRD). Grant: ITMS 26240220030: Research and development of new biotherapeutic methods and its application in some illnesses treatment.

CLINICAL LEECH THERAPY STUDIES IN WESTERN EUROPE AND USA

Detlev Koeppen

Legal and financial restrictions constrain the performance of clinical leech therapy studies with respect to the number of studies and their target indications. In several European countries, leech therapy is hardly known or is performed in legally not clearly defined areas. In USA, the use of leeching is limited to rescue therapy measure in reconstructive surgery. The limited financial resources of leech distributors do not allow the performance of extensive clinical studies.

The existing studies focus on osteoarthritis, epicondylitis and venous congestion.

Methodology and results of these studies are presented. Despite of the limitations of clinical study performance in Western Europe and USA, the existing results look encouraging and justify further preclinical and clinical research.

LEECH FARMING AT THE UNIVERSITY OF TEHRAN, PROSPECTIVE AND CHALLENGE

Masoumeh Malek

School of Biology and Center of Excellence in Phylogeny of living organisms, College of Science, University of Tehran, Iran

Since 2008, study of leech ecology and breeding were started at University of Tehran. After field and laboratory study of Hirudo orientalis Utevsky & Trontelj, 2005, leech farm was established at the Science and Education field of the University of Tehran. Adult leeches were fed with blood sausage, kept on mosses.  Cocoons were collected and kept on wet moss until juveniles were hatched. During these activities, we faced several problems, including feeding juveniles, preventing the leeches from scape, Drain fly, Psychodid fly larva in the cocoons. Different strategies for solving these problems will be discussed. Because of high demand for leech therapy in the traditional medicine, we aim to produce 200000 leeches annually.   

DO SALIVARY PROTEINS FROM HEMATOPHAGOUS LEECHES REACH PHARMACOLOGICALLY EFFECTIVE CONCENTRATIONS IN THE HOST?

Sarah Lemke, Christian Müller, Elisabeth Lipke, Gabriele Uhl, Jan-Peter Hildebrandt

Animal Physiology and Biochemistry, Ernst Moritz Arndt-University, Biotechnikum, Rathenaustraße 49 a, D - 17489 Greifswald

Saliva of hematophagous leeches (Hirudo verbana) contains bioactive substances which facilitate feeding and storage of ingested blood, but may also exert beneficial effects in the host. As leeches transfer the entire mass of stored salivary proteins into the host's wound during feeding, we determined the resulting concentrations of salivary proteins in the circulatory system of the host. We used serial sections (5 μm) of leech tissue from the anterior body for 3D reconstruction of a block of leech tissue, determined the mean volume of an individual gland cell (67,000 µm³) and the total number of salivary gland cells present in one animal (37,000). Using protein staining in homogenates of reference tissues and staining intensities of tissue sections of reference as well as leech tissues, we determined the total mass of salivary protein in one leech (average of 1.2 mg). Analyzing protein extracts of unfed and fed leeches by 2D electrophoresis, we calculated the amounts of individual salivary proteins which are transferred to the host during feeding. Assuming a mean plasma volume in humans of 5 l, the maximally achievable molar concentrations of salivary proteins in host plasma were calculated for 23 different proteins (3 - 236 pmol/l). As hirudin, the only salivary agent that has been pharmacologically characterized as yet, has been shown to inhibit human thrombin with an IC₅₀of 1 pmol/l, we concluded that many other substances in leech saliva may reach concentrations in host plasma which may be sufficiently high to achieve systemic pharmacological effects.

PHARMACY OF THE MEDICINAL LEECH SALIVA IS THE SOURCE OF NOVEL NATURAL MULTIFUNCTIONAL DRUGS

Baskova I.P.

Biological Faculty of Moscow State University, e-mail: saliva1@yandex.ru

The medicinal leech pharmacy is localized in salivary cells , which disseminated among the muscle tissue of the anterior leech body. By using some methods of proteomic analysis there were detected in secretion of salivary cells (SSC) more than 100 high molecular weight (mol. 10 – 97 kDa) and about 150 low molecular weight (200-700 Da) substances (Baskova et al.., 2004, 2008). Now it is described more than 20 proteins and low molecular weight compounds of SSC. Their composition is depended on the seasons and on the species of the medicinal leech (Hirudo verbana, H. medicinalis, and H. orientalis). Among them there are the substances, which non synthesized in salivary cells, but detected in their secretions. Mammalian blood, the important component of the leech nutrition, is the sours of cholesterol, its derivatives and of some steroid hormones. It is the way of medicinal leech adaptation to feeding by blood of the warm-blooded animals.

Different methods of proteomic analysis demonstrated the presence in the SSC of numerous unknown substances of low- and high molecular weight. The medicinal leech Pharmacy is full of the real biologically active components, which have been manifesting as the humoral agents of hirudotherapy, effective method for treatment of the diseases related to different divisions of medicine. Identification of the medicinal leech pharmaceutical compounds would help to understand the mechanisms of these treatment, but it will be possible only after the discovering genome of this unique animal.

HIRUDOTHERAPY: A UNIQUE METHOD IN TREATMENT OF THE CERTAIN EYE DISEASES

Tatiana Semyonovna Semikova, MD, PhD

The S.N. Fyodorov IRTC "Eye Microsurgery” (IRTC EM), Moscow

Purpose: To substantiate and extend indications for hirudotherapy application in ophthalmology. Medical leeches are traditionally used in treatment of acute glaucoma attacks and choroidal inflammations.

Materials and methods: An opportunity to extend the range of hirudotherapy application in treatment of various eye pathologies appeared basing on the mechanics of leech effect.

The medical effect of leeches is determined by their salivary glands properties: its secretion penetrates into human body when the leech bites through the coverlet and is incoming into the body during the whole period of bloodsucking. At that the secretion blocks the hemostatic thrombus formation in situ of the vascular wall damage realizing the optimal mechanisms of inhibition of platelet-vascular and plasma links of human homeostasis.

Enzymes of the leech’s saliva possess the following properties: anticoagulative, antithrombotic, anti-inflammatory, immunopotentiating, bacteriostatic, bactericidal and analgesic. Bioactive compounds being a part of the leech’s saliva secretion provide a successful process of blood microcirculation rendering nonspecific influence on the organism as a whole increasing phagocytic activity of the blood neutrophils.  Leeches bites cause stimulation of the certain reflexogenous zones on the surface of the body that produces favourable influence on the patient’s organism.

At the IRTC "Eye Microsurgery” leeches have found its application not only for glaucoma treatment both for arresting of the acute attack and for pre-op preparation. Hirudotherapy is widely used for elimination of the post-op complications, for macular edema diagnostics, inflammatory eye diseases treatment, iridocyclitis, chorioretinitis, dacryocystitis, sclerite, retrobulbar neuritis, hemorrhages of various etiology, endocrine ophthalmopathy, degenerative choroidal diseases, dry and wet forms of maculopathy, for acceleration of intraocular tumors resolution after brachytherapy, for elimination of complications related to radiation exposure.

Hirudotherapy creates practically no complications and side effects; it significantly decreases the period of recovery in inflammatory diseases; it makes the intraocular melanomas resolution period after brachytherapy 2-3 times shorter; it serves as a prevention of such post-op complications as secondary glaucoma and glaucoma-related pain syndrome, retinal detachment, hemorrhages into the vitreous body and retina. Leeches help to diagnose the macular edema and to treat it. Hirudotherapy can be conducted both in hospital and under outpatient conditions.

Conclusions: Hirudotherapy is a method of high efficiency for various eye pathologies treatment; it can be used both independently and in combination with the other methods in glaucoma, inflammatory, degenerative, tumor diseases and in ocular trauma.