[Localization of leucyl-aminopeptidase (LAP) in the leech (Hirudo medicinalis L.) and horse leech (Haemopis sanguisuga L.)].
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Leeches, fed on swine infected with hog cholera, contained virus for as long as 87 days after their infective blood meals. In three instances, infected leeches apparently transmitted hog cholera virus to susceptible swine in the process of normal feeding. Myxoma virus persisted in leeches for as long as 154 days after the ingestion of a blood meal from rabbits with myxomatosis. Leeches fed consecutively, first on swine with hog cholera, and later on rabbits with myxomatosis, acquired both viruses. In such dually infected leeches, the hog cholera virus persisted for as long as 122 days and the myxoma virus for as long as 110 days, the longest periods tested. Leeches fed consecutively, first on rabbits with myxomatosis, and later on swine with hog cholera, acquired only the myxoma virus. Hog cholera virus could not be demonstrated in such dually fed leeches. Myxoma and hog cholera viruses appeared to be present in about equivalent amounts in the anterior and posterior thirds of the bodies of infected leeches. Myxoma and hog cholera viruses were present in the bloody gut contents of infected leeches but were not demonstrable in the body tissues of these leeches. It seems from the findings presented that leeches are not biological carriers of either myxoma or hog cholera virus but instead carry these two agents mechanically in their gastrointestinal tracts. In doing this, they appear to protect the viruses from various deleterious chemical and physical influences to which they would have been exposed in the open. It is speculated that leeches could play a role in nature in perpetuating the blood-borne viruses of certain diseases in which close association with bodies of fresh water is of epidemiological importance.
Tissues threatened by venous congestion often can be saved by timely leech therapy. Methods to restimulate sated leeches, particularly emergently, are only poorly described in the nineteenth-century literature. Sated leeches were purged of their blood meals by (1) posterior crop incision, (2) hypertonic saline (3 percent) immersion, (3) gentle finger pressure emesis, or (4) wood ash exposure. Their ability to reattach and refeed with or without serotonin stimulation was evaluated. All 20 leeches (100 percent) purged by posterior crop incision reattached, with 75 percent refeeding. After purging again, 87 percent of these refed leeches reattached, with 46 percent refeeding for a third time. Those leeches which did not initially refeed were exposed to serotonin 10 microM with 100 percent reattaching and 40 percent refeeding. None of the leeches purged by hypertonic saline immersion regurgitation reattached or refed. A single leech purged by finger pressure emesis reattached (20 percent) but did not refeed. After exposure to serotonin, two (40 percent) of each saline and finger pressure group reattached, with neither of the hypertonic saline group refeeding, while both finger pressure-purged leeches refed, consuming a meal 38 percent (+/- 29 percent) of original meal. None of those leeches which refed would reattach or refeed a third time. None of the wood ash-purged leeches reattached or refed even with serotonin exposure. The best method of purging leeches of their blood meals for emergent reuse is by posterior crop incision. Additional refeeding behavior can be achieved by immersion in serotonin 10 microM for 20 minutes.
This study evaluates the effect of hyperbaric oxygen and medicinal leeching on axial skin flaps subjected to total venous occlusion. Axial epigastric skin flaps (3 x 6 cm) were elevated on their vascular pedicles in 40 male Wistar rats. Total venous occlusion was achieved by division of all veins draining the skin flap. Arterial inflow was left intact. Animals were randomly assigned to one of five groups: sham (n = 8); control, total venous occlusion only (n = 8); occlusion with hyperbaric oxygen (n = 8); occlusion with leeching (n = 8); occlusion with leeching and hyperbaric oxygen (n = 8). The hyperbaric oxygen protocol consisted of 90-minute treatments, twice daily, with 100% O2 at 2.5 atmospheres absolute for 4 days. The leeching protocol consisted of placing medicinal leeches on the congested flaps for 15 minutes, once daily, for 4 days. Laser Doppler measurements of flap perfusion were recorded preoperatively, postoperatively, and on postoperative days 1 and 3. The percentage of flap necrosis was evaluated on postoperative day 3. Mean percentage necrosis and mean laser Doppler readings were compared between both groups. The flaps in the sham group demonstrated 99 percent survival, whereas the flaps in the occlusion-only group demonstrated 100 percent necrosis. The flaps in the occlusion with oxygen, the occlusion with leeching, and the occlusion with oxygen and leeching groups demonstrated 1, 25, and 67 percent survival, respectively. Sham laser Doppler readings remained within normal limits. Laser Doppler readings in the occlusion-only and the occlusion with oxygen groups decreased to negligible levels on postoperative day 1, and on postoperative day 3 no perfusion was demonstrated. In both the occlusion with leeching and the occlusion with leeching and oxygen groups, there was also a significant decrease in laser Doppler measurements after surgery, but perfusion remained stable throughout the remainder of the study. This study demonstrates that hyperbaric oxygen alone is not an effective treatment for skin flaps compromised by total venous occlusion. The combination of leeching and hyperbaric oxygen treatment of total venous occlusion results in a significant increase in flap survival above that found with leeching alone. It appears that hyperbaric oxygen is effective because of the venous outflow provided by leeching as demonstrated by laser Doppler flow readings.
Aeromonas hydrophila infections are a recognized complication of postoperative leech application, and can occur with measurable frequency in populations of patients treated with leeches. We review 11 previously reported leech-related Aeromonas infections and analyze seven unreported cases. These infections range from minor wound complications to extensive tissue loss and sepsis. Often, these infections followed leech application to tissue with questionable arterial perfusion. Onset of clinical infection in these patients ranged from within 24 hours of leech application to 10 days or more after leech application. Late infections may represent bacterial invasion from colonized necrotic tissue. Based on these observations, we recommend that leech applications be restricted to tissue with arterial perfusion to minimize contamination of necrotic tissue. We also recommend that patients treated with leeches receive antibiotics effective against Aeromonas hydrophila before leech application. Patients treated with leeches and discharged with eschars or open wounds might benefit from oral antibiotic therapy until wound closure. These precautions may minimize or eliminate this complication of leech use.
OBJECTIVE: To review the use of the medicinal leech, Hirudo medicinalis, in salvaging the failing, venous-congested flap. A protocol for the use of leeches is presented. Four illustrative cases of failing flaps (pectoralis major, midline forehead, and temporalis) are presented. STUDY DESIGN: Literature review comprised of MEDLINE search 1965 to present. Retrospective review of four cases involving the management of the failing, venous-congested flap. METHODS: A retrospective review of four cases of failing, venous-congested flaps was performed. RESULTS: The authors' experience, as well as the data from the reviewed medical literature, demonstrates the importance of early intervention in order to salvage the failing, venous-congested flap. Leeches are an immediate and efficacious treatment option. CONCLUSIONS: 1. Review of the literature indicates that the survival of the compromised, venous-congested flap is improved by early intervention with the medicinal leech. H medicinalis injects salivary components that inhibit both platelet aggregation and the coagulation cascade. The flap is decongested initially as the leech extracts blood and is further decongested as the bite wound oozes after the leech detaches. 2. When a flap begins to fail, salvage of that flap demands early recognition of reversible processes, such as venous congestion. The surgeon must be familiar with the use of leeches and should consider their use early, since flaps demonstrate significantly decreased survival after 3 hours if venous congestion is not relieved. In the four cases presented, a standardized protocol facilitated early leech use and provided for the psychological preparation of the patient, availability of leeches, and an antibiotic prophylaxis regimen. 3. The complications associated with leech use can be minimized with antibiotic therapy, wound care, and hematocrit monitoring. 4. The use of the medicinal leech for salvage of the venous-congested flap is a safe, efficacious, economical, and well-tolerated intervention.
BACKGROUND: Medicinal leech, Hirudo medicinalis, has been used in plastic and reconstructive surgery, to relieve venous congestion and to improve the microrevascularization of flaps. In many countries, wild leeches are still provided from local markets and utilised with antibiotic prophylaxies. In this research, results of identification of bacteria in the transport fluid is reported, oral and intestinal floras and the antibiograms of the identified microorganisms are investigated. Also, to avoid possible infections, the ability of hypochloric acid, a disinfectant, to suppress the relevant microorganisms without changing the life style and behavior of leeches in terms of sucking function, is investigated. METHODS: Bacterial identifications and antibiograms of oral and intestinal flora and transport medium were performed for 10 leeches. The optimum concentration of hypochloric acid which eliminated microorganisms without affecting the viability and sucking function of the leeches were determined by dilution of hypochloric acid to 100, 50, 25, 12.5, 6.25 ppm concentrations in different groups of 25 leeches. Finally, 20 leeches were applied atraumatically to the bleeding areas of rats, the duration of suction was determined and compared statistically between the leeches treated and not treated with hypochloric acid solution. RESULTS: Aeromonas hydrophilia was the most commonly identified microorganism and found to be resistant to first generation cephalosporins, frequently used in prophylaxis at surgical wards. In the next stages of the study, the leeches were subjected to a series of diluted hypochloric acid solutions. Although disinfection of the transport material and suppression of the oral flora of hirudo medicinalis were successful in 100, 50, 25, 12.5, 6.25 ppm concentrations; 12.5 ppm solution was the greatest concentration in which hirudo medicinalis could survive and sucking function was not affected significantly. CONCLUSIONS: External decontamination of wild leeches with 12.5 ppm hypochloric acid enables bacterial suppression without causing negative effects on leech sucking function and life.
BACKGROUND: Leech therapy was commonly used in traditional medicine for treating localized pain. Clinically significant pain relief after leech therapy for osteoarthritis of the knee has been demonstrated by preliminary data. OBJECTIVE: To evaluate the effectiveness of leech therapy for symptomatic relief of osteoarthritis of the knee. DESIGN: Randomized, controlled trial. SETTING: Outpatient department for integrative medicine of an academic teaching hospital. PATIENTS: 51 patients with osteoarthritis of the knee (leech therapy: 24 patients, mean age [+/-SD], 62.5 +/- 10.2 years; topical diclofenac therapy: 27 patients, mean age [+/-SD], 65.5 +/- 6.7 years). INTERVENTION: A single treatment with 4 to 6 locally applied leeches (leech therapy group) or a 28-day topical diclofenac regimen (control group). MEASUREMENTS: Mean of the pain, function, and stiffness subscores of the Western Ontario and McMaster Universities Osteoarthritis Index and physical sum score of the Medical Outcomes Study 36-Item Short-Form Health Survey with group comparisons at days 3, 7, 28, and 91. RESULTS: The primary end point, pain at day 7, was reduced from a mean (+/-SD) of 53.5 +/- 13.7 to 19.3 +/- 12.2 after leech therapy compared with 51.5 +/- 16.8 to 42.4 +/- 19.7 with topical diclofenac (estimated group difference, -23.9 [95% CI, -32.8 to -15.1]; P < 0.001). Although the difference between group pain scores was no longer significant after day 7, differences for function, stiffness, and total symptoms remained significant in favor of leech therapy until the end of study and for quality of life until day 28. Results were not affected by outcome expectation. CONCLUSIONS: Leech therapy helps relieve symptoms in patients with osteoarthritis of the knee. The potential of leech therapy for treating osteoarthritis and the pharmacologic properties of leech saliva remain to be clarified.
OBJECTIVES: The purpose of this study was to localize in leeches the renin-like enzyme previously characterized as well as the leech angiotensin-converting like enzyme (ACE). METHODS: Immunocytochemical as well as whole mount experiments were performed with an antibody raised against a fragment of the leech renin-like enzyme (VLWAAEKTQLDTGSS) and with anti-leech ACE. RESULTS: Anti-leech renin stains the vascular pole of the glomerulus and the afferent arteriole of the rat kidney. Immunostaining of leech sections revealed labeling in neurons and glial cells of the central nervous system (CNS), immunocytes and the nephridial canal, canaliculi and the periphery of the ciliated funnel, as well as the epithelium lining nephridia. Co-localization between antibodies raised against this fragment and a fragment of leech angiotensin-converting enzyme was demonstrated in neurons and glial cells of the leech CNS, as in vertebrates MAIN FINDING: Leech renin is localized in leeches like in vertebrate in the excretory system and in the nervous system. CONCLUSIONS: Our findings suggest the presence of a renin-angiotensin system involved in osmoregulation in leeches.
OBJECTIVE: To compare pharmacodynamics of decoction and powder of Leech; To identify effect of pharmacodynamics of Leech with new methods of process and ultrmicro-pulverization. METHOD: The size of ultramicor-powder of Leech and new method processed Leech is determined by BT-1500 size distributive instrument. It is compared that the pharmacodynamics of Leech decoction and powder as well as ultramicro-powder with new method processed of 1/2 decoction dose by anticoagulation and antithrombus action of mice. The safety of different samples is identified. RESULT: The pharmacodynamics of samples is as ultramicro-powder of Leech with new method processed > ultrmicro-powder of Leech > powder of Leech > decoction of Leech. CONCLUSION: The ultramicro-powder of Leech with new method processed has better pharmacodynamic effect and the smell of Leech is improved.
Medicinal leeches (Hirudo medicinalis) are a standard treatment for venous congestion, a complication that can occur after reconstructive surgery. If the cause of venous congestion cannot be surgically corrected, then medicinal leeches are used to temporarily increase perfusion levels and maintain physiologic requirements within the congested tissue. Leeches increase perfusion within congested tissue by actively drawing off blood as a bloodmeal. Furthermore, the leech bite continues to bleed and relieve congestion after detachment because of the anticoagulation effects of leech saliva left behind in the bite. In a porcine model, a 10 x 10 cm cutaneous flank flap was congested by clamping the venae comitantes. Four medicinal leeches were allowed to attach to the congested flap, and parameters of active feeding and passive bleeding after detachment were recorded. The average bloodmeal volume for the medicinal leeches was 2.45 ml. Average passive bleeding for the first 2 and 4 hours after leech detachment totaled 2.21 and 2.50 ml, respectively, with 90 percent of passive bleeding occurring within 5 hours after detachment. Laser Doppler imaging indicated that the spatial arrangement of surface perfusion increases were localized to a 1.6-cm-diameter circle around the leech head (bite) and corresponded well with the visual return of normal skin tones to the same area. This study provides a realistic and quantitative estimate of the spatial and volumetric characteristics of leech feeding and passive bleeding using a clinically relevant model of acute, severe congestion.
1. Serotonin is sequestered by a limited population of identified neurons in the 32 ganglia of the leech nervous system. A major fraction of the serotonin in each ganglion is contained in the paired Retzius cells, colossal effector neurons whose size varies longitudinally. The 5 other classes of identified serotonin-containing neurons, one effector cell and 4 interneurons, are approximately twice as numerous in anterior as in posterior ganglia. 2. We dissected 6 longitudinal samples from the ventral nerve cords of hungry Hirudo medicinalis, and measured their serotonin content using high pressure liquid chromatography with electrochemical detection. A consistent neurochemical pattern emerged in which segmental ganglia 2-4 had the highest quantity of serotonin: 18.51 pmol per ganglion. The anterior cerebral ganglion contained 14.78 pmol, and the content of the 4 posterior samples, segmental ganglia 7-10, 12-15, 17-20 and the caudal ganglion, decreased continuously from 16.35, 15.08, 10.75 to 2.51 pmol per ganglion, respectively. Morphometric analyses indicated that this pattern of ganglionic serotonin correlated primarily with longitudinal variations in the number of serotonin neurons per ganglion and secondarily with volume of the Retzius cells. Retzius cell volume correlated highly with the mass of their innervated body segments both of which are largest in mid-body domains. 3. Serotonin expresses leech feeding, and its ganglionic levels are a potentially useful index of behavioral state. We measured serotonin in the ganglionic samples from hungry and satiated leeches. The samples from recently fed animals contained 28% less serotonin than those from hungry ones. The amounts of serotonin in the cerebral and all the segmental samples from satiated leeches were significantly lower than equivalent samples of hungry animals. A similar pattern of depletion was seen in leeches which fed for a prolonged period (90 to 120 min) rather than the normal period of 30 min. 4. The effects of ingestion on serotonin-containing neurons was examined with the glyoxylic acid-induced histochemical fluorescence. The levels of fluorescence in all serotonin neurons in fed leeches were consistently lower than those in equivalent neurons in hungry animals, corroborating the ganglionic decrease in serotonin in satiated leeches. 5. To examine effects of body wall distension on serotonin levels, hungry leeches were fed to satiation, and half of them were relaxed by removing their distending blood meals. After 6 weeks, ganglionic serotonin in leeches with relaxed bodies was 21% higher than in those with distended bodies. 6. Ingestive behavior depletes serotonin from leech neurons and body wall distension appears to interfere with its synthesis.(ABSTRACT TRUNCATED AT 400 WORDS)
Leeches were studied for their efficacy to improve survival of venous compromised rat skin flaps. In 22 rats, bilateral epigastric island flaps were created and subjected to 6 h of venous occlusion. One flap in each animal was randomised to leech treatment, while the contralateral flap served as its own control. Flap survival, leech feeding time, weight gained by the leech and bleeding time from leech bites were measured. The area of flap survival was significantly increased in leech treated flaps compared to contralateral controls (n = 22 pairs, p = 0.03; Wilcoxon signed rank test). Weight gained in the feeding leech averaged 1.3 +/- 0.2 g (n = 18). Leech feeding time was 107 +/- 13 min (n = 18). Bleeding time from each leech bite averaged 79 +/- 12 min (n = 18). Hence, the extent of flap necrosis resulting from venous impairment can be partly diminished by leech treatment until definitive surgical venous revascularisation.
In a 58-year-old hospitalized woman with gonarthrosis a leech therapy was applied to both knee joints. In the evening of the following day she observed strong pruritus in the area of the leech bites; in addition a maculopapular exanthema appeared on the torso and her lower extremities. The allergic reaction lasted four days. Administration of antihistamines only led to a slight improvement of the symptoms. A full restitution could only be achieved after a systemic dose of glucocorticoids on the fourth day after leech therapy. Eight days before beginning of the leech therapy a five-day antibiotic therapy with trimethoprim and sulfamethoxazole (Cotrim forte) had been administered to treat an uncomplicated urinary infection. Allergic reactions are well-known complications of these antibiotics and of leech therapy. The four-day duration of the allergic reaction after leech therapy, however, was untypical. In order to explain these symptoms, a prick test and an epicutaneous test for the antibiotic components were executed five weeks after the leech therapy. Furthermore, a second leech therapy was administered and a lymphocyte transformation test (LTT) was carried out. The results of the LTT showed a sensitization for sulfamethoxazole and a possible sensitization for trimethoprim, the results of the epicutaneous test showed a positive reaction to sodium lauryl sulfate, a component of the antibiotic. In the area of the leech bites a clear local skin reaction was observed. These results suggest a drug exanthema, in all probability triggered by the leech therapy.
Venous congestion is a constant threat in the survival of free flaps, pedicled flaps, tissue replantations and traumatized tissue. Leech therapy has proved effective in salvaging much of these compromised tissues. To be effective, leeches must be both readily available and hungry, requirements not always easily met. Our investigation seeks to establish a means to return sated leeches to their previous unfed, hungry state for reuse. Sated leeches were purged of their blood meals by placement in 3% hypertonic saline or by gentle finger pressure, then exposed to serotonin 0 microM (control), 10 microM, 30 microM or 90 microM for 20 min. Subsequent rebiting/refeeding was observed and analysed. Leeches bathed in serotonin rebit or reattached at nearly four times the rate of unexposed leeches; 30% vs 8% respectively. Biting, however, is not refeeding. Overall, 41 leeches were treated with serotonin with four (10%) refeeding. Those that refed consumed a significantly smaller blood meal than the initial feeding; 50% +/- 47% SD vs 348% +/- 143% SD of original body weight. None of the control group refed. As a method for routinely reusing leeches, serotonin bathing cannot be recommended. In the immediate postoperative period with the sudden emergence of venous congestion requiring leech therapy, but with an inadequate number available, this 20% [corrected] refeed rate after 10 microM serotonin exposure could potentially determine the success or failure of the flap/replantation until fresh leeches are made available.
Behaviour has been shown to be a sensitive indicator of chemically induced stress and pathology in aquatic organisms. However, ecotoxicological investigations on medicinal leech are restricted because of scarce leech resources in natural waters. We used artificially bred medicinal leech (Hirudo medicinalis L.) of two different ages: young (1-2 weeks old) and adult leeches (1 year old). Animals were exposed to: (1) waters of Lake Drukshiai-the cooler of Ignalina Nuclear Power Plant, (2) sediments of the river Nemunas and (3) solution of heavy metal model mixture (HMMM). The following behavioural responses of leech were investigated: mobility (number of moving individuals within certain periods of time), avoidance response (number of individuals escaping the tested water or sediments) changes in body shape (contractions of some muscles, abnormal position of suckers) and feeding activity (longevity of attachment process, interruptions of feeding bouts, size of blood meal). Mobility of young leeches was increased in the tested waters of Lake Drukshiai, whereas adult leeches showed no changes of this index. Avoidance response as well as impaired feeding activity (prolonged attachment process or completely suppressed attachment reflex, decreased size of blood meal) was observed in young leeches exposed to the waters from Lake Drukshiai and in adult animals exposed to all three kind of trials mentioned above. Avoidance response and changes in mobility were recorded during the first hour of exposure to tested samples; therefore, these responses can be used as an express method for water and sediment pollution assessment. Impaired feeding activity was recorded after 1-3-week exposure: this response may be used in assessing a chronic toxicity of pollution. Medicinal leech, due to its sensitivity, simplicity of measured indices and ease of laboratory maintenance, can be used as a test organism in ecotoxicity studies.
Aeromonas hydrophila infections are a recognized complication of the use of medicinal leeches. The authors performed an experiment designed to find a safe and practical way to sterilize the leech gut of pathogenic organisms. Leeches were incubated for a 12-hour period in solutions of antibiotic effective against A. hydrophila. The incubations in the antibiotic solutions failed to eradicate pathogenic bacteria from the gut of the leeches. The authors examined cultures of bacteria isolated from the guts of the commonly used Hirudo medicinalis (European leech) and found a wide variety of pathogenic organisms. A. hydrophila is widely believed to be the most common enteric pathogen, but the authors found A. sobria more frequently in their experiment. They also cultured the guts of the leech H. michaelseni recently used clinically in South Africa. A. caviae was the most common pathogen encountered in these leeches. A. caviae and A. sobria cause a spectra of disease similar to A. hydrophila. The authors endorse the current recommendation that all patients who have leech therapy for congested flaps or replants receive broad-spectrum prophylactic antibiotics. This appears to be the safest and simplest way to prevent leech-related infections.