Science.gov

Sample records for leeches

  1. Annelida, Euhirudinea (leeches)

    EPA Science Inventory

    Worldwide, there are over 600 species of leeches described which occur in freshwater, marine, estuarine, and moist-terrestrial ecosystems. Leeches are included in the Class Clitellata, Subclass Hirudinida, and Superorder Euhirudinea. Seven of the ten families of leeches occur in...

  2. Annelida, Euhirudinea (leeches)

    EPA Science Inventory

    Worldwide, there are over 600 species of leeches described which occur in freshwater, marine, estuarine, and moist-terrestrial ecosystems. Leeches are included in the Class Clitellata, Subclass Hirudinida, and Superorder Euhirudinea. Seven of the ten families of leeches occur in...

  3. Leech Therapeutic Applications

    PubMed Central

    Abdualkader, A. M.; Ghawi, A. M.; Alaama, M.; Awang, M.; Merzouk, A.

    2013-01-01

    Hematophagous animals including leeches have been known to possess biologically active compounds in their secretions, especially in their saliva. The blood-sucking annelids, leeches have been used for therapeutic purposes since the beginning of civilization. Ancient Egyptian, Indian, Greek and Arab physicians used leeches for a wide range of diseases starting from the conventional use for bleeding to systemic ailments, such as skin diseases, nervous system abnormalities, urinary and reproductive system problems, inflammation, and dental problems. Recently, extensive researches on leech saliva unveiled the presence of a variety of bioactive peptides and proteins involving antithrombin (hirudin, bufrudin), antiplatelet (calin, saratin), factor Xa inhibitors (lefaxin), antibacterial (theromacin, theromyzin) and others. Consequently, leech has made a comeback as a new remedy for many chronic and life-threatening abnormalities, such as cardiovascular problems, cancer, metastasis, and infectious diseases. In the 20th century, leech therapy has established itself in plastic and microsurgery as a protective tool against venous congestion and served to salvage the replanted digits and flaps. Many clinics for plastic surgery all over the world started to use leeches for cosmetic purposes. Despite the efficacious properties of leech therapy, the safety, and complications of leeching are still controversial. PMID:24019559

  4. [Medicinal leeches and hirudotherapy].

    PubMed

    Gödekmerdan, Ahmet; Arusan, Suat; Bayar, Batu; Sağlam, Naim

    2011-01-01

    Leeches have been used in the treatment of certain diseases since ancient times. There are a few species of medicinal leeches. However, H. medicinalis and H. verbana are known in Turkey. Currently, Turkey is one of the world's most important leech-exporting countries. Secretion of the salivary glands of medical leeches contains more than 100 bioactive substances. These secretions include vasodilators, bacteriostatic, analgesic, anti-inflammatory and anticoagulants, anti-edematous, which eliminate microcirculatory disorders, restore the damaged vascular permeability of tissues and organs, eliminate hypoxia, reduce blood pressure, increase immune system activity, resolving the cause of pain and improve the bioenergetic status of the organism. U.S. Food and Drug Administration (Food and Drug Authority of USA-FDA) have allowed the sale of leeches in this country, the use for plastic surgery, the general purposes and microsurgery in 2004. Turkey is the richest source in terms of the medical leech. Better evaluation of this valuable commodity and more effective use of hirudotherapy in modern medical practice as supportive treatment is very important.

  5. Ocular leech infestation

    PubMed Central

    Lee, Yueh-Chang; Chiu, Cheng-Jen

    2015-01-01

    This case report describes a female toddler with manifestations of ocular leech infestation. A 2-year-old girl was brought to our outpatient clinic with a complaint of irritable crying after being taken to a stream in Hualien 1 day previous, where she played in the water. The parents noticed that she rubbed her right eye a lot. Upon examination, the girl had good fix and follow in either eye. Slit-lamp examination showed conjunctival injection with a moving dark black–brown foreign body partly attached in the lower conjunctiva. After applying topical anesthetics, the leech, measuring 1 cm in length, was extracted under a microscope. The patient began using topical antibiotic and corticosteroid agents. By 1 week after extraction, the patient had no obvious symptoms or signs, except for a limited subconjunctival hemorrhage, and no corneal/scleral involvement was observed. PMID:25784786

  6. Satellite stabilization using space leeches

    NASA Technical Reports Server (NTRS)

    Walker, Michael W.; Kim, Dong-Min

    1990-01-01

    A control algorithm for satellite stabilization using a space leech is presented. The space leech is assumed to have n reaction wheels with known moments of inertia about their axis of rotation. All mass properties of the satellite are assumed to be unknown. The algorithm brings the satellite to a specified attitude trajectory. Simulations were performed to demonstrate the controller. The model parameters and specific algorithm used and the results obtained are presented.

  7. LEECHES (ANNELIDA: EUHIRUDINEA) OF NORTHERN ARKANSAS

    EPA Science Inventory

    Twenty-one lotic and lentic environments throughout central and northern Arkansas were surveyed for the presence of leeches during June 2004, and April, July - October, 2005. Fourteen species of leeches (Desserobdella cryptobranchii, Desserobdella phalera, Desserobdella picta, H...

  8. LEECHES (ANNELIDA: EUHIRUDINEA) OF NORTHERN ARKANSAS

    EPA Science Inventory

    Twenty-one lotic and lentic environments throughout central and northern Arkansas were surveyed for the presence of leeches during June 2004, and April, July - October, 2005. Fourteen species of leeches (Desserobdella cryptobranchii, Desserobdella phalera, Desserobdella picta, H...

  9. The medicinal leech in Jewish writings.

    PubMed

    Rosner, F

    1999-12-01

    Leeches were widely used for healing throughout antiquity and the Middle Ages. The rabbinic responsa literature acknowledges the use of leeching for the prevention and treatment of certain illnesses, and classic Jewish sources including the Bible, Talmud, and Codes of Jewish Law describe leeches and mention their medicinal use. Although the swallowing of a leech is considered dangerous and may lead to abdominal swelling, the Talmud describes an oral concoction containing leeches in wine for patients with enlarged spleens. It is also documented that engorged leeches placed in salt quickly discharge their blood and can be used again. Modern plastic and microsurgery have rediscovered the medicinal leech, Hirudo medicinalis, to reduce venous congestion. Clearly, the value of this annelid worm lies in its secretion of an anticoagulant known as hirudin, which has several advantages over heparin. One cannot help but marvel at the wisdom and ingenuity of our forefathers.

  10. Leeching in the history--a review.

    PubMed

    Munshi, Younis; Ara, Irfat; Rafique, Huma; Ahmad, Zahoor

    2008-07-01

    Leeching has enjoyed a good reputation in the past where it had been used in various ailments from fevers to flatulence. The present day scientists have limited its role and the leeching nowadays is being used only in microsurgeries to relieve the venous congestion. This study was designed to explore the possibility of revival of the leech therapy (leeching) which is still being used traditionally as therapeutic agent in various ailments. Leeching is not the outcome of the medieval period but has been in use during the times when there was no concept of the disease and medicament. The earliest clearly documented record of leeches being used for remedial purpose appears in a painting in an Egyptian Tomb of around 1500 BC. The journey of the leech therapy reached its zenith in 17th and 18th century AD in Europe while as during the Arab era the leeches were used medicinally but only for the bloodletting. During the 17th and 18th century AD there was shortage of leeches in certain European countries due to its rigorous use. During early 20th century AD when germ theory was put forward and medical fraternity believed that every disease has its origin from germs and the era of antibacterial therapy gained a pace, the leech therapy was considered the myth of the past. It was in 1970's that the leech therapy was revived by only limited to the microsurgeries to relieve venous congestions. During the 21st century there were certain studies when the leeches were tried in certain ailments like arthritis etc. and the Food and Drug Authority of USA (FDA) gave permission for sale and use of leeches in USA but limited its use in microsurgeries and plastic surgeries only. The aim of this study was to explore different diseases where the role of leech therapy can be seen and clinical trials can be started in this direction. Although the researchers in Regional Research Institute of Unani Medicine, Srinagar, Kashmir, India working under the aegis of Central Council for Research in

  11. [Design of an electronic leech].

    PubMed

    Lomelí-Mejía, Pedro Alejandro; Domínguez-Rubio, René; Espinosa-Gutiérrez, Alejandro; Lecona-Butrón, Hugo; Rodríguez-Reyna, Reynaldo Manuel

    2014-07-01

    This research presents the development of a continual suction electromechanical device (CSED) which emulates the feeding characteristics of a medicinal leech to drain body fluids. After the research, design and building of the device, its performance in normal conditions with fluids of different viscosity was evaluated. Finally, the device was submitted to a test of blood draining in three adult male rabbits NZW with a weight of three kilograms, obtaining drain.

  12. Ciprofloxacin-resistant Aeromonas hydrophila cellulitis following leech therapy.

    PubMed

    Giltner, Carmen L; Bobenchik, April M; Uslan, Daniel Z; Deville, Jaime G; Humphries, Romney M

    2013-04-01

    We report a case of surgical site infection with ciprofloxacin-resistant Aeromonas hydrophila following leech therapy. Antimicrobial and genetic analyses of leech and patient isolates demonstrated that the resistant isolates originated from the leech gut microbiota. These data suggest that ciprofloxacin monotherapy as a prophylaxis regimen prior to leech therapy may not be effective in preventing infection.

  13. Ciprofloxacin-Resistant Aeromonas hydrophila Cellulitis following Leech Therapy

    PubMed Central

    Giltner, Carmen L.; Bobenchik, April M.; Uslan, Daniel Z.; Deville, Jaime G.

    2013-01-01

    We report a case of surgical site infection with ciprofloxacin-resistant Aeromonas hydrophila following leech therapy. Antimicrobial and genetic analyses of leech and patient isolates demonstrated that the resistant isolates originated from the leech gut microbiota. These data suggest that ciprofloxacin monotherapy as a prophylaxis regimen prior to leech therapy may not be effective in preventing infection. PMID:23363826

  14. Leeches run cold, then hot

    PubMed Central

    Petersen, Ann M.; Chin, Wendy; Feilich, Kara L.; Jung, Grace; Quist, Jessica L.; Wang, Jasmine; Ellerby, David J.

    2011-01-01

    Food processing is costly, potentially limiting the energy and time devoted to other essential functions such as locomotion or reproduction. In ectotherms, post-prandial thermophily, the selection of a warm environmental temperature after feeding, may be advantageous in minimizing the duration of this elevated cost. Although present in many vertebrate taxa, this behaviour had not previously been observed in invertebrates. Sanguivorous leeches ingest large blood meals that are costly to process and limit mobility until excess fluid can actively be expelled to reduce body volume. When presented with a temperature gradient from 10°C to 30°C, leeches select a temperature that is significantly warmer (24.3 ± 0.9°C, n = 6) than their acclimation temperature (Ta, 21°C). Unfed leeches preferred temperatures that were significantly cooler than ambient (12.8 ± 0.9°C, n = 6). This behavioural strategy is consistent with minimizing the time course of elevated post-feeding energy costs and reducing energy expenditure during fasting. Our observations raise the possibility that thermoregulatory behaviour of this type is an unrecognized feature of other invertebrate taxa. PMID:21551223

  15. Quantitative characterization and classification of leech behavior.

    PubMed

    Mazzoni, Alberto; Garcia-Perez, Elizabeth; Zoccolan, Davide; Graziosi, Sergio; Torre, Vincent

    2005-01-01

    This paper describes an automatic system for the analysis and classification of leech behavior. Three colored beads were attached to the dorsal side of a free moving or pinned leech, and color CCD camera images were taken of the animal. The leech was restrained to moving in a small tank or petri dish, where the water level can be varied. An automatic system based on color processing tracked the colored beads over time, allowing real-time monitoring of the leech motion for several hours. At the end of each experimental session, six time series (2 for each bead) describing the leech body motion were obtained. A statistical analysis based on the speed and frequency content of bead motion indicated the existence of several stereotypical patterns of motion, corresponding to different leech behaviors. The identified patterns corresponded to swimming, pseudo-swimming, crawling, exploratory behavior, stationary states, abrupt movements, and combinations of these behaviors. The automatic characterization of leech behavior demonstrated here represents an important step toward understanding leech behavior and its properties. This method can be used to characterize the behavior of other invertebrates and also for some small vertebrates.

  16. Statistics of decision making in the leech.

    PubMed

    Garcia-Perez, Elizabeth; Mazzoni, Alberto; Zoccolan, Davide; Robinson, Hugh P C; Torre, Vincent

    2005-03-09

    Animals continuously decide among different behaviors, but, even in invertebrates, the mechanisms underlying choice and decision are unknown. In this article, leech spontaneous behavior was tracked and quantified for up to 12 h. We obtained a statistical characterization, in space and time domains, of the decision processes underlying selection of behavior in the leech. We found that the spatial distribution of leech position in a uniform environment is isotropic (the same in all directions), but this isotropy is broken in the presence of localized external stimuli. In the time domain, transitions among behaviors can be described by a Markov process, the structure of which (allowed states and transitions) is highly conserved across individuals. Finally, a wide range of recurrent, deterministic motifs was identified in the apparently irregular and unstructured exploratory behavior. These results provide a rigorous description of the inner dynamics that control the spontaneous and continuous flow of behavioral decisions in the leech.

  17. The Effect of Anchoring Sutures on Medicinal Leech Mortality

    PubMed Central

    Davila, Victor J.; Hoppe, Ian C.; Landi, Rocco; Ciminello, Frank S.

    2009-01-01

    Objective: The implementation of leech therapy for surgical flaps is not always logistically easy or comfortable for patients or healthcare providers. We examine different methods of placing sutures in the medicinal leech, Hirudo medicinalis, to make the implementation of leech therapy easier. Methods: Sixteen leeches were randomly divided into 3 groups: a control group, a deep anchoring suture group, and a superficial anchoring suture group. The leeches were observed to determine if either of these methods had an adverse effect on survival compared with the control group. Results: No difference in survival time was observed across the different groups. Conclusion: The placement of anchoring sutures in leeches can ease the implementation of leech therapy by allowing for greater control of the leeches and thus increased patient comfort. PMID:19696874

  18. Building exploration with leeches Hirudo verbana.

    PubMed

    Adamatzky, Andrew; Sirakoulis, Georgios Ch

    2015-08-01

    Safe evacuation of people from building and outdoor environments, and search and rescue operations, always will remain actual in course of all socio-technological developments. Modern facilities offer a range of automated systems to guide residents towards emergency exists. The systems are assumed to be infallible. But what if they fail? How occupants not familiar with a building layout will be looking for exits in case of very limited visibility where tactile sensing is the only way to assess the environment? Analogous models of human behaviour, and socio-dynamics in general, are provided to be fruitful ways to explore alternative, or would-be scenarios. Crowd, or a single person, dynamics could be imitated using particle systems, reaction-diffusion chemical medium, electro-magnetic fields, or social insects. Each type of analogous model offer unique insights on behavioural patterns of natural systems in constrained geometries. In this particular paper we have chosen leeches to analyse patterns of exploration. Reasons are two-fold. First, when deprived from other stimuli leeches change their behavioural modes in an automated regime in response to mechanical stimulation. Therefore leeches can give us invaluable information on how human beings might behave under stress and limited visibility. Second, leeches are ideal blueprints of future soft-bodied rescue robots. Leeches have modular nervous circuitry with a rich behavioral spectrum. Leeches are multi-functional, fault-tolerant with autonomous inter-segment coordination and adaptive decision-making. We aim to answer the question: how efficiently a real building can be explored and whether there any dependencies on the pathways of exploration and geometrical complexity of the building. In our case studies we use templates made on the floor plan of real building. Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.

  19. Reproductive strategies of the kangaroo leech, Marsupiobdella africana (Glossiphoniidae)

    PubMed Central

    Kruger, Natasha; Du Preez, Louis

    2015-01-01

    The Kangaroo Leech, Marsupiobdella africana, is a hermaphroditic organism, with insemination taking place by the planting of a spermatophore on another leech. Spermatophores are mostly planted on the anterior of the recipient leech, but not always. Several spermatophores may be planted by different leeches on a single recipient. The spermatophore consists of two side by side lobes. Within minutes from planting of the spermatophore, the contents are squeezed out and into the body of the recipient. Sperm are believed to find the way to the ova by following chemical cues. Kangaroo Leeches display advanced parental care by transferring fertilized eggs from the reproductive opening to a brood pouch on the ventral side. Fully developed leeches may copulate after detaching from the amphibian host Xenopus laevis, or from the Cape River Crab Potamonautes perlatus with which it maintains a phoretic association. PMID:25830114

  20. External decontamination of wild leeches with hypochloric acid

    PubMed Central

    Aydin, Atakan; Nazik, Hasan; Kuvat, Samet Vasfi; Gurler, Nezahat; Ongen, Betigul; Tuncer, Serdar; Hocaoglu, Emre; Kesim, Sinan Nur

    2004-01-01

    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

  1. The eyes have it: long-distance dispersal by an intraorbital leech parasite of birds.

    PubMed

    Siddall, Mark E; Rood-Goldman, Rebecca; Barrio, Amalie; Barboutis, Christos

    2013-12-01

    A leech was found parasitizing the ocular orbit of a common redstart captured during a faunistic survey of Antikythira in the Aegean Sea during the spring migration of 2012. Morphological and molecular characterizations placed the leech in the mucous-membrane specific leech family Praobdellidae and definitively as the species Parapraobdella lineata. This is the first record of any leech parasitizing a passerine bird, Phoenicurus phoenicurus , and the first of a praobdellid leech on any avian host.

  2. 33 CFR 207.330 - Mississippi River between Winnibigoshish and Pokegama dams, Leech River between outlet of Leech...

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 33 Navigation and Navigable Waters 3 2011-07-01 2011-07-01 false Mississippi River between Winnibigoshish and Pokegama dams, Leech River between outlet of Leech Lake and Mississippi River, and Pokegama... OF THE ARMY, DEPARTMENT OF DEFENSE NAVIGATION REGULATIONS § 207.330 Mississippi River between...

  3. 33 CFR 207.330 - Mississippi River between Winnibigoshish and Pokegama dams, Leech River between outlet of Leech...

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 33 Navigation and Navigable Waters 3 2013-07-01 2013-07-01 false Mississippi River between Winnibigoshish and Pokegama dams, Leech River between outlet of Leech Lake and Mississippi River, and Pokegama... OF THE ARMY, DEPARTMENT OF DEFENSE NAVIGATION REGULATIONS § 207.330 Mississippi River between...

  4. 33 CFR 207.330 - Mississippi River between Winnibigoshish and Pokegama dams, Leech River between outlet of Leech...

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 33 Navigation and Navigable Waters 3 2012-07-01 2012-07-01 false Mississippi River between Winnibigoshish and Pokegama dams, Leech River between outlet of Leech Lake and Mississippi River, and Pokegama... OF THE ARMY, DEPARTMENT OF DEFENSE NAVIGATION REGULATIONS § 207.330 Mississippi River between...

  5. 33 CFR 207.330 - Mississippi River between Winnibigoshish and Pokegama dams, Leech River between outlet of Leech...

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 33 Navigation and Navigable Waters 3 2010-07-01 2010-07-01 false Mississippi River between Winnibigoshish and Pokegama dams, Leech River between outlet of Leech Lake and Mississippi River, and Pokegama... OF THE ARMY, DEPARTMENT OF DEFENSE NAVIGATION REGULATIONS § 207.330 Mississippi River between...

  6. 33 CFR 207.330 - Mississippi River between Winnibigoshish and Pokegama dams, Leech River between outlet of Leech...

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 33 Navigation and Navigable Waters 3 2014-07-01 2014-07-01 false Mississippi River between Winnibigoshish and Pokegama dams, Leech River between outlet of Leech Lake and Mississippi River, and Pokegama... OF THE ARMY, DEPARTMENT OF DEFENSE NAVIGATION REGULATIONS § 207.330 Mississippi River between...

  7. A rare case of adverse effects caused by leech bite

    PubMed Central

    Lok, Ugur; Bozkurt, Selim; Okur, Mehmet; Gulacti, Umut; Hatipoglu, Sinan

    2013-01-01

    Patient: Female, 42 Final Diagnosis: Leech bite Symptoms: — Medication: — Clinical Procedure: — Specialty: Infectious diseases Objective: Unusual clinical course Background: Hirudo medicinalis (H. Medicinalis), also referred to as the medical leech, not only has been used by alternative medicine since ancient times, but it has also been used in modern medical care in the last century. This report introduces and discusses some rare complications from leech bite, beyond external skin hemorrhage. Case Report: A 42-year-old woman was referred to our emergency room with painful and itchy lesions on her feet and legs. Because of her knee and leg pain, about 14 leeches were adhered onto both her legs. There were left rubor, tumor and some other areas of itching on leech-adhered regions. In physical examination, the right popliteal region and the same leg posterior area lesions were erythematous, edematous, and plaque-like shaped, as well as ecchymotic and hemorrhagic areas at the centre of the lesions. Conclusions: In this article we present the case of a woman referred to our emergency department with skin findings related to a leech bite. We could not find any report about regional subcutaneous hematoma and cutaneous reactions in the literature. H. medicinalis may cause ecchymosis and various skin lesions, not only external skin bleeding. Physicians should keep in mind leech bite in patients with itchy lesions on the calves, knees, and feet. PMID:23826465

  8. Developmental biology of the leech Helobdella

    PubMed Central

    WEISBLAT, DAVID A.; KUO, DIAN-HAN

    2015-01-01

    Glossiphoniid leeches of the genus Helobdella provide experimentally tractable models for studies in evolutionary developmental biology (Evo-Devo). Here, after a brief rationale, we will summarize our current understanding of Helobdella development and highlight the near term prospects for future investigations, with respect to the issues of: D quadrant specification; the transition from spiral to bilaterally symmetric cleavage; segmentation, and the connections between segmental and non-segmental tissues; modifications of BMP signaling in dorsoventral patterning and the O-P equivalence group; germ line specification and genome rearrangements. The goal of this contribution is to serve as a summary of, and guide to, published work. PMID:25690960

  9. Leeches--the good, the bad and the wiggly.

    PubMed

    O'Dempsey, Tim

    2012-11-01

    Leeches have been making a remarkable come-back in medical practice in recent years, particularly in reconstructive surgery. This paper opens with a brief review of the historical aspects of the medicinal use of leeches and their recent renaissance in modern medical practice. We then embark on a journey through the murky waters of leech infestation and the potentially life-threatening complications arising from close encounters with these extraordinary creatures. Finally, we get to grips with the tricky issue of safely apprehending and removing uninvited hangers-on.

  10. Micromere lineages in the glossiphoniid leech Helobdella

    NASA Technical Reports Server (NTRS)

    Huang, Francoise Z.; Kang, Dongmin; Ramirez-Weber, Felipe-Andres; Bissen, Shirley T.; Weisblat, David A.

    2002-01-01

    In leech embryos, segmental mesoderm and ectoderm arise from teloblasts by lineages that are already relatively well characterized. Here, we present data concerning the early divisions and the definitive fate maps of the micromeres, a group of 25 small cells that arise during the modified spiral cleavage in leech (Helobdella robusta) and contribute to most of the nonsegmental tissues of the adult. Three noteworthy results of this work are as follows. (1) The c"' and dm' clones (3d and 3c in traditional nomenclature) give rise to a hitherto undescribed network of fibers that run from one end of the embryo to the other. (2) The clones of micromeres b" and b"' (2b and 3b in traditional nomenclature) die in normal development; the b" clone can be rescued to assume the normal c" fate if micromere c" or its clone are ablated in early development. (3) Two qualitative differences in micromere fates are seen between H. robusta (Sacramento) and another Helobdella sp. (Galt). First, in Helobdella sp. (Galt), the clone of micromere b" does not normally die, and contributes a subset of the cells arising exclusively from c" in H. robusta (Sacramento). Second, in Helobdella sp. (Galt), micromere c"' makes no definitive contribution, whereas micromere dm' gives rise to cells equivalent to those arising from c"' and dm' in H. robusta (Sacramento).

  11. Calcium dynamics and compartmentalization in leech neurons.

    PubMed

    Andjelic, Sofija; Torre, Vincent

    2005-12-01

    Calcium dynamics in leech neurons were studied using a fast CCD camera. Fluorescence changes (DeltaF/F) of the membrane impermeable calcium indicator Oregon Green were measured. The dye was pressure injected into the soma of neurons under investigation. DeltaF/F caused by a single action potential (AP) in mechanosensory neurons had approximately the same amplitude and time course in the soma and in distal processes. By contrast, in other neurons such as the Anterior Pagoda neuron, the Annulus Erector motoneuron, the L motoneuron, and other motoneurons, APs evoked by passing depolarizing current in the soma produced much larger fluorescence changes in distal processes than in the soma. When APs were evoked by stimulating one distal axon through the root, DeltaF/F was large in all distal processes but very small in the soma. Our results show a clear compartmentalization of calcium dynamics in most leech neurons in which the soma does not give propagating action potentials. In such cells, the soma, while not excitable, can affect information processing by modulating the sites of origin and conduction of AP propagation in distal excitable processes.

  12. Micromere lineages in the glossiphoniid leech Helobdella

    NASA Technical Reports Server (NTRS)

    Huang, Francoise Z.; Kang, Dongmin; Ramirez-Weber, Felipe-Andres; Bissen, Shirley T.; Weisblat, David A.

    2002-01-01

    In leech embryos, segmental mesoderm and ectoderm arise from teloblasts by lineages that are already relatively well characterized. Here, we present data concerning the early divisions and the definitive fate maps of the micromeres, a group of 25 small cells that arise during the modified spiral cleavage in leech (Helobdella robusta) and contribute to most of the nonsegmental tissues of the adult. Three noteworthy results of this work are as follows. (1) The c"' and dm' clones (3d and 3c in traditional nomenclature) give rise to a hitherto undescribed network of fibers that run from one end of the embryo to the other. (2) The clones of micromeres b" and b"' (2b and 3b in traditional nomenclature) die in normal development; the b" clone can be rescued to assume the normal c" fate if micromere c" or its clone are ablated in early development. (3) Two qualitative differences in micromere fates are seen between H. robusta (Sacramento) and another Helobdella sp. (Galt). First, in Helobdella sp. (Galt), the clone of micromere b" does not normally die, and contributes a subset of the cells arising exclusively from c" in H. robusta (Sacramento). Second, in Helobdella sp. (Galt), micromere c"' makes no definitive contribution, whereas micromere dm' gives rise to cells equivalent to those arising from c"' and dm' in H. robusta (Sacramento).

  13. NEW HOST DATA FOR THE LEECH OLIGOBDELLA BIANNULATE (EUHIRUDINEA: GLOSSIPHONIIDAE)

    EPA Science Inventory

    The leech Oligobdella biannulata is a relatively rare species, endemic to mountain streams of the Southern Blue Ridge Physiographic Zone, exclusive of Virginia. Oligobdella biannulata was originally thought to be host specific to Desmognathus quadramaculatus. However, the host ...

  14. Leech infestation and its association with water drinking habits.

    PubMed

    Raza, Syed Nusrat; Shabbir, Syed Muhammad Asad

    2006-03-01

    To assess the common presentations of leech infestation in leech endemic areas of Pakistan and to establish its association with unsafe water drinking habits of the individual. A cohort study. CMH, Kohat, between 1st February 1997 and 30th April 2002. Fourteen patients with leech infestation who reported to ENT department of CMH, Kohat, during the above period were selected for study. A specific comparison of their water drinking habits was made with a control group of 42 normal individuals who had come from the same leech endemic area as the first group and belonged to the same socioeconomic class. Twelve cases (83.4%) in the first group had unsafe water drinking habit i.e. they drank water directly from marshes without seeing its contents substituting cupped palm of hand for a drinking cup. This was in contrast to the control group where only 6 individuals (14%) gave a history of unsafe water drinking habits. Nose was the most common ENT site of leech infestation (71%) with epistaxis being the most prominent symptom. Other sites included hypopharynx (14%), nasopharynx (7 %) and oropharynx (7 %). All the 14 cases with leech infestation were males, 26.09 years being the mean age. The association between patient s unsafe water drinking habits and leech infestation in ENT region was statistically proved with odds ratio being 36. Epistaxis or any other related symptom must be taken with suspicion in leech endemic area. This condition is closely related to unsafe water drinking habits, therefore, the incidence of this infestation can be significantly reduced by educating the individuals to adopt safe water drinking habits.

  15. Intermediate filament genes as differentiation markers in the leech Helobdella.

    PubMed

    Kuo, Dian-Han; Weisblat, David A

    2011-10-01

    The intermediate filament (IF) cytoskeleton is a general feature of differentiated cells. Its molecular components, IF proteins, constitute a large family including the evolutionarily conserved nuclear lamins and the more diverse collection of cytoplasmic intermediate filament (CIF) proteins. In vertebrates, genes encoding CIFs exhibit cell/tissue type-specific expression profiles and are thus useful as differentiation markers. The expression of invertebrate CIFs, however, is not well documented. Here, we report a whole-genome survey of IF genes and their developmental expression patterns in the leech Helobdella, a lophotrochozoan model for developmental biology research. We found that, as in vertebrates, each of the leech CIF genes is expressed in a specific set of cell/tissue types. This allows us to detect earliest points of differentiation for multiple cell types in leech development and to use CIFs as molecular markers for studying cell fate specification in leech embryos. In addition, to determine the feasibility of using CIFs as universal metazoan differentiation markers, we examined phylogenetic relationships of IF genes from various species. Our results suggest that CIFs, and thus their cell/tissue-specific expression patterns, have expanded several times independently during metazoan evolution. Moreover, comparing the expression patterns of CIF orthologs between two leech species suggests that rapid evolutionary changes in the cell or tissue specificity of CIFs have occurred among leeches. Hence, CIFs are not suitable for identifying cell or tissue homology except among very closely related species, but they are nevertheless useful species-specific differentiation markers.

  16. On exploration of geometrically constrained space by medicinal leeches Hirudo verbana.

    PubMed

    Adamatzky, Andrew

    2015-04-01

    Leeches are fascinating creatures: they have simple modular nervous circuitry yet exhibit a rich spectrum of behavioural modes. Leeches could be ideal blue-prints for designing flexible soft robots which are modular, multi-functional, fault-tolerant, easy to control, capable for navigating using optical, mechanical and chemical sensorial inputs, have autonomous inter-segmental coordination and adaptive decision-making. With future designs of leech-robots in mind we study how leeches behave in geometrically constrained spaces. Core results of the paper deal with leeches exploring a row of rooms arranged along a narrow corridor. In laboratory experiments we find that rooms closer to ends of the corridor are explored by leeches more often than rooms in the middle of the corridor. Also, in series of scoping experiments, we evaluate leeches capabilities to navigating in mazes towards sources of vibration and chemo-attraction. We believe our results lay foundation for future developments of robots mimicking behaviour of leeches.

  17. Nonassociative learning in the leech Hirudo medicinalis.

    PubMed

    Zaccardi, M L; Traina, G; Cataldo, E; Brunelli, M

    2001-11-29

    In the present study we examined nonassociative learning of the induction of swimming which was evoked by weak electrical stimulation in the leech Hirudo medicinalis. The behavioural response to stimuli applied repeatedly to the body wall at an inter-trial interval (ITI) of 1 min decreased, eventually ceased, and then recovered spontaneously. More rapid reduction of the behavioural response occurred in repeated training sessions. This decrement of response conformed to the operational definition of habituation. Moreover, a noxious stimulus (i.e. brushing on the skin) facilitated the decremented response (dishabituation). In addition, we compared response decrement in naive animals with decrement in dishabituated and in sensitized animals. The analysis of the best fitting functions of the habituation, the habituation of dishabituation and the habituation of sensitization revealed interesting differences in these processes.

  18. [Review on scientific connotation of leech processed under high temperature].

    PubMed

    Ma, Li; Ma, Lin; Wang, Shu-bin; Wang, Xuan; Yue, Lin

    2015-10-01

    Animal medicines mainly contain protein which was organic molecule with quaternary structure and had the property of thermal denaturation. When suffering from heat for a consistent time, the native conformation of protein would be destroyed. After denaturation the biological activity of protein will lose and some physicochemical and biochemical properties will be changed. Leech was a classical animal medicine in the views of traditional Chinese medicine (TCM) which had the functions of breaking stagnant and eliminating blood stasis. In the usage history, it was processed for a long time. No matter stir-frying leech with talc powder embodied in Chinese Pharmacopoeia or stir-baking with wine as a distinctive method in Beijing district, the process procedure was basically performed under high temperature. The purposes and intentions of process are mostly limited to technology conditions at specific historical period. In this article, based on existing processing procedure and its character of Leech, the changes of active components and pharmacological activities before and after processing under high temperature were summarized. The results demonstrate that the protein of leech would be denaturated; some active peptide such as hirudin were partly or totally destroyed; some toxic mineral elements, such as Pb, Hg, Cd, were decreased; at the same time, heating can promote some chemical components transforming into hypoxanthine which had the function of antihypertensive, antiasthmatic and antalgic. Consequently, after processed under high temperature, the purpose of decreasing toxicity and alleviating the strong property was achieved. Pharmacological changes of leech processed under high temperature were mainly manifested in the anticoagulant and antithrombotic activity, etc. Based on current processing research status about animal medicine leech, future research methods and directions on scientific connotation of leech processed under high temperature were put forward in

  19. Poly-paraphyly of Hirudinidae: many lineages of medicinal leeches

    PubMed Central

    Phillips, Anna J; Siddall, Mark E

    2009-01-01

    Background Medicinal leeches became infamous for their utility in bloodletting popularized in the 19th century, and have seen a recent resurgence in post-operative treatments for flap and replantation surgeries, and in terms of characterization of salivary anticoagulants. Notorious throughout the world, the quintessential leech family Hirudinidae has been taken for granted to be monophyletic, as has the non-bloodfeeding family Haemopidae. Results This study is the first to evaluate molecular evidence from hirudinid and haemopid leeches in a manner that encompasses the global scope of their taxonomic distributions. We evaluated the presumed monophyly of the Hirudinidae and assessed previous well-accepted classification schemes. The Hirudinidae were found not to be monophyletic, falling instead into two distinct and unrelated clades. Members of the non-bloodfeeding family Haemopidae were scattered throughout the tree and among traditional hirudinid genera. A combination of nuclear 18S rDNA and 28S rDNA with mitochondrial 12S rDNA and cytochrome c oxidase I were analyzed with Parsimony and with Bayesian methods. Conclusion The family Hirudinidae must be refined to include only the clade containing Hirudo medicinalis (European medicinal leech) and related leeches irrespective of bloodfeeding behavior. A second clade containing Macrobdella decora (North American medicinal leech) and its relatives may yet be recognized in Semiscolecidae in order to avoid paraphyly. The African distribution of species from each of the divergent hirudinid clades suggests that a deep divergence took place in the history of the medicinal leeches hundreds of millions of years ago. PMID:19811660

  20. The leech and the physician: biology, etymology, and medical practice with Hirudinea medicinalis.

    PubMed

    Mory, R N; Mindell, D; Bloom, D A

    2000-07-01

    The history of the word "leech" and the practice of leeching reveal interconnected social histories. We give the linguistic and medical histories of the word, and explore its biology and clinical history. Our historical account extends from the earliest known record of leeching to current research. Despite historical variation in its reputation as a therapeutic technique, leeching remains useful today in a number of applications. Further investigation may well disclose even more uses for the leech, particularly for its enzymes with anesthetic, anticoagulant, and antimetastatic properties.

  1. Bed bugs, leeches and hookworm larvae in the skin.

    PubMed

    Heukelbach, Jorg; Hengge, Ulrich R

    2009-01-01

    Bed bugs, leeches, and hookworm-related cutaneous larva migrans are skin infestations that are usually considered of minor importance because they produce discomfort rather than cause or transmit disease. Bed bugs have been increasing tremendously in high-income countries in recent years, causing distress to affected individuals and economic loss. Infestation by land leeches causes mainly unpleasant skin reactions, whereas infestation by aquatic leeches may be more dangerous, leading to anemia and in severe cases, to death. Cutaneous larva migrans produces an intense pruritus that can be exasperating for the patient and cause sleep disturbance. An overview is given of these three infestations with a discussion of the causative agents, transmission, clinical manifestations, diagnosis, and treatment.

  2. Spontaneous electrical activity and behavior in the leech hirudo medicinalis.

    PubMed

    Garcia-Perez, Elizabeth; Mazzoni, Alberto; Torre, Vincent

    2007-01-01

    In the absence of external stimuli, animals explore the environment by performing irregular movements, but the neuronal mechanisms underlying this arrhythmic motion are largely unknown. In this paper, we studied the relationship between the spontaneous neuronal activity in the leech (Hirudo medicinalis) and its behavior. We analyzed the electrical activity of isolated ganglia, chains of two connected ganglia, and semi-intact preparations. The spontaneous electrical activity in ganglia was characterized by the occurrence of irregular bursts of spikes with variable duration and size. Properties of these bursts were modified by synaptic inputs arriving from the neighboring ganglia and from the two primitive brains located in the head and tail. In fact, in semi-intact preparations, unusually large bursts of spikes occurring spontaneously were recorded and caused the leech to move even in the absence of any external sensory stimulation. These large bursts appear to act as internal triggers controlling the spontaneous leech behavior and determining the duration of stereotypical motor patterns.

  3. Leech therapy for epidermoid cysts and review of the literature

    PubMed Central

    Rasi, Abbas; Faghihi, Alireza; Jalali, Mirhadi Aziz; Zamanian, Abbas; Ghaffarpour, Gholamhossein

    2014-01-01

    Hirudo medicinalis sucks blood directly through the external mammalian skin. We recently observed a healthy 64-year-old Iranian man, who presented with numerous asymptomatic multilobular oval-to-round well-defined 0.5 to 1.5 cm cystic lesions with central umbilication (central black eschar) over the upper portion of his chest. We made the diagnosis of epidermoid cyst, giant comedone and leech bite on the basis of the constellation of clinical features. The patient was treated with oral ciprofloxacin at a dose of 2 g daily, and 2% topical erythromycin solution. Despite improvement, the evidence of cystic lesions persisted. There was no history of similar lesions in any other family member. There was no history of trauma. The patient was not using any topical or systemic medication. Two weeks before his visit, he had a history of leech therapy under the supervision of a general practitioner. His medical history was significant for leech therapy of the lesions, five days previously. He was followed up for another two weeks and after disappearance of the inflammation, with the patient under local anesthesia, the well-circumscribed mass was completely evacuated with a sharp curette and comedone extractor. The patient was subsequently lost to follow-up. Conclusion: Considering the efficacy of leeches, it would be favorable to breed a germ-free leech. In Iran, the use of the leeches in surgery, in recent years, has been infrequent. It appears that the positive effects of this ancient remedy may now be explained through scientific methods, promising potentially even more uses of this admirable creature in medicine. PMID:24804186

  4. Bronchial Leech Infestation in a 15-Year-Old Female

    PubMed Central

    Moslehi, Mohammad Ashkan; Imanieh, Mohammad Hadi

    2016-01-01

    Foreign body aspiration (FBA) is a common incidence in young children. Leeches are rarely reported as FBA at any age. This study describes a 15-year-old female who presented with hemoptysis, hematemesis, coughs, melena, and anemia seven months prior to admission. Chest X-ray showed a round hyperdensity in the right lower lobe. A chest computed tomography (CT) demonstrated an area of consolidation and surrounding ground glass opacities in the right lower lobe. Hematological investigations revealed anemia. Finally, bronchoscopy was performed and a 5 cm leech was found within the right B7-8 bronchus and removed by forceps and a Dormia basket. PMID:27752380

  5. Bronchial Leech Infestation in a 15-Year-Old Female.

    PubMed

    Moslehi, Mohammad Ashkan; Imanieh, Mohammad Hadi; Adib, Ali

    2016-01-01

    Foreign body aspiration (FBA) is a common incidence in young children. Leeches are rarely reported as FBA at any age. This study describes a 15-year-old female who presented with hemoptysis, hematemesis, coughs, melena, and anemia seven months prior to admission. Chest X-ray showed a round hyperdensity in the right lower lobe. A chest computed tomography (CT) demonstrated an area of consolidation and surrounding ground glass opacities in the right lower lobe. Hematological investigations revealed anemia. Finally, bronchoscopy was performed and a 5 cm leech was found within the right B7-8 bronchus and removed by forceps and a Dormia basket.

  6. Ribosomal RNA characterization in the leech Hirudo medicinalis.

    PubMed

    Macchi, M; Durante, M; Bernardi, R; Traina, G

    2008-09-01

    In the present study the ribosomal RNA of the leech Hirudo medicinalis has been characterized at the aim of identifying possible analogies with other invertebrates. Upon electrophoresis on denaturating gels, ribosomal RNA fraction of H. medicinalis exhibited a remarkable thermal instability by dissociating into two hydrogen-bonded components when heated at 60 degrees C, at variance with the behaviour of the rat rRNA, which does not show this process. This result suggests a functional role in leech ribosome organisation that requires deeper structural studies.

  7. Evaluation of performance characteristics of the medicinal leech (Hirudo medicinalis) for the treatment of venous congestion.

    PubMed

    Conforti, Michael L; Connor, Nadine P; Heisey, Dennis M; Hartig, Gregory K

    2002-01-01

    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.

  8. Entrainment of leech swimming activity by the ventral stretch receptor.

    PubMed

    Yu, Xintian; Friesen, W Otto

    2004-11-01

    Rhythmic animal movements originate in CNS oscillator circuits; however, sensory inputs play an important role in shaping motor output. Our recent studies demonstrated that leeches with severed nerve cords swim with excellent coordination between the two ends, indicating that sensory inputs are sufficient for maintaining intersegmental coordination. In this study, we examined the neuronal substrates that underlie intersegmental coordination via sensory mechanisms. Among the identified sensory neurons in the leech, we found the ventral stretch receptor (VSR) to be the best candidate for our study because of its sensitivity to tension in longitudinal muscle. Our experiments demonstrate that (1) the membrane potential of the VSR is depolarized during swimming and oscillates with an amplitude of 1.5-5.0 mV, (2) rhythmic currents injected into the VSR can entrain ongoing swimming over a large frequency range (0.9-1.8 Hz), and (3) large current pulses injected into the VSR shift the phase of the swimming rhythm. These results suggest that VSRs play an important role in generating and modulating the swim rhythm. We propose that coordinated swimming in leech preparations with severed nerve cords results from mutual entrainment between the two ends of the leech mediated by stretch receptors.

  9. Neuro-immune lessons from an annelid: The medicinal leech.

    PubMed

    Tasiemski, Aurélie; Salzet, Michel

    2017-01-01

    An important question that remains unanswered is how the vertebrate neuroimmune system can be both friend and foe to the damaged nervous tissue. Some of the difficulty in obtaining responses in mammals probably lies in the conflation in the central nervous system (CNS), of the innate and adaptive immune responses, which makes the vertebrate neuroimmune response quite complex and difficult to dissect. An alternative strategy for understanding the relation between neural immunity and neural repair is to study an animal devoid of adaptive immunity and whose CNS is well described and regeneration competent. The medicinal leech offers such opportunity. If the nerve cord of this annelid is crushed or partially cut, axons grow across the lesion and conduction of signals through the damaged region is restored within a few days, even when the nerve cord is removed from the animal and maintained in culture. When the mammalian spinal cord is injured, regeneration of normal connections is more or less successful and implies multiple events that still remain difficult to resolve. Interestingly, the regenerative process of the leech lesioned nerve cord is even more successful under septic than under sterile conditions suggesting that a controlled initiation of an infectious response may be a critical event for the regeneration of normal CNS functions in the leech. Here are reviewed and discussed data explaining how the leech nerve cord sensu stricto (i.e. excluding microglia and infiltrated blood cells) recognizes and responds to microbes and mechanical damages. Copyright © 2016 Elsevier Ltd. All rights reserved.

  10. A leech capable of surviving exposure to extremely low temperatures.

    PubMed

    Suzuki, Dai; Miyamoto, Tomoko; Kikawada, Takahiro; Watanabe, Manabu; Suzuki, Toru

    2014-01-01

    It is widely considered that most organisms cannot survive prolonged exposure to temperatures below 0°C, primarily because of the damage caused by the water in cells as it freezes. However, some organisms are capable of surviving extreme variations in environmental conditions. In the case of temperature, the ability to survive subzero temperatures is referred to as cryobiosis. We show that the ozobranchid leech, Ozobranchus jantseanus, a parasite of freshwater turtles, has a surprisingly high tolerance to freezing and thawing. This finding is particularly interesting because the leach can survive these temperatures without any acclimation period or pretreatment. Specifically, the leech survived exposure to super-low temperatures by storage in liquid nitrogen (-196°C) for 24 hours, as well as long-term storage at temperatures as low as -90°C for up to 32 months. The leech was also capable of enduring repeated freeze-thaw cycles in the temperature range 20°C to -100°C and then back to 20°C. The results demonstrated that the novel cryotolerance mechanisms employed by O. jantseanus enable the leech to withstand a wider range of temperatures than those reported previously for cryobiotic organisms. We anticipate that the mechanism for the observed tolerance to freezing and thawing in O. jantseanus will prove useful for future studies of cryopreservation.

  11. Serotonin modulates muscle function in the medicinal leech Hirudo verbana

    PubMed Central

    Gerry, Shannon P.; Ellerby, David J.

    2011-01-01

    The body wall muscles of sanguivorous leeches power mechanically diverse behaviours: suction feeding, crawling and swimming. These require longitudinal muscle to exert force over an extremely large length range, from 145 to 46 per cent of the mean segmental swimming length. Previous data, however, suggest that leech body wall muscle has limited capacity for force production when elongated. Serotonin (5-HT) alters the passive properties of the body wall and stimulates feeding. We hypothesized that 5-HT may also have a role in allowing force production in elongated muscle by changing the shape of the length–tension relationship (LTR). LTRs were measured from longitudinal muscle strips in vitro in physiological saline with and without the presence of 10 µM 5-HT. The LTR was much broader than previously measured for leech muscle. Rather than shifting the LTR, 5-HT reduced passive muscle tonus and increased active stress at all lengths. In addition to modulating leech behaviour and passive mechanical properties, 5-HT probably enhances muscle force and work production during locomotion and feeding. PMID:21561963

  12. Cannibalism in a population of the medicinal leech (Hirudo medicinalis L.).

    PubMed

    Kutschera, U; Roth, M

    2005-01-01

    Medicinal leeches (Hirudo medicinalis L.) were maintained in large ponds in a commercial leech farm at Biebertal, Germany. The feeding of hungry adult leeches was performed on representative individuals that were placed on cloth soaked with mammalian blood obtained from a local butchery (pig, Sus scrofa). In a second set of experiments, cane toads (Bufo marinus) were used as host organisms. The leeches rapidly attached to the toads, explored the body and sucked blood. After feeding, the fully engorged leeches were placed into the pond or an aquarium. In this artificial habitat, the satiated leeches were attacked by hungry conspecifics, sucked off and killed. This observation demonstrates that H. medicinalis must be classified as a cannibalistic annelid.

  13. Presence of the leech Placobdella costata in the south of the Iberian Peninsula.

    PubMed

    Romero, David; Duarte, Jesús; Narváez-Ledesma, Lucía; Farfán, Miguel Angel; Real, Raimundo

    2014-06-01

    Placobdella costata is a leech specific to freshwater turtle Emys orbicularis. Both genera are native to North America and have co-evolved and undergone dispersion through the Palearctic. The leech is present throughout the Mediterranean area, always associated with E. orbicularis. Their only known presence in the Iberian Peninsula is in the north and center of the peninsula. Here we present the first description of the leech in southern Spain (Andalusia) in association with a small fragmented population of fresh-water turtles in which E. orbicularis and Mauremys leprosa coexist. Unusually, the leech was found attached to the carapace of a male M. leprosa.

  14. [Study on the processing of leech by FTIR and 2D-IR correlation spectroscopy].

    PubMed

    Li, Bing-Ning; Wu, Yan-Wen; Ouyang, Jie; Sun, Su-Qin; Chen, Shun-Cong

    2011-04-01

    The chemical differences of traditional Chinese medicine leech before and after processing were analyzed by FTIR and two-dimensional correlation infrared (2D-IR) spectroscopy. The result showed that the leech was high in protein, with characteristic peaks of amide I, II bands. Comparing the IR spectra of samples, the primary difference was that the characteristic peak of fresh leech was at 1 543 cm(-1), while that of crude and processed leech was at 1 535 cm(-1). A 2D-IR spectrum with heating perturbation was used to track the processing dynamics of leech In the 2D-IR correlation spectra, fresh leech exhibited stronger automatic peaks of the amide I and II bands than that of processed leech, which indicates that the protein components of the fresh leech were more sensitive to heat perturbation than the processed one. Moreover, the result of FTIR and 2D-IR correlation spectra validated that the 3-dimensional structure of protein was damaged and hydrogen bonds were broken after processing, which resulted in the inactivation of protein. The fatty acids and cholesterol components of leech were also oxidized in this process.

  15. An improved anti-leech mechanism based on session identifier

    NASA Astrophysics Data System (ADS)

    Zhang, Jianbiao; Zhu, Tong; Zhang, Han; Lin, Li

    2011-12-01

    With the rapid development of information technology and extensive requirement of network resource sharing, plenty of resource hotlinking phenomenons appear on the internet. The hotlinking problem not only harms the interests of legal websites but also leads to a great affection to fair internet environment. The anti-leech technique based on session identifier is highly secure, but the transmission of session identifier in plaintext form causes some security flaws. In this paper, a proxy hotlinking technique based on session identifier is introduced firstly to illustrate these security flaws; next, this paper proposes an improved anti-leech mechanism based on session identifier, the mechanism takes the random factor as the core and detects hotlinking request using a map table that contains random factor, user's information and time stamp; at last the paper analyzes the security of mechanism in theory. The result reveals that the improved mechanism has the merits of simple realization, high security and great flexibility.

  16. An improved anti-leech mechanism based on session identifier

    NASA Astrophysics Data System (ADS)

    Zhang, Jianbiao; Zhu, Tong; Zhang, Han; Lin, Li

    2012-01-01

    With the rapid development of information technology and extensive requirement of network resource sharing, plenty of resource hotlinking phenomenons appear on the internet. The hotlinking problem not only harms the interests of legal websites but also leads to a great affection to fair internet environment. The anti-leech technique based on session identifier is highly secure, but the transmission of session identifier in plaintext form causes some security flaws. In this paper, a proxy hotlinking technique based on session identifier is introduced firstly to illustrate these security flaws; next, this paper proposes an improved anti-leech mechanism based on session identifier, the mechanism takes the random factor as the core and detects hotlinking request using a map table that contains random factor, user's information and time stamp; at last the paper analyzes the security of mechanism in theory. The result reveals that the improved mechanism has the merits of simple realization, high security and great flexibility.

  17. Linear and nonlinear measures predict swimming in the leech

    PubMed

    Cellucci; Brodfuehrer; Acera-Pozzi; Dobrovolny; Engler; Los; Thompson; Albano

    2000-10-01

    Stimulation of a trigger interneuron of an isolated nerve cord preparation of the medicinal leech, Hirudo medicinalis, sometimes leads to swimming; sometimes it does not. We investigate signals transmitted in the ventral cord of the leech after stimulation and seek quantitative measures that would make it possible to distinguish signals that predict swimming from those that do not. We find that a number of linear as well as nonlinear measures provide statistically significant distinctions between the two kinds of signals. The linear measures are the time dependence of (i) the standard deviation and (ii) the autocorrelation function at a small time delay. The nonlinear measures are (i) a measure of nonlinear predictability and (ii) the time dependence of a measure of the size of the embedded signal trajectory. Calculations using surrogate data suggest that the differences between the two classes of signals are dynamical as well as statistical.

  18. Proteinase inhibitors from the medicinal leech Hirudo medicinalis.

    PubMed

    Baskova, I P; Zavalova, L L

    2001-07-01

    The medicinal leech Hirudo medicinalis produces various types of proteinase inhibitors: bdellins (inhibitors of trypsin, plasmin, and acrosin), hirustasin (inhibitor of tissue kallikrein, trypsin, alpha-chymotrypsin, and granulocyte cathepsin G), tryptase inhibitor, eglins (inhibitors of alpha-chymotrypsin, subtilisin, and chymasin and the granulocyte proteinases elastase and cathepsin G), inhibitor of factor Xa, hirudin (thrombin inhibitor), inhibitor of carboxypeptidase, and inhibitor of complement component C1s. This review summarizes data on their primary and tertiary structures, action mechanisms, and biological activities.

  19. Spontaneous Electrical Activity and Behavior in the Leech Hirudo Medicinalis

    PubMed Central

    Garcia-Perez, Elizabeth; Mazzoni, Alberto; Torre, Vincent

    2007-01-01

    In the absence of external stimuli, animals explore the environment by performing irregular movements, but the neuronal mechanisms underlying this arrhythmic motion are largely unknown. In this paper, we studied the relationship between the spontaneous neuronal activity in the leech (Hirudo medicinalis) and its behavior. We analyzed the electrical activity of isolated ganglia, chains of two connected ganglia, and semi-intact preparations. The spontaneous electrical activity in ganglia was characterized by the occurrence of irregular bursts of spikes with variable duration and size. Properties of these bursts were modified by synaptic inputs arriving from the neighboring ganglia and from the two primitive brains located in the head and tail. In fact, in semi-intact preparations, unusually large bursts of spikes occurring spontaneously were recorded and caused the leech to move even in the absence of any external sensory stimulation. These large bursts appear to act as internal triggers controlling the spontaneous leech behavior and determining the duration of stereotypical motor patterns. PMID:18958236

  20. Leeches as Sensor-bioindicators of River Contamination by PCBs

    PubMed Central

    Macova, Stanislava; Harustiakova, Danka; Kolarova, Jitka; Machova, Jana; Zlabek, Vladimir; Vykusova, Blanka; Randak, Tomas; Velisek, Josef; Poleszczuk, Gorzyslaw; Hajslova, Jana; Pulkrabova, Jana; Svobodova, Zdenka

    2009-01-01

    The aim of the study was to evaluate the use of leeches of the genus Erpobdella as a means of assessing polychlorinated biphenyl contamination of watercourses. The River Skalice, heavily contaminated with PCBs, was selected as a model. The source of contamination was a road gravel processing factory in Rožmitál pod Třemšínem from which an estimated 1 metric ton of PCBs leaked in 1986. Levels of PCB were measured in leeches collected between 1992 to 2003 from 11 sites covering about 50 km of the river (the first sampling site upstream to the source of contamination and 10 sites downstream). The PCB indicator congeners IUPA no. 28, 52, 101, 118, 138, 153, and 180 were measured. Levels were highest at the four sampling sites nearest the source of pollution. The highest values of PCB congeners were found in 1992. PCB content decreased from 1992 to 2003 and with distance from the source. The study indicated that leeches of the genus Erpobdella are a suitable bioindicator of contamination in the surface layer of river sediments. PMID:22573988

  1. [Leech hirudo medicinalis: a therapeutic alternative available in Mexico].

    PubMed

    Cornejo-Esquerra, Agustín; Talleri-de-Andrea, Giancarlo; de Jesús Blanco-Favela, José; Ramos-Mora, Alberto; Villarán-Muñoz, Benjamín

    2009-01-01

    Throughout history a diversity of therapeutic properties have been attributed to the leech Hirudo medicinalis, however, during the past twenty years its use has proven to be a practical and effective method for the relief of venous engorgement in reconstructive surgery. In Mexico this practice has been limited due to the lack of supply and to the time-consuming process of importation. Based on this need, the animal facilities of the Occidental Biomedical Research Center-IMSS in Guadalajara, Jalisco, has initiated a breeding and reproduction program of this species with the aim at improving its availability as a therapeutic device. The salivary gland of Hirudo medicinalis contains one of the most potent natural anticoagulants, known as hirudin, a histamine-like vasodilator, platelet aggregation inhibitors and highly specific proteases, all of which favor venous flow and turn this species into the most useful in the clinical setting. Nevertheless, it is important to know the potential risks of leech therapy since it has been associated with considerable morbidity when adequate precautions are not taken. This review exposes relevant aspects of the medicinal leech, the indications, mechanism of action and the protocol for this therapy.

  2. Occurrence of three leech species (Annelida: Hirudinida) on fishes in the Kentucky River

    EPA Science Inventory

    Leeches were collected from six fish species distributed among four of ten sites sampled. The leech species observed were Myzobdella reducta (Meyer, 1940) and Myzobdella lugubris Leidy, 1851 of the family Piscicolidae and Placobdella pediculata Hemingway, 1908 of the family Gloss...

  3. Johann Friedrich Dieffenbach: successful use of leeches in plastic surgery in the 1820s.

    PubMed

    Sawyer, R T

    2000-04-01

    Johann Friedrich Dieffenbach (1792-1847) regularly and successfully utilised leeches in sophisticated plastic surgery in Berlin in the 1820s and 1830s, well before anaesthesia, antisepsis and antibiotics. Inexplicably, it took nearly another 150 years before the use of leeches in this context was revived.

  4. Occurrence of three leech species (Annelida: Hirudinida) on fishes in the Kentucky River

    EPA Science Inventory

    Leeches were collected from six fish species distributed among four of ten sites sampled. The leech species observed were Myzobdella reducta (Meyer, 1940) and Myzobdella lugubris Leidy, 1851 of the family Piscicolidae and Placobdella pediculata Hemingway, 1908 of the family Gloss...

  5. OCCURRENCE OF TWO LEECH SPECIES (ANNELIDA: HIRUDINEA) ON FISHES IN THE KENTUCKY RIVER

    EPA Science Inventory

    Little is known specifically on the feeding relationships between parasitic leeches and fish in North America. During an electrofishing survey conducted on the main stem of the Kentucky River in the summer of 2000, the presence of leeches was documented on six species of fish. ...

  6. Leech-Repellent Property of Eastern Red-Spotted Newts, Notophthalmus viridescens.

    PubMed

    Pough, F H

    1971-12-10

    Eastern red-spotted newts (Notophthalmus viridescens, Salamandridae) are rarely attacked by leeches. This protection is not shared by related salamandrids or by Ambystoma (Ambystomatidae). Tetrodotoxin is not the repelent. The immunity of Notophthalmus to leech parasitism is probably most significant in its aquatic stages, although the terrestrial efts are also protected.

  7. [Hirudo medicinalis-leech applications in plastic and reconstructive microsurgery--a literature review].

    PubMed

    Knobloch, K; Gohritz, A; Busch, K; Spies, M; Vogt, P M

    2007-04-01

    Medical leech therapy has enjoyed a renaissance in the world of reconstructive microsurgery during recent years. Especially venous congestion is decreased using hirudo medicinalis application such as following replantation of amputated fingers or congested flaps. They provide a temporary relief to venous engorgement whilst venous drainage is re-established. Living in symbiosis with Aeromonas hydrophila, who can digest the sixfold blood meal related to their body weight, and a broad number of anticoagulant agents such as the thrombin inhibitor hirudin, apyrase as well as collagenase, hyaluronidase, Factor Xa inhibitor and fibrinase I and II, leeches decrease venous congestion. Laser Doppler flowmetry could demonstrate a significant increase in superficial skin perfusion following leech application 16 mm around the biting zone. Following the initial blood meal accounting for about 2.5 ml, the anticoagulant effect of the various leeches enzymes follows within the next 5-6 hours, which both account for the beneficial effects. Infection associated with leech therapy is a documented complication of leech application, with reported incidences ranging from 2.4 to 20 % and a chinolone antibiotic is currently recommended to face the potential Aeromonas hydrophila infection. Anemia is a second adverse effect during medicinal leech application which has to be taken account with repetitive blood samples. Besides the successful applications of leeches in various applications in plastic and reconstructive microsurgery, randomized-controlled trials are pending to elucidate the value of hirudo medicinalis according to evidence-based criteria above from case series and case studies.

  8. OCCURRENCE OF TWO LEECH SPECIES (ANNELIDA: HIRUDINEA) ON FISHES IN THE KENTUCKY RIVER

    EPA Science Inventory

    Little is known specifically on the feeding relationships between parasitic leeches and fish in North America. During an electrofishing survey conducted on the main stem of the Kentucky River in the summer of 2000, the presence of leeches was documented on six species of fish. ...

  9. From Stimulation to Undulation: A Neuronal Pathway for the Control of Swimming in the Leech

    NASA Astrophysics Data System (ADS)

    Brodfuehrer, Peter D.; Friesen, W. Otto

    1986-11-01

    Initiation and performance of the swimming movement in the leech (Hirudo medicinalis) are controlled by neurons organized at at least four functional levels--sensory neurons, gating neurons, oscillator neurons, and motor neurons. A paired neuron, designated as Tr1, in the subesophageal ganglion of the leech has now been shown to define a fifth level, interposed between sensory and gating neurons. Cell Tr1 is activated by pressure and nociceptive mechanosensory neurons, which mediate body-wall stimulus--evoked swimming activity in intact leeches. In the isolated leech nervous system, brief stimulation of cell Tr1 elicits sustained activation of the gating neurons and triggers the onset of swimming activity. The synaptic interactions between all five levels of control are direct. Discovery of the Tr1 cells thus completes the identification of a synaptic pathway by which mechanosensory stimulation leads to the swimming movements of the leech.

  10. Diverse molecular data demonstrate that commercially available medicinal leeches are not Hirudo medicinalis.

    PubMed

    Siddall, Mark E; Trontelj, Peter; Utevsky, Serge Y; Nkamany, Mary; Macdonald, Kenneth S

    2007-06-22

    The European medicinal leech is one of vanishingly few animal species with direct application in modern medicine. In addition to the therapeutic potential held by many protease inhibitors purified from leech saliva, and notwithstanding the historical association with quackery, Hirudo medicinalis has been approved by the United States Food and Drug Administration as a prescription medical device. Accurate annotation of bioactive compounds relies on precise species determination. Interpretations of developmental and neurophysiological characteristics also presuppose uniformity within a model species used in laboratory settings. Here, we show, with mitochondrial sequences and nuclear microsatellites, that there are at least three species of European medicinal leech, and that leeches marketed as H. medicinalis are actually Hirudo verbana. Beyond the obvious need for reconsideration of decades of biomedical research on this widely used model organism, these findings impact regulatory statutes and raise concerns for the conservation status of European medicinal leeches.

  11. Development of a mechanical device to replace medicinal leech (Hirudo medicinalis) for treatment of venous congestion.

    PubMed

    Conforti, Michael L; Connor, Nadine P; Heisey, Dennis M; Vanderby, Ray; Kunz, David; Hartig, Gregory K

    2002-01-01

    Medicinal leeches are used to treat venous congestion, a complication of reconstructive surgery. Despite substantial drawbacks of leeching, little progress has been made to develop a device that would replace the leech for this purpose. The goal of this study was to develop and test mechanical prototypes for the treatment of venous congestion. We tested four prototypes (1, 2, 3a, and 3b) using congested fasciocutaneous flaps in swine. Blood removed by each prototype was measured for up to 4 hours. On average, the four prototypes removed 609%, 644%, 853%, and 811% more blood, respectively, from congested flaps versus a leech. Prototypes 3a and 3b, which allowed for innovative subcutaneous chemical (3a and 3b) and mechanical (3b) anticoagulation at the bleeding wound, sustained high levels of blood removal for up to 4 hours. Thus, a mechanical device can potentially replace the use of leeches for treating venous congestion.

  12. Diverse molecular data demonstrate that commercially available medicinal leeches are not Hirudo medicinalis

    PubMed Central

    Siddall, Mark E; Trontelj, Peter; Utevsky, Serge Y; Nkamany, Mary; Macdonald, Kenneth S

    2007-01-01

    The European medicinal leech is one of vanishingly few animal species with direct application in modern medicine. In addition to the therapeutic potential held by many protease inhibitors purified from leech saliva, and notwithstanding the historical association with quackery, Hirudo medicinalis has been approved by the United States Food and Drug Administration as a prescription medical device. Accurate annotation of bioactive compounds relies on precise species determination. Interpretations of developmental and neurophysiological characteristics also presuppose uniformity within a model species used in laboratory settings. Here, we show, with mitochondrial sequences and nuclear microsatellites, that there are at least three species of European medicinal leech, and that leeches marketed as H. medicinalis are actually Hirudo verbana. Beyond the obvious need for reconsideration of decades of biomedical research on this widely used model organism, these findings impact regulatory statutes and raise concerns for the conservation status of European medicinal leeches. PMID:17426015

  13. Detection and selective avoidance of near ultraviolet radiation by an aquatic annelid: the medicinal leech

    PubMed Central

    Jellies, John

    2014-01-01

    Medicinal leeches are aquatic predators that inhabit surface waters during daylight and also leave the water where they might be exposed to less screened light. Whereas the leech visual system has been shown to respond to visible light, leeches in the genus Hirudo do not appear to be as negatively phototactic as one might expect in order to avoid potential ultraviolet radiation (UVR)-induced damage. I used high intensity light emitting diodes to test the hypothesis that leeches could detect and specifically avoid near UVR (395–405 nm). Groups of unfed juvenile leeches exhibited a robust negative phototaxis to UVR, but had no behavioral response to blue or red and only a slight negative phototaxis to green and white light. Individual leeches also exhibited a vigorous negative phototaxis to UVR; responding in 100% of trials compared with modest negative responses to visible light (responding in ~8% of the trials). The responses in fed and unfed leeches were comparable for UVR stimuli. The responses depended upon the stimulus site: leeches shortened away from UV light to the head, and extended away from UV light to the tail. Electrophysiological nerve recordings showed that the cephalic eyes responded vigorously to UVR. Additionally, individual leech photoreceptors also showed strong responses to UVR, and a higher-order neuron associated with shortening and rapid behavioral responses, the S-cell, was activated by UVR, on both the head and tail. These results demonstrate that the leech can detect UVR and is able to discriminate behaviorally between UVR and visible light. PMID:24265432

  14. Detection and selective avoidance of near ultraviolet radiation by an aquatic annelid: the medicinal leech.

    PubMed

    Jellies, John

    2014-03-15

    Medicinal leeches are aquatic predators that inhabit surface waters during daylight and also leave the water where they might be exposed to less screened light. Whereas the leech visual system has been shown to respond to visible light, leeches in the genus Hirudo do not appear to be as negatively phototactic as one might expect in order to avoid potential ultraviolet radiation (UVR)-induced damage. I used high intensity light emitting diodes to test the hypothesis that leeches could detect and specifically avoid near UVR (395-405 nm). Groups of unfed juvenile leeches exhibited a robust negative phototaxis to UVR, but had no behavioral response to blue or red and only a slight negative phototaxis to green and white light. Individual leeches also exhibited a vigorous negative phototaxis to UVR; responding in 100% of trials compared with modest negative responses to visible light (responding in ~8% of the trials). The responses in fed and unfed leeches were comparable for UVR stimuli. The responses depended upon the stimulus site: leeches shortened away from UV light to the head, and extended away from UV light to the tail. Electrophysiological nerve recordings showed that the cephalic eyes responded vigorously to UVR. Additionally, individual leech photoreceptors also showed strong responses to UVR, and a higher-order neuron associated with shortening and rapid behavioral responses, the S-cell, was activated by UVR, on both the head and tail. These results demonstrate that the leech can detect UVR and is able to discriminate behaviorally between UVR and visible light.

  15. Central nervous system regeneration: from leech to opossum

    PubMed Central

    Mladinic, M; Muller, K J; Nicholls, J G

    2009-01-01

    A major problem of neurobiology concerns the failure of injured mammalian spinal cord to repair itself. This review summarizes work done on two preparations in which regeneration can occur: the central nervous system of an invertebrate, the leech, and the spinal cord of an immature mammal, the opossum. The aim is to understand cellular and molecular mechanisms that promote and prevent regeneration. In the leech, an individual axon regrows successfully to re-establish connections with its synaptic target, while avoiding other neurons. Functions that were lost are thereby restored. Moreover, pairs of identified neurons become re-connected with appropriate synapses in culture. It has been shown that microglial cells and nitric oxide play key roles in leech CNS regeneration. In the opossum, the neonatal brain and spinal cord are so tiny that they survive well in culture. Fibres grow across spinal cord lesions in neonatal animals and in vitro, but axon regeneration stops abruptly between postnatal days 9 and 12. A comprehensive search has been made in spinal cords that can and cannot regenerate to identify genes and establish their locations. At 9 days, growth-promoting genes, their receptors and key transcription molecules are up-regulated. By contrast at 12 days, growth-inhibitory molecules associated with myelin are prominent. The complete sequence of the opossum genome and new methods for transfecting genes offer ways to determine which molecules promote and which inhibit spinal cord regeneration. These results lead to questions about how basic research on mechanisms of regeneration could be ‘translated’ into effective therapies for patients with spinal cord injuries. PMID:19525562

  16. Central nervous system regeneration: from leech to opossum.

    PubMed

    Mladinic, M; Muller, K J; Nicholls, J G

    2009-06-15

    A major problem of neurobiology concerns the failure of injured mammalian spinal cord to repair itself. This review summarizes work done on two preparations in which regeneration can occur: the central nervous system of an invertebrate, the leech, and the spinal cord of an immature mammal, the opossum. The aim is to understand cellular and molecular mechanisms that promote and prevent regeneration. In the leech, an individual axon regrows successfully to re-establish connections with its synaptic target, while avoiding other neurons. Functions that were lost are thereby restored. Moreover, pairs of identified neurons become re-connected with appropriate synapses in culture. It has been shown that microglial cells and nitric oxide play key roles in leech CNS regeneration. In the opossum, the neonatal brain and spinal cord are so tiny that they survive well in culture. Fibres grow across spinal cord lesions in neonatal animals and in vitro, but axon regeneration stops abruptly between postnatal days 9 and 12. A comprehensive search has been made in spinal cords that can and cannot regenerate to identify genes and establish their locations. At 9 days, growth-promoting genes, their receptors and key transcription molecules are up-regulated. By contrast at 12 days, growth-inhibitory molecules associated with myelin are prominent. The complete sequence of the opossum genome and new methods for transfecting genes offer ways to determine which molecules promote and which inhibit spinal cord regeneration. These results lead to questions about how basic research on mechanisms of regeneration could be 'translated' into effective therapies for patients with spinal cord injuries.

  17. Medicinal use of leeches in the texts of ancient Greek, Roman and early Byzantine writers.

    PubMed

    Papavramidou, N; Christopoulou-Aletra, H

    2009-09-01

    Blood-letting was a common therapeutic method in antiquity; many means were used to draw blood, including the application of leeches. In this paper, ancient Greek, Roman and Byzantine authors up to the 7th century AD were studied, a research that provided us with references that may be divided into two groups: those related to the medicinal use of leeches, and those related to cases in which leeches were swallowed and had to be removed. In the first group, detailed descriptions of the method of usage and of the diseases requiring leeching were found. In the second group, brief reference is made to the problems caused by swallowing leeches, and to the methods used to expel them from the human organism. The earliest references to the medicinal use of leeches may be found in the writings of Theocritus (3rd century BC), Nicander (2nd century BC) and Horace (1st century BC, while the phenomenon of swallowing a leech is first mentioned in one of the Epidaurian 'iamata' dating to the 4th century BC.

  18. Cholesterol and its derivatives, are the principal steroids isolated from the leech species Hirudo medicinalis.

    PubMed

    Zipser, B; Bradford, J J; Hollingsworth, R I

    1998-08-01

    Steroids were isolated from the blood-sucking leech species Hirudo medicinalis and their structure was studied with one- and two-dimensional NMR spectroscopy (DQF-COSY and HMQC), GC-MS and ESI-MS spectrometry. Fractionating leech lipid using silicic acid chromatography led to the isolation of cholesterol in an early chloroform-eluted peak. Only minor traces of cholest-4-en-3-one, 4 beta-methylcholesterol, and sitosterol were present. The subsequent acetone-eluted fraction contained steroidtriols that were further purified by preparative TLC; these included cholest-7-ene-3,5,6 triol, cholest-4,7-diene-3,6,15 triol and to a lesser amount, cholestane-3,5,6 triol. A developmental study on cholesterol content in the leech showed that it is also the principal steroid in embryonic and freshly hatched leeches prior to feeding. The abundance of cholesterol, comprising approximately 5% of the total leech lipid, suggests that H. medicinalis, a blood sucking leech, has adapted itself fully to its mammalian host in terms of its steroid content. It remains to be seen whether lipids are directly transferred from the host to the parasite or whether leeches have evolved mechanisms to synthesize their own steroids.

  19. Multiplexed Population Coding of Stimulus Properties by Leech Mechanosensory Cells.

    PubMed

    Pirschel, Friederice; Kretzberg, Jutta

    2016-03-30

    Sensory coding has long been discussed in terms of a dichotomy between spike timing and rate coding. However, recent studies found that in primate mechanoperception and other sensory systems, spike rates and timing of cell populations complement each other. They simultaneously carry information about different stimulus properties in a multiplexed way. Here, we present evidence for multiplexed encoding of tactile skin stimulation in the tiny population of leech mechanoreceptors, consisting of only 10 cells of two types with overlapping receptive fields. Each mechanoreceptor neuron of the leech varies spike count and response latency to both touch intensity and location, leading to ambiguous responses to different stimuli. Nevertheless, three different stimulus estimation techniques consistently reveal that the neuronal population allows reliable decoding of both stimulus properties. For the two mechanoreceptor types, the transient responses of T (touch) cells and the sustained responses of P (pressure) cells, the relative timing of the first spikes of two mechanoreceptors encodes stimulus location, whereas summed spike counts represent touch intensity. Differences between the cell types become evident in responses to combined stimulus properties. The best estimation performance for stimulus location is obtained from the relative first spike timing of the faster and temporally more precise T cells. Simultaneously, the sustained responses of P cells indicate touch intensity by summed spike counts and stimulus duration by the duration of spike responses. The striking similarities of these results with previous findings on primate mechanosensory afferents suggest multiplexed population coding as a general principle of somatosensation. Multiplexing, the simultaneous encoding of different stimulus properties by distinct neuronal response features, has recently been suggested as a mechanism used in several sensory systems, including primate somatosensation. While a

  20. [Extract of Hirudo medicinalis leeches: biogenic substance for the creation of effective drugs].

    PubMed

    Nikonov, G I; Titova, E A; Lebedeva, A O

    2015-01-01

    Investigations of the extract of medicinal leech Girulux established parameters that enable one to verify the authenticity of related drugs using the characteristics of their UV spectra the and anticoagulant and antithrombin activity. The limits of pH must range from 5.5 to 7.5. Pilot-scale series of medical leech extract are in compliance with terms of "microbiological purity" quality characteristics. The natural storage has been used to assess the temporal stability of leech extract. The obtained characteristics showed that the activity of extract was retained after 3-year storage.

  1. Characterization and optimization of lyophilization and storage conditions of Leech saliva extract from the tropical leech Hirudinaria manillensis.

    PubMed

    Abdualkader, Abdualrahman Mohammed; Ghawi, Abbas Mohammad; Alaama, Mohamed; Awang, Mohamed; Merzouk, Ahmed

    2013-05-01

    The medicinal Malaysian leeches have been used in traditional medicine to treat many different ailments. In this study, leech saliva extract (LSE) was collected from the medicinal Malaysian leech Hirudinaria manillensis. Gel electrophoresis of LSE was carried out to estimate the peptide and protein molecular weights of its content. Results showed that LSE contains more than 60 peptides and proteins with molecular masses ranging from 1.9-250kDa. Thrombin time assay in vitro was employed to assess the collected LSE antithrombin activity. First, to study its stability, LSE was lyophilized under the following different conditions: pre-freezing temperature, type of container and lyophilization cycle. Pre-freezed LSE sample at -20°C and lyophilized for 24 hours retained about 100-95% of its original biological activities. Second, the LSE antithrombin activity was monitored for a period of six months. Storage temperature, type of the container and photosensitivity effects on antithrombin activity of the lyophilized (solid state) and non-lyophilized (liquid state) were investigated. Results showed that storage temperature drastically affected the biological activity of LSE with -20 °C as the optimum temperature. Samples stored at ambient temperature and +4 °C were light photosensitive and adversely affected when stored in polypropylene tubes. Lyophilized samples were more stable than non-lyophilized ones over the period of study. To sum up, in order to have a biologically active stock of LSE, it has to be lyophilized for no more than 24 hours following freezing at -20°C and has to be stored at -20°C in glass tubes protected from light.

  2. Control of epileptiform bursting in the leech heart interneuron

    NASA Astrophysics Data System (ADS)

    Barnett, William; Anquez, Martin; Harris, Torrey; Cymbalyuk, Gennady

    2009-11-01

    The network controlling heartbeat in the medicinal leech contains leech heart interneurons (HNs). We modeled them under specific pharmacological conditions. The Ca^2+ currents were blocked by Co^2+. The K^+ currents, apart from the non-inactivating current, IK2, were blocked by 4AP. The hyperpolarization-activated current, Ih, was blocked by Cs^+. Under these conditions, epileptiform bursting characterized by long interburst intervals (IBI) has been shown. We considered three distinct cases. Model 1 included IK2, Ih, and the fast Na^+ current, INa. Model 2 was characterized by INa, IK2, and the persistent Na^+ current, INaP. Model 3 consisted of INa, IK2, Ih, and INaP. We also investigated the bi-stability of bursting and silence as the leak conductance, gleak, was varied. We showed that in 1 and 3, model HNs demonstrated bi-stability of silence and bursting. We analyzed how IBI and burst duration are controlled by the manipulation of Ih and INaP. In 1, as V1/2 of Ih decreased, IBI grew towards infinity one over the square root of the parameter difference. In 2, we showed that as gNaP decreased from 6.156 nS to 6.155 nS, IBI grew in accordance with the one over square root law. The system underwent a saddle-node bifurcation just below 6.155 nS. Supported by NSF PHY-0750456.

  3. Host Matters: Medicinal Leech Digestive-Tract Symbionts and Their Pathogenic Potential

    PubMed Central

    Marden, Jeremiah N.; McClure, Emily A.; Beka, Lidia; Graf, Joerg

    2016-01-01

    Digestive-tract microbiota exert tremendous influence over host health. Host-symbiont model systems are studied to investigate how symbioses are initiated and maintained, as well as to identify host processes affected by resident microbiota. The medicinal leech, Hirudo verbana, is an excellent model to address such questions owing to a microbiome that is consistently dominated by two species, Aeromonas veronii and Mucinivorans hirudinis, both of which are cultivable and have sequenced genomes. This review outlines current knowledge about the dynamics of the H. verbana microbiome. We discuss in depth the factors required for A. veronii colonization and proliferation in the leech crop and summarize the current understanding of interactions between A. veronii and its annelid host. Lastly, we discuss leech usage in modern medicine and highlight how leech-therapy associated infections, often attributable to Aeromonas spp., are of growing clinical concern due in part to an increased prevalence of fluoroquinolone resistant strains. PMID:27790190

  4. STUDIES ON RARE AND POORLY KNOWN LEECHES (ANNELIDA: HIRUDINEA: GLOSSIPHONIIDAE) IN EASTERN NORTH AMERICA

    EPA Science Inventory

    Three taxa within the leech family Glossiphoniidae, Actinobdella inequiannulata, Placobdella hollensis, and Theromyzon spp., though widespread in eastern North America, remain poorly known with respect to their biology and systematics. All three taxa have been collected in New E...

  5. Prolonged venous bleeding due to traditional treatment with leech bite: a case report

    PubMed Central

    2011-01-01

    Introduction The medicinal leech, Hirudo medicinalis, has been used in the treatment of many diseases for thousands of years. In Turkey, it is used most commonly in the management of venous diseases of lower extremities. Case presentation A 25-year-old Turkish woman presented to our emergency room with bleeding from her left leg. She had been treated for varicose veins in her lower extremities with leeches about 24 hours before admission to the emergency room. The bleeding was controlled by applying pressure with sterile gauze upon the wound, and she was discharged. She returned after four hours having started bleeding again. Hemostasis was achieved by vein ligation under local anesthesia. Conclusions Leech bite should be evaluated as a special injury. Prolonged bleeding can be seen after leech bites. In such cases, hemostasis either with local pressure or ligation of the bleeding vessel is mandatory. PMID:21548915

  6. STUDIES ON RARE AND POORLY KNOWN LEECHES (ANNELIDA: HIRUDINEA: GLOSSIPHONIIDAE) IN EASTERN NORTH AMERICA

    EPA Science Inventory

    Three taxa within the leech family Glossiphoniidae, Actinobdella inequiannulata, Placobdella hollensis, and Theromyzon spp., though widespread in eastern North America, remain poorly known with respect to their biology and systematics. All three taxa have been collected in New E...

  7. A new species of glossiphoniid leech from Rana pretiosa (Amphibia: Ranidae) in Oregon.

    PubMed

    Siddall, Mark E; Bowerman, Jay

    2006-08-01

    A new species of ectoparasitic glossiphoniid leech was found feeding on frogs in the Nature Center Pond and elsewhere in Deschutes County, Oregon. The new species of Placobdella resembles the southern alligator leech, Placobdella multilineata Moore, 1953, notwithstanding their vast geographic separation in North America. The new species is readily distinguished by possessing subdivided annuli, by its papillation and pigmentation patterns as well as by the arrangement of ovarian tissues. There is strong evidence of nocturnality and of the potential for parasitizing humans.

  8. Elimination of symbiotic Aeromonas spp. from the intestinal tract of the medicinal leech, Hirudo medicinalis, using ciprofloxacin feeding.

    PubMed

    Mumcuoglu, K Y; Huberman, L; Cohen, R; Temper, V; Adler, A; Galun, R; Block, C

    2010-06-01

    The use of the medicinal leech (Hirudo medicinalis) in promoting venous drainage in tissues whose vitality is threatened by venous congestion and obstruction, especially in plastic and reconstructive surgery, has been complicated by infections caused by Aeromonas spp. These are leech endosymbionts for which patients undergoing hirudotherapy frequently receive systemic chemoprophylaxis. In order to evaluate the possibility of rendering leeches safe for use on patients, H. medicinalis were fed artificially with a 2 g/L arginine solution (used as a phagostimulant) supplemented with ciprofloxacin (100 mg/L). Aeromonads were detected in 57 out of 80 control leeches (71.3%), but in none of the 56 leeches treated with ciprofloxacin (p <0.001). Treated leeches survived for up to 4 months. Tested weekly, 61% of these leeches took human blood for at least 4 weeks after treatment and all remained negative for aeromonads. All water samples in which leeches were kept before treatment were contaminated with Aeromonas spp.; none were detected in any of the NaCl/arginine solutions with which treated animals were fed. Molecular characterization of two phenotypically distinct isolates using gyrB sequencing showed that one clustered tightly with A. veronii and the other was closely related to A. media. Other environmental bacteria and fungi were isolated from 26.5% of treated leeches that had taken a blood meal 1-4 weeks after treatment. Ciprofloxacin reduced the number of leech-associated aeromonads to undetectable levels for extended periods. Most treated leeches were ready to take a blood meal after treatment, suggesting the possibility of using ciprofloxacin-treated leeches instead of chemoprophylaxis in patients undergoing hirudotherapy.

  9. First isolation of a giant virus from wild Hirudo medicinalis leech: Mimiviridae isolation in Hirudo medicinalis.

    PubMed

    Boughalmi, Mondher; Pagnier, Isabelle; Aherfi, Sarah; Colson, Philippe; Raoult, Didier; La Scola, Bernard

    2013-11-27

    Giant viruses and amoebae are common in freshwater, where they can coexist with other living multicellular organisms. We screened leeches from the species Hirudo medicinalis for giant viruses. We analyzed five H. medicinalis obtained from Tunisia (3) and France (2). The leeches were decontaminated and then dissected to remove internal parts for co-culture with Acanthamoeba polyphaga. The genomes of isolated viruses were sequenced on a 454 Roche instrument, and a comparative genomics analysis was performed. One Mimivirus was isolated and the strain was named Hirudovirus. The genome assembly generated two scaffolds, which were 1,155,382 and 25,660 base pairs in length. Functional annotations were identified for 47% of the genes, which corresponds to 466 proteins. The presence of Mimividae in the same ecological niche as wild Hirudo may explain the presence of the mimivirus in the digestive tract of the leech, and several studies have already shown that viruses can persist in the digestive tracts of leeches fed contaminated blood. As leeches can be used medically and Mimiviruses have the potential to be an infectious agent in humans, patients treated with leeches should be surveyed to investigate a possible connection.

  10. New Records of Lake Baikal Leech Fauna: Species Diversity and Spatial Distribution in Chivyrkuy Gulf

    PubMed Central

    Kaygorodova, Irina A.; Pronin, Nikolay M.

    2013-01-01

    The study of several Lake Baikal leech collections offered us the possibility to determine species diversity in the Chivyrkuy Gulf, the biggest one in the lake. As a result, the first information on the Chivyrkuy Hirudinea fauna (Annelida, Clitellata) has been revealed. There are two orders and four families of leeches in the Chivyrkuy Gulf: order Rhynchobdellida (families Glossiphoniidae and Piscicolidae) and order Arhynchobdellida (families Erpobdellidae and Haemopidae). In total, 22 leech species and 2 subspecies belonging to 11 genera were identified. Of these, 4 taxa belong to the family Glossiphoniidae (G. concolor, A. hyalina, A. heteroclita f. papillosa, and A. heteroclita f. striata) recorded in Baikal for the first time. Representatives of 8 unidentified species (Glossophinia sp., Baicaloclepsis sp., Baicalobdella sp., Piscicola sp. 1, Piscicola sp. 2, Erpobdella sp. 1, Erpobdella sp. 2, and Erpobdella sp. 3) have been also recorded. The checklist gives a contemporary overview of the species composition of leech parasites, their hosts, and distribution within the Chivyrkuy Gulf. The analysis of spatial distribution has shown that the leech species diversity is correlated with the biological productivity of the bay. The most diverse community of leech species is detected in the eutrophic zone of the lake. PMID:23844382

  11. The use of medical leeches for venous congestion. A review and case report.

    PubMed

    Buote, N J

    2014-01-01

    The medicinal leech, Hirudo medicinalis, has been used for hundreds of years in human medicine for a variety of diseases, most recently including venous congestion following reconstructive surgeries (skin flaps and reimplantations), excessive lingual and periorbital swelling, and non-traditional treatments for osteoarthritis, compartment syndrome and sialoadenitis. The treatment of venous congestion in animals using leeches has been mentioned anecdotally, but the only published report pertains to the use of leeches in a cat suffering from polycythemia vera. We report the use of medical grade leeches in a one-year-old male castrated Domestic Shorthaired cat presenting with severe swelling of the paw after sustaining a constrictive injury from a bandage. The limb use had become compromised and the swelling was not responsive to compression bandages so leech therapy was instituted for four days and the swelling dramatically improved. The patient's limb use improved back to normal and the constrictive wound went on to heal without complication. The use of leeches in this case allowed for resolution of severe venous congestion and a full return to function in this patient with no continued skin loss. The mechanism of action of hirudotherapy for venous congestion is to allow for an alternative egress of pooled venous blood leading to a reduction in capillary pressure and increases in arterial reperfusion of capillary beds. The human and veterinary applications, method, and potential complications with hirudotherapy are discussed in this review.

  12. Detection of Infectious Pancreatic Necrosis Virus from the Leeches Hemiclepsis marginata and Hirudo medicinalis.

    PubMed

    Salimi, Behnam; Abdi, Kazem

    2016-12-01

    Leeches have been reported to harbor several important fish pathogens, including spring viremia of carp virus, infectious hematopoietic necrosis virus (IHNV), and viral hemorrhagic septicemia virus (VHSV), and also may contain blood protozoa. In the present study, leeches were collected from water bodies located in Kurdistan province, Iran. The specimens were tested for IHNV, VHSV, and infectious pancreatic necrosis virus (IPNV) using the PCR method. The results showed that two different species of leeches, Hemiclepsis marginata and Hirudo medicinalis, were infected by IPNV among the seven species studied. The infected leeches were found in areas that were polluted with untreated sewage coming from upstream fish farms culturing Rainbow Trout Oncorhynchus mykiss. In addition, the fish at fish farms in the vicinity had been infected with IPNV 9 months previously. Our results showed that the virus causing infectious pancreatic necrosis is present in the leeches H. marginata and H. medicinalis, suggesting that leeches are a potential source of IPNV in fish farms. Received October 14, 2015; accepted June 1, 2016.

  13. First Isolation of a Giant Virus from Wild Hirudo medicinalis Leech: Mimiviridae isolation in Hirudo medicinalis

    PubMed Central

    Boughalmi, Mondher; Pagnier, Isabelle; Aherfi, Sarah; Colson, Philippe; Raoult, Didier; La Scola, Bernard

    2013-01-01

    Giant viruses and amoebae are common in freshwater, where they can coexist with other living multicellular organisms. We screened leeches from the species Hirudo medicinalis for giant viruses. We analyzed five H. medicinalis obtained from Tunisia (3) and France (2). The leeches were decontaminated and then dissected to remove internal parts for co-culture with Acanthamoeba polyphaga. The genomes of isolated viruses were sequenced on a 454 Roche instrument, and a comparative genomics analysis was performed. One Mimivirus was isolated and the strain was named Hirudovirus. The genome assembly generated two scaffolds, which were 1,155,382 and 25,660 base pairs in length. Functional annotations were identified for 47% of the genes, which corresponds to 466 proteins. The presence of Mimividae in the same ecological niche as wild Hirudo may explain the presence of the mimivirus in the digestive tract of the leech, and several studies have already shown that viruses can persist in the digestive tracts of leeches fed contaminated blood. As leeches can be used medically and Mimiviruses have the potential to be an infectious agent in humans, patients treated with leeches should be surveyed to investigate a possible connection. PMID:24287596

  14. Electronic neuron within a ganglion of a leech (Hirudo medicinalis)

    NASA Astrophysics Data System (ADS)

    Aliaga, J.; Busca, N.; Minces, V.; Mindlin, G. B.; Pando, B.; Salles, A.; Sczcupak, L.

    2003-06-01

    We report the construction of an electronic device that models and replaces a neuron in a midbody ganglion of the leech Hirudo medicinalis. In order to test the behavior of our device, we used a well-characterized synaptic interaction between the mechanosensory, sensitive to pressure, (P) cell and the anteropagoda (because of the action potential shape) (AP) neuron. We alternatively stimulated a P neuron and our device connected to the AP neuron, and studied the response of the latter. The number and timing of the AP spikes were the same when the electronic parameters were properly adjusted. Moreover, after changes in the depolarization of the AP cell, the responses under the stimulation of both the biological neuron and the electronic device vary in a similar manner.

  15. Electronic neuron within a ganglion of a leech (Hirudo medicinalis).

    PubMed

    Aliaga, J; Busca, N; Minces, V; Mindlin, G B; Pando, B; Salles, A; Sczcupak, L

    2003-06-01

    We report the construction of an electronic device that models and replaces a neuron in a midbody ganglion of the leech Hirudo medicinalis. In order to test the behavior of our device, we used a well-characterized synaptic interaction between the mechanosensory, sensitive to pressure, (P) cell and the anteropagoda (because of the action potential shape) (AP) neuron. We alternatively stimulated a P neuron and our device connected to the AP neuron, and studied the response of the latter. The number and timing of the AP spikes were the same when the electronic parameters were properly adjusted. Moreover, after changes in the depolarization of the AP cell, the responses under the stimulation of both the biological neuron and the electronic device vary in a similar manner.

  16. Detection of Viral Hemorrhagic Septicemia virus (VHSV) from the leech Myzobdella lugubris Leidy, 1851.

    PubMed

    Faisal, Mohamed; Schulz, Carolyn A

    2009-09-28

    The leech Myzobdella lugubris is widespread in the Lake Erie Watershed, especially Lake St. Clair. However, its role in pathogen transmission is not fully understood. In this same watershed, several widespread fish mortalities associated with the Viral Hemorrhagic Septicemia virus (VHSV) were recorded. Viral Hemorrhagic Septicemia is an emerging disease in the Great Lakes Basin that is deadly to the fish population, yet little is known about its mode of transmission. To assess the potential role of M. lugubris in VHSV transmission, leeches were collected from Lake St. Clair and Lake Erie and pooled into samples of five. Cell culture and reverse transcriptase polymerase chain reaction (RT-PCR) were used to determine the presence of the virus and its identity. Results showed that 57 of the 91 pooled leech samples were positive by cell culture for VHSV and 66 of the 91 pooled leech samples were positive by RT-PCR for the VHSV. Two representative virus isolates were sequenced for further genetic confirmation and genotype classification. VHSV detected within M. lugubris was homologous to the Great Lakes strain of VHSV genotype IVb. This is the first record of the VHSV being detected from within a leech, specifically M. lugubris, and suggests the potential of M. lugubris being involved in VHSV transmission.

  17. An Unusual Cause of Bleeding on the Floor of Mouth: Leech Infestation.

    PubMed

    Kantekin, Yunus; Sarı, Kamran; Özkırış, Mahmut; Kapusuz Gencer, Zeliha

    2015-12-01

    Leech infestation is a very rare phenomenon in humans. It mostly occurs in humans when rural untreated water is drunk or while swimming in streams or lakes. When leeches adhere to the mucous membrane, they ingest blood. Thus, they can sometimes cause severe anemia that may require blood transfusion. We report a case that was referred to emergency service with bleeding in the floor of the mouth. A 10-year-old child was referred to the emergency service of a city hospital with a complaint of swelling in the floor of the mouth and spitting of blood. The patient was promptly taken to the operating room. Using local anesthesia, a surgical incision was made, and a moving, dark brown foreign body was removed from the floor of the mouth and identified as a leech. Leech endoparasitism should be considered as a cause of unexplained anemia due to bleeding from the throat. Accordingly, leech infestation must be considered in differential diagnosis when a patient complains of spitting of blood, hoarseness, or dysphagia.

  18. Specialized brain regions and sensory inputs that control locomotion in leeches

    PubMed Central

    Mullins, Olivia J.; Brodfuehrer, Peter D.; Jusufović, Saša; Hackett, John T.; Friesen, W. Otto

    2011-01-01

    Locomotor systems are often controlled by specialized cephalic neurons and undergo modulation by sensory inputs. In many species, dedicated brain regions initiate and maintain behavior and set the duration and frequency of the locomotor episode. In the leech, removing the entire head brain enhances swimming, but the individual roles of its components, the supra- and subesophageal ganglia, in the control of locomotion are unknown. Here we describe the influence of these two structures and that of the tail brain on rhythmic swimming in isolated nerve cord preparations and in nearly-intact leeches suspended in an aqueous, “swim-enhancing” environment. We found that, in isolated preparations, swim episode duration and swim burst frequency are greatly increased when the supraesophageal ganglion is removed, but the subesophageal ganglion is intact. The prolonged swim durations observed with the anterior-most ganglion removed were abolished by removal of the tail ganglion. Experiments on the nearly intact leeches show that, in these preparations, the subesophageal ganglion acts to decrease cycle period but, unexpectedly, also decreases swim duration. These results suggest that the supraesophageal ganglion is the primary structure that constrains leech swimming; however, the control of swim duration in the leech is complex, especially in the intact animal. PMID:22037913

  19. The medical use of leeches in contemporary Spain: between science and tradition.

    PubMed

    Vallejo, Jose Ramon; Gonzalez, Jose Antonio

    2015-01-01

    In Spain leeches have been used both in popular and scientific medicine throughout its history. In this study we analyze the historical fluctuations of leech therapy. At the start of the 20th century it was still being used in in scientific medicine, as can be seen in the treatment administered to Germán Gamazo, a minister during the reign of Alfonso XII and the regency of Archduchess Maria Christina of Austria, during a serious illness in 1901. Leech therapy was to fall dramatically into disuse and was to survive only in folk medicine, with leeches losing their reputation as a therapeutic agent. The data obtained is the result of a systematic review of the literature and of the major databases in the fields of folklore, ethnography, social anthropology and medical anthropology. Leeches have been used in Spanish folk medicine to treat ailments and disorders in up to 11 categories of the International Statistical Classification of Diseases and Related Health Problems (ICD-10), particularly in the treatment of diseases of the circulatory, respiratory and musculoskeletal systems. According to the available literature, they were part of the folk therapeutic arsenal, at least until the seventies of the last century. Our study also provides information about the medicinal use, commerce and consumption of these animals in recent years.

  20. Cloning, characterisation and expression of the alpha-tubulin genes of the leech, Hirudo medicinalis.

    PubMed

    Fedorov, A; Johnston, H; Korneev, S; Blackshaw, S; Davies, J

    1999-02-04

    We have isolated two alpha-tubulin cDNAs from the leech, Hirudo medicinalis. Both encode putative proteins of 451 amino-acids which differ from each other at only two positions. Southern blotting suggests that there are only two alpha-tubulin genes in the leech. The genes contain two introns and, because of the extremely high homology of the nucleotide sequence from the second intron to the end of the genes, we have inferred that a gene conversion event about 9.5 million years ago has homogenised the Hirudo alpha-tubulin sequences. Using in situ hybridisation to tissue sections, we have shown that the two genes are probably expressed in all neurons of the leech ganglia and that their spatial distribution remains unchanged during neuronal regeneration. The deduced amino-acid sequences of the leech alpha-tubulins show that they have greatest similarity to those from a platyhelminth, echiuran and mollusc with rather less to arthropod alpha-tubulins. The protein sequences of the leech alpha-tubulins have been compared with representatives of those from across all phyla to determine if any specific feature labels certain isotypes of tubulin for neuronal expression.

  1. Diversity and selective pressures of anticoagulants in three medicinal leeches (Hirudinida: Hirudinidae, Macrobdellidae)

    PubMed Central

    Kvist, Sebastian; Min, Gi-Sik; Siddall, Mark E

    2013-01-01

    Although medicinal leeches have long been used as treatment for various ailments because of their potent anticoagulation factors, neither the full diversity of salivary components that inhibit coagulation, nor the evolutionary selection acting on them has been thoroughly investigated. Here, we constructed expressed sequence tag libraries from salivary glands of two species of medicinal hirudinoid leeches, Hirudo verbana and Aliolimnatis fenestrata, and identified anticoagulant-orthologs through BLASTx searches. The data set then was augmented by the addition of a previously constructed EST library from the macrobdelloid leech Macrobdella decora. The identified orthologs then were compared and contrasted with well-characterized anticoagulants from a variety of leeches with different feeding habits, including non-sanguivorous species. Moreover, four different statistical methods for predicting signatures of positive and negative evolutionary pressures were used for 10 rounds each to assess the level and type of selection acting on the molecules as a whole and on specific sites. In total, sequences showing putative BLASTx-orthology with five and three anticoagulant-families were recovered in the A. fenestrata and H. verbana EST libraries respectively. Selection pressure analyses predicted high levels of purifying selection across the anticoagulant diversity, although a few isolated sites showed signatures of positive selection. This study represents a first attempt at mapping the anticoagulant repertoires in a comparative fashion across several leech families. PMID:23610634

  2. Detection of Viral Hemorrhagic Septicemia virus (VHSV) from the leech Myzobdella lugubris Leidy, 1851

    PubMed Central

    Faisal, Mohamed; Schulz, Carolyn A

    2009-01-01

    The leech Myzobdella lugubris is widespread in the Lake Erie Watershed, especially Lake St. Clair. However, its role in pathogen transmission is not fully understood. In this same watershed, several widespread fish mortalities associated with the Viral Hemorrhagic Septicemia virus (VHSV) were recorded. Viral Hemorrhagic Septicemia is an emerging disease in the Great Lakes Basin that is deadly to the fish population, yet little is known about its mode of transmission. To assess the potential role of M. lugubris in VHSV transmission, leeches were collected from Lake St. Clair and Lake Erie and pooled into samples of five. Cell culture and reverse transcriptase polymerase chain reaction (RT-PCR) were used to determine the presence of the virus and its identity. Results showed that 57 of the 91 pooled leech samples were positive by cell culture for VHSV and 66 of the 91 pooled leech samples were positive by RT-PCR for the VHSV. Two representative virus isolates were sequenced for further genetic confirmation and genotype classification. VHSV detected within M. lugubris was homologous to the Great Lakes strain of VHSV genotype IVb. This is the first record of the VHSV being detected from within a leech, specifically M. lugubris, and suggests the potential of M. lugubris being involved in VHSV transmission. PMID:19785752

  3. Allium sativum L.: the anti-immature leech (Limnatis nilotica) activity compared to Niclosomide.

    PubMed

    Bahmani, Mahmoud; Abbasi, Javad; Mohsenzadegan, Ava; Sadeghian, Sirous; Ahangaran, Majid Gholami

    2013-03-01

    This study was carried out to determine the effects of methanolic extracts of Allium sativum L. on Limnatis nilotica compared with Niclosomide. In this experimental study in September 2010, a number of leeches (70 in total) from the southern area of Ilam province were prepared, and the effects of methanolic extract of A. sativum L. with Niclosomide as the control drug were compared and distilled water was evaluated as the placebo group which investigated L. nilotica using anti-leech assay. The average time of paralysis and death of L. nilotica for Niclosomide (1,250 mg/kg) and the methanol extract of A. sativum L. (600 μg/ml) were 6.22 ± 2.94 and 68.44 ± 28.39 min, respectively. Distilled water and garlic tablets at a dose of 400 mg were determined as the inert group. In this research, the attraction time of the leeches' death among different treatments is significant. In this study, it was determined that Niclosomide, with an intensity of 4+, and methanolic extracts of A. sativum L., with an intensity of 3+, have a good anti-leech effect and can be shown to be effective in cases of leech biting, while distilled water was negative.

  4. Bugs as drugs, part two: worms, leeches, scorpions, snails, ticks, centipedes, and spiders.

    PubMed

    Cherniack, E Paul

    2011-03-01

    In this second of a two-part series analyzing the evidence for the use of organisms as medicine, the use of a number of different "bugs" (worms, leeches, snails, ticks, centipedes, and spiders) is detailed. Several live organisms are used as treatments: leeches for plastic surgery and osteoarthritis and the helminths Trichuris suis and Necator americanus for inflammatory bowel disease. Leech saliva is the source of a number of anticoagulants, including the antithrombin agent hirudin and its synthetic analogues, which have been approved for human use. Predatory arthropods, such as certain species of snails, spiders, scorpions, centipedes, and ticks provide a trove of potential analgesic peptides in their venom. A synthetic analogue of a snail venom peptide, ziconotide, has been approved for human use and is used as an alternative to opioids in severe pain cases. Arthropods, such as ticks, have venom that contains anticoagulants and centipede venom has a protein that corrects abnormalities in lipid metabolism.

  5. 3H-GABA uptake selectively labels identifiable neurons in the leech central nervous system

    SciTech Connect

    Cline, H.T.

    1983-04-10

    Segmental ganglia of the leech ventral nerve cord synthesize the neurotransmitter gamma-aminobutyric acid (GABA) when incubated in the presence of the precursor glutamate, suggesting that there may be GABA-ergic neurons in the leech nerve cord. GABA-accumulating neurons of the two taxonomically distant leech species, Haementeria ghilianii and Hirudo medicinalis, have been labeled by taking advantage of their high-affinity uptake system for the neurotransmitter. Autoradiography of sectioned segmental ganglia previously exposed to 3H-GABA reveals a reproducible pattern of about thirty 3H-GABA-labeled neuronal cell bodies per ganglion. The majority of 3H-GABA-labeled neuronal cell bodies are bilaterally paired, although some apparently unpaired cell bodies also accumulate label. Neuronal processes were reproducibly labeled by GABA uptake and could be traced in the neuropil through commissures and fiber tracts into the segmental nerve roots and interganglionic connectives, respectively.

  6. Triassic leech cocoon from Antarctica contains fossil bell animal

    NASA Astrophysics Data System (ADS)

    Bomfleur, Benjamin; Kerp, Hans; Taylor, Thomas N.; Moestrup, Øjvind; Taylor, Edith L.

    2012-12-01

    Our understanding of the evolution of life on Earth is limited by the imperfection of the fossil record. One reason for this imperfect record is that organisms without hard parts, such as bones, shells, and wood, have a very low potential to enter the fossil record. Occasionally, however, exceptional fossil deposits that preserve soft-bodied organisms provide a rare glimpse of the true biodiversity during past periods of Earth history. We here present an extraordinary find of a fossil ciliate that is encased inside the wall layer of a more than 200 Ma leech cocoon from Antarctica. The microfossil consists of a helically contractile stalk that attaches to a main body with a peristomial feeding apparatus and a large C-shaped macronucleus. It agrees in every aspect with the living bell animals, such as Vorticella. Vorticellids and similar peritrichs are vital constituents of aquatic ecosystems worldwide, but so far have lacked any fossil record. This discovery offers a glimpse of ancient soft-bodied protozoan biotas, and also highlights the potential of clitellate cocoons as microscopic "conservation traps" comparable to amber.

  7. Excitotoxicity of lathyrus sativus neurotoxin in leech retzius neurons.

    PubMed

    Cemerikic, D; Nedeljkov, V; Lopicic, S; Dragovic, S; Beleslin, B

    2001-01-01

    The effects of Lathyrus sativus neurotoxin were studied on the cell membrane potential and cellular cation composition in Retzius nerve cells of the leech Haemopis sanguisuga, with ion-selective microelectrodes using liquid ion-exchangers. Bath application of 10(-4) mol/l Lathyrus sativus neurotoxin for 3 min depolarized the cell membrane potential and decreased the input resistance of directly polarized membrane in Retzius neurons. At the same time the cellular Na+ activity increased and cellular K+ activity decreased with slow but complete recovery, while the intracellular Ca2+ concentration was not changed. Na+-free Ringer solutions inhibited the depolarizing effect of the neurotoxin on the cell membrane potential. Zero-Ca2+ Ringer solution or Ni2+-Ringer solution had no influence on the depolarizing effect of the neurotoxin on the cell membrane potential. It is obvious that the increase in membrane conductance and depolarization of the cell membrane potential are due to an influx of Na+ into the cell accompanied by an efflux of K+ from the cell.

  8. Sibling competition in a brood-tending leech

    PubMed Central

    Burd, Martin; Govedich, Fredric R; Bateson, Laura

    2006-01-01

    Conflict among siblings over parental investment, particularly over parental feeding, is a feature of family life in many kinds of animals. In some bird species, the size of prey items provided to juveniles has been implicated as a cause of aggressive competition among sibling chicks, because prey size determines whether dominance allows monopolization of parental offerings. Our experiment was meant to test the generality of this factor in creating intrafamilial conflict. We investigated sibling competition in relation to prey size using the carnivorous, brood-tending leech Helobdella papillornata. We equalized the total amount of food available to H. papillornata broods, but varied the size of individual prey items. Competition, measured by disparity in body size at independence, was more intense in broods provisioned with small items than in broods receiving large items, but similar between broods receiving large items and broods fed ad libitum. These patterns suggest that the intensity of conflict did not depend only on the total food amount, but was enhanced by small prey size. Our results indicate that conflict over the provision of parental resources to offspring can have a similar basis across very dissimilar organisms. PMID:16959636

  9. Triassic leech cocoon from Antarctica contains fossil bell animal.

    PubMed

    Bomfleur, Benjamin; Kerp, Hans; Taylor, Thomas N; Moestrup, Øjvind; Taylor, Edith L

    2012-12-18

    Our understanding of the evolution of life on Earth is limited by the imperfection of the fossil record. One reason for this imperfect record is that organisms without hard parts, such as bones, shells, and wood, have a very low potential to enter the fossil record. Occasionally, however, exceptional fossil deposits that preserve soft-bodied organisms provide a rare glimpse of the true biodiversity during past periods of Earth history. We here present an extraordinary find of a fossil ciliate that is encased inside the wall layer of a more than 200 Ma leech cocoon from Antarctica. The microfossil consists of a helically contractile stalk that attaches to a main body with a peristomial feeding apparatus and a large C-shaped macronucleus. It agrees in every aspect with the living bell animals, such as Vorticella. Vorticellids and similar peritrichs are vital constituents of aquatic ecosystems worldwide, but so far have lacked any fossil record. This discovery offers a glimpse of ancient soft-bodied protozoan biotas, and also highlights the potential of clitellate cocoons as microscopic "conservation traps" comparable to amber.

  10. Triassic leech cocoon from Antarctica contains fossil bell animal

    PubMed Central

    Bomfleur, Benjamin; Kerp, Hans; Taylor, Thomas N.; Moestrup, Øjvind; Taylor, Edith L.

    2012-01-01

    Our understanding of the evolution of life on Earth is limited by the imperfection of the fossil record. One reason for this imperfect record is that organisms without hard parts, such as bones, shells, and wood, have a very low potential to enter the fossil record. Occasionally, however, exceptional fossil deposits that preserve soft-bodied organisms provide a rare glimpse of the true biodiversity during past periods of Earth history. We here present an extraordinary find of a fossil ciliate that is encased inside the wall layer of a more than 200 Ma leech cocoon from Antarctica. The microfossil consists of a helically contractile stalk that attaches to a main body with a peristomial feeding apparatus and a large C-shaped macronucleus. It agrees in every aspect with the living bell animals, such as Vorticella. Vorticellids and similar peritrichs are vital constituents of aquatic ecosystems worldwide, but so far have lacked any fossil record. This discovery offers a glimpse of ancient soft-bodied protozoan biotas, and also highlights the potential of clitellate cocoons as microscopic “conservation traps” comparable to amber. PMID:23213234

  11. Channel modulation by tyrosine phosphorylation in an identified leech neuron.

    PubMed Central

    Aniksztejn, L; Catarsi, S; Drapeau, P

    1997-01-01

    1. We have examined the effects of tyrosine phosphorylation on a spontaneously active cation channel that also participates in the modulation of pressure-sensitive (P) neurons in the leech. Cation channel activity in cell-attached or isolated, inside-out membrane patches from P cells in culture was monitored before and after treatments that altered the level of tyrosine phosphorylation. 2. In cell-attached recordings from intact P cells, bath application of genistein, an inhibitor of tyrosine kinases, resulted in a 6.6 +/- 2.6-fold increase in channel activity with no change in the mean open time or amplitude. Daidzein, an inactive form of genistein, was without effect. Addition of pervanadate, a membrane-permeant inhibitor of tyrosine phosphatases, had no effect on its own and blocked the effect of subsequent addition of genistein. 3. In inside-out P cell membrane patch recordings, exposure to a catalytically active fragment of a tyrosine phosphatase resulted in a 10.3 +/- 3.6-fold increase in channel activity with no change in the mean open time or amplitude. Orthovanadate had no effect on channel activity and, when added with the phosphatase, prevented the increase in activity. 4. Our results demonstrate that the basal activity of cation channels is increased by tyrosine dephosphorylation, suggesting a constitutive modulation of channel activity under resting conditions. PMID:9023773

  12. [Inheritance of reproductive traits of medicinal leeches (Hirudo medicinalis L.)].

    PubMed

    Utevskaia, O M; Atramentova, L A

    2002-01-01

    The phenotype variability and inheritance of reproductive traits were investigated in the medicinal leech. Distribution parameters were determined for the following traits: batch size (X = 4.3 +/- 0.2, sigma = 1.7, CV = 40%, As = 0.23 +/- 0.25, Ex = 0.19 +/- 0.51), number of juveniles in a cocoon (X = 10.9 +/- 0.3, sigma = 4.6, CV = 42%, As = 0.31 +/- 0.15, Ex = 0.23 +/- 0.30), and juvenile weight (X = 32.0 +/- 0.3, sigma = 14.9, CV = 47%, As = 1.38 +/- 0.05, Ex = 3.32 +/- 0.11. A nonlinear negative correlation between the number of juveniles in a cocoon and their weight was found (correlation ratio R = 0.86). It was shown that the environmental variance dominated over the genotypic one in the structure of phenotypic variance of the traits studied. The genetic variability is determined mainly by additive gene interactions and, to a small extent, intralocus dominance. The narrow-sense heritability, h2, for batch size was 0.35-0.40; for the number of juveniles in a cocoon, 0.35; for juvenile weight, 0.42.

  13. Leeches (Hirudinida: Piscicolidae)--parasites of Antarctic fish from Channichthyidae family.

    PubMed

    Bielecki, Aleksander; Rokicka, Magdalena; Ropelewska, Ewa; Dziekońska-Rynko, Janina

    2008-01-01

    There has hitherto been very few research projects focusing on ectoparasites of Antarctic fishes. The presently reported study provides data on the prevalence and the intensity of leeches (Hirudinida: Piscicolidae) infecting fishes. The materials were collected in December-February 1986/87 off the Elephant Island, South Georgia, Joinville Island, and South Shetlands. The following leech taxa were recorded in the Antarctic fishes of the family Channichthyidae: Trulliobdella capitis (Brinkmann, 1947); Cryobdella antarctica Epstein, 1970; Nototheniobdella sawyeri Utevsky, 1997; and Cryobdella sp. The above findings constitute new geographic records from off Elephant and Joinville Island and South Georgia.

  14. Microsurgical replantation and postoperative leech treatment of a large severed nasal segment.

    PubMed

    Stemann Andersen, Peter; Elberg, Jens Jørgen

    2012-12-01

    The survival of a microsurgically replanted segment of nose in a 41-year-old woman was facilitated by the assistance of the medicinal leech Hirudo medicinalis. An arterial microanastomosis was made to a severed partial segment of nose with no possibility of recreating a venous anastomosis. The resulting venous congestion was treated with nine days of treatment with a medical leech until venous neovascularisation had been achieved. At follow-up six months after discharge there was a well-heeled nasal segment and a satisfying functional - as well as cosmetic - result.

  15. The physiology and mechanics of undulatory swimming: a student laboratory exercise using medicinal leeches.

    PubMed

    Ellerby, David J

    2009-09-01

    The medicinal leech is a useful animal model for investigating undulatory swimming in the classroom. Unlike many swimming organisms, its swimming performance can be quantified without specialized equipment. A large blood meal alters swimming behavior in a way that can be used to generate a discussion of the hydrodynamics of swimming, muscle mechanics, hydrostatic skeletons, and the physiological features that allow leeches to deal with the volume increase and osmotic load imposed by the meal. Analyses can be carried out at a range of levels tailored to suit a particular class.

  16. Evidence for an endocannabinoid system in the central nervous system of the leech Hirudo medicinalis.

    PubMed

    Matias, I; Bisogno, T; Melck, D; Vandenbulcke, F; Verger-Bocquet, M; De Petrocellis, L; Sergheraert, C; Breton, C; Di Marzo, V; Salzet, M

    2001-03-05

    In invertebrates, like Hydra and sea urchins, evidence for a functional cannabinoid system was described. The partial characterization of a putative CB1 cannabinoid receptor in the leech Hirudo medicinalis led us to investigate the presence of a complete endogenous cannabinoid system in this organism. By using gas chromatography-mass spectrometry, we demonstrate the presence of the endocannabinoids anandamide (N-arachidonoylethanolamine, 21.5+/-0.7 pmol/g) and 2-arachidonoyl-glycerol (147.4+/-42.7 pmol/g), and of the biosynthetic precursor of anandamide, N-arachidonylphosphatidyl-ethanolamine (16.5+/-3.3 pmol/g), in the leech central nervous system (CNS). Anandamide-related molecules such as N-palmitoylethanolamine (32.4+/-1.6 pmol/g) and N-linolenoylethanolamine (5.8 pmol/g) were also detected. We also found an anandamide amidase activity in the leech CNS cytosolic fraction with a maximal activity at pH 7 and little sensitivity to typical fatty acid amide hydrolase (FAAH) inhibitors. Using an antiserum directed against the amidase signature sequence, we focused on the identification and the localization of the leech amidase. Firstly, leech nervous system protein extract was subjected to Western blot analysis, which showed three immunoreactive bands at ca. approximately 42, approximately 46 and approximately 66 kDa. The former and latter bands were very faint and were also detected in whole homogenates from the coelenterate Hydra vulgaris, where the presence of CB1-like receptors, endocannabinoids and a FAAH-like activity was reported previously. Secondly, amidase immunocytochemical detection revealed numerous immunoreactive neurons in the CNS of three species of leeches. In addition, we observed that leech amidase-like immunoreactivity matches to a certain extent with CB1-like immunoreactivity. Finally, we also found that stimulation by anandamide of this receptor leads, as in mammals, to inhibition of cAMP formation, although this effect appeared to be occurring

  17. Free Flap Salvage after Recurrent Venous Thrombosis by Means of Large-Scale Treatment with Medical Leeches

    PubMed Central

    Fujiki, Masahide; Arikawa, Masaki; Kagaya, Yu; Miyamoto, Shimpei

    2016-01-01

    Summary: An anterolateral thigh flap was salvaged using 110 medical leeches in the absence of an available vein for reanastomosis. After surgical construction of the flap for full-thickness thoracic wall reconstruction, the patient developed complete venous occlusion. Specifically, the anastomotic vein developed complete occlusion, and the internal jugular vein had a thrombus. In addition, because the lung was posterior to the flap, the angiogenic area of the flap was very small. When the medical leeches were first applied, the flap showed prominent venous congestion. However, the congestion began to resolve by day 6 of leech use, leading to complete survival of the flap. PMID:28293509

  18. Medicinal Leech Therapy for Glans Penis Congestion After Primary Bladder Exstrophy-Epispadias Repair in an Infant: A Case Report.

    PubMed

    Wagenheim, Gavin N; Au, Jason; Gargollo, Patricio C

    2016-01-01

    Many postoperative complications have been reported after repair of classic bladder exstrophy. We present a case of medicinal leech therapy for glans penis congestion following exstrophy repair in an infant. A 2-week-old male with classic bladder exstrophy underwent complete primary repair. On postoperative day 1, he developed rapidly worsening glans penis venous congestion. Medicinal leech therapy was instituted with antibiotics and blood transfusions to maintain a hematocrit >30%. After 24 hours, venous congestion improved and therapy was discontinued. The patient's remaining hospital course was uncomplicated. Medicinal leeches are an effective therapy to relieve glans penis venous congestion. Copyright © 2015 Elsevier Inc. All rights reserved.

  19. [Lysozyme activity of the salivary gland secretion of the medicinal leech H. verbana, H. medicinalis and H. orientalis].

    PubMed

    Baskova, I P; Kharitonova, O V; Zavalova, L L

    2011-01-01

    Salivary gland secretions of three species of the medicinal leech differ in the level of lysozyme peptidoglycan-lysing activity. Using the synthetic fluorogenic substrate, 4-methyl-umbelliferyl tetra N-acetyl-beta-chitotetraosid, the glycosidase activity (as one of peptidoglycan-lysing activities) of salivary gland secretion of three species of the medicinal leech was quantitatively evaluated in comparison with egg lysozyme. It is supposed, that lysozyme activity of the leech secretions is determined not only by 5 isoforms of destabilase-lysozyme, but by some other enzymes which can utilize this substrate. These may be lysozymes other than i- (invertebrate) lysozymes (such as destabilase-lysozyme, or related enzymes).

  20. Avulsion of the auricle in an anticoagulated patient: is leeching contraindicated? A review and a case.

    PubMed

    Mommsen, Jens; Rodríguez-Fernández, Javier; Mateos-Micas, Mario; Vázquez-Bouso, Olga; Gumbao-Grau, Victor; Forteza-Gonzalez, Gabriel

    2011-06-01

    Amputation of the auricle is a periodic occurrence leading to disfigurement if not treated properly. Venous stasis is a common complication in reattachments and requires decongestant and anticoagulant treatment. Today, leech therapy is the treatment of choice. Common problems are that it is not available everywhere and that it is usually contraindicated in anticoagulated patients. The peculiarities of leech therapy and the various aspects of surgical management are reviewed. A case of a partial amputation of the auricle in a patient under concomitant anticoagulation therapy with warfarin is presented. The amputated part was reattached in another hospital without microvascular anastomosis. The patient presented to our department with early signs of venous congestion. Leech therapy was started 35 hours after trauma, and the patient continued his anticoagulation therapy. With this treatment, 90% of the amputated part was rescued. The anticoagulation therapy of the patient may have played an important role in the first hours after reattachment, preventing capillary thrombosis and in consequence facilitating the minimal oxygenation necessary. The claim that anticoagulation therapy is a contraindication to leeching should be questioned in cases of reattachments in well-controllable locations without arterial anastomosis.

  1. High-yield novel leech hyaluronidase to expedite the preparation of specific hyaluronan oligomers

    PubMed Central

    Jin, Peng; Kang, Zhen; Zhang, Na; Du, Guocheng; Chen, Jian

    2014-01-01

    Hyaluronidases (HAases), particularly leech HAases, have attracted intense attention due to their broad applications in medical treatments and great potential for the enzymatic production of hyaluronan oligosaccharides. However, little is known about this third interesting family of HAases. Here, we applied the random amplification of cDNA ends polymerase chain reaction (RACE-PCR) approach to identify the first leech HAase-encoding gene. By combining protein engineering and high-density culture, we achieved high-level production (8.42 × 105 U ml−1) in the yeast Pichia pastoris secretory expression system. Compared with the commercial bovine testicular HAase, the recombinant leech HAase exhibited superior enzymatic properties. Furthermore, analysis of the hydrolytic process suggested that this novel enzyme adopts a nonprocessive endolytic mode, yielding a narrow-spectrum of specific HA oligosaccharides with different incubation times. Large-scale production of this novel leech HAase will not only greatly promote medical applications but also facilitate the enzymatic production of specific HA oligosaccharides. PMID:24667183

  2. LIFE HISTORY AND DISTRIBUTION OF THE LEECH OLIGOBDELLA BIANNULATA (MOORE, 1900) (EUHIRUDINEA: GLOSSIPHONNIDAE)

    EPA Science Inventory

    Oligobdella biannulata (Moore, 1900) is a rare, endemic leech species originally described from a mountain stream near Blowing Rock, North Carolina. Specimens of O. biannulata were collected seasonally from Fall 1999, to Summer 2002, with new county records in North Carolina and ...

  3. Avulsion of the Auricle in an Anticoagulated Patient: Is Leeching Contraindicated? A Review and a Case

    PubMed Central

    Mommsen, Jens; Rodríguez-Fernández, Javier; Mateos-Micas, Mario; Vázquez-Bouso, Olga; Gumbao-Grau, Victor; Forteza-Gonzalez, Gabriel

    2011-01-01

    Amputation of the auricle is a periodic occurrence leading to disfigurement if not treated properly. Venous stasis is a common complication in reattachments and requires decongestant and anticoagulant treatment. Today, leech therapy is the treatment of choice. Common problems are that it is not available everywhere and that it is usually contraindicated in anticoagulated patients. The peculiarities of leech therapy and the various aspects of surgical management are reviewed. A case of a partial amputation of the auricle in a patient under concomitant anticoagulation therapy with warfarin is presented. The amputated part was reattached in another hospital without microvascular anastomosis. The patient presented to our department with early signs of venous congestion. Leech therapy was started 35 hours after trauma, and the patient continued his anticoagulation therapy. With this treatment, 90% of the amputated part was rescued. The anticoagulation therapy of the patient may have played an important role in the first hours after reattachment, preventing capillary thrombosis and in consequence facilitating the minimal oxygenation necessary. The claim that anticoagulation therapy is a contraindication to leeching should be questioned in cases of reattachments in well-controllable locations without arterial anastomosis. PMID:22655116

  4. LIFE HISTORY AND DISTRIBUTION OF THE LEECH OLIGOBDELLA BIANNULATA (MOORE, 1900) (EUHIRUDINEA: GLOSSIPHONNIDAE)

    EPA Science Inventory

    Oligobdella biannulata (Moore, 1900) is a rare, endemic leech species originally described from a mountain stream near Blowing Rock, North Carolina. Specimens of O. biannulata were collected seasonally from Fall 1999, to Summer 2002, with new county records in North Carolina and ...

  5. The Physiology and Mechanics of Undulatory Swimming: A Student Laboratory Exercise Using Medicinal Leeches

    ERIC Educational Resources Information Center

    Ellerby, David J.

    2009-01-01

    The medicinal leech is a useful animal model for investigating undulatory swimming in the classroom. Unlike many swimming organisms, its swimming performance can be quantified without specialized equipment. A large blood meal alters swimming behavior in a way that can be used to generate a discussion of the hydrodynamics of swimming, muscle…

  6. Experimental infection of yellow stingrays Urobatis jamaicensis with the marine leech Branchellion torpedinis.

    PubMed

    Marancik, David P; Dove, Alistair D; Camus, Alvin C

    2012-10-10

    Infestations of elasmobranchs by the marine leech Branchellion torpedinis can be problematic in aquaria and negatively affect host health. To better characterize the extent and pathogenesis of disease, 12 yellow stingrays Urobatis jamaicensis were infected with 1 or 3 leeches for 14 d. Leeches were associated with anorexia, extensive cutaneous ulceration, decreased host packed cell volume (PCV) and serum total solids (TS), and mortality in 3 rays. Average decrease in host PCV positively correlated with ulcer size and parasite:host ratio. Average decrease in host serum TS positively correlated with parasite:host ratio. Blood chemistry and total white blood cell counts revealed no significant trends. Additional necropsy findings included gill and splenic pallor, pericardial edema, perirenal edema, and decreased hepatocellular lipid deposits. Microscopic evaluation of leeches demonstrated host erythrocytes and proteinaceous fluid within parasite intestines, confirming active blood feeding. Results indicate B. torpedinis has the potential to cause significant disease in elasmobranchs, including death in as few as 5 d, and identifies ulcer size and parasite:host ratio as risk factors for disease. Elucidation of this host-parasite interaction helps characterize host response to parasites and facilitate care of parasitized elasmobranchs in aquarium and wild settings.

  7. High contrast imaging at the LBT: the LEECH exoplanet imaging survey

    NASA Astrophysics Data System (ADS)

    Skemer, Andrew J.; Hinz, Philip; Esposito, Simone; Skrutskie, Michael F.; Defrère, Denis; Bailey, Vanessa; Leisenring, Jarron; Apai, Daniel; Biller, Beth; Bonnefoy, Mickaël.; Brandner, Wolfgang; Buenzli, Esther; Close, Laird; Crepp, Justin; De Rosa, Robert J.; Desidera, Silvano; Eisner, Josh; Fortney, Jonathan; Henning, Thomas; Hofmann, Karl-Heinz; Kopytova, Taisiya; Maire, Anne-Lise; Males, Jared R.; Millan-Gabet, Rafael; Morzinski, Katie; Oza, Apurva; Patience, Jenny; Rajan, Abhijith; Rieke, George; Schertl, Dieter; Schlieder, Joshua; Su, Kate; Vaz, Amali; Ward-Duong, Kimberly; Weigelt, Gerd; Woodward, Charles E.; Zimmerman, Neil

    2014-07-01

    In Spring 2013, the LEECH (LBTI Exozodi Exoplanet Common Hunt) survey began its ~130-night campaign from the Large Binocular Telescope (LBT) atop Mt Graham, Arizona. This survey benefits from the many technological achievements of the LBT, including two 8.4-meter mirrors on a single fixed mount, dual adaptive secondary mirrors for high Strehl performance, and a cold beam combiner to dramatically reduce the telescope's overall background emissivity. LEECH neatly complements other high-contrast planet imaging efforts by observing stars at L' (3.8 μm), as opposed to the shorter wavelength near-infrared bands (1-2.4 μm) of other surveys. This portion of the spectrum offers deep mass sensitivity, especially around nearby adolescent (~0.1-1 Gyr) stars. LEECH's contrast is competitive with other extreme adaptive optics systems, while providing an alternative survey strategy. Additionally, LEECH is characterizing known exoplanetary systems with observations from 3-5μm in preparation for JWST.

  8. The Physiology and Mechanics of Undulatory Swimming: A Student Laboratory Exercise Using Medicinal Leeches

    ERIC Educational Resources Information Center

    Ellerby, David J.

    2009-01-01

    The medicinal leech is a useful animal model for investigating undulatory swimming in the classroom. Unlike many swimming organisms, its swimming performance can be quantified without specialized equipment. A large blood meal alters swimming behavior in a way that can be used to generate a discussion of the hydrodynamics of swimming, muscle…

  9. ERPOBDELLA LAHONTANA (ANNELIDA: HIRUDINEA: ARHYNCHOBDELLIDA: ERPOBDELLIDAE), A NEW SPECIES OF FRESHWATER LEECH FROM NORTH AMERICA

    EPA Science Inventory

    New species of a leech, Erpobdella lahontana, is described from the Lahontan Basin in California and Nevada of the western United States. This species has four pairs of eyes, the preatrial loops of male paired ducts extend to ganglion XI, and the male and female gonopores are loc...

  10. Behavioral choice across leech species: chacun à son goût

    PubMed Central

    Gaudry, Q.; Ruiz, N.; Huang, T.; Kristan, W. B.; Kristan, W. B.

    2010-01-01

    At any one time, animals are simultaneously bombarded with many sensory stimuli, but they typically choose to respond to only a few of them. We used multidimensional analysis to determine the behavioral responses of six species of leeches to stimulation, as the responses are affected by species identity, diet, behavioral state and stimulus location. Our results show that each of the species tested while not feeding displayed remarkably similar behaviors in response to tactile stimulation of the surface of the body. When not feeding, stimulus location was the most reliable factor in determining behavioral response. While feeding, the three sanguivorous (bloodsucking) species tested ignored stimulation, whereas the three carnivorous leeches abandoned feeding in favor of locomotory responses, regardless of phylogenetic relationships. In the sanguivorous leeches, feeding abolished all mechanically elicited responses and mechanical stimulation in turn had no effect on feeding. We also show that the behavioral hierarchy of leeches was fixed and unchanging even in species that can consume both a carnivorous and a sanguivorous diet. PMID:20348348

  11. Intracellular Recording, Sensory Field Mapping, and Culturing Identified Neurons in the Leech, Hirudo medicinalis

    PubMed Central

    Titlow, Josh; Majeed, Zana R.; Nicholls, John G; Cooper, Robin L.

    2013-01-01

    The freshwater leech, Hirudo medicinalis, is a versatile model organism that has been used to address scientific questions in the fields of neurophysiology, neuroethology, and developmental biology. The goal of this report is to consolidate experimental techniques from the leech system into a single article that will be of use to physiologists with expertise in other nervous system preparations, or to biology students with little or no electrophysiology experience. We demonstrate how to dissect the leech for recording intracellularly from identified neural circuits in the ganglion. Next we show how individual cells of known function can be removed from the ganglion to be cultured in a Petri dish, and how to record from those neurons in culture. Then we demonstrate how to prepare a patch of innervated skin to be used for mapping sensory or motor fields. These leech preparations are still widely used to address basic electrical properties of neural networks, behavior, synaptogenesis, and development. They are also an appropriate training module for neuroscience or physiology teaching laboratories. PMID:24299987

  12. The Leech Nervous System: A Valuable Model to Study the Microglia Involvement in Regenerative Processes

    PubMed Central

    Le Marrec-Croq, Françoise; Drago, Francesco; Vizioli, Jacopo; Sautière, Pierre-Eric; Lefebvre, Christophe

    2013-01-01

    Microglia are intrinsic components of the central nervous system (CNS). During pathologies in mammals, inflammatory processes implicate the resident microglia and the infiltration of blood cells including macrophages. Functions of microglia appear to be complex as they exhibit both neuroprotective and neurotoxic effects during neuropathological conditions in vivo and in vitro. The medicinal leech Hirudo medicinalis is a well-known model in neurobiology due to its ability to naturally repair its CNS following injury. Considering the low infiltration of blood cells in this process, the leech CNS is studied to specify the activation mechanisms of only resident microglial cells. The microglia recruitment is known to be essential for the usual sprouting of injured axons and does not require any other glial cells. The present review will describe the questions which are addressed to understand the nerve repair. They will discuss the implication of leech factors in the microglial accumulation, the identification of nerve cells producing these molecules, and the study of different microglial subsets. Those questions aim to better understand the mechanisms of microglial cell recruitment and their crosstalk with damaged neurons. The study of this dialog is necessary to elucidate the balance of the inflammation leading to the leech CNS repair. PMID:23878582

  13. Intracellular recording, sensory field mapping, and culturing identified neurons in the leech, Hirudo medicinalis.

    PubMed

    Titlow, Josh; Majeed, Zana R; Nicholls, John G; Cooper, Robin L

    2013-11-04

    The freshwater leech, Hirudo medicinalis, is a versatile model organism that has been used to address scientific questions in the fields of neurophysiology, neuroethology, and developmental biology. The goal of this report is to consolidate experimental techniques from the leech system into a single article that will be of use to physiologists with expertise in other nervous system preparations, or to biology students with little or no electrophysiology experience. We demonstrate how to dissect the leech for recording intracellularly from identified neural circuits in the ganglion. Next we show how individual cells of known function can be removed from the ganglion to be cultured in a Petri dish, and how to record from those neurons in culture. Then we demonstrate how to prepare a patch of innervated skin to be used for mapping sensory or motor fields. These leech preparations are still widely used to address basic electrical properties of neural networks, behavior, synaptogenesis, and development. They are also an appropriate training module for neuroscience or physiology teaching laboratories.

  14. Molecular characterization and embryonic expression of innexins in the leech Hirudo medicinalis.

    PubMed

    Dykes, Iain M; Macagno, Eduardo R

    2006-04-01

    Gap junctions are direct intercellular channels that permit the passage of ions and small signaling molecules. The temporal and spatial regulation of gap junctional communication is, thus, one mechanism by which cell interactions, and hence cell properties and cell fate, may be regulated during development. The nervous system of the leech, Hirudo medicinalis, is a particularly advantageous system in which to study developmental mechanisms involving gap junctions because interactions between identified cells may be studied in vivo in both the embryo and the adult. As in most invertebrates, gap junctions in the leech are composed of innexin proteins, which are distantly related to the vertebrate pannexins and are encoded by a multi-gene family. We have cloned ten novel leech innexins and describe the expression of these, plus two other previously reported members of this gene family, in the leech embryo between embryonic days 6 and 12, a period during which the main features of the central nervous system are established. Four innexins are expressed in neurons and two in glia, while several innexins are expressed in the excretory, circulatory, and reproductive organs. Of particular interest is Hm-inx6, whose expression appears to be restricted to the characterized S cell and two other neurons putatively identified as presynaptic to this cell. Two other innexins also show highly restricted expressions in neurons and may be developmentally regulated.

  15. Identification, Isolation and Expansion of Myoendothelial Cells Involved in Leech Muscle Regeneration

    PubMed Central

    Grimaldi, Annalisa; Banfi, Serena; Gerosa, Laura; Tettamanti, Gianluca; Noonan, Douglas M.; Valvassori, Roberto; de Eguileor, Magda

    2009-01-01

    Adult skeletal muscle in vertebrates contains myoendothelial cells that express both myogenic and endothelial markers, and which are able to differentiate into myogenic cells to contribute to muscle regeneration. In spite of intensive research efforts, numerous questions remain regarding the role of cytokine signalling on myoendothelial cell differentiation and muscle regeneration. Here we used Hirudo medicinalis (Annelid, leech) as an emerging new model to study myoendothelial cells and muscle regeneration. Although the leech has relative anatomical simplicity, it shows a striking similarity with vertebrate responses and is a reliable model for studying a variety of basic events, such as tissue repair. Double immunohistochemical analysis were used to characterize myoendothelial cells in leeches and, by injecting in vivo the matrigel biopolymer supplemented with the cytokine Vascular Endothelial Growth Factor (VEGF), we were able to isolate this specific cell population expressing myogenic and endothelial markers. We then evaluated the effect of VEGF on these cells in vitro. Our data indicate that, similar to that proposed for vertebrates, myoendothelial cells of the leech directly participate in myogenesis both in vivo and in vitro, and that VEGF secretion is involved in the recruitment and expansion of these muscle progenitor cells. PMID:19876402

  16. ERPOBDELLA LAHONTANA (ANNELIDA: HIRUDINEA: ARHYNCHOBDELLIDA: ERPOBDELLIDAE), A NEW SPECIES OF FRESHWATER LEECH FROM NORTH AMERICA

    EPA Science Inventory

    New species of a leech, Erpobdella lahontana, is described from the Lahontan Basin in California and Nevada of the western United States. This species has four pairs of eyes, the preatrial loops of male paired ducts extend to ganglion XI, and the male and female gonopores are loc...

  17. Effects of 5-hydroxytryptamine on the dorsal muscle of the leech (hirudo medicinalis)

    PubMed Central

    Schain, R. J.

    1961-01-01

    5-Hydroxytryptamine has an inhibiting effect on the leech muscle. It reduces the contractions produced by acetylcholine or nicotine and accelerates the relaxation of the muscle when these substances are washed out. This acceleration of relaxation allows a more rapid assay of acetylcholine in this preparation. PMID:13747232

  18. An extract from medical leech improve the function of endothelial cells in vitro and attenuates atherosclerosis in ApoE null mice by reducing macrophages in the lesions.

    PubMed

    Wang, Yao; Zhao, Xin; Wang, Yun-Shan; Song, Shu-Liang; Liang, Hao; Ji, Ai-Guo

    2014-12-05

    Medicinal leech has been widely used as a traditional Chinese medicine in cardiovascular diseases. However, its pharmaceutical effect is not fully revealed. The goal of this study was to determine whether a leech extract has the effect of anti-atherosclerosis in ApoE −/− mice and the mechanism of this effect. In vivo experiments: ApoE −/− mice fed on high-cholesterol diet were separated into 5 groups. Control group was administrated with normal water; leech extract of low dose treatment group was given a leech extract of 0.02 g/kg/d; leech extract of medium dose treatment group was given a leech extract of 0.1 g/kg/d; leech extract of high dose treatment group was given a leech extract of 0.5 g/kg/d; simvastatin group was given simvastatin of 10 mg/kg/d. Leech extract significantly reduced atherosclerotic lesions in aortic root compared with control group. And the number of macrophage in or around the atherosclerosis plaque is significantly reduced in the leech extract groups compared with control group. In vitro experiments: human endothelial cell line, EA.hy926, was induced with TNF-α to perform endothelial dysfunction. Control group: EA.hy926 cells with no special treatment; TNF-α group: EA.hy926 cells were induced by 10 ng/ml TNF-α for 6 h; leech extract only group: EA.hy926 cells were treated with 200 mg/ml leech extract only; leech extract and TNF-α group: 200 mg/ml leech extract was applied before TNF-α induction. Protein and mRNA level were detected in each group, leech extract can decrease the expression of intercellular adhesion factor (ICAM-1) and monocyte chemotactic protein (MCP-1) compared with TNF-α group. Furthermore, it showed less adhesion and migration of THP-1 cells to EA.hy926 cells in the adhesion assay and transwell assay. The NF-κB translation to nucleus was blocked by leech extract in the NF-κB translocation assay. Leech extract could obviously attenuate the area of atherosclerosis lesion in ApoE −/− mice. And this

  19. Infections following the application of leeches: two case reports and review of the literature

    PubMed Central

    2012-01-01

    Introduction Since the 1980s, leeches have been ingeniously used in the management of venous flap congestion. The presence of anticoagulative substances in their saliva improves the blood drainage. Their digestive tract contains several bacterial species, the main ones being Aeromonas hydrophila and Aeromonas veronii biovar sobria, which contribute to the digestion of ingested blood. These bacteria can be the cause of infections. Case presentation We report two cases of septicemia related to Aeromonas veronii biovar sobria that presented after leeches had been applied to congested transverse rectus abdominis myocutaneous flaps for delayed mammary reconstructions. Patient number 1 was a 55-year-old Caucasian woman who underwent a delayed breast reconstruction procedure. On the sixth postoperative day she showed a clinical presentation of septicemia. Aeromonas veronii biovar sobria was identified in the patient’s skin and blood bacteriological samples. Her fever ceased after 4 days of antibiotic treatment. Patient number 2 was a 56-year-old Caucasian woman who underwent a delayed breast reconstruction procedure. On the seventh postoperative day we noticed that she showed a clinical presentation of septicemia. Aeromonas veronii biovar sobria was identified in the patient’s blood cultures and local bacteriological samples. An antibiogram showed resistance to amoxicillin/clavulanic acid. Her fever ceased on the eleventh postoperative day after 4 days of antibiotic treatment. Conclusion The rate of infection after application of leeches is not negligible. The concentration of Aeromonas inside the digestive tracts of leeches largely decreases when the patient is under antibiotic therapy. These germs are sensitive to third-generation cephalosporins and fluoroquinolones and resistant to amoxicillin/clavulanic acid. We recommend preventive treatment based on classical measures of asepsis and on oral antibioprophylaxy with a fluoroquinolone during the whole period of

  20. BIOLOGY AND OCCURRENCE OF THE LEECH, ACTINOBDELLA INEQUIANNULATA (ANNELIDA: HIRUDINEA: GLOSSIPHONIIDAE) PARASITIC ON TWO SPECIES OF SUCKERS

    EPA Science Inventory

    Actinobdella inequiannulata was found on the white sucker, Catostomus commersoni, and less frequently on the longnose sucker, Catostomus catostomus, in Algonquin Provincial Park, Ontario, Canada. This study established the presence of only one species of leech, Actinobdela inequ...

  1. Cloning and expression of a cDNA coding for the anticoagulant hirudin from the bloodsucking leech, Hirudo medicinalis.

    PubMed Central

    Harvey, R P; Degryse, E; Stefani, L; Schamber, F; Cazenave, J P; Courtney, M; Tolstoshev, P; Lecocq, J P

    1986-01-01

    Cloned cDNAs have been isolated that encode a variant of hirudin, a potent thrombin inhibitor that is secreted by the salivary glands of the medicinal leech, Hirudo medicinalis. This variant probably corresponds to a form that has been purified from leech heads but differs in amino acid sequence from the hirudin purified from whole leeches. There are at least three hirudin transcripts detectable in leech RNAs that are different in size, site of synthesis, inducibility by starvation, and relationship to hirudin activity. The new hirudin variant predicted by the cDNA and the heterodisperse transcription products suggest a hirudin protein family. The hirudin cDNA was expressed in Escherichia coli under the control of the bacteriophage lambda PL promoter. The recombinant product is biologically active, inhibiting the cleavage by thrombin of fibrinogen and a synthetic tripeptide substrate. Images PMID:3513162

  2. BIOLOGY AND OCCURRENCE OF THE LEECH, ACTINOBDELLA INEQUIANNULATA (ANNELIDA: HIRUDINEA: GLOSSIPHONIIDAE) PARASITIC ON TWO SPECIES OF SUCKERS

    EPA Science Inventory

    Actinobdella inequiannulata was found on the white sucker, Catostomus commersoni, and less frequently on the longnose sucker, Catostomus catostomus, in Algonquin Provincial Park, Ontario, Canada. This study established the presence of only one species of leech, Actinobdela inequ...

  3. [Infection with Aeromonas hydrophila after use of leeches (Hirudo medicinalis) in a free microvascular osteo-(myo-)cutaneous flap--suggestions for successful management].

    PubMed

    Kalbermatten, D F; Rieger, U M; Uike, K; Erba, P; Laifer, G; Hintermann, B; Pierer, G

    2007-04-01

    Medical leeches (Hirudo medicinalis) are an important therapeutic option in the treatment of venous congestion of flaps. In a case report an infection with Aeromonas hydrophila of a free microvascular osteo-(myo)-cutaneous flap after leech application for venous congestion is described and the current literature reviewed. Infection associated with leech therapy is a documented complication of leech application, with reported incidences ranging from 2.4 to 20 %. In some cases an infection of the wound developed with Aeromonas hydrophila, a Gram-negative rod that lives symbiotically in the intestines of the leech. Because of the risk of graft loss, early diagnosis and immediate initiation of an empirical intravenous antibiotic therapy with Piperacillin/Tacobactam or a third or fourth generation cephalosporins are essential even before results for sensitivity testing are received. An alternative is a short-term preemptive therapy with Cotrimoxazol or Ciprofloxacin during leech application.

  4. A new leech species (Hirudinida: Erpobdellidae: Erpobdella) from a cave in the West Azerbaijan province of Iran.

    PubMed

    Cichocka, Joanna M; Bielecki, Aleksander; Kur, Jarosław; Pikuła, Dorota; Kilikowska, Adrianna; Biernacka, Beata

    2015-09-09

    Erpobdella borisi n. sp. is a predatory leech inhabiting cave waters in Iran. Probably, it is either a troglobiont or troglophile. The leech has no eyes, and the complete mid-body somite is divided unequally into five annuli. Results of phylogenetic analysis based on morphological characters and COI gene sequence indicate the species to be closely related to Erpobdella japonica, E. octoculata and E. testacea.

  5. Proteins and peptides of the salivary gland secretion of medicinal leeches Hirudo verbana, H. medicinalis, and H. orientalis.

    PubMed

    Baskova, I P; Kostrjukova, E S; Vlasova, M A; Kharitonova, O V; Levitskiy, S A; Zavalova, L L; Moshkovskii, S A; Lazarev, V N

    2008-03-01

    The protein and peptide composition of medicinal leech salivary gland secretion (SGS) was analyzed in preparations obtained in July from three species--Hirudo verbana, H. medicinalis, and H. orientalis. Two-dimensional electrophoresis (molecular mass 10-150 kD and pI 3-10) revealed no distinctions in the distribution of over 100 silver-stained proteins. Differences were noted only in intensity of 10 protein spots at 30-90 kD and pI 4.7-7.5. Mass spectrometric profiling of SGS of the three leech species using the Zip-Tip/golden chip scheme and cation-exchanging chips CM-10 revealed over 50 components in SGS of each of the three leech species. It was noted that 30-40% of the individual masses of the SGS of each leech species fall within the masses present in SGS of at least one of the two other species. This rather small part of the total mass may be indicative of a high polymorphism of amino acid sequences or a high frequency of posttranslational modifications of the SGS proteins and peptides. Calculation of Jacquard's coefficient showed that H. medicinalis and H. orientalis are closest to each other in SGS composition, which is consistent with data in the literature on the phylogenetic relationship between these two species of medicinal leech. Comparison of detected molecular masses with those of six known biologically active compounds produced by medicinal leeches revealed their uneven distribution in SGS of each of the three medicinal leech species. This opens prospects for using certain species of medicinal leech for targeted therapy of various pathologies.

  6. Basic morphofunctional features of pharmaceutic leech (Hirudo verbana Carena, 1820) tissues in various forms of response after hirudotherapeutic procedures.

    PubMed

    Frolov, Aleksandr K; Litvinenko, Raisa A

    2015-01-01

    It is analyzed morphofunctional features of Hirudo verbana tissues on histological sections in order to determine the possible causes of their partial death after hirudotherapeutic procedure. In the study was taken 4 groups of leeches: hungry (control), healthy well-fed, well-fed that vomited blood and well-fed dead. Morphofunctional changes in midgut of healthy well-fed H. verbana compared to the hungry are the changes of epithelium height through stretching eaten blood. In well-fed that vomited blood and well-fed dead leeches in the intestinal epithelium were observed degenerative processes that are accompanied by a decrease in the height of the epithelium, its partial desquamation. Botryoidal tissue of hungry leeches is in an inactive state, in healthy well-fed ones it responds with an increased physiological vascularization, activation of some botryoidal granulocytes with an increase in their average size, on receipt of the products of digestion. Activation of some botryoidal granulocytes, without a significant increase in their sizes, in well-fed leeches that vomited eaten blood and well-fed dead leeches, with increased infiltration of the adjacent connective tissue lymphocyte- and macrophage-like cells, amebocytes and granulocytes indicates the development of pathological processes, which are resulting in immunological conflict between eaten human blood and leeches' tissue microenvironment.

  7. Model of intersegmental coordination in the leech heartbeat neuronal network.

    PubMed

    Hill, Andrew A V; Masino, Mark A; Calabrese, Ronald L

    2002-03-01

    We have created a computational model of the timing network that paces the heartbeat of the medicinal leech, Hirudo medicinalis. The rhythmic activity of this network originates from two segmental oscillators located in the third and fourth midbody ganglia. In the intact nerve cord, these segmental oscillators are mutually entrained to the same cycle period. Although experiments have shown that the segmental oscillators are coupled by inhibitory coordinating interneurons, the underlying mechanisms of intersegmental coordination have not yet been elucidated. To help understand this coordination, we have created a simple computational model with two variants: symmetric and asymmetric. In the symmetric model, neurons within each segmental oscillator called oscillator interneurons, inhibit the coordinating interneurons. In contrast, in the asymmetric model only the oscillator interneurons of one segmental oscillator inhibit the coordinating interneurons. In the symmetric model, when two segmental oscillators with different inherent periods are coupled, the faster one leads in phase, and the period of the coupled system is equal to the period of the faster oscillator. This behavior arises because, during each oscillation cycle, the oscillator interneurons of the faster segmental oscillator begin to burst before those of the slower oscillator, thereby terminating spike activity in the coordinating interneurons. Thus there is a brief period of time in each cycle when the oscillator interneurons of the slower segmental oscillator are relieved of inhibition from the coordinating interneurons. This "removal of synaptic inhibition" allows, within certain limits, the slower segmental oscillator to be sped to the period of the faster one. Thus the symmetric model demonstrates a plausible biophysical mechanism by which one segmental oscillator can entrain the other. In general the asymmetric model, in which only one segmental oscillator has the ability to inhibit the

  8. Earthquake relocation near the Leech River Fault, southern Vancouver Island

    NASA Astrophysics Data System (ADS)

    Li, G.; Liu, Y.; Regalla, C.

    2015-12-01

    The Leech River Fault (LRF), a northeast dipping thrust, extends across the southern tip of Vancouver Island in Southwest British Columbia, where local tectonic regime is dominated by the subduction of the Juan de Fuca plate beneath the North American plate at the present rate of 40-50 mm/year. British Columbia geologic map (Geoscience Map 2009-1A) shows that this area also consists of many crosscutting minor faults in addition to the San Juan Fault north of the LRF. To investigate the seismic evidence of the subsurface structures of these minor faults and of possible hidden active structures in this area, precise earthquake locations are required. In this study, we relocate 941 earthquakes reported by Canadian National Seismograph Network (CNSN) catalog from 2000 to 2015 within a 100km x 55km study area surrounding the LRF. We use HypoDD [Waldhauser, F., 2001] double-difference relocation method by combining P/S phase arrivals provided by the CNSN at 169 stations and waveform data with correlation coefficient values greater than 0.7 at 50 common stations and event separation less than 10km. A total of 900 out of the 931 events satisfy the above relocation criteria. Velocity model used is a 1-D model extracted from the Ramachandran et al. (2005) model. Average relative location errors estimated by the bootstrap method are 546.5m (horizontal) and 1128.6m (in depth). Absolute errors reported by SVD method for individual clusters are ~100m in both dimensions. We select 5 clusters visually according to their epicenters (see figure). Cluster 1 is parallel to the LRF and a thrust FID #60. Clusters 2 and 3 are bounded by two faults: FID #75, a northeast dipping thrust marking the southwestern boundary of the Wrangellia terrane, and FID #2 marking the northern boundary. Clusters 4 and 5, to the northeast and northwest of Victoria respectively, however, do not represent the surface traces of any mapped faults. The depth profile of Cluster 5 depicts a hidden northeast

  9. Morphological and molecular characterization of a marine fish trypanosome from South Africa, including its development in a leech vector

    PubMed Central

    2014-01-01

    Background Trypanosomes are ubiquitous blood parasites of marine and freshwater fishes, typically transmitted by aquatic leeches. Phylogenetic studies have been dominated by examples derived from freshwater fishes, with few marine representatives. Furthermore, life cycle studies on marine fish trypanosomes have focused on those of the northern hemisphere. In this investigation, we have examined the life cycle and molecular taxonomy of a marine fish trypanosome from South Africa. Methods To locate trypanosome stages, leeches were removed from fishes captured on the west and south coasts of South Africa, and fish blood films and leech squashes were Giemsa-stained and screened; leeches were also examined histologically. To determine whether trypanosome stages in fishes and leeches were of the same genotype, DNA was extracted from Giemsa-stained fish blood films and leech squashes, and from fish whole blood. Fragments of the 18S rRNA gene were amplified by PCR using trypanosome-specific primers and sequenced. Resulting sequence data were compared with each other and with published trypanosome 18S rDNA sequences, and used for phylogenetic analysis. Results Trypanosomes were detected in blood films from fishes of the families Clinidae, Blenniidae and Gobiidae. The flagellates ranged in size and staining properties within the films and across fish hosts. In squashes and histological sections of adult and juvenile leeches, identified as Zeylanicobdella arugamensis, trypanosome developmental stages were predominantly slender epimastigotes. Sequence data showed that trypanosomes derived from fishes were identical, irrespective of whether they were small or large forms; sequences derived largely from leech epimastigotes were also identical to those obtained from fish trypanosomes. Fish and leech trypanosome sequences fell into a marine fish aquatic clade, and aligned most closely with two trypanosome sequences from marine fishes off Norway. Conclusions Combined morphological

  10. Morphological and molecular characterization of a marine fish trypanosome from South Africa, including its development in a leech vector.

    PubMed

    Hayes, Polly M; Lawton, Scott P; Smit, Nico J; Gibson, Wendy C; Davies, Angela J

    2014-01-24

    Trypanosomes are ubiquitous blood parasites of marine and freshwater fishes, typically transmitted by aquatic leeches. Phylogenetic studies have been dominated by examples derived from freshwater fishes, with few marine representatives. Furthermore, life cycle studies on marine fish trypanosomes have focused on those of the northern hemisphere. In this investigation, we have examined the life cycle and molecular taxonomy of a marine fish trypanosome from South Africa. To locate trypanosome stages, leeches were removed from fishes captured on the west and south coasts of South Africa, and fish blood films and leech squashes were Giemsa-stained and screened; leeches were also examined histologically. To determine whether trypanosome stages in fishes and leeches were of the same genotype, DNA was extracted from Giemsa-stained fish blood films and leech squashes, and from fish whole blood. Fragments of the 18S rRNA gene were amplified by PCR using trypanosome-specific primers and sequenced. Resulting sequence data were compared with each other and with published trypanosome 18S rDNA sequences, and used for phylogenetic analysis. Trypanosomes were detected in blood films from fishes of the families Clinidae, Blenniidae and Gobiidae. The flagellates ranged in size and staining properties within the films and across fish hosts. In squashes and histological sections of adult and juvenile leeches, identified as Zeylanicobdella arugamensis, trypanosome developmental stages were predominantly slender epimastigotes. Sequence data showed that trypanosomes derived from fishes were identical, irrespective of whether they were small or large forms; sequences derived largely from leech epimastigotes were also identical to those obtained from fish trypanosomes. Fish and leech trypanosome sequences fell into a marine fish aquatic clade, and aligned most closely with two trypanosome sequences from marine fishes off Norway. Combined morphological and molecular methods indicate that the

  11. A glutamine-rich factor affects stem cell genesis in leech.

    PubMed

    Hohenstein, Kristi A; Lang, Shirley A; Nuthulaganti, Tej; Shain, Daniel H

    2010-01-01

    Leech embryogenesis is a model for investigating cellular and molecular processes of development. Due to the unusually large size of embryonic stem cells (teloblasts; 50 - 300 μm) in the glossiphoniid leech, Theromyzon tessulatum, and the presence of identifiable stem cell precursors (proteloblasts), we previously isolated a group of genes up-regulated upon stem cell birth. In the current study, we show that one of these genes, designated Tpr (Theromyzon proliferation), is required for normal stem cell genesis; specifically, transient Tpr knockdown experiments conducted with antisense oligonucleotides and monitored by semi-quantitative RT-PCR, caused abnormal proteloblast proliferation leading to embryonic death, but did not overtly affect neuroectodermal or mesodermal stem cell development once these cells were born. Tpr encodes a large, glutamine-rich (~34%) domain that shares compositional similarity with strong transcriptional enhancers, many of which have been linked with trinucleotide repeat disorders (e.g., Huntingtons).

  12. Functional morphology of suction discs and attachment performance of the Mediterranean medicinal leech (Hirudo verbana Carena)

    PubMed Central

    Eberhard, Laura; Gallenmüller, Friederike; Speck, Thomas

    2016-01-01

    Medicinal leeches use their suction discs for locomotion, adhesion to the host and, in the case of the anterior disc, also for blood ingestion. The biomechanics of their suction-based adhesion systems has been little understood until now. We investigated the functional morphology of the anterior and posterior suckers of Hirudo verbana by using light and scanning electron microscopy. Furthermore, we analysed the adhesion qualitatively and quantitatively by conducting behavioural and mechanical experiments. Our high-speed video analyses provide new insights into the attachment and detachment processes and we present a detailed description of the leech locomotion cycle. Pull-off force measurements of the anterior and posterior suction organs on seven different substrates under both aerial and water-submersed conditions reveal a significant influence of the surrounding medium, the substrate surface roughness and the tested organ on attachment forces and tenacities. PMID:27075001

  13. New species of rhynchobdellid leech (Hirudinea: Glossiphoniidae): a parasite of turtles from Chiapas, Mexico.

    PubMed

    López-Jiménez, Serapio; Oceguera-Figueroa, Alejandro

    2009-12-01

    A new species of rhynchobdellid leech, Placobdella ringueleti n. sp., is described based on the examination of 25 specimens. Leeches were collected from body appendages of freshwater turtles (Kinosternon leucostomum, Dermatemys mawii, and Staurotypus triporcatus ) from Chiapas, Mexico. The new species resembles other members of the genus in the ocular morphology, bilobated ovaries, and presence of 1 pair of elongated mycetomes, but it is distinguished from them by the dorsal papillar pattern, by having a continuous longitudinal mid-dorsal stripe along the dorsal surface and papillae on the dorsal surface of the posterior sucker. Placobdella ringueleti represents the third species of the genus in Mexico. The type locality of the new species is part of the “El Ocote” Biosphere Reserve, Chiapas, Mexico.

  14. A mathematical model of motorneuron dynamics in the heartbeat of the leech

    NASA Astrophysics Data System (ADS)

    Buono, Pietro-Luciano; Palacios, A.

    2004-02-01

    The heartbeat of the medicinal leech is driven by direct contact between two arrays of motorneurons and two lateral blood vessels. At any given time, motorneurons exhibit one of two alternating states so that, on one side of the animal, the heart beats in a rear-to-front fashion (peristaltic), while on the other side the heart beats synchronously. Every 20 heartbeats, approximately, the two sides switch modes. It is known that the heartbeat rhythm is generated through burst of oscillatory activity produced by a central pattern generator (CPG) network of neurons. However, to the best of our knowledge, how the CPG activity is translated into peristaltic and synchronous rhythms in the motorneurons is yet unknown. In this work, we use symmetric systems of differential equations, accompanied with computational simulations, to investigate possible mechanisms for generating the motorneuron activity that characterizes the heartbeat of leeches and in particular the switching scenario.

  15. Annotated checklist of the leech species diversity in the Maloe More Strait of Lake Baikal, Russia

    PubMed Central

    Kaygorodova, Irina A.

    2015-01-01

    Abstract In this paper, the very first checklist of the freshwater leeches of Maloe More Strait, a special part of Lake Baikal, is presented. It includes 14 free-living and parasitic species, of which four species belong to endemic Baikal genera – two species from Baicalobdella and one species each from Baicaloclepsis and Codonobdella. The checklist highlights six potentially new morphological species recorded for the first time in the area. The exact systematic position is stated for all leech species. Each species from the list is provided with information on taxonomic synonymy, data on its geographic distribution, and ecological characteristics. New species records are additionally provided with brief morphological characteristics and photos of their external morphology. PMID:26798292

  16. Alpha-conotoxin ImI disrupts central control of swimming in the medicinal leech.

    PubMed

    Wagenaar, Daniel A; Gonzalez, Ruben; Ries, David C; Kristan, William B; French, Kathleen A

    2010-11-26

    Medicinal leeches (Hirudo spp.) swim using a metachronal, front-to-back undulation. The behavior is generated by central pattern generators (CPGs) distributed along the animal's midbody ganglia and is coordinated by both central and peripheral mechanisms. Here we report that a component of the venom of Conus imperialis, α-conotoxin ImI, known to block nicotinic acetyl-choline receptors in other species, disrupts swimming. Leeches injected with the toxin swam in circles with exaggerated dorsoventral bends and reduced forward velocity. Fictive swimming in isolated nerve cords was even more strongly disrupted, indicating that the toxin targets the CPGs and central coordination, while peripheral coordination partially rescues the behavior in intact animals. Copyright © 2010 Elsevier Ireland Ltd. All rights reserved.

  17. Identification and quantification of nucleosides and nucleobases in Geosaurus and Leech by hydrophilic-interaction chromatography.

    PubMed

    Chen, Pei; Li, Wei; Li, Qin; Wang, Yinghua; Li, Zhenguo; Ni, Yefeng; Koike, Kazuo

    2011-09-15

    A simple hydrophilic-interaction chromatography (HILIC) method was developed for the identification and quantification of 14 nucleosides and nucleobases, namely cytosine, uracil, cytidine, guanine, hypoxanthine, xanthine, uridine, thymine, inosine, guanosine, thymidine, 2'-deoxyadenosine, 2'-deoxyinosine and 2'-deoxyuridine in two traditional Chinese medicines, Geosaurus and Leech. The separation was achieved on a TSKgel Amide-80 column (150 mm × 2.0 mm, 3.0 μm) with a mixture of acetonitrile and 10 mM aqueous ammonium acetate as the mobile phase at a flow rate of 0.2 mL/min. The temperature was set at 30°C and UV detection wavelength was set at 260 nm. All calibration curves showed good linearity (R(2)>0.9957) within the test ranges. The overall intra- and inter-day RSD ranged from 0.4 to 3.4% and from 0.7 to 3.3%, respectively. The LOD and LOQ were in the range of 0.07-30.49 ng/mL and 0.26-60.98 ng/mL, respectively. The repeatability of the method was in the range of 2.2-5.8% for Geosaurus and 1.4-5.5% for Leech. The recoveries of the samples were in the range of 91.4-100.9% for Geosaurus, and 91.9-99.3% for Leech. The established method was applied successfully for the analysis of nucleosides and nucleobases in 22 commercially available samples collected from different regions in China and Japan. Our data showed that HILIC had advantages as a useful tool for the study of the bioactive components in Geosaurus and Leech as well as their quality control, and could therefore be used for the determination of the analytes in pharmaceutical products and biological fluids.

  18. Dynamics and reproducibility of a moderately complex sensory-motor response in the medicinal leech.

    PubMed

    Garcia-Perez, Elizabeth; Zoccolan, Davide; Pinato, Giulietta; Torre, Vincent

    2004-09-01

    Local bending, a motor response caused by mechanical stimulation of the leech skin, has been shown to be remarkably reproducible, in its initial phase, despite the highly variable firing of motoneurons sustaining it. In this work, the reproducibility of local bending was further analyzed by monitoring it over a longer period of time and by using more intact preparations, in which muscle activation in an entire body segment was studied. Our experiments showed that local bending is a moderately complex motor response, composed of a sequence of four different phases, which were consistently identified in all leeches. During each phase, longitudinal and circular muscles in specific areas of the body segment acted synergistically, being co-activated or co-inhibited depending on their position relative to the stimulation site. Onset and duration of the first phase were reproducible across different trials and different animals as a result of the massive co-activation of excitatory motoneurons sustaining it. The other phases were produced by the inhibition of excitatory and activation of inhibitory motoneurons, and also by the intrinsic relaxation dynamics of leech muscles. As a consequence, their duration and relative timing was variable across different preparations, whereas their order of appearance was conserved. These results suggest that, during local bending, the leech neuromuscular system 1) operates a reduction of its available degrees of freedom, by simultaneously recruiting groups of otherwise antagonistic muscles and large populations of motoneurons; and 2) ensures reliability and effectiveness of this escape reflex, by guaranteeing the reproducibility of its crucial initial phase.

  19. Human recombinant RNASET2-induced inflammatory response and connective tissue remodeling in the medicinal leech.

    PubMed

    Baranzini, Nicolò; Pedrini, Edoardo; Girardello, Rossana; Tettamanti, Gianluca; de Eguileor, Magda; Taramelli, Roberto; Acquati, Francesco; Grimaldi, Annalisa

    2017-01-09

    In recent years, several studies have demonstrated that the RNASET2 gene is involved in the control of tumorigenicity in ovarian cancer cells. Furthermore, a role in establishing a functional cross-talk between cancer cells and the surrounding tumor microenvironment has been unveiled for this gene, based on its ability to act as an inducer of the innate immune response. Although several studies have reported on the molecular features of RNASET2, the details on the mechanisms by which this evolutionarily conserved ribonuclease regulates the immune system are still poorly defined. In the effort to clarify this aspect, we report here the effect of recombinant human RNASET2 injection and its role in regulating the innate immune response after bacterial challenge in an invertebrate model, the medicinal leech. We found that recombinant RNASET2 injection induces fibroplasias, connective tissue remodeling and the recruitment of numerous infiltrating cells expressing the specific macrophage markers CD68 and HmAIF1. The RNASET2-mediated chemotactic activity for macrophages has been further confirmed by using a consolidated experimental approach based on injection of the Matrigel biomatrice (MG) supplemented with recombinant RNASET2 in the leech body wall. One week after injection, a large number of CD68(+) and HmAIF-1(+) macrophages massively infiltrated MG sponges. Finally, in leeches challenged with lipopolysaccharides (LPS) or with the environmental bacteria pathogen Micrococcus nishinomiyaensis, numerous macrophages migrating to the site of inoculation expressed high levels of endogenous RNASET2. Taken together, these results suggest that RNASET2 is likely involved in the initial phase of the inflammatory response in leeches.

  20. A new lineage of trypanosomes from Australian vertebrates and terrestrial bloodsucking leeches (Haemadipsidae).

    PubMed

    Hamilton, P B; Stevens, J R; Gidley, J; Holz, P; Gibson, W C

    2005-04-01

    Little is known about the trypanosomes of indigenous Australian vertebrates and their vectors. We surveyed a range of vertebrates and blood-feeding invertebrates for trypanosomes by parasitological and PCR-based methods using primers specific to the small subunit ribosomal RNA (SSU rRNA) gene of genus Trypanosoma. Trypanosome isolates were obtained in culture from two common wombats, one swamp wallaby and an Australian bird (Strepera sp.). By PCR, blood samples from three wombats, one brush-tailed wallaby, three platypuses and a frog were positive for trypanosome DNA. All the blood-sucking invertebrates screened were negative for trypanosomes both by microscopy and PCR, except for specimens of terrestrial leeches (Haemadipsidae). Of the latter, two Micobdella sp. specimens from Victoria and 18 Philaemon sp. specimens from Queensland were positive by PCR. Four Haemadipsa zeylanica specimens from Sri Lanka and three Leiobdella jawarerensis specimens from Papua New Guinea were also PCR positive for trypanosome DNA. We sequenced the SSU rRNA and glycosomal glyceraldehyde phosphate dehydrogenase (gGAPDH) genes in order to determine the phylogenetic positions of the new vertebrate and terrestrial leech trypanosomes. In trees based on these genes, Australian vertebrate trypanosomes fell in several distinct clades, for the most part being more closely related to trypanosomes outside Australia than to each other. Two previously undescribed wallaby trypanosomes fell in a clade with Trypanosoma theileri, the cosmopolitan bovid trypanosome, and Trypanosoma cyclops from a Malaysian primate. The terrestrial leech trypanosomes were closely related to the wallaby trypanosomes, T. cyclops and a trypanosome from an Australian frog. We suggest that haemadipsid leeches may be significant and widespread vectors of trypanosomes in Australia and Asia.

  1. [Predicting efficiency of selection on reproductive traits in the medicinal leech Hirudo medicinalis L].

    PubMed

    Utevskaia, O M; Atramentova, L A

    2002-03-01

    A set of selection measures for increasing reproduction efficiency in Hirudo medicinalis has been developed. The optimal values of reproductive traits corresponding to the highest progeny number were determined and recommended. The probability of correlated selection response in traits "number of threads in a cocoon" and "weight of threads" was estimated. Based on earlier results on phenotypic variation and heritability of reproductive traits in medicinal leech, efficiency of different selection modes was predicted.

  2. Land Leeches of the g. Haemadipsa (Haemadipsoidea: Haemadipsidae). I. Conditions Essential to Laboratory Colonization.

    DTIC Science & Technology

    1982-05-01

    segmented aquatic and terrestrial worms that belong to the Order Hirudinea, ( Annelida : Clitellata, Scriban and Autrum (2)) and are most closely related to the...WORTH, C.B. Description and discussion of the biting of an Indian land leech. ( Annelida : Hirudinea). J Bombay Nat Hist Soc 50:423-426, 1951 39...Haemadlpsidae and their Suborder, the Gnathobdellae (after 2-6,60). Phylum: Annelida Class: Clitellata Order: Ifirudinea (=Hirudiniformes) Suborder 1

  3. Multiple Changes in Peptide and Lipid Expression Associated with Regeneration in the Nervous System of the Medicinal Leech

    PubMed Central

    Meriaux, Céline; Arafah, Karim; Tasiemski, Aurélie; Wisztorski, Maxence; Bruand, Jocelyne; Boidin-Wichlacz, Céline; Desmons, Annie; Debois, Delphine; Laprévote, Olivier; Brunelle, Alain; Gaasterland, Terry; Macagno, Eduardo; Fournier, Isabelle; Salzet, Michel

    2011-01-01

    Background The adult medicinal leech central nervous system (CNS) is capable of regenerating specific synaptic circuitry after a mechanical lesion, displaying evidence of anatomical repair within a few days and functional recovery within a few weeks. In the present work, spatiotemporal changes in molecular distributions during this phenomenon are explored. Moreover, the hypothesis that neural regeneration involves some molecular factors initially employed during embryonic neural development is tested. Results Imaging mass spectrometry coupled to peptidomic and lipidomic methodologies allowed the selection of molecules whose spatiotemporal pattern of expression was of potential interest. The identification of peptides was aided by comparing MS/MS spectra obtained for the peptidome extracted from embryonic and adult tissues to leech transcriptome and genome databases. Through the parallel use of a classical lipidomic approach and secondary ion mass spectrometry, specific lipids, including cannabinoids, gangliosides and several other types, were detected in adult ganglia following mechanical damage to connected nerves. These observations motivated a search for possible effects of cannabinoids on neurite outgrowth. Exposing nervous tissues to Transient Receptor Potential Vanilloid (TRPV) receptor agonists resulted in enhanced neurite outgrowth from a cut nerve, while exposure to antagonists blocked such outgrowth. Conclusion The experiments on the regenerating adult leech CNS reported here provide direct evidence of increased titers of proteins that are thought to play important roles in early stages of neural development. Our data further suggest that endocannabinoids also play key roles in CNS regeneration, mediated through the activation of leech TRPVs, as a thorough search of leech genome databases failed to reveal any leech orthologs of the mammalian cannabinoid receptors but revealed putative TRPVs. In sum, our observations identify a number of lipids and

  4. Experimental Investigation on the Morphology and Adhesion Mechanism of Leech Posterior Suckers

    PubMed Central

    Dong, Wenhao

    2015-01-01

    The posterior sucker of a leech represents a fascinating natural system that allows the leech to adhere to different terrains and substrates. However, the mechanism of adhesion and desorption has not yet to be elucidated. In order to better understand how the adhesion is performed, we analyzed the surface structure, adsorption movements, the muscles’ distribution, physical characteristics, and the adsorption force of the leech posterior suckers by experimental investigation. Three conclusions can be drawn based on the obtained experimental results. First, the adhesion by the posterior sucker is wet adhesion, because the surface of the posterior sucker is smooth and the sealing can only be achieved on wet surfaces. Second, the deformation texture, consisting of soft collagen tissues and highly ductile epidermal tissues, plays a key role in adhering to rough surfaces. Finally, the adhesion and desorption is achieved by the synergetic operation of six muscle fibers working in different directions. Concrete saying, directional deformation of the collagen/epithermal interface driven by spatially-distributed muscle fibers facilitates the excretion of fluids in the sucker venter, thus allowing liquid sealing. Furthermore, we found that the adhesion strength is directly related to the size of the contact surface which is generated and affected by the sucker deformation. Such an underlying physical mechanism offers potential cues for developing innovative bio-inspired artificial adhesion systems. PMID:26536352

  5. De novo transcriptome assembly databases for the central nervous system of the medicinal leech.

    PubMed

    Hibsh, Dror; Schori, Hadas; Efroni, Sol; Shefi, Orit

    2015-01-01

    The study of non-model organisms stands to benefit greatly from genetic and genomic data. For a better understanding of the molecular mechanisms driving neuronal development, and to characterize the entire leech Hirudo medicinalis central nervous system (CNS) transcriptome we combined Trinity for de-novo assembly and Illumina HiSeq2000 for RNA-Seq. We present a set of 73,493 de-novo assembled transcripts for the leech, reconstructed from RNA collected, at a single ganglion resolution, from the CNS. This set of transcripts greatly enriches the available data for the leech. Here, we share two databases, such that each dataset allows a different type of search for candidate homologues. The first is the raw set of assembled transcripts. This set allows a sequence-based search. A comprehensive analysis of which revealed 22,604 contigs with high e-values, aligned versus the Swiss-Prot database. This analysis enabled the production of the second database, which includes correlated sequences to annotated transcript names, with the confidence of BLAST best hit.

  6. Highly variable spike trains underlie reproducible sensorimotor responses in the medicinal leech.

    PubMed

    Zoccolan, Davide; Pinato, Giulietta; Torre, Vincent

    2002-12-15

    The nervous system of the leech is a particularly suitable model to investigate neural coding of sensorimotor responses because it allows both observation of behavior and the simultaneous measurement of a large fraction of its underlying neuronal activity. In this study, we used a combination of multielectrode recordings, videomicroscopy, and computation of the optical flow to investigate the reproducibility of the motor response caused by local mechanical stimulation of the leech skin. We analyzed variability at different levels of processing: mechanosensory neurons, motoneurons, muscle activation, and behavior. Spike trains in mechanosensory neurons were very reproducible, unlike those in motoneurons. The motor response, however, was reproducible because of two distinct biophysical mechanisms. First, leech muscles contract slowly and therefore are poorly sensitive to the jitter of motoneuron spikes. Second, the motor response results from the coactivation of a population of motoneurons firing in a statistically independent way, which reduces the variability of the population firing. These data show that reproducible spike trains are not required to sustain reproducible behaviors and illustrate how the nervous system can cope with unreliable components to produce reliable action.

  7. Decision points: the factors influencing the decision to feed in the medicinal leech.

    PubMed

    Gaudry, Quentin; Kristan, William B

    2012-01-01

    The decision to feed is a complex task that requires making several small independent choices. Am I hungry? Where do I look for food? Is there something better I'd rather be doing? When should I stop? With all of these questions, it is no wonder that decisions about feeding depend on several sensory modalities and that the influences of these sensory systems would be evident throughout the nervous system. The leech is uniquely well suited for studying these complicated questions due to its relatively simple nervous system, its exceptionally well-characterized behaviors and neural circuits, and the ease with which one can employ semi-intact preparations to study the link between physiology and decision-making. We will begin this review by discussing the cellular substrates that govern the decision to initiate and to terminate a bout of feeding. We will then discuss how feeding temporarily blocks competing behaviors from being expressed while the animal continues to feed. Then we will review what is currently known about how feeding affects long-term behavioral choices of the leech. Finally, we conclude with a short discussion of the advantages of the leech's decision-making circuit's design and how this design might be applicable to all decision circuits.

  8. Small bite, large impact-saliva and salivary molecules in the medicinal leech, Hirudo medicinalis

    NASA Astrophysics Data System (ADS)

    Hildebrandt, Jan-Peter; Lemke, Sarah

    2011-12-01

    Blood-sucking leeches have been used for medical purposes in humans for hundreds of years. Accordingly, one of the most prominent species has been named Hirudo medicinalis by Carl Linne in 1758. Feeding on vertebrate blood poses some serious problems to blood-sucking ectoparasites, as they have to penetrate the body surface of the host and to suppress the normal reactions of the host to such injuries (swelling, pain, inflammation) to remain undetected during the feeding period. Furthermore, the parasites have to take measures to inhibit the normal reactions in host tissues to blood vessel damage, namely hemostasis and blood coagulation (platelet aggregation and activation, activation of thrombin and formation of fibrin clots). During evolution, leeches have acquired the ability to control these processes in their hosts by transferring various bioactive substances to the host. These substances are supposedly produced in unicellular salivary gland cells and injected into the wound at the feeding site through tiny salivary ductule openings in the jaws that the leech uses to slice open the host body surface and to cut blood vessels in the depth of the wound. This review summarizes current knowledge about the salivary gland cells and the biological effects of individual saliva components as well as hints to the potential usefulness of some of these compounds for medical purposes.

  9. Small bite, large impact-saliva and salivary molecules in the medicinal leech, Hirudo medicinalis.

    PubMed

    Hildebrandt, Jan-Peter; Lemke, Sarah

    2011-12-01

    Blood-sucking leeches have been used for medical purposes in humans for hundreds of years. Accordingly, one of the most prominent species has been named Hirudo medicinalis by Carl Linne in 1758. Feeding on vertebrate blood poses some serious problems to blood-sucking ectoparasites, as they have to penetrate the body surface of the host and to suppress the normal reactions of the host to such injuries (swelling, pain, inflammation) to remain undetected during the feeding period. Furthermore, the parasites have to take measures to inhibit the normal reactions in host tissues to blood vessel damage, namely hemostasis and blood coagulation (platelet aggregation and activation, activation of thrombin and formation of fibrin clots). During evolution, leeches have acquired the ability to control these processes in their hosts by transferring various bioactive substances to the host. These substances are supposedly produced in unicellular salivary gland cells and injected into the wound at the feeding site through tiny salivary ductule openings in the jaws that the leech uses to slice open the host body surface and to cut blood vessels in the depth of the wound. This review summarizes current knowledge about the salivary gland cells and the biological effects of individual saliva components as well as hints to the potential usefulness of some of these compounds for medical purposes.

  10. De novo transcriptome assembly databases for the central nervous system of the medicinal leech

    PubMed Central

    Hibsh, Dror; Schori, Hadas; Efroni, Sol; Shefi, Orit

    2015-01-01

    The study of non-model organisms stands to benefit greatly from genetic and genomic data. For a better understanding of the molecular mechanisms driving neuronal development, and to characterize the entire leech Hirudo medicinalis central nervous system (CNS) transcriptome we combined Trinity for de-novo assembly and Illumina HiSeq2000 for RNA-Seq. We present a set of 73,493 de-novo assembled transcripts for the leech, reconstructed from RNA collected, at a single ganglion resolution, from the CNS. This set of transcripts greatly enriches the available data for the leech. Here, we share two databases, such that each dataset allows a different type of search for candidate homologues. The first is the raw set of assembled transcripts. This set allows a sequence-based search. A comprehensive analysis of which revealed 22,604 contigs with high e-values, aligned versus the Swiss-Prot database. This analysis enabled the production of the second database, which includes correlated sequences to annotated transcript names, with the confidence of BLAST best hit. PMID:25977819

  11. Reciprocal immune benefit based on complementary production of antibiotics by the leech Hirudo verbana and its gut symbiont Aeromonas veronii.

    PubMed

    Tasiemski, Aurélie; Massol, François; Cuvillier-Hot, Virginie; Boidin-Wichlacz, Céline; Roger, Emmanuel; Rodet, Franck; Fournier, Isabelle; Thomas, Frédéric; Salzet, Michel

    2015-12-04

    The medicinal leech has established a long-term mutualistic association with Aeromonas veronii, a versatile bacterium which can also display free-living waterborne and fish- or human-pathogenic lifestyles. Here, we investigated the role of antibiotics in the dynamics of interaction between the leech and its gut symbiont Aeromonas. By combining biochemical and molecular approaches, we isolated and identified for the first time the antimicrobial peptides (AMPs) produced by the leech digestive tract and by its symbiont Aeromonas. Immunohistochemistry data and PCR analyses evidenced that leech AMP genes are induced in the gut epithelial cells when Aeromonas load is low (starved animals), while repressed when Aeromonas abundance is the highest (post blood feeding). The asynchronous production of AMPs by both partners suggests that these antibiotic substances (i) provide them with reciprocal protection against invasive bacteria and (ii) contribute to the unusual simplicity of the gut microflora of the leech. This immune benefit substantially reinforces the evidence of an evolutionarily stable association between H. verbana and A. veronii. Altogether these data may provide insights into the processes making the association with an Aeromonas species in the digestive tract either deleterious or beneficial.

  12. Prevalence, Abundance, and Intensity of Implanted Spermatophores in the Leech Haementeria officinalis (Glossiphoniidae: Hirudinida) from Guanajuato, Mexico.

    PubMed

    Salas-Montiel, Ricardo; Phillips, Anna J; Contreras-Mirón, Samantha; Oceguera-Figueroa, Alejandro

    2017-02-01

    Fertilization through hypodermic implantation of spermatophores has been recorded in at least 4 groups of leeches: Glossiphoniidae, Piscicolidae, Ozobranchidae, and Erpobdelliformes. In Piscicola respirans (Piscicolidae), vector tissue responsible for sperm transfer from a specialized region of the body to the ovaries has led to the non-random attachment of spermatophores on the body surface of the recipient leech. It has been suggested that in glossiphoniid leeches, spermatophores are implanted in any part of the body surface of the recipient leech without a clear pattern or preference for region. In order to determine if the donor leech implants its spermatophores in a specific area of the conspecific recipient's body, we surveyed 81 specimens of Haementeria officinalis (Clitellata: Glossiphoniidae) from a wild population in Guanajuato, Mexico, and recorded the distribution of the spermatophores over the recipient's body surface. We describe for the first time a spermatophore of H. officinalis using scanning electron and light microscopy. Spermatophores were found attached dorsally between somites XVII and XXI 59.57% of the time, and the rest were found in other parts of the body, including on the ventral surface. The non-specific attachment for spermatophores does not support the presence of specialized tissue responsible for sperm transfer and instead attributes the placement of implantation to mechanical characteristics of the copulation process.

  13. Reciprocal immune benefit based on complementary production of antibiotics by the leech Hirudo verbana and its gut symbiont Aeromonas veronii

    PubMed Central

    Tasiemski, Aurélie; Massol, François; Cuvillier-Hot, Virginie; Boidin-Wichlacz, Céline; Roger, Emmanuel; Rodet, Franck; Fournier, Isabelle; Thomas, Frédéric; Salzet, Michel

    2015-01-01

    The medicinal leech has established a long-term mutualistic association with Aeromonas veronii, a versatile bacterium which can also display free-living waterborne and fish- or human-pathogenic lifestyles. Here, we investigated the role of antibiotics in the dynamics of interaction between the leech and its gut symbiont Aeromonas. By combining biochemical and molecular approaches, we isolated and identified for the first time the antimicrobial peptides (AMPs) produced by the leech digestive tract and by its symbiont Aeromonas. Immunohistochemistry data and PCR analyses evidenced that leech AMP genes are induced in the gut epithelial cells when Aeromonas load is low (starved animals), while repressed when Aeromonas abundance is the highest (post blood feeding). The asynchronous production of AMPs by both partners suggests that these antibiotic substances (i) provide them with reciprocal protection against invasive bacteria and (ii) contribute to the unusual simplicity of the gut microflora of the leech. This immune benefit substantially reinforces the evidence of an evolutionarily stable association between H. verbana and A. veronii. Altogether these data may provide insights into the processes making the association with an Aeromonas species in the digestive tract either deleterious or beneficial. PMID:26635240

  14. Effect of ischemia preconditioning and leech therapy on cutaneous pedicle flaps subjected to prolonged ischemia in a mouse model.

    PubMed

    Moosavian, Hamid Reza; Mirghazanfari, Sayid Mahdi; Moghaddam, Katayoun Gohari

    2014-10-01

    We sought to determine the effect of ischemic preconditioning (IPC) and hirudotherapy (leech therapy) on cutaneous pedicle flaps after they underwent prolonged ischemia (global ischemia) in a mouse model. Twenty cutaneous pedicle flaps were elevated in 20 mice, and the animals were randomized into four groups: sham, control, IPC and leech (5 flaps in each group). Except in the sham group, all flaps were subjected to global ischemia for 5 h via pedicle clamping. The control group did not receive any treatment before or after global ischemia. In the IPC group, global ischemia was preceded by three 10-min episodes of ischemia, each followed by 10 min of reperfusion. In the leech therapy group, after global ischemia, hirudotherapy was performed. Flap survival area and histopathological changes were evaluated on the 10th day after surgery. Flap survival areas were significantly higher in both the IPC and leech groups than in the control group and were significantly higher in the leech group than in the IPC group (p < 0.05). In conclusion IPC and hirudotherapy had definite effects on the survival area of cutaneous pedicle flaps that underwent prolonged ischemia in a mouse model.

  15. Calreticulin contributes to C1q-dependent recruitment of microglia in the leech Hirudo medicinalis following a CNS injury.

    PubMed

    Le Marrec-Croq, Francoise; Bocquet-Garcon, Annelise; Vizioli, Jacopo; Vancamp, Christelle; Drago, Francesco; Franck, Julien; Wisztorski, Maxence; Salzet, Michel; Sautiere, Pierre-Eric; Lefebvre, Christophe

    2014-04-19

    The medicinal leech is considered as a complementary and appropriate model to study immune functions in the central nervous system (CNS). In a context in which an injured leech's CNS can naturally restore normal synaptic connections, the accumulation of microglia (immune cells of the CNS that are exclusively resident in leeches) has been shown to be essential at the lesion to engage the axonal sprouting. HmC1q (Hm for Hirudo medicinalis) possesses chemotactic properties that are important in the microglial cell recruitment by recognizing at least a C1q binding protein (HmC1qBP alias gC1qR). Recombinant forms of C1q were used in affinity purification and in vitro chemotaxis assays. Anti-calreticulin antibodies were used to neutralize C1q-mediated chemotaxis and locate the production of calreticulin in leech CNS. A newly characterized leech calreticulin (HmCalR) has been shown to interact with C1q and participate to the HmC1q-dependent microglia accumulation. HmCalR, which has been detected in only some microglial cells, is consequently a second binding protein for HmC1q, allowing the chemoattraction of resident microglia in the nerve repair process. These data give new insight into calreticulin/C1q interaction in an immune function of neuroprotection, suggesting another molecular target to use in investigation of microglia reactivity in a model of CNS injury.

  16. Evidence for a novel chemotactic C1q domain-containing factor in the leech nerve cord.

    PubMed

    Tahtouh, Muriel; Croq, Françoise; Vizioli, Jacopo; Sautiere, Pierre-Eric; Van Camp, Christelle; Salzet, Michel; Daha, Mohamed R; Pestel, Joël; Lefebvre, Christophe

    2009-02-01

    In vertebrates, central nervous system (CNS) protection is dependent on many immune cells including microglial cells. Indeed, activated microglial cells are involved in neuroinflammation mechanisms by interacting with numerous immune factors. Unlike vertebrates, some lophotrochozoan invertebrates can fully repair their CNS following injury. In the medicinal leech Hirudo medicinalis, the recruitment of microglial cells at the lesion site is essential for sprouting of injured axons. Interestingly, a new molecule homologous to vertebrate C1q was characterized in leech, named HmC1q (for H. medicinalis) and detected in neurons and glial cells. In chemotaxis assays, leech microglial cells were demonstrated to respond to human C1q. The chemotactic activity was reduced when microglia was preincubated with signaling pathway inhibitors (Pertussis Toxin or wortmannin) or anti-human gC1qR antibody suggesting the involvement of gC1qR in C1q-mediated migration in leech. Assays using cells preincubated with NO chelator (cPTIO) showed that C1q-mediated migration was associated to NO production. Of interest, by using anti-HmC1q antibodies, HmC1q released in the culture medium was shown to exhibit a similar chemotactic effect on microglial cells as human C1q. In summary, we have identified, for the first time, a molecule homologous to mammalian C1q in leech CNS. Its chemoattractant activity on microglia highlights a new investigation field leading to better understand leech CNS repair mechanisms.

  17. Historical Article: Hirudo medicinalis: ancient origins of, and trends in the use of medicinal leeches throughout history.

    PubMed

    Whitaker, I S; Rao, J; Izadi, D; Butler, P E

    2004-04-01

    Blood letting and the therapeutic use of Hirudo medicinalis date back to ancient Egypt and the beginning of civilisation. Their popularity has varied over the years, reaching such a peak in Europe between 1825 and 1850 that supplies were exhausted. Towards the end of the century they fell out of favour and, during this period, the leech, once used by the physicians of emperors and influential academic surgeons, became associated with lay therapists and quackery. Leeches have enjoyed a renaissance in reconstructive microsurgery during the last 15 years, having been used by maxillofacial [Br. J. Oral Maxillofac. Surg 41 (2003) 44] and other reconstructive surgeons to aid salvage of compromised microvascular free tissue transfers [Laryngoscope 108 (1998) 1129; Br. J. Plast. Surg. 34 (1984) 358], replanted digits [Int. J. Microsurg. 3 (1981) 265], ears [Ann. Plast. Surg. 43 (1999) 427], lips [Plast. Reconstr. Surg. 102 (1998) 358; J. Reconstr. Microsurg. 9 (1993) 327] and nasal tips [Br. J. Oral Maxillofac. Surg. 36 (1998) 462]. Peer-reviewed evidence suggests that the survival of compromised, venous-congested tissues is improved by early application of a leech [J. Reconstr. Microsurg. 12 (1996) 165; Arch. Otolaryngol. Head Neck Surg. 114 (1988) 1395; Br. J. Plast. Surg. 45 (1992) 235]. Leeches have also recently been used to treat a wide range of conditions, including periorbital haematomas [Br. J. Ophthalmol. 75 (1991) 755], severe macroglossia [Otolaryngol. Head Neck Surg. 125 (2001) 649; J. Laryngol. Otol. 109 (1995) 442] and purpura fulminans [Ann. Plast. Surg. 35 (1995) 300]. The first medicinal leech farm, Biopharm, was set up in Swansea in 1981 by Dr Roy Sawyer, and now supplies leeches to hospitals all over the world. In this paper, we summarise the history of treatment with Hirudo medicinalis from its origin to the present day, and take a brief look at the possible future of the annelid.

  18. A homologous form of human interleukin 16 is implicated in microglia recruitment following nervous system injury in leech Hirudo medicinalis.

    PubMed

    Croq, Françoise; Vizioli, Jacopo; Tuzova, Marina; Tahtouh, Muriel; Sautiere, Pierre-Eric; Van Camp, Christelle; Salzet, Michel; Cruikshank, William W; Pestel, Joel; Lefebvre, Christophe

    2010-11-01

    In contrast to mammals, the medicinal leech Hirudo medicinalis can completely repair its central nervous system (CNS) after injury. This invertebrate model offers unique opportunities to study the molecular and cellular basis of the CNS repair processes. When the leech CNS is injured, microglial cells migrate and accumulate at the site of lesion, a phenomenon known to be essential for the usual sprouting of injured axons. In the present study, we demonstrate that a new molecule, designated HmIL-16, having functional homologies with human interleukin-16 (IL-16), has chemotactic activity on leech microglial cells as observed using a gradient of human IL-16. Preincubation of microglial cells either with an anti-human IL-16 antibody or with anti-HmIL-16 antibody significantly reduced microglia migration induced by leech-conditioned medium. Functional homology was demonstrated further by the ability of HmIL-16 to promote human CD4+ T cell migration which was inhibited by antibody against human IL-16, an IL-16 antagonist peptide or soluble CD4. Immunohistochemistry of leech CNS indicates that HmIL-16 protein present in the neurons is rapidly transported and stored along the axonal processes to promote the recruitment of microglial cells to the injured axons. To our knowledge, this is the first identification of a functional interleukin-16 homologue in invertebrate CNS. The ability of HmIL-16 to recruit microglial cells to sites of CNS injury suggests a role for HmIL-16 in the crosstalk between neurons and microglia in the leech CNS repair.

  19. Humoral response of captive zebra sharks Stegostoma fasciatum to salivary gland proteins of the leech Branchellion torpedinis.

    PubMed

    Marancik, David P; Leary, John H; Fast, Mark M; Flajnik, Martin F; Camus, Alvin C

    2012-10-01

    Parasitism by the marine leech Branchellion torpedinis is known to cause disease and mortality in captive elasmobranchs and is difficult to control when inadvertently introduced into public aquaria. Preliminary characterization of the salivary gland transcriptome of B. torpedinis has identified anticoagulants, proteases, and immunomodulators that may be secreted into host tissues to aid leech feeding. This retrospective study examined antigen-specific serum IgM responses in captive zebra sharks Stegostoma fasciatum to leech salivary gland extract. Antibody response was examined by ELISA and Western blot assays in 20 serum samples from six zebra sharks, with a 5 year history of leech infection, and 18 serum samples from 8 captive bred zebra sharks, with no history of leech exposure. ELISA demonstrated significantly higher serum IgM titers to salivary gland extract in exposed zebra sharks compared to the non-exposed population. No obvious trends in antibody titers were appreciated in exposed zebra sharks over a four-year period. One-dimensional and two-dimensional Western blot assays revealed IgM targeted specific salivary gland proteins within the 40, 55, 70 and 90 kD range. Antigenic proteins identified by liquid chromatography-tandem mass spectrometry and de novo peptide sequencing include a secreted disintegrin, metalloproteinase and thrombospondin motif containing protein (ADAMTS), tubulin, aldehyde dehydrogenase and two unknown proteins. Humoral immune responses to leech salivary gland proteins warrants further investigation as there may be options to exploit immune mechanisms to reduce parasite burdens in aquaria. Copyright © 2012 Elsevier Ltd. All rights reserved.

  20. Occurrence of the Leech, Pontobdella muricata Linnaeus, on Elasmobranch Species in the Northern and Central Adriatic Sea.

    PubMed

    Bolognini, Luca; Leoni, Simone; Polidori, Piero; Grati, Fabio; Scarcella, Giuseppe; Pellini, Giulio; Domenichetti, Filippo; Ferrà, Carmen; Fabi, Gianna

    2016-12-01

    This study provides a parasitological analysis of the elasmobranch species caught in the northern and central Adriatic Sea. Sixty-two marine leeches were recorded on 747 individuals of Raja clavata Linnaeus, 1758 (thornback ray), Myliobatis aquila Linnaeus, 1758 (common eagle ray), and Torpedo marmorata Risso, 1810 (marbled torpedo ray) caught in 56 hauls over a 5 yr period. All leeches were identified as Pontobdella muricata, which is a typical ectoparasite of benthic elasmobranchs. The prevalence of infection ranged from 7.11% on R. clavata to 12.00% on M. aquila. The intensity of infection, the preferential sites of attachment to the host, and the periodicity of infection were evaluated.

  1. Establishment of segment polarity in the ectoderm of the leech Helobdella

    NASA Technical Reports Server (NTRS)

    Seaver, E. C.; Shankland, M.

    2001-01-01

    The segmented ectoderm and mesoderm of the leech arise via a stereotyped cell lineage from embryonic stem cells called teloblasts. Each teloblast gives rise to a column of primary blast cell daughters, and the blast cells generate descendant clones that serve as the segmental repeats of their particular teloblast lineage. We have examined the mechanism by which the leech primary blast cell clones acquire segment polarity - i.e. a fixed sequence of positional values ordered along the anteroposterior axis of the segmental repeat. In the O and P teloblast lineages, the earliest divisions of the primary blast cell segregate anterior and posterior cell fates along the anteroposterior axis. Using a laser microbeam, we ablated single cells from both o and p blast cell clones at stages when the clone was two to four cells in length. The developmental fate of the remaining cells was characterized with rhodamine-dextran lineage tracer. Twelve different progeny cells were ablated, and in every case the ablation eliminated the normal descendants of the ablated cell while having little or no detectable effect on the developmental fate of the remaining cells. This included experiments in which we specifically ablated those blast cell progeny that are known to express the engrailed gene, or their lineal precursors. These findings confirm and extend a previous study by showing that the establishment of segment polarity in the leech ectoderm is largely independent of cell interactions conveyed along the anteroposterior axis. Both intercellular signaling and engrailed expression play an important role in the segment polarity specification of the Drosophila embryo, and our findings suggest that there may be little or no conservation of this developmental mechanism between those two organisms.

  2. Establishment of segment polarity in the ectoderm of the leech Helobdella

    NASA Technical Reports Server (NTRS)

    Seaver, E. C.; Shankland, M.

    2001-01-01

    The segmented ectoderm and mesoderm of the leech arise via a stereotyped cell lineage from embryonic stem cells called teloblasts. Each teloblast gives rise to a column of primary blast cell daughters, and the blast cells generate descendant clones that serve as the segmental repeats of their particular teloblast lineage. We have examined the mechanism by which the leech primary blast cell clones acquire segment polarity - i.e. a fixed sequence of positional values ordered along the anteroposterior axis of the segmental repeat. In the O and P teloblast lineages, the earliest divisions of the primary blast cell segregate anterior and posterior cell fates along the anteroposterior axis. Using a laser microbeam, we ablated single cells from both o and p blast cell clones at stages when the clone was two to four cells in length. The developmental fate of the remaining cells was characterized with rhodamine-dextran lineage tracer. Twelve different progeny cells were ablated, and in every case the ablation eliminated the normal descendants of the ablated cell while having little or no detectable effect on the developmental fate of the remaining cells. This included experiments in which we specifically ablated those blast cell progeny that are known to express the engrailed gene, or their lineal precursors. These findings confirm and extend a previous study by showing that the establishment of segment polarity in the leech ectoderm is largely independent of cell interactions conveyed along the anteroposterior axis. Both intercellular signaling and engrailed expression play an important role in the segment polarity specification of the Drosophila embryo, and our findings suggest that there may be little or no conservation of this developmental mechanism between those two organisms.

  3. Distributed motor pattern underlying whole-body shortening in the medicinal leech.

    PubMed

    Arisi, I; Zoccolan, D; Torre, V

    2001-11-01

    Whole-body shortening was studied in the leech, Hirudo medicinalis, by a combination of videomicroscopy and multielectrode recordings. Video microscopy was used to monitor the animal behavior and muscle contraction. Eight suction pipettes were used to obtain simultaneous electrical recordings from fine roots emerging from ganglia. This vital escape reaction was rather reproducible. The coefficient of variation of the animal contraction during whole-body shortening was between 0.2 and 0.3. The great majority of all leech longitudinal motoneurons were activated during this escape reaction, in particular motoneurons 3, 4, 5, 8, 107, 108, and L. The firing pattern of all these motoneurons was poorly reproducible from trial to trial, and the coefficient of variation of their firing varied between 0.3 and 1.5 for different motoneurons. The electrical activity of pairs of coactivated motoneurons did not show any sign of correlation over a time window of 100 ms. Only the left and right motoneurons L in the same ganglion had a correlated firing pattern, resulting from their strong electrical coupling. As a consequence of the low correlation between coactivated motoneurons, the global electrical activity during whole-body shortening became reproducible with a coefficient of variation below 0.3 during maximal contraction. These results indicate that whole-body shortening is mediated by the coactivation of a large fraction of all leech motoneurons, i.e., it is a distributed process, and that coactivated motoneurons exhibit a significant statistical independence. Probably due to this statistical independence this vital escape reaction is smooth and reproducible.

  4. Isolation of infectious hematopoietic necrosis virus from a leech (Piscicola salmositica) and a copepod (Salmincola sp.), ectoparasites of sockeye salmon Oncorhynchus nerka

    USGS Publications Warehouse

    Mulcahy, D.; Klaybor, D.; Batts, W.N.

    1990-01-01

    ectious hematopoietic necrosis (IHN) virus was isolated from freshwater leeches Piscicola salmositica and copepods Salmincola sp. removed from the gills of spawning sockeye salmon Oncorhynchus nerka. This is the first report of the isolation of IHN virus from an animal other than salmonid fishes. High levels of IHN virus were also found in leeches taken from the bottom gravel of the spawning area. The prevalence of IHN virus in samples of individual leeches was as high as 100 "/o and the virus was isolated from 95 % of pooled samples of copepods. The highest level of virus was 8.7 X lo5 pfu (plaque forming units) g-' in the copepod and 1.5 X 10"fu g-' in the leech. The level of virus in leeches removed from fish gills was sometimes higher than the level of virus in the gill tissue itself. Virus persisted for at least 16 d in leeches held in the laboratory without feeding. Transmission of IHN virus by leeches probably increases the infection rate of spawning sockeye salmon.

  5. Kinetics of the inhibition of human leucocyte elastase by eglin from the leech Hirudo medicinalis.

    PubMed Central

    Baici, A; Seemüller, U

    1984-01-01

    The rate constants for the inhibition of human leucocyte elastase by eglin from the leech Hirudo medicinalis were determined by using a pre-steady-state kinetic approach. kon and koff for complex-formation and dissociation were 1 X 10(6)M-1 X S-1 and 8 X 10(-4)S-1 respectively. Ki was calculated as the ratio koff/kon = 8 X 10(-10)M, the binding of eglin to elastase was reversible and the inhibition mechanism was of the fully competitive type. The mechanistic properties of the system and the biological significance of the rate constants are discussed. PMID:6562888

  6. iDNA from terrestrial haematophagous leeches as a wildlife surveying and monitoring tool - prospects, pitfalls and avenues to be developed.

    PubMed

    Schnell, Ida Bærholm; Sollmann, Rahel; Calvignac-Spencer, Sébastien; Siddall, Mark E; Yu, Douglas W; Wilting, Andreas; Gilbert, M Thomas P

    2015-01-01

    Invertebrate-derived DNA (iDNA) from terrestrial haematophagous leeches has recently been proposed as a powerful non-invasive tool with which to detect vertebrate species and thus to survey their populations. However, to date little attention has been given to whether and how this, or indeed any other iDNA-derived data, can be combined with state-of-the-art analytical tools to estimate wildlife abundances, population dynamics and distributions. In this review, we discuss the challenges that face the application of existing analytical methods such as site-occupancy and spatial capture-recapture (SCR) models to terrestrial leech iDNA, in particular, possible violations of key assumptions arising from factors intrinsic to invertebrate parasite biology. Specifically, we review the advantages and disadvantages of terrestrial leeches as a source of iDNA and summarize the utility of leeches for presence, occupancy, and spatial capture-recapture models. The main source of uncertainty that attends species detections derived from leech gut contents is attributable to uncertainty about the spatio-temporal sampling frame, since leeches retain host-blood for months and can move after feeding. Subsequently, we briefly address how the analytical challenges associated with leeches may apply to other sources of iDNA. Our review highlights that despite the considerable potential of leech (and indeed any) iDNA as a new survey tool, further pilot studies are needed to assess how analytical methods can overcome or not the potential biases and assumption violations of the new field of iDNA. Specifically we argue that studies to compare iDNA sampling with standard survey methods such as camera trapping, and those to improve our knowledge on leech (and other invertebrate parasite) physiology, taxonomy, and ecology will be of immense future value.

  7. Purification and Characterization of a Novel Kazal-Type Trypsin Inhibitor from the Leech of Hirudinaria manillensis

    PubMed Central

    Lai, Yanmei; Li, Bowen; Liu, Weihui; Wang, Gan; Du, Canwei; Ombati, Rose; Lai, Ren; Long, Chengbo; Li, Hongyuan

    2016-01-01

    Kazal-type serine proteinase inhibitors are found in a large number of living organisms and play crucial roles in various biological and physiological processes. Although some Kazal-type serine protease inhibitors have been identified in leeches, none has been reported from Hirudinaria manillensis, which is a medically important leech. In this study, a novel Kazal-type trypsin inhibitor was isolated from leech H. manillensis, purified and named as bdellin-HM based on the sequence similarity with bdellin-KL and bdellin B-3. Structural analysis revealed that bdellin-HM was a 17,432.8 Da protein and comprised of 149 amino acid residues with six cysteines forming three intra-molecular disulfide bonds. Bdellin-HM showed similarity with the Kazal-type domain and may belong to the group of “non-classical” Kazal inhibitors according to its CysI-CysII disulfide bridge position. Bdellin-HM had no inhibitory effect on elastase, chymotrypsin, kallikrein, Factor (F) XIIa, FXIa, FXa, thrombin and plasmin, but it showed a potent ability to inhibit trypsin with an inhibition constant (Ki) of (8.12 ± 0.18) × 10−9 M. These results suggest that bdellin-HM from the leech of H. manillensis plays a potent and specific inhibitory role towards trypsin. PMID:27455325

  8. Physiological, pharmacological and behavioral evidence for a TRPA1 channel that can elicit defensive responses in the medicinal leech.

    PubMed

    Summers, Torrie; Wang, Yanqing; Hanten, Brandon; Burrell, Brian D

    2015-10-01

    Transient receptor potential ankyrin subtype 1 (TRPA1) channels are chemosensitive to compounds such as allyl isothiocyanate (AITC, the active component of mustard oil) and other reactive electrophiles and may also be thermodetectors in many animal phyla. In this study, we provide the first pharmacological evidence of a putative TRPA1-like channel in the medicinal leech. The leech's polymodal nociceptive neuron was activated by both peripheral and central application of the TRPA1 agonist AITC in a concentration-dependent manner. Responses to AITC were inhibited by the selective TRPA1 antagonist HC030031, but also by the TRPV1 antagonist SB366791. Other TRPA1 activators - N-methylmaleimide (NMM) and cinnamaldehyde (CIN) - also activated this nociceptive neuron, although HC030031 only inhibited the effects of NMM. The polymodal nociceptive neurons responded to moderately cold thermal stimuli (<17°C) and these responses were blocked by HC030031. AITC sensitivity was also found in the pressure-sensitive sensory neurons and was blocked by HC030031, but not by SB366791. AITC elicited a nocifensive withdrawal of the posterior sucker in a concentration-dependent manner that could be attenuated with HC030031. Peripheral application of AITC in vivo also produced swimming-like behavior that was attenuated by HC030031. These results suggest the presence of a TRPA1-like channel in the medicinal leech nervous system that responds to cold temperatures and may interact with the leech TRPV-like channel. © 2015. Published by The Company of Biologists Ltd.

  9. More than just one: multiplicity of Hirudins and Hirudin-like Factors in the Medicinal Leech, Hirudo medicinalis.

    PubMed

    Müller, Christian; Mescke, Katharina; Liebig, Stephanie; Mahfoud, Hala; Lemke, Sarah; Hildebrandt, Jan-Peter

    2016-02-01

    Blood-sucking leeches like the medicinal leech, Hirudo medicinalis, have been used for medical purposes since ancient times. During feeding, medicinal leeches transfer a broad range of bioactive substances into the host's wound to prevent premature hemostasis and blood coagulation. Hirudin is probably the best known of these substances. Despite its long history of investigation, recombinant production and clinical use, there still exist conflicting data regarding the primary structure of hirudin. Entirely unclear is the potential biological significance of three different subtypes and many isoforms of hirudins that have been characterized so far. Furthermore, there is only incomplete information on their cDNA sequences and no information at all on gene structures and DNA sequences are available in the databases. Our efforts to fill these gaps revealed the presence of multiple hirudin-encoding genes in the genome of Hirudo medicinalis. We have strong evidence for the expression of all three subtypes of hirudin within individual leeches and for the expression of additional hirudins or hirudin-like factors that may have different biological functions and may be promising candidates for new drugs.

  10. Up-regulation of neurohemerythrin expression in the central nervous system of the medicinal leech, Hirudo medicinalis, following septic injury.

    PubMed

    Vergote, David; Sautière, Pierre-Eric; Vandenbulcke, Franck; Vieau, Didier; Mitta, Guillaume; Macagno, Eduardo R; Salzet, Michel

    2004-10-15

    We report here some results of a proteomic analysis of changes in protein expression in the leech Hirudo medicinalis in response to septic injury. Comparison of two-dimensional protein gels revealed several significant differences between normal and experimental tissues. One protein found to be up-regulated after septic shock was identified, through a combination of Edman degradation, mass spectrometry, and molecular cloning, as a novel member of the hemerythrin family, a group of non-heme-iron oxygen transport proteins found in four invertebrate phyla: sipunculids, priapulids, brachiopods, and annelids. We found by in situ hybridization and immunocytochemistry that the new leech protein, which we have called neurohemerythrin, is indeed expressed in the leech central nervous system. Both message and protein were detected in the pair of large glia within the ganglionic neuropile, in the six packet glia that surround neuronal somata in each central ganglion, and in the bilateral pair of glia that separate axonal fascicles in the interganglionic connective nerves. No expression was detected in central neurons or in central nervous system microglia. Expression was also observed in many other, non-neuronal tissues in the body wall. Real-time PCR experiments suggest that neurohemerythrin is up-regulated posttranscriptionaly. We consider potential roles of neurohemerythrin, associated with its ability to bind oxygen and iron, in the innate immune response of the leech nervous system to bacterial invasion.

  11. Modulation of Ca2+ influx in leech Retzius neurons. I. Effect of extracellular pH.

    PubMed

    Hochstrate, P; Dierkes, P W; Kilb, W; Schlue, W R

    2001-11-01

    We investigated the cytosolic free Ca2+ concentration ([Ca2+]i) of leech Retzius neurons in situ while varying the extracellular and intracellular pH as well as the extracellular ionic strength. Changing these parameters had no significant effect on [Ca2+]i when the membrane potential of the cells was close to its resting value. However, when the cells were depolarized by raising the extracellular K+ concentration or by applying the glutamatergic agonist kainate, extracellular pH and ionic strength markedly affected [Ca2+]i, whereas intracellular pH changes appeared to have virtually no effect. An extracellular acidification decreased [Ca2+]i, while alkalinization or reduction of the ionic strength increased it. Correspondingly, [Ca2+]i also increased when the kainate-induced extracellular acidification was reduced by raising the pH-buffering capacity. At low extracellular pH, the membrane potential to which the cells must be depolarized to evoke a detectable [Ca2+]i increase was shifted to more positive values, and it moved to more negative values at high pH. We conclude that in leech Retzius neurons extracellular pH, but not intracellular pH, affects [Ca2+]i by modulating Ca2+ influx through voltage-dependent Ca2+ channels. The results suggest that this modulation is mediated primarily by shifts in the surface potential at the extracellular side of the plasma membrane.

  12. Expression and function of an even-skipped homolog in the leech Helobdella robusta.

    PubMed

    Song, Mi Hye; Huang, Françoise Z; Chang, Gwendolen Y; Weisblat, David A

    2002-08-01

    We have identified homologs of the Drosophila pair-rule gene even-skipped in the glossiphoniid leeches Helobdella robusta and Theromyzon trizonare. In leech embryos, segments arise sequentially from five pairs of embryonic stem cells (teloblasts) that undergo iterated divisions to generate columns (bandlets) of segmental founder cells (primary blast cells), which in turn generate segmentally iterated sets of definitive progeny. In situ hybridization revealed that Hro-eve is expressed in the teloblasts and primary blast cells, and that these transcripts appear to be associated with mitotic chromatin. In more advanced embryos, Hro-eve is expressed in segmentally iterated sets of cells in the ventral nerve cord. Lineage analysis revealed that neurons expressing Hro-eve arise from the N teloblast. To assess the function of Hro-eve, we examined embryos in which selected blastomeres had been injected with antisense Hro-eve morpholino oligonucleotide (AS-Hro-eve MO), concentrating on the primary neurogenic (N teloblast) lineage. Injection of AS-Hro-eve MO perturbed the normal patterns of teloblast and blast cell divisions and disrupted gangliogenesis. These results suggest that Hro-eve is important in regulating early cell divisions through early segmentation, and that it also plays a role in neuronal differentiation.

  13. New host and geographical records for the leech Acanthobdella peledina Grube 1851 (Hirudinea, Acanthobdellidae)

    USGS Publications Warehouse

    Hauck, A. K.; Fallon, Michael J.; Burger, Carl V.

    1979-01-01

    A total of four leeches (Acanthobdella peledina), parasitizing four specimens of the least cisco (Coregonus sardinella), were found during July and August 1977. The hosts and parasites were collected during a fishery survey by the U.S. Fish and Wildlife Service in the North Slope waters of Naval Petroleum Reserve, Alaska. Two host fishes were collected from the Chipp River (70035' latitude, 155012' longitude) and two from an unnamed, landlocked lake (69054' latitude, 153o23' longitude). The Chipp River collection site is about 130 km southeast of Barrow, and the unnamed lake about 200 km southeast of Barrow. The leeches, which were fixed in situ with neutral formalin, appeared to have penetrated the integument and were embedded in subcutaneous tissues and white muscle at the base of the pelvic fins. The specimens were cylindrical and about 23 mm long and 2-3 mm wide. Color before fixation was olive-green. The five anterior segments each had four pairs of hooked setae at the ventral surface. Our identification was based on a description in Bykhovskaya-Pavlovskaya et al., 1962, Key to parasites of freshwater fish of the USSR (Transl. from Russian), NTIS TT-64-11040.

  14. Spatial-specific action of serotonin within the leech midbody ganglion.

    PubMed

    Calviño, María Ana; Szczupak, Lidia

    2008-06-01

    Serotonin is a conspicuous neuromodulator in the nervous system of many vertebrates and invertebrates. In previous experiments performed in the leech nervous system, we compared the effect of the amine released from endogenous sources [using selective serotonin reuptake inhibitors (SSRIs), e.g. fluoxetine] with that of bath-applied serotonin. The results suggested that the amine does not reach all its targets in a uniform way, but produces the activation of an interneuronal pathway that generated specific synaptic responses on different neurons. Taking into account that the release of the amine is often regulated at the presynaptic level, we have investigated whether autoreceptor antagonists mimic the SSRIs effect. We found that methiothepin (100 microM) produced similar effects than fluoxetine. To further test the hypothesis that endogenous serotonin produce its effect by acting locally at specific sites, we analyzed the effect of iontophoretic applications of serotonin. We found a site in the neuropil of the leech ganglia where serotonin application mimicked the effect of the SSRIs and the 5-HT antagonist. The results further support the view that the effect of serotonin exhibits a spatial specificity that can be relevant to understand its modulatory actions.

  15. Toxicity induced by cumene hydroperoxide in leech Retzius nerve cells: the protective role of glutathione.

    PubMed

    Jovanovic, Zorica; Jovanovic, Svetlana

    2013-01-01

    In the present study, we studied the ability of glutathione (GSH) to detoxify exogenously applied cumene hydroperoxide (CHP). Exposure of leech Retzius nerve cells to CHP (1.5 mM) induced a marked prolongation of the spontaneous spike potential of these cells. Early after depolarization, and a cardiac-like action potential with a rapid depolarization followed by a sustained depolarization or plateau, which is terminated by a rapid repolarization were recorded. GSH (0.2 mM) significantly inhibited the effects of CHP on the duration of the action potential and suppressed CHP-induced spontaneous repetitive activity. Voltage-clamp recordings showed that CHP (1.5 mM) caused significant changes in the outward potassium currents. The fast and slow steady part of the potassium outward current was reduced by 46% and 39%, respectively. GSH applied in a concentration of 0.2 mM partially blocked the effect of CHP on the calcium-activated potassium currents. The fast and slow calcium-activated potassium currents were suppressed by about 20% and 15%, respectively. These results suggest that the neurotoxic effect of CHP on spontaneous spike electrogenesis and calcium-activated potassium currents of leech Retzius nerve cells was reduced in the presence of GSH.

  16. Decision Points: The Factors Influencing the Decision to Feed in the Medicinal Leech

    PubMed Central

    Gaudry, Quentin; Kristan, William B.

    2012-01-01

    The decision to feed is a complex task that requires making several small independent choices. Am I hungry? Where do I look for food? Is there something better I’d rather be doing? When should I stop? With all of these questions, it is no wonder that decisions about feeding depend on several sensory modalities and that the influences of these sensory systems would be evident throughout the nervous system. The leech is uniquely well suited for studying these complicated questions due to its relatively simple nervous system, its exceptionally well-characterized behaviors and neural circuits, and the ease with which one can employ semi-intact preparations to study the link between physiology and decision-making. We will begin this review by discussing the cellular substrates that govern the decision to initiate and to terminate a bout of feeding. We will then discuss how feeding temporarily blocks competing behaviors from being expressed while the animal continues to feed. Then we will review what is currently known about how feeding affects long-term behavioral choices of the leech. Finally, we conclude with a short discussion of the advantages of the leech’s decision-making circuit’s design and how this design might be applicable to all decision circuits. PMID:22783162

  17. Molecular mechanisms of short-term habituation in the leech Hirudo medicinalis.

    PubMed

    Zaccardi, Maria Luisa; Mozzachiodi, Riccardo; Traina, Giovanna; Brunelli, Marcello; Scuri, Rossana

    2012-04-01

    Although habituation is ubiquitous in the animal kingdom, its underlying mechanisms remain poorly understood. In this study, we began to explore the molecular cascades underlying short-term habituation in the leech Hirudo medicinalis. In H. medicinalis, a training paradigm, consisting of low-frequency repetitive electrical stimulation of the skin, produces a gradual increase in the latency to swim that spontaneously recovers within 20-30 min. As first step in determining the molecular pathways in short-term habituation, we examined the role of Ca(2+). Both Ca(2+) influx through voltage-gated channels and Ca(2+) release from intracellular stores were found to contribute to short-term habituation. The analysis of the downstream targets of elevated cytosolic Ca(2+) revealed that the activation of the phosholipase A(2) was required for the induction of short-term habituation. Finally, we reported that the recruitment of arachidonic acid metabolites, generated by the 5-lipoxygenase pathway, was also necessary for the induction of swim induction habituation. These results provide the framework for a comprehensive characterization of the molecular underpinnings of habituation. This outcome will allow us to compare the mechanisms of habituation with those underlying other forms of nonassociative learning in the leech, such as sensitization and dishabituation, and, more in general, with those governing habituation in different vertebrate and invertebrate model systems. Copyright © 2012 Elsevier B.V. All rights reserved.

  18. The phosphoinositide signaling cascade is involved in photoreception in the leech Hirudo medicinalis.

    PubMed

    Ukhanov, K; Walz, B

    The effects of BAPTA, heparin, and neomycin on electrical light responses were studied in the photoreceptors of Hirudo medicinalis. Light activation produces a fast increase in intracellular Ca2+ concentration (Cai) as detected with the fluorescent Ca2+ indicator calcium green-5N. Chelating intracellular calcium by injections of 10 mmol(-1) BAPTA suppresses spontaneous quantum bumps, reduces light sensitivity by more than 2 log(10) units, and substantially increases the latent period of light responses. BAPTA strongly inhibits the plateau phase of responses to long steps of light. Injections of 45-100 mg ml(-1) of heparin act in a similar manner to BAPTA, affecting the latency of the light responses even more. De-N-sulfated heparin, an inactive analog, is almost ineffective at the same concentration compared with heparin. Heparin diminishes the light-induced Cai elevation significantly, whereas de-N-sulfated heparin does not. Intracellular injections of 50-100 mmol l(-1) of the aminoglycoside neomycin, which inhibits phospholipase-C-mediated inositol 1,4,5-trisphosphate formation, acts similar to BAPTA and heparin. Pressure injections of the hydrolysis resistant analog of inositol 1,4,5-trisphosphate, inositol 2,4,5-trisphosphate, strongly depolarize leech photoreceptors and mimic an effect of light adaptation. These results suggest a close similarity between phototransduction mechanisms in leech photoreceptors and existing models for visual transduction in other invertebrate microvillar photoreceptors.

  19. Molecular basis of gap junctional communication in the CNS of the leech Hirudo medicinalis.

    PubMed

    Dykes, Iain M; Freeman, Fiona M; Bacon, Jonathan P; Davies, Jane A

    2004-01-28

    Gap junctions are intercellular channels that allow the passage of ions and small molecules between cells. In the nervous system, gap junctions mediate electrical coupling between neurons. Despite sharing a common topology and similar physiology, two unrelated gap junction protein families exist in the animal kingdom. Vertebrate gap junctions are formed by members of the connexin family, whereas invertebrate gap junctions are composed of innexin proteins. Here we report the cloning of two innexins from the leech Hirudo medicinalis. These innexins show a differential expression in the leech CNS: Hm-inx1 is expressed by every neuron in the CNS but not in glia, whereas Hm-inx2 is expressed in glia but not neurons. Heterologous expression in the paired Xenopus oocyte system demonstrated that both innexins are able to form functional homotypic gap junctions. Hm-inx1 forms channels that are not strongly gated. In contrast, Hm-inx2 forms channels that are highly voltage-dependent; these channels demonstrate properties resembling those of a double rectifier. In addition, Hm-inx1 and Hm-inx2 are able to cooperate to form heterotypic gap junctions in Xenopus oocytes. The behavior of these channels is primarily that predicted from the properties of the constituent hemichannels but also demonstrates evidence of an interaction between the two. This work represents the first demonstration of a functional gap junction protein from a Lophotrochozoan animal and supports the hypothesis that connexin-based communication is restricted to the deuterostome clade.

  20. Glial hyperpolarization upon nerve root stimulation in the leech Hirudo medicinalis.

    PubMed

    Schmidt, J; Prinz, P; Deitmer, J W

    1999-07-01

    Hyperpolarizing responses in neuropil glial cells evoked by nerve root stimulation were studied in the central nervous system of the leech Hirudo medicinalis using intracellular recording and extracellular stimulation techniques. From a mean resting potential of -60.5 +/- 1.0, the glial membrane was hyperpolarized by -8.6 +/- 0.8 mV, via stimulation of the dorsal posterior nerve root in an isolated ganglion. Nerve root stimulation evoked biphasic or depolarizing responses in glial cells with resting potentials around -70 mV (Rose CR, Deitmer JW. J. Neurophysiol. 73:125-131, 1995). The hyperpolarizing response was reduced by the ionotropic glutamate receptor antagonist CNQX (50 microM) to 58% of its initial amplitude. In 15 mM Ca2+/15 mM Mg(2+)-saline the hyperpolarization was reduced by 44%. The hyperpolarization that persisted in high-divalent cation saline was not affected by CNQX. Bath-applied glutamate (500 microM) and kainate (2 microM) elicited glial hyperpolarizations that were sensitive to CNQX and 10 mM Mg2+/1 mM Ca(2+)-saline. The 5-HT-antagonist methysergide did not affect the hyperpolarizations evoked by nerve root stimulation. The results show that in the leech glial membrane responses to neuronal activity include not only depolarizations, as shown previously, but also hyperpolarizations, which are mediated by direct and indirect neuron-glial communication pathways. In the indirect pathway, glutamate is a transmitter between neurons.

  1. A new molecular logic for BMP-mediated dorsoventral patterning, in the leech Helobdella

    PubMed Central

    Kuo, Dian-Han; Weisblat, David A.

    2011-01-01

    Summary BMP signaling is broadly implicated in dorsoventral (DV) patterning of bilaterally symmetric animals [1–3], and its role in axial patterning apparently predates the birth of Bilateria [4–7]. In fly and vertebrate embryos, BMPs and their antagonists (primarily Sog/chordin) diffuse and interact to generate signaling gradients that pattern fields of cells [8–10]. Work in other species reveals diversity in essential facets of this ancient patterning process, however. Here, we report that BMP signaling patterns the DV axis of segmental ectoderm in the leech Helobdella, a clitellate annelid (Super-phylum Lophotrochozoa) featuring stereotyped developmental cell lineages; but the detailed mechanisms of DV patterning in Helobdella differ markedly from fly and vertebrates. In Helobdella, BMP2/4s are expressed broadly, rather than in dorsal territory, while a dorsally expressed BMP5-8 specifies dorsal fate by short-range signaling. A BMP antagonist, gremlin, is up-regulated by BMP5-8 in dorsolateral territory, rather than ventral, and yet the BMP-antagonizing activity of gremlin is required for normal ventral cell fates. Gremlin promotes ventral fates without disrupting dorsal fates by selectively inhibiting BMP2/4s, not BMP5-8. Thus, DV patterning in the development of the leech revealed unexpected evolutionary plasticity of the ‘conserved’ BMP patterning system, presumably reflecting its adaptation to different modes of embryogenesis. PMID:21782437

  2. Field and experimental evidence of a new caiman trypanosome species closely phylogenetically related to fish trypanosomes and transmitted by leeches

    PubMed Central

    Fermino, Bruno R.; Paiva, Fernando; Soares, Priscilla; Tavares, Luiz Eduardo R.; Viola, Laerte B.; Ferreira, Robson C.; Botero-Arias, Robinson; de-Paula, Cátia D.; Campaner, Marta; Takata, Carmen S.A.; Teixeira, Marta M.G.; Camargo, Erney P.

    2015-01-01

    Trypanosoma terena and Trypanosoma ralphi are known species of the South American crocodilians Caiman crocodilus, Caiman yacare and Melanosuchus niger and are phylogenetically related to the tsetse-transmitted Trypanosoma grayi of the African Crocodylus niloticus. These trypanosomes form the Crocodilian clade of the terrestrial clade of the genus Trypanosoma. A PCR-survey for trypanosomes in caiman blood samples and in leeches taken from caimans revealed unknown trypanosome diversity and frequent mixed infections. Phylogenies based on SSU (small subunit) of rRNA and gGAPDH (glycosomal Glyceraldehyde Phosphate Dehydrogenase) gene sequences revealed a new trypanosome species clustering with T. terena and T. ralphi in the crocodilian clade and an additional new species nesting in the distant Aquatic clade of trypanosomes, which is herein named Trypanosoma clandestinus n. sp. This new species was found in Caiman yacare, Caiman crocodilus and M. niger from the Pantanal and Amazonian biomes in Brazil. Large numbers of dividing epimastigotes and unique thin and long trypomastigotes were found in the guts of leeches (Haementeria sp.) removed from the mouths of caimans. The trypanosomes recovered from the leeches had sequences identical to those of T. clandestinus of caiman blood samples. Experimental infestation of young caimans (Caiman yacare) with infected leeches resulted in long-lasting T. clandestinus infections that permitted us to delineate its life cycle. In contrast to T. terena, T. ralphi and T. grayi, which are detectable by hemoculturing, microscopy and standard PCR of caiman blood, T. clandestinus passes undetected by these methods due to very low parasitemia and could be detected solely by the more sensitive nested PCR method. T. clandestinus n. sp. is the first crocodilian trypanosome known to be transmitted by leeches and positioned in the aquatic clade closest to fish trypanosomes. Our data show that caimans can host trypanosomes of the aquatic or

  3. The European land leech: biology and DNA-based taxonomy of a rare species that is threatened by climate warming.

    PubMed

    Kutschera, U; Pfeiffer, I; Ebermann, E

    2007-12-01

    The European land leech Xerobdella lecomtei was discovered in 1868 and is one of the rarest animals on Earth. During the 1960s, several individuals of these approx. 40 mm long, cold-adapted terrestrial annelids that inhabit the moist soils of birch forests around Graz, Austria, were investigated. Only one original research paper has been published on the biology of this species. Between 2001 and 2005, we re-investigated the morphology of preserved specimens and searched for living individuals in their natural habitat that appeared to be intact. We found only one juvenile individual (length approx. 10 mm), indicating that this local leech population became largely extinct over the past four decades. The feeding behaviour of our 'lonesome George of the annelids' was studied and is described here in detail. After its death, the Xerobdella individual was used for chemical extraction and molecular studies (deoxyribonucleic acid [DNA] barcoding, based on one gene, the mitochondrial cytochrome c oxidase subunit I). In addition, novel DNA barcodes for a land leech from Madagascar and a recently discovered species from Europe were obtained. Our phylogenetic tree shows that X. lecomtei is not a member of the tropical land leeches (family Haemadipsidae), as previously thought, but represents a separate line of descent (family Xerobdellidae). The decline of the local leech population around Graz correlates with a rise in average summer temperatures of +3 degrees C between 1961 and 2004. This warming led to a drastic reduction in the moisture content of the soil where X. lecomtei lives. We suggest that human-induced climate change without apparent habitat destruction can lead to the extinction of populations of cold-adapted species that have a low colonization ability.

  4. Helminth and leech community structure in tadpoles and caudatan larvae of two amphibian species from Western Nebraska.

    PubMed

    Rhoden, Heather R; Bolek, Matthew G

    2012-04-01

    Currently no comparative studies exist on helminth and leech community structure among sympatric anuran tadpoles and salamander larvae. During June-August 2007-2009, we examined 50 bullfrog tadpoles, Rana catesbeiana , 50 barred tiger salamander larvae, Ambystoma mavortium , and 3 species of snails from Nevens Pond, Keith County, Nebraska for helminth and leech infections. The helminth and leech compound community of this larval amphibian assemblage consisted of at least 7 species, 4 in bullfrog tadpoles and 4 in barred tiger salamander larvae. Bullfrog tadpoles were infected with 2 species of nematodes ( Gyrinicola batrachiensis and Spiroxys sp.) and 2 types of metacercariae ( Telorchis sp. and echinostomatids), whereas barred tiger salamander larva were infected with 1 species of leech ( Placobdella picta ), 2 species of adult trematodes ( Telorchis corti and Halipegus sp.), and 1 species of an unidentified metacercaria. The component community of bullfrog tadpoles was dominated by helminths acquired through active penetration, or incidentally ingested through respiratory currents, or both, whereas the component community of larval salamanders was dominated by helminths acquired through ingestion of intermediate hosts (χ²  =  3,455.00, P < 0.00001). Differences in amphibian larval developmental time (2-3 yr for bullfrog tadpoles versus 2-5 mo for salamander larvae), the ephemeral nature of intermediate hosts in Nevens Pond, and the ability of bullfrog tadpole to eliminate echinostome infections had significant effects on mean helminth species richness among amphibian species and years (t  =  12.31, P < 0.0001; t  =  2.09, P  =  0.04). Differences in herbivorous and carnivorous diet and time to metamorphosis among bullfrog tadpoles and barred tiger salamander larvae were important factors in structuring helminth communities among the larval stages of these 2 sympatric amphibian species, whereas size was important in structuring helminth and leech

  5. The European land leech: biology and DNA-based taxonomy of a rare species that is threatened by climate warming

    NASA Astrophysics Data System (ADS)

    Kutschera, U.; Pfeiffer, I.; Ebermann, E.

    2007-12-01

    The European land leech Xerobdella lecomtei was discovered in 1868 and is one of the rarest animals on Earth. During the 1960s, several individuals of these approx. 40 mm long, cold-adapted terrestrial annelids that inhabit the moist soils of birch forests around Graz, Austria, were investigated. Only one original research paper has been published on the biology of this species. Between 2001 and 2005, we re-investigated the morphology of preserved specimens and searched for living individuals in their natural habitat that appeared to be intact. We found only one juvenile individual (length approx. 10 mm), indicating that this local leech population became largely extinct over the past four decades. The feeding behaviour of our ‘lonesome George of the annelids’ was studied and is described here in detail. After its death, the Xerobdella individual was used for chemical extraction and molecular studies (deoxyribonucleic acid [DNA] barcoding, based on one gene, the mitochondrial cytochrome c oxidase subunit I). In addition, novel DNA barcodes for a land leech from Madagascar and a recently discovered species from Europe were obtained. Our phylogenetic tree shows that X. lecomtei is not a member of the tropical land leeches (family Haemadipsidae), as previously thought, but represents a separate line of descent (family Xerobdellidae). The decline of the local leech population around Graz correlates with a rise in average summer temperatures of +3°C between 1961 and 2004. This warming led to a drastic reduction in the moisture content of the soil where X. lecomtei lives. We suggest that human-induced climate change without apparent habitat destruction can lead to the extinction of populations of cold-adapted species that have a low colonization ability.

  6. Field and experimental evidence of a new caiman trypanosome species closely phylogenetically related to fish trypanosomes and transmitted by leeches.

    PubMed

    Fermino, Bruno R; Paiva, Fernando; Soares, Priscilla; Tavares, Luiz Eduardo R; Viola, Laerte B; Ferreira, Robson C; Botero-Arias, Robinson; de-Paula, Cátia D; Campaner, Marta; Takata, Carmen S A; Teixeira, Marta M G; Camargo, Erney P

    2015-12-01

    Trypanosoma terena and Trypanosoma ralphi are known species of the South American crocodilians Caiman crocodilus, Caiman yacare and Melanosuchus niger and are phylogenetically related to the tsetse-transmitted Trypanosoma grayi of the African Crocodylus niloticus. These trypanosomes form the Crocodilian clade of the terrestrial clade of the genus Trypanosoma. A PCR-survey for trypanosomes in caiman blood samples and in leeches taken from caimans revealed unknown trypanosome diversity and frequent mixed infections. Phylogenies based on SSU (small subunit) of rRNA and gGAPDH (glycosomal Glyceraldehyde Phosphate Dehydrogenase) gene sequences revealed a new trypanosome species clustering with T. terena and T. ralphi in the crocodilian clade and an additional new species nesting in the distant Aquatic clade of trypanosomes, which is herein named Trypanosoma clandestinus n. sp. This new species was found in Caiman yacare, Caiman crocodilus and M. niger from the Pantanal and Amazonian biomes in Brazil. Large numbers of dividing epimastigotes and unique thin and long trypomastigotes were found in the guts of leeches (Haementeria sp.) removed from the mouths of caimans. The trypanosomes recovered from the leeches had sequences identical to those of T. clandestinus of caiman blood samples. Experimental infestation of young caimans (Caiman yacare) with infected leeches resulted in long-lasting T. clandestinus infections that permitted us to delineate its life cycle. In contrast to T. terena, T. ralphi and T. grayi, which are detectable by hemoculturing, microscopy and standard PCR of caiman blood, T. clandestinus passes undetected by these methods due to very low parasitemia and could be detected solely by the more sensitive nested PCR method. T. clandestinus n. sp. is the first crocodilian trypanosome known to be transmitted by leeches and positioned in the aquatic clade closest to fish trypanosomes. Our data show that caimans can host trypanosomes of the aquatic or

  7. Ammonia-independent sodium uptake mediated by Na(+) channels and NHEs in the freshwater ribbon leech Nephelopsis obscura.

    PubMed

    Quijada-Rodriguez, Alex R; Schultz, Aaron G; Wilson, Jonathan M; He, Yuhe; Allen, Garett J P; Goss, Greg G; Weihrauch, Dirk

    2017-09-15

    Freshwater organisms actively take up ions from their environment to counter diffusive ion losses due to inhabiting hypo-osmotic environments. The mechanisms behind active Na(+) uptake are quite well understood in freshwater teleosts; however, the mechanisms employed by invertebrates are not. Pharmacological and molecular approaches were used to investigate Na(+) uptake mechanisms and their link to ammonia excretion in the ribbon leech Nephelopsis obscura At the molecular level, we identified a Na(+) channel and a Na(+)/H(+) exchanger (NHE) in the skin of N. obscura, where the NHE was up-regulated when acclimated to extremely low [Na(+)] (0.05 mmol l(-1), pH 5) conditions. Additionally, we found that leeches in dilute freshwater environments use both a vacuolar-type H(+)-ATPase (VHA)-assisted uptake via a Na(+) channel and a NHE-based mechanisms for Na(+) uptake. Immunolocalization of VHA and Na(+)/K(+)-ATPase (NKA) indicated at least two cell types present within leech skin, VHA(+) and VHA(-) cells, where the VHA(+) cells are probably involved in Na(+) uptake. NKA was present throughout the epithelium. We also found that increasing ammonia excretion by decreasing water pH, ammonia loading leeches or exposing leeches to high environmental ammonia does not affect Na(+) uptake, providing indications that an NHE-Rh metabolon is not present and that ammonia excretion and Na(+) uptake are not coupled in N. obscura To our knowledge, this is the first study showing the mechanisms of Na(+) uptake and their links to ammonia excretion in a freshwater invertebrate, where results suggest an ammonia-independent Na(+) uptake mechanism relying on both Na(+) channels and NHEs. © 2017. Published by The Company of Biologists Ltd.

  8. OA01.40. A clinical study to evaluate the efficacy of leech therapy and panchatikta ghrita in the management of psoriasis)

    PubMed Central

    Gond, Pushpa; Rani, Rekha; Shringi, M. K.

    2012-01-01

    Purpose: Modern medical science treats psoriasis with PUVA, corticosteroid, anti-mitotic drugs which gives serious side effects like liver and kindney failure etc. There is a need to discover safe and effective medicine without any side effects for Psoriasis and the role of Leech Therapy (Shodhan) and Panchatikta Ghrita (Shaman Karma) is evaluated in this study. Method: 30 patients were included who matched the clinical signs and symptoms of psoriasis. These patients were randomised into three groups. Group A Only on leech therapy, Group B-Only on panchatikta ghrita and Group C On both leech therapy and panchatikta ghrita Result: Group A showed 45% improvement and group B showed 47% improvement, while group C reported 65% improvement. Conclusion: It can be concluded that shodan(leech application) along with shaman (panchatikta ghrita) is effective in the management of psoriasis as it is safe, cost effective and free from any side effects.

  9. Encoding of Tactile Stimuli by Mechanoreceptors and Interneurons of the Medicinal Leech

    PubMed Central

    Kretzberg, Jutta; Pirschel, Friederice; Fathiazar, Elham; Hilgen, Gerrit

    2016-01-01

    For many animals processing of tactile information is a crucial task in behavioral contexts like exploration, foraging, and stimulus avoidance. The leech, having infrequent access to food, developed an energy efficient reaction to tactile stimuli, avoiding unnecessary muscle movements: The local bend behavior moves only a small part of the body wall away from an object touching the skin, while the rest of the animal remains stationary. Amazingly, the precision of this localized behavioral response is similar to the spatial discrimination threshold of the human fingertip, although the leech skin is innervated by an order of magnitude fewer mechanoreceptors and each midbody ganglion contains only 400 individually identified neurons in total. Prior studies suggested that this behavior is controlled by a three-layered feed-forward network, consisting of four mechanoreceptors (P cells), approximately 20 interneurons and 10 individually characterized motor neurons, all of which encode tactile stimulus location by overlapping, symmetrical tuning curves. Additionally, encoding of mechanical force was attributed to three types of mechanoreceptors reacting to distinct intensity ranges: T cells for touch, P cells for pressure, and N cells for strong, noxious skin stimulation. In this study, we provide evidences that tactile stimulus encoding in the leech is more complex than previously thought. Combined electrophysiological, anatomical, and voltage sensitive dye approaches indicate that P and T cells both play a major role in tactile information processing resulting in local bending. Our results indicate that tactile encoding neither relies on distinct force intensity ranges of different cell types, nor location encoding is restricted to spike count tuning. Instead, we propose that P and T cells form a mixed type population, which simultaneously employs temporal response features and spike counts for multiplexed encoding of touch location and force intensity. This hypothesis

  10. Dextran backfill tracers combined with Lucifer yellow injections for neuroanatomic studies of the leech head ganglion.

    PubMed

    Daberkow, D P; Vaughan, D K

    1996-08-01

    Several neuronal tracing substances were applied to the cut ends of leech cephalic nerves and the resulting backfills into the subesophageal ganglion (sbEG) were mapped. A 12 h incubation in 3 kDa dextrans conjugated either to a fluorochrome or to biotin (subsequently tagged with peroxidase) was satisfactory. In separate experiments, possible targets of cephalic nerve afferents (R3 Retzius neurons) were injected with Lucifer Yellow (LY) to visualize their projections. Comparison of the LY-R3 Retzius neuron map with that of the dextran-backfilled D1 nerve revealed extensive overlap in the sbEG. Experiments were performed combining the two protocols, confirming this observation. Moreover, confocal microscopy placed D1 nerve processes in close proximity to R3 Retzius neuron processes, suggesting that they could make synaptic contact with one another in the sbEG. With modifications, this method could be used to identify such contacts using electron microscopy.

  11. Effect of a nonspiking neuron on motor patterns of the leech.

    PubMed

    Rodriguez, Mariano J; Alvarez, Rodrigo J; Szczupak, Lidia

    2012-04-01

    Premotor and motoneurons could play regulatory roles in motor control. We have investigated the role of a premotor nonspiking (NS) neuron of the leech nervous system in two locomotive patterns: swimming and crawling. The NS neuron is coupled through rectifying electrical junctions to all the excitatory motoneurons examined. In addition, activation of motoneurons evokes chemically mediated inhibitory responses in NS. During swimming and crawling, the NS membrane potential (Vm(NS)) oscillated phase locked to the motor output. Hyperpolarization or depolarization of NS had no effect on swimming, but hyperpolarization of NS slowed down the crawling activity and decreased the motoneuron firing frequency. Depolarization of NS increased the motoneuron activity, and, at stages where the crawling pattern was fading, depolarization of NS reinstated it. Future work should determine if NS is actually a member of the central pattern generator or a regulatory element.

  12. Development of swimming in the medicinal leech, the gradual acquisition of a behavior.

    PubMed

    French, K A; Chang, J; Reynolds, S; Gonzalez, R; Kristan, W B; Kristan, W B

    2005-09-01

    Observing the development of behavior provides an assay for the developmental state of an embryo's nervous system. We have previously described the development of behaviors that were largely confined to one or a few segments. We now extend the work to a kinematic analysis of the development of swimming, a behavior that requires coordination of the entire body. When leech embryos first begin to swim they make little forward progress, but within several days they swim as effectively as adults. This increase in efficacy depends on changes in body shape and on improved intersegmental coordination of the swim central pattern generator. These kinematic details suggest how the swim central pattern generating circuit is assembled during embryogenesis.

  13. Cathepsin L and cystatin B gene expression discriminates immune coelomic cells in the leech Theromyzon tessulatum.

    PubMed

    Lefebvre, Christophe; Vandenbulcke, Franck; Bocquet, Béatrice; Tasiemski, Aurélie; Desmons, Annie; Verstraete, Mathilde; Salzet, Michel; Cocquerelle, Claude

    2008-01-01

    Previous studies evidenced that cystatin B-like gene is specifically expressed and induced in large circulating coelomic cells following bacterial challenge in the leech Theromyzon tessulatum. In order to understand the role of that cysteine proteinase inhibitor during immune response, we investigated the existence of members of cathepsin family. We cloned a cathepsin L-like gene and studied its tissue distribution. Immunohistochemical studies using anti-cathepsin L and anti-cystatin B antibodies and ultrastructural results demonstrated the presence of three distinct coelomic cell populations: (1) the chloragocytes, which were initially defined as large coelomocytes, (2) the granular amoebocytes and (3) small coelomic cells. Among those cells, while chloragocytes contain cystatin B and cathepsin L, granular amoebocytes contain only cathepsin L and the third cell population contains neither cathepsin nor inhibitor. Finally, results evidenced that cathepsin L immunopositive granular amoebocytes are chemoattracted to the site of injury and phagocyte bacteria.

  14. Widespread inhibition proportional to excitation controls the gain of a leech behavioral circuit

    PubMed Central

    Kristan, William B.

    2014-01-01

    SUMMARY Changing gain in a neuronal system has important functional consequences, but the underlying mechanisms have been elusive. Models have suggested a variety of neuronal and systems properties to accomplish gain control. Here, we show that the gain of the neuronal network underlying local bending behavior in leeches depends on widespread inhibition. Using behavioral analysis, intracellular recordings, and voltage-sensitive dye imaging, we compared the effects of blocking just the known lateral inhibition with blocking all GABAergic inhibition. This revealed an additional source of inhibition, which was widespread and increased in proportion to increasing stimulus intensity. In a model of the input/output functions of the three-layered local bending network, we showed that inhibiting all interneurons in proportion to the stimulus strength produces the experimentally observed change in gain. This relatively simple mechanism for controlling behavioral gain could be prevalent in vertebrate as well as invertebrate nervous systems. PMID:18215624

  15. Leech segmental repeats develop normally in the absence of signals from either anterior or posterior segments

    NASA Technical Reports Server (NTRS)

    Seaver, E. C.; Shankland, M.

    2000-01-01

    We have investigated whether the development of segmental repeats is autonomous in the embryo of the leech Helobdella robusta. The segmental tissues of the germinal band arise from progeny of five stem cells called teloblasts. Asymmetric divisions of the teloblasts form chains of segment founder cells (called primary blast cells) that divide in a stereotypical manner to produce differentiated descendants. Using two distinct techniques, we have looked for potential interactions between neighboring blast cell clones along the anterior-posterior axis. In one technique, we prevented the birth of primary blast cells by injection of DNase I into the teloblast, thereby depriving the last blast cell produced before the ablation of its normal posterior neighbors. We also ablated single blast cells with a laser microbeam, which allowed us to assess potential signals acting on either more anterior or more posterior primary blast cell clones. Our results suggest that interactions along the anterior-posterior axis between neighboring primary blast cell clones are not required for development of normal segmental organization within the blast cell clone. We also examined the possibility that blast cells receive redundant signals from both anterior and posterior neighboring clones and that either is sufficient for normal development. Using double blast cell laser ablations to isolate a primary blast cell clone by removal of both its anterior and its posterior neighbor, we found that the isolated clone still develops normally. These results reveal that the fundamental segmental repeat in the leech embryo, the primary blast cell clone, can develop normally in the apparent absence of signals from adjacent repeats along the anterior-posterior axis.

  16. Leech segmental repeats develop normally in the absence of signals from either anterior or posterior segments

    NASA Technical Reports Server (NTRS)

    Seaver, E. C.; Shankland, M.

    2000-01-01

    We have investigated whether the development of segmental repeats is autonomous in the embryo of the leech Helobdella robusta. The segmental tissues of the germinal band arise from progeny of five stem cells called teloblasts. Asymmetric divisions of the teloblasts form chains of segment founder cells (called primary blast cells) that divide in a stereotypical manner to produce differentiated descendants. Using two distinct techniques, we have looked for potential interactions between neighboring blast cell clones along the anterior-posterior axis. In one technique, we prevented the birth of primary blast cells by injection of DNase I into the teloblast, thereby depriving the last blast cell produced before the ablation of its normal posterior neighbors. We also ablated single blast cells with a laser microbeam, which allowed us to assess potential signals acting on either more anterior or more posterior primary blast cell clones. Our results suggest that interactions along the anterior-posterior axis between neighboring primary blast cell clones are not required for development of normal segmental organization within the blast cell clone. We also examined the possibility that blast cells receive redundant signals from both anterior and posterior neighboring clones and that either is sufficient for normal development. Using double blast cell laser ablations to isolate a primary blast cell clone by removal of both its anterior and its posterior neighbor, we found that the isolated clone still develops normally. These results reveal that the fundamental segmental repeat in the leech embryo, the primary blast cell clone, can develop normally in the apparent absence of signals from adjacent repeats along the anterior-posterior axis.

  17. Forskolin induces NMDA receptor-dependent potentiation at a central synapse in the leech.

    PubMed

    Grey, Kathryn B; Burrell, Brian D

    2008-05-01

    In vertebrate hippocampal neurons, application of forskolin (an adenylyl cyclase activator) and rolipram (a phosphodiesterase inhibitor) is an effective technique for inducing chemical long-term potentiation (cLTP) that is N-methyl-d-aspartate (NMDA) receptor (NMDAR)-dependent. However, it is not known whether forskolin induces a similar potentiation in invertebrate synapses. Therefore, we examined whether forskolin plus rolipram treatment could induce potentiation at a known glutamatergic synapse in the leech (Hirudo sp.), specifically between the pressure (P) mechanosensory and anterior pagoda (AP) neurons. Perfusion of isolated ganglia with forskolin (50 muM) in conjunction with rolipram (0.1 muM) in Mg(2+)-free saline significantly potentiated the P-to-AP excitatory postsynaptic potential. Application of 2-amino-5-phosphonovaleric acid (APV, 100 muM), a competitive NMDAR antagonist, blocked the potentiation, indicating P-to-AP potentiation is NMDAR-dependent. Potentiation was blocked by injection of bis-(o-aminophenoxy)-N,N,N',N'-tetraacetic acid (BAPTA, 1 mM) into the postsynaptic cell, but not by BAPTA injection into the presynaptic neuron, indicating a requirement for postsynaptic elevation of intracellular Ca(2+). Application of db-cAMP mimicked the potentiating effects of forskolin, and Rp-cAMP, an inhibitor of protein kinase A, blocked forskolin-induced potentiation. Potentiation was also blocked by autocamtide-2-related inhibitory peptide (AIP), indicating a requirement for activation of Ca(2+)/calmodulin-dependent kinase II (CaMKII). Finally, potentiation was blocked by botulinum toxin, suggesting that trafficking of glutamate receptors also plays a role in this form of synaptic plasticity. These experiments demonstrate that techniques used to induce cLTP in vertebrate synapses also induce NMDAR-dependent potentiation in the leech CNS and that many of the cellular processes that mediate LTP are conserved between vertebrate and invertebrate phyla.

  18. Acetyl-L-carnitine affects nonassociative learning processes in the leech Hirudo medicinalis.

    PubMed

    Ristori, C; Cataldo, E; Zaccardi, M L; Traina, G; Calvani, M; Lombardo, P; Scuri, R; Brunelli, M

    2006-11-03

    Acetyl-L-carnitine is a natural molecule widely distributed in vertebrate and invertebrate nervous system. It is known to have significant effects on neuronal activity playing a role as neuroprotective and anti-nociceptive agent, as well as neuromodulatory factor. About its capability of affecting learning processes the available data are controversial. In the present study, we utilized the simplified model system of the leech Hirudo medicinalis to analyze the effects of acetyl-L-carnitine, assessing whether and how it might affect elementary forms of nonassociative learning processes. In leeches with the head ganglion disconnected from the first segmental ganglion, repetitive application of weak electrical shocks onto the caudal portion of the body wall induces habituation of swim induction whereas brush strokes on the dorsal skin produces sensitization or dishabituation when the nociceptive stimulus is delivered on previously habituated animals. Herein, the effects of different concentrations of acetyl-L-carnitine (2 mM - 0.05 mM) have been tested at different times on both sensitization and dishabituation. The results show that a single treatment of acetyl-L-carnitine blocked the onset of sensitization in a dose- and time-dependent manner. In fact, the most effective concentration able to block this process was 2 mM, which induced its major effects 11 days after the treatment, whereas 0.05 mM was unable to affect the sensitization process at all considered time points. On the contrary, acetyl-L-carnitine did not completely abolish dishabituation at the tested concentrations and at every time point. Finally, acetyl-L-carnitine also impaired the habituation of swim induction, but only 11 days after treatment.

  19. Homolog of allograft inflammatory factor-1 induces macrophage migration during innate immune response in leech.

    PubMed

    Schorn, Tilo; Drago, Francesco; Tettamanti, Gianluca; Valvassori, Roberto; de Eguileor, Magda; Vizioli, Jacopo; Grimaldi, Annalisa

    2015-03-01

    Allograft inflammatory factor-1 (AIF-1) is a 17-kDa cytokine-inducible calcium-binding protein that, in vertebrates, plays an important role in the allograft immune response. Its expression is mostly limited to the monocyte/macrophage lineage. Until recently, AIF-1 was assumed to be a novel molecule involved in inflammatory responses. To clarify this aspect, we have investigated the expression of AIF-1 after bacterial challenge and its potential role in regulating the innate immune response in an invertebrate model, the medicinal leech (Hirudo medicinalis). Analysis of an expressed sequence tag library from the central nervous system of Hirudo revealed the presence of the gene Hmaif-1/alias Hmiba1, showing high homology with vertebrate aif-1. Immunohistochemistry with an anti-HmAIF-1 polyclonal antibody revealed the constitutive presence of this protein in spread CD68(+) macrophage-like cells. A few hours after pathogen (bacterial) injection into the body wall, the amount of these immunopositive cells co-expressing HmAIF-1 and the common leucocyte marker CD45 increased at the injected site. Moreover, the recombinant protein HmAIF-1 induced massive angiogenesis and was a potent chemoattractant for macrophages. Following rHmAIF-1 stimulation, macrophage-like cells co-expressed the macrophage marker CD68 and the surface glycoprotein CD45, which, in vertebrates, seems to have a role in the integrin-mediated adhesion of macrophages and in the regulation of the functional responsiveness of cells to chemoattractants. CD45 is therefore probably involved in leech macrophage-like cell activation and migration towards an inflammation site. We have also examined its potential effect on HmAIF-1-induced signalling.

  20. Flavobacterium cutihirudinis sp. nov., isolated from the skin of the medical leech Hirudo verbana.

    PubMed

    Glaeser, S P; Galatis, H; Martin, K; Kämpfer, P

    2013-08-01

    A Gram-staining-negative, non-endospore-forming, yellow-pigmented strain (E89(T)) was isolated from the skin of the medical leech Hirudo verbana obtained from a leech farm located in Biebertal, Germany. 16S rRNA gene sequencing analysis showed that the isolate was grouped in the genus Flavobacterium. Strain E89(T) was most closely related to Flavobacterium chilense LM-09-Fp(T) (98.2 %), Flavobacterium chungangense CJ7(T) (98.1 %), and Flavobacterium oncorhynchi 631-08(T) (98.1 %). 16S rRNA gene sequence similarities to all other species of the genus Flavobacterium were ≤ 97.4 %. A menaquinone of the type MK-6 was found to be the predominant respiratory quinone and the polar lipid profile consisted of the major compounds phosphatidylethanolamine, phosphatidylserine, two unidentified aminolipids, one unknown phospholipid and two unknown lipids. The fatty acid profile was composed of iso-C15 : 0, C15 : 0, summed feature 3 (C16 : 1ω7c and/or iso-C15 : 0 2-OH) found in major amounts and several hydroxylated fatty acids in smaller amounts, among them iso-C15 : 0 3-OH and iso-C17 : 0 3-OH. All these data support the allocation of the isolate in the genus Flavobacterium. Physiological/biochemical characterization and DNA-DNA hybridizations with the type strains of the most closely related species allowed a clear phenotypic and genotypic differentiation of the strain. Based on these data, strain E89(T) represents a novel species of the genus Flavobacterium, for which the name Flavobacterium cutihirudinis sp. nov. is proposed. The type strain is E89(T) (= DSM 25795(T) = LMG 26922(T) = CIP 110374(T)).

  1. Sublethal effects of crude oil on a cold-water marine leech, Johanssonia arcitca, following chronic exposure

    SciTech Connect

    Khan, R.A. ); Kiceniuk, J.W. )

    1989-10-01

    Petroleum polyaromatic hydrocarbons (PAH) are known to affect many species of animals. Mortality and/or morbidity from acute toxicity are usually restricted to the immediate vicinity of a spill, seep or discharge. Subtle effects are sometimes not recognized unless long term observations are made. Several studies have shown that some species of invertebrates tend to accumulate and retain petroleum hydrocarbons for varying periods, sometimes with detrimental effects. In a previous communication, it was noted that water soluble fractions (WSF) of a crude oil distributed digestion and affected reproduction of a hematophagous marine leech, Johanssonia arctica. This leech is a benthic, cold-adapted species that is widely distributed on the continental shelf off eastern Canada especially on the Grand Banks where petroleum reserves have been recently discovered. The present study provides additional evidence that crude oil fractions not only alters egg production but also hatching of eggs and survival of the progeny.

  2. From the Worm in a Bottle of Mezcal: iDNA Confirmation of a Leech Parasitizing the Antillean Manatee.

    PubMed

    Pérez-Flores, J; Rueda-Calderon, H; Kvist, S; Siddall, M E; Oceguera-Figueroa, A

    2016-10-01

    Invertebrate-derived ingested DNA (iDNA) is quickly proving to be a valuable, non-invasive tool for monitoring vertebrate species of conservation concern. Using the DNA barcoding locus, we successfully identified both the blood-feeding leech Haementeria acuecueyetzin and its blood meal-the latter is shown to be derived from the Caribbean manatee, Trichechus manatus . DNA amplification was successful despite the fact that the specimen was fixed in Mezcal (a beverage distilled from agave). We report the first confirmed case of a leech feeding on a manatee, the first record of H. acuecueyetzin for the State of Chiapas and, to our knowledge, the first case of successful DNA amplification of a biological sample fixed in Mezcal other than the caterpillar "worms" more commonly found in that beverage.

  3. Identification and characterization of a novel neuropeptide (neuropeptide Y-HS) from leech salivary gland of Haemadipsa sylvestris.

    PubMed

    Liu, Wei-Hui; Chen, Yan; Bai, Xue-Wei; Yao, Hui-Min; Zhang, Xu-Guang; Yan, Xiu-Wen; Lai, Ren

    2016-09-01

    The present study was designed to identify immunomodulatory components from the leech salivary gland of Haemadipsa sylvestris. The Sephadex G-50, Resource(TM) S column chromatography and reverse-phase high performance liquid chromatography (RP-HPLC) were used to isolate and purify the salivary gland extracts (SGE). Structural analysis of isolated compounds was based on Edman degradation and matrix assisted laser desorption ionization time-of-flight mass spectrometer (MALDI-TOF-MS). The cDNA encoding the precursor of the compound was cloned from the cDNA library of the salivary gland of H. sylvestris. The levels of inflammatory mediators, including tumor necrosis factor-α (TNF-α), interferon γ (IFN-γ), interleukin-6 (IL-6), and monocyte chemotactic protein-1 (MCP-1) were assayed using an enzyme-linked immunosorbent assay (ELISA). The effects on cell proliferation and cell viability were observed using MTT assay. A novel neuropeptide Y (Neuropeptide Y-HS) from the leech salivary gland of H. sylvestris was purified and characterized. It was composed of 36 amino acid residues and the amino acid sequence was determined to be FLEPPERPAVFTSVEQMKSYIKALNDYYLLLGRPRF-NH2, containing an amidated C-terminus. It showed significant inhibitory effects on the production of inflammatory cytokines including TNF-α, IFN-γ, IL-6, and MCP-1. Neuropeptide Y was identified from leeches for the first time. The presence of neuropeptide Y-HS in leech salivary gland may help get blood meal from hosts and inhibit inflammation. Copyright © 2016 China Pharmaceutical University. Published by Elsevier B.V. All rights reserved.

  4. Cultural Resources Investigation of the Reservoir Shorelines: Gull Lake, Leech Lake, Pine River, and Lake Pokegama. Volume 1,

    DTIC Science & Technology

    1979-06-01

    nominations to the National Register of Historic Places. Examples of such sites and structures on Leech Lake are the Chase Hotel in the city of Walker and...Laboratory, University of Minnesota. A Anonymous 1925 Vacation Days; a Complete Guide to the Hotels and Vacation Places in Minnesota. Minneapolis Civic...Summer. Participated in a field school at the University of the Americas, 2holula, Puebla , Mexico. 1972 wuiner. Participated in a field school

  5. Be ready at any time: postprandial synthesis of salivary proteins in salivary gland cells of the haematophagous leech Hirudo verbana.

    PubMed

    Lemke, Sarah; Müller, Christian; Hildebrandt, Jan-Peter

    2016-04-15

    Sanguivorous leeches are ectoparasites having access to body fluids of potential hosts only infrequently. During feeding, salivary proteins are released from unicellular salivary glands into the wound. These substances, among them anti-coagulants, anti-inflammatory or anti-microbial agents, allow these animals proper feeding and long-term storage of host blood in their crops for several months. Using histological, protein biochemical and molecular techniques, we investigated whether synthesis of salivary proteins and refilling of salivary gland cells occur immediately after feeding or later when stored nutrients in the crop are getting scarce. The results of the histological analyses showed that gland cell area was significantly smaller right after feeding when compared with those in unfed animals. This parameter recovered quickly and reached the control level at 1 week after feeding. 2D gel electrophoresis and analysis of the abundance of individual proteins in extracts of leech tissues revealed that a subset of proteins that had been present in extracts of unfed animals virtually disappeared during feeding, but re-appeared within 1 week of feeding (most probably secretory proteins) while another subset did not change during the experimental period (most probably housekeeping proteins). Semi-quantitative PCR analysis of hirudin cDNA prepared from leech RNA samples revealed that the amount of hirudin transcripts increased immediately after feeding, peaked at 5 days after feeding and declined to control values thereafter. Our results indicate that bloodsucking leeches synthesize salivary proteins and refill their salivary gland cell reservoirs within a week of a blood meal to be prepared for another feeding opportunity. © 2016. Published by The Company of Biologists Ltd.

  6. Calreticulin contributes to C1q-dependent recruitment of microglia in the leech Hirudo medicinalis following a CNS injury

    PubMed Central

    Le Marrec-Croq, Françoise; Bocquet-Garcon, Annelise; Vizioli, Jacopo; Vancamp, Christelle; Drago, Francesco; Franck, Julien; Wisztorski, Maxence; Salzet, Michel; Sautiere, Pierre-Eric; Lefebvre, Christophe

    2014-01-01

    Background The medicinal leech is considered as a complementary and appropriate model to study immune functions in the central nervous system (CNS). In a context in which an injured leech’s CNS can naturally restore normal synaptic connections, the accumulation of microglia (immune cells of the CNS that are exclusively resident in leeches) has been shown to be essential at the lesion to engage the axonal sprouting. HmC1q (Hm for Hirudo medicinalis) possesses chemotactic properties that are important in the microglial cell recruitment by recognizing at least a C1q binding protein (HmC1qBP alias gC1qR). Material/Methods Recombinant forms of C1q were used in affinity purification and in vitro chemotaxis assays. Anti-calreticulin antibodies were used to neutralize C1q-mediated chemotaxis and locate the production of calreticulin in leech CNS. Results A newly characterized leech calreticulin (HmCalR) has been shown to interact with C1q and participate to the HmC1q-dependent microglia accumulation. HmCalR, which has been detected in only some microglial cells, is consequently a second binding protein for HmC1q, allowing the chemoattraction of resident microglia in the nerve repair process. Conclusions These data give new insight into calreticulin/C1q interaction in an immune function of neuroprotection, suggesting another molecular target to use in investigation of microglia reactivity in a model of CNS injury. PMID:24747831

  7. Phoretic interaction between the kangaroo leech Marsupiobdella africana (Hirudinea: Glossiphoniidae) and the cape river crab Potamonautes perlatus (Decapoda: Potamonautidae).

    PubMed

    Badets, Mathieu; Preez, Louis Du

    2014-04-01

    The South African leech Marsupiobdella africana is a temporary ectoparasite of the amphibian Xenopus laevis, has a phoretic association with a freshwater crab Potamonautes perlatus, and exhibits advanced parental care by incubating its offspring in a brood pouch. Because phoretic associations are usually regarded to favor the phoront's dispersion, its occurrence within the biology of a parasitic species reflects an intimate context of interactions. In addition to phoresy, attachment to the crab may confer other advantages pertaining to offspring development and predator avoidance, dispersion and the parasitic life cycle. Two ponds where amphibian and crab hosts co-occur were sampled twice a month for a period of 1 year. The population dynamics of the leeches and their use of specific microhabitats as attachment sites on the crabs were also investigated. Results indicate a direct relationship between intra-specific variation in the sex ratio among captured crab hosts and the number of leeches recruited over time. The attachments to specific microhabitats on the hard surfaces of the host suggest a proximal proximate anti-predatory strategy. Finally, the importance of oxygen accessibility for the offspring development has been investigated experimentally. Results revealed a remarkable network of interactions linking all partners of this system raising the question as to whether the crabs merely act as a vehicle or play a role within the parasitic life cycle.

  8. Comparative Mitogenomics of Leeches (Annelida: Clitellata): Genome Conservation and Placobdella-Specific trnD Gene Duplication.

    PubMed

    Oceguera-Figueroa, Alejandro; Manzano-Marín, Alejandro; Kvist, Sebastian; Moya, Andrés; Siddall, Mark E; Latorre, Amparo

    2016-01-01

    Mitochondrial DNA sequences, often in combination with nuclear markers and morphological data, are frequently used to unravel the phylogenetic relationships, population dynamics and biogeographic histories of a plethora of organisms. The information provided by examining complete mitochondrial genomes also enables investigation of other evolutionary events such as gene rearrangements, gene duplication and gene loss. Despite efforts to generate information to represent most of the currently recognized groups, some taxa are underrepresented in mitochondrial genomic databases. One such group is leeches (Annelida: Hirudinea: Clitellata). Herein, we expand our knowledge concerning leech mitochondrial makeup including gene arrangement, gene duplication and the evolution of mitochondrial genomes by adding newly sequenced mitochondrial genomes for three bloodfeeding species: Haementeria officinalis, Placobdella lamothei and Placobdella parasitica. With the inclusion of three new mitochondrial genomes of leeches, a better understanding of evolution for this organelle within the group is emerging. We found that gene order and genomic arrangement in the three new mitochondrial genomes is identical to previously sequenced members of Clitellata. Interestingly, within Placobdella, we recovered a genus-specific duplication of the trnD gene located between cox2 and atp8. We performed phylogenetic analyses using 12 protein-coding genes and expanded our taxon sampling by including GenBank sequences for 39 taxa; the analyses confirm the monophyletic status of Clitellata, yet disagree in several respects with other phylogenetic hypotheses based on morphology and analyses of non-mitochondrial data.

  9. A new species of leech of the genus Placobdella (Hirudinida, Glossiphoniidae) from the American alligator (Alligator mississippiensis) in Mississippi, USA.

    PubMed

    Richardson, Dennis J; Moser, William E; Hammond, Charlotte I; Lazo-Wasem, Eric A; McAllister, Chris T; Pulis, Eric E

    2017-01-01

    To date, the only species of leech reported from the American Alligator, Alligator mississippiensis is Placobdella multilineata. Seven specimens of a previously undescribed species of Placobdella were collected from the feet and lower jaw of a single female alligator from the Pascagoula River Wildlife Management Area, George County, Mississippi. The new species was named Placobdella siddalli Richardson & Moser, sp. n., in honor of the contributions of Dr. Mark Siddall to our understanding of the biology of leeches. Placobdella siddalli Richardson & Moser is similar to other papillated members of the genus Placobdella, but differs from Placobdella ali Hughes & Siddall, 2007, Placobdella rugosa (Verrill, 1874), Placobdella multilineata Moore, 1953, and Placobdella papillifera (Verrill, 1872) in coloration, papillation, ventral striping, and in the possession of a relatively large caudal sucker. In addition, molecular comparison of 626 nucleotides of CO-I between the new species and other papillated leeches (P. ali, P. multilineata, Placobdella ornata, P. papillifera, P. rugosa) revealed interspecific differences of 14.0-18.0% (88-113 nucleotides).

  10. Comparative Mitogenomics of Leeches (Annelida: Clitellata): Genome Conservation and Placobdella-Specific trnD Gene Duplication

    PubMed Central

    Moya, Andrés; Siddall, Mark E.; Latorre, Amparo

    2016-01-01

    Mitochondrial DNA sequences, often in combination with nuclear markers and morphological data, are frequently used to unravel the phylogenetic relationships, population dynamics and biogeographic histories of a plethora of organisms. The information provided by examining complete mitochondrial genomes also enables investigation of other evolutionary events such as gene rearrangements, gene duplication and gene loss. Despite efforts to generate information to represent most of the currently recognized groups, some taxa are underrepresented in mitochondrial genomic databases. One such group is leeches (Annelida: Hirudinea: Clitellata). Herein, we expand our knowledge concerning leech mitochondrial makeup including gene arrangement, gene duplication and the evolution of mitochondrial genomes by adding newly sequenced mitochondrial genomes for three bloodfeeding species: Haementeria officinalis, Placobdella lamothei and Placobdella parasitica. With the inclusion of three new mitochondrial genomes of leeches, a better understanding of evolution for this organelle within the group is emerging. We found that gene order and genomic arrangement in the three new mitochondrial genomes is identical to previously sequenced members of Clitellata. Interestingly, within Placobdella, we recovered a genus-specific duplication of the trnD gene located between cox2 and atp8. We performed phylogenetic analyses using 12 protein-coding genes and expanded our taxon sampling by including GenBank sequences for 39 taxa; the analyses confirm the monophyletic status of Clitellata, yet disagree in several respects with other phylogenetic hypotheses based on morphology and analyses of non-mitochondrial data. PMID:27176910

  11. Segmental control of midbody peristalsis during the consummatory phase of feeding in the medicinal leech, Hirudo medicinalis.

    PubMed

    Wilson, R J; Kleinhaus, A L

    2000-06-01

    The ingestive behavior of the medicinal leech includes peristalsis-like movements of the midbody that are organized into both rostral-to-caudal and caudal-to-rostral waves (C. M. Lent, K. H. Fliegner, E. Freedman, & M. H. Dickenson, 1988). The neuronal control of this behavior is unknown. Using surgical manipulations and electromyograms, the authors show that (a) the head and tail ganglia are not necessary for this behavior; (b) the circuit is distributed, with components reiterated along the length of the leech; (c) excitatory signals transmitted from rostral segments via the nerve cord can initiate peristalsis in "empty" caudal segments; (d) inhibitory signals from caudal segments limit the frequency of peristalsis; and (e) stretch of the gut and/or body wall is sufficient to produce peristalsis in the absence of heat or chemical cues. These results are compared with peristalsis in the digestive tract of mammals. The leech may be a good model for studying peristalsis-like behaviors at the cellular level.

  12. Construction of a medicinal leech transcriptome database and its application to the identification of leech homologs of neural and innate immune genes

    PubMed Central

    2010-01-01

    Background The medicinal leech, Hirudo medicinalis, is an important model system for the study of nervous system structure, function, development, regeneration and repair. It is also a unique species in being presently approved for use in medical procedures, such as clearing of pooled blood following certain surgical procedures. It is a current, and potentially also future, source of medically useful molecular factors, such as anticoagulants and antibacterial peptides, which may have evolved as a result of its parasitizing large mammals, including humans. Despite the broad focus of research on this system, little has been done at the genomic or transcriptomic levels and there is a paucity of openly available sequence data. To begin to address this problem, we constructed whole embryo and adult central nervous system (CNS) EST libraries and created a clustered sequence database of the Hirudo transcriptome that is available to the scientific community. Results A total of ~133,000 EST clones from two directionally-cloned cDNA libraries, one constructed from mRNA derived from whole embryos at several developmental stages and the other from adult CNS cords, were sequenced in one or both directions by three different groups: Genoscope (French National Sequencing Center), the University of Iowa Sequencing Facility and the DOE Joint Genome Institute. These were assembled using the phrap software package into 31,232 unique contigs and singletons, with an average length of 827 nt. The assembled transcripts were then translated in all six frames and compared to proteins in NCBI's non-redundant (NR) and to the Gene Ontology (GO) protein sequence databases, resulting in 15,565 matches to 11,236 proteins in NR and 13,935 matches to 8,073 proteins in GO. Searching the database for transcripts of genes homologous to those thought to be involved in the innate immune responses of vertebrates and other invertebrates yielded a set of nearly one hundred evolutionarily conserved

  13. Phylogenetic and morphological characterization of trypanosomes from Brazilian armoured catfishes and leeches reveal high species diversity, mixed infections and a new fish trypanosome species.

    PubMed

    Lemos, Moara; Fermino, Bruno R; Simas-Rodrigues, Cíntia; Hoffmann, Luísa; Silva, Rosane; Camargo, Erney P; Teixeira, Marta M G; Souto-Padrón, Thaïs

    2015-11-06

    Several Trypanosoma species transmitted by leeches infect marine and freshwater fish worldwide. To date, all South American fish trypanosome species identified have been based on unreliable morphological parameters. We recently isolated and cultured trypanosomes from the Brazilian armoured catfishes Hypostomus luetkeni and H. affinis. Here, we report the first phylogenetic analyses of South American (Brazilian) trypanosomes isolated from fish, and from leeches removed from these fish. We also analysed morphologically and morphometrically the different forms of fish, leech and cultured trypanosomes. V7V8 SSU rRNA and gGAPDH sequences were used for phylogenetic analysis of Brazilian fish and leech trypanosomes. Trypanosomes from cultures, fish blood and leech samples were also characterized morphologically and morphometrically by light and electron microscopy. In blood smears from fish high trypanosome prevalence (90-100 %) and parasitemia (0.9-1.0x10(2)) were observed. Phylogenetic relationships using SSU rRNA and gGAPDH showed that, despite relevant sequence divergence, all Brazilian fish (and derived cultures) and leech trypanosomes clustered together into a single clade. The Brazilian clade clustered with European, North American and African fish trypanosomes. Based on sequence analysis, we uncovered a new species of Brazilian fish trypanosome, Trypanosoma abeli n. sp. Trypanosoma abeli cultures contained pleomorphic epimastigotes, small trypomastigotes and rare sphaeromastigotes. Ultrastructural features of T. abeli included a cytostome-cytopharynx complex in epi- and trypomastigotes, a compact rod-like kinetoplast, lysosome-related organelles (LROs) and multivesicular bodies. Trypanosomes found in fish blood smears and leech samples were highly pleomorphic, in agreement with sequence data suggesting that catfishes and leeches often have mixed trypanosome infections. Trypanosoma abeli n. sp. is the first trypanosome from South American fishes isolated in

  14. Microglia of medicinal leech (Hirudo medicinalis) express a specific activation marker homologous to vertebrate ionized calcium-binding adapter molecule 1 (Iba1/alias aif-1).

    PubMed

    Drago, Francesco; Sautière, Pierre-Eric; Le Marrec-Croq, Françoise; Accorsi, Alice; Van Camp, Christelle; Salzet, Michel; Lefebvre, Christophe; Vizioli, Jacopo

    2014-10-01

    The Ionized calcium-Binding Adapter molecule 1 (Iba1), also known as Allograft Inflammatory Factor 1 (AIF-1), is a 17 kDa cytokine-inducible protein, produced by activated macrophages during chronic transplant rejection and inflammatory reactions in Vertebrates. In mammalian central nervous system (CNS), Iba1 is a sensitive marker associated with activated macrophages/microglia and is upregulated following neuronal death or brain lesions. The medicinal leech Hirudo medicinalis is able to regenerate its CNS after injury, leading to a complete functional repair. Similar to Vertebrates, leech neuroinflammatory processes are linked to microglia activation and recruitment at the lesion site. We identified a gene, named Hmiba1, coding a 17.8 kDa protein showing high similarity with Vertebrate AIF-1. The present work constitutes the first report on an Iba1 protein in the nervous system of an invertebrate. Immunochemistry and gene expression analyses showed that HmIba1, like its mammalian counterpart, is modulated in leech CNS by mechanical injury or chemical stimuli (ATP). We presently demonstrate that most of leech microglial cells migrating and accumulating at the lesion site specifically expressed the activation marker HmIba1. While the functional role of Iba1, whatever species, is still unclear in reactive microglia, this molecule appeared as a good selective marker of activated cells in leech and presents an interesting tool to investigate the functions of these cells during nerve repair events. © 2014 Wiley Periodicals, Inc.

  15. Distribution and functional properties of glutamate receptors in the leech central nervous system.

    PubMed

    Dierkes, P W; Hochstrate, P; Schlue, W R

    1996-06-01

    1. The effect of kainate and other glutamatergic agonists on the membrane potential (Em), the intracellular Na+ activity (aNai), and the intracellular free Ca2+ concentration ([Ca2+]i) of identified leech neurons and neuropile glial cells was measured with conventional and ion-sensitive microelectrodes, as well as with the use of the iontophoretically injected fluorescent indicators sodium-binding benzofuran isophthalate and Fura-2. 2. In Retzius neurons, AE, L, 8, and 101 motoneurons, and in the unclassified 50 neurons (Leydig cells) and AP neurons, as well as in neuropile glial cells, bath application of 100 microM kainate evoked a marked membrane depolarization and an increase in aNai and [Ca2+]i. The kainate-induced aNai increase persisted in solutions with high Mg2+ concentration in which synaptic transmission is blocked. 3. A membrane depolarization as well as an increase in aNai and [Ca2+]i was also evoked by L-glutamate, quisqualate, and L-alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionate (AMPA). The agonist-induced [Ca2+]i increase was inhibited by 6,7-dinitroquinoxaline-2,3-dione (DNQX). 4. In Ca(2+)-free solution, the kainate-induced [Ca2+]i increase was abolished in the neurons and in neuropile glial cells, whereas membrane depolarization and aNai increase were unchanged. In Na(+)-free solution, kainate had no effect on Em, aNai, or [Ca2+]i in the neurons. 5. In the mechanosensory T, P, and N neurons, kainate induced considerably smaller membrane depolarizations than in the other neurons or in neuropile glial cells, and it had no significant effect on aNai or [Ca2+]i. 6. It is concluded that in leech segmental ganglia the majority of the neurons and the neuropile glial cells, but probably not the mechanosensory neurons, possess glutamate receptors of the AMPA-kainate type. In the neurons, the [Ca2+]i increase caused by glutamatergic agonists is due to Ca2+ influx through voltage-dependent Ca2+ channels that are activated by the agonist

  16. The Ozobranchus leech as a mechanical vector for the fibropapilloma-associated turtle herpes virus found latently infecting skin tumors on Hawaiian green turtles (Chelonia mydas)

    USGS Publications Warehouse

    Greenblatt, R.J.; Work, T.M.; Balazs, G.; Sutton, C.A.; Casey, R.N.; Casey, J.W.

    2004-01-01

    Fibropapillomatosis (FP) of marine turtles is a neoplastic disease of ecological concern. A fibropapilloma-associated turtle herpesvirus (FPTHV) is consistently present, usually at loads exceeding one virus copy per tumor cell. DNA from an array of parasites of green turtles (Chelonia mydas) was examined with quantitative PCR (qPCR) to determine whether any carried viral loads are sufficient to implicate them as vectors for FPTHV. Marine leeches (Ozobranchus spp.) were found to carry high viral DNA loads; some samples approached 10 million copies per leech. Isopycnic sucrose density gradient/qPCR analysis confirmed that some of these copies were associated with particles of the density of enveloped viruses. The data implicate the marine leech Ozobranchus as a mechanical vector for FPTHV. Quantitative RT-PCR analysis of FPTHV gene expression indicated that most of the FPTHV copies in a fibropapilloma have restricted DNA polymerase expression, suggestive of latent infection.

  17. The Ozobranchus leech is a candidate mechanical vector for the fibropapilloma-associated turtle herpesvirus found latently infecting skin tumors on Hawaiian green turtles (Chelonia mydas)

    USGS Publications Warehouse

    Greenblatt, R.J.; Work, Thierry M.; Balazs, G.; Sutton, C.A.; Casey, R.N.; Casey, J.W.

    2004-01-01

    Fibropapillomatosis (FP) of marine turtles is a neoplastic disease of ecological concern. A fibropapilloma-associated turtle herpesvirus (FPTHV) is consistently present, usually at loads exceeding one virus copy per tumor cell. DNA from an array of parasites of green turtles (Chelonia mydas) was examined with quantitative PCR (qPCR) to determine whether any carried viral loads are sufficient to implicate them as vectors for FPTHV. Marine leeches (Ozobranchus spp.) were found to carry high viral DNA loads; some samples approached 10 million copies per leech. Isopycnic sucrose density gradient/qPCR analysis confirmed that some of these copies were associated with particles of the density of enveloped viruses. The data implicate the marine leech Ozobranchus as a mechanical vector for FPTHV. Quantitative RT-PCR analysis of FPTHV gene expression indicated that most of the FPTHV copies in a fibropapilloma have restricted DNA polymerase expression, suggestive of latent infection.

  18. A Tale of Transmission: Aeromonas veronii Activity within Leech-Exuded Mucus

    PubMed Central

    Ott, Brittany M.; Dacks, Andrew M.; Ryan, Kenneth J.

    2016-01-01

    Transmission, critical to the establishment and persistence of host-associated microbiotas, also exposes symbionts to new environmental conditions. With horizontal transmission, these different conditions represent major lifestyle shifts. Yet genome-wide analyses of how microbes adjust their transcriptomes toward these dramatic shifts remain understudied. Here, we provide a comprehensive and comparative analysis of the global transcriptional profiles of a symbiont as it shifts between lifestyles during transmission. The gammaproteobacterium Aeromonas veronii is transmitted from the gut of the medicinal leech to other hosts via host mucosal castings, yet A. veronii can also transition from mucosal habitancy to a free-living lifestyle. These three lifestyles are characterized by distinct physiological constraints and consequently lifestyle-specific changes in the expression of stress-response genes. Mucus-bound A. veronii had the greatest expression in terms of both the number of loci and levels of transcription of stress-response mechanisms. However, these bacteria are still capable of proliferating within the mucus, suggesting the availability of nutrients within this environment. We found that A. veronii alters transcription of loci in a synthetic pathway that obtains and incorporates N-acetylglucosamine (NAG; a major component of mucus) into the bacterial cell wall, enabling proliferation. Our results demonstrate that symbionts undergo dramatic local adaptation, demonstrated by widespread transcriptional changes, throughout the process of transmission that allows them to thrive while they encounter new environments which further shape their ecology and evolution. PMID:26896136

  19. Maintenance of Fura-2 fluorescence in glial cells and neurons of the leech central nervous system.

    PubMed

    Munsch, T; Deitmer, J W

    1995-04-01

    Identified glial cells and neurones of the leech central nervous system (CNS) were injected iontophoretically with the calcium indicator dye Fura-2 to measure intracellular Ca2+, while simultaneously recording the membrane potential using a double-barrelled theta-type microelectrode. Both glial cells and neurones responded with Ni(2+)-sensitive Ca2+ transients upon membrane depolarization, indicating Ca2+ influx through voltage-gated Ca2+ channels. In contrast to neurones, the glial cells showed a rapid loss of fluorescence with a half-time of 6.3 +/- 1.8 min (n = 6) after dye injection. Both kinetics and amplitudes of the stimulus-induced Ca2+ transients were affected by this rapid dye loss. The anion exchange inhibitor probenicid (2 mM) significantly reduced, but did not prevent, the loss of Fura-2 fluorescence, suggesting that some dye left the glial cell via an anion exchanger. In order to compensate this fluorescence loss, we injected Fura-2 throughout the experiment. Under this condition, similar Ca2+ transients could be elicited repeatedly for more than 1 h. In Retzius neurones single injections of Fura-2 yielded enough intracellularly trapped dye to allow measurement of intracellular Ca2+ for up to 30 min after the end of injection without large decrease in absolute fluorescence.

  20. Asymmetric cell divisions in the early embryo of the leech Helobdella robusta.

    PubMed

    Weisblat, David A

    2007-01-01

    The small glossiphoniid leech Helobdella robusta is among the best-studied representatives of the super-phylum Lophotrochozoa in terms of early development. The Helobdella embryo undergoes a modified version of spiral cleavage, characterized by stereotyped cell lineages comprising multiple examples of equal and unequal divisions, many of which are well-conserved with respect to those of other clitellate annelids, such as the oligochaete Tubifex. Here, we review the early development of Helobdella, focusing on the variety of unequal cell divisions. We then summarize an experimental analysis of the mechanisms underlying the unequal first cleavage in Helobdella, concluding that the unequal first cleavages in Helobdella and Tubifex proceed by different mechanisms. This result demonstrates the evolvability of the basic cell biological mechanisms underlying well-conserved developmental processes. Finally, we propose a model in which the unequal second cleavage in Helobdella may be regulated by the polarized distribution of PAR protein homologs, convergent with the unequal first cleavage of the nematode Caenorhabditis elegans (super-phylum Ecdysozoa).

  1. Riluzole suppresses postinhibitory rebound in an excitatory motor neuron of the medicinal leech.

    PubMed

    Angstadt, James D; Simone, Amanda M

    2014-08-01

    Postinhibitory rebound (PIR) is an intrinsic property often exhibited by neurons involved in generating rhythmic motor behaviors. Cell DE-3, a dorsal excitatory motor neuron in the medicinal leech exhibits PIR responses that persist for several seconds following the offset of hyperpolarizing stimuli and are suppressed in reduced Na(+) solutions or by Ca(2+) channel blockers. The long duration and Na(+) dependence of PIR suggest a possible role for persistent Na(+) current (I NaP). In vertebrate neurons, the neuroprotective agent riluzole can produce a selective block of I NaP. This study demonstrates that riluzole inhibits cell DE-3 PIR in a concentration- and Ca(2+)-dependent manner. In 1.8 mM Ca(2+) solution, 50-100 µM riluzole selectively blocked the late phase of PIR, an effect similar to that of the neuromodulator serotonin. However, 200 µM riluzole blocked both the early and late phases of PIR. Increasing extracellular Ca(2+) to 10 mM strengthened PIR, but high riluzole concentrations continued to suppress both phases of PIR. These results indicate that riluzole may suppress PIR via a nonspecific inhibition of Ca(2+) conductances and suggest that a Ca(2+)-activated nonspecific current (I(CAN)), rather than I NaP, may underlie the Na(+)-dependent component of PIR.

  2. Using optical flow to characterize sensory-motor interactions in a segment of the medicinal leech.

    PubMed

    Zoccolan, Davide; Torre, Vincent

    2002-03-15

    Activation of motoneurons innervating leech muscles causes the appearance of a two-dimensional vector field of deformations on the skin surface that can be fully characterized using a new technique (Zoccolan et al., 2001) based on the computation of the optical flow, the two-dimensional vector field describing the point displacements on the skin. These vector fields are characterized by their origin (i.e., the singular point) and by four elementary components that combine linearly: expansion (or compression), rotation, longitudinal shear, and oblique shear. All motoneurons can be classified and recognized according to the components of the deformations they elicit: longitudinal motoneurons give rise almost exclusively to longitudinal negative shear, whereas circular motoneurons give rise to both positive longitudinal shear and significant negative expansion. Oblique motoneurons induce strong oblique shear, in addition to longitudinal shear and negative expansion. Vector fields induced by the contraction of longitudinal, circular, and oblique fibers superimpose linearly. Skin deformations can therefore be attributed rather reliably to the contraction of distinct longitudinal, circular, and oblique muscle fibers. We compared the deformation patterns produced by touching the skin with those produced by intracellular stimulation of P, T, and N cells: vector fields resulting from the activation of P cells were almost identical to those produced by mechanical stimulation. Therefore, motor responses triggered by light or moderate touch are almost entirely mediated by excitation of P cells, with minor contributions from T and N cells.

  3. Transmission at a 'direct' electrical connexion mediated by an interneurone in the leech.

    PubMed Central

    Muller, K J; Scott, S A

    1981-01-01

    1. Touch sensory neurones in the leech excite a rapidly conducting interneurone called the S-cell. Although the electrical synaptic connexion between the two cells is monosynaptic by physiological criteria, intracellular staining reveals that the touch cells and the S-cell do not make contact, but instead are linked by a pair of small interneurones. 2. The electrical coupling between touch cells and S-cells rectifies, in that depolarizing current but not hyperpolarizing current passes from the touch cell into the S-cell. The rectifying junction is between the touch cells and coupling interneurones, while the connexion between coupling interneurones and the S-cell passes current in both directions. 3. Selective destruction of the coupling interneurones by intracellular injection of a protease interrupts the disynaptic electrical connexion between touch and S-cells. 4. The touch cell's geometry and membrane properties account for the failure of impulses that are generated in certain portions of the receptive field in the skin to propagate beyond the first branch-points of the touch cell axon within the ganglion. Conduction block at branch-points is used to examine physiologically the spatial distribution of contacts between the touch cell and the coupling interneurones. In addition, it is shown that under natural conditions branch-point failure presynaptically reduces the effectiveness of the electrical synaptic connexions. Images Plate 1 Plate 2 Plate 3 Plate 4 PMID:6267257

  4. Embryonic electrical connections appear to pre-figure a behavioral circuit in the leech CNS.

    PubMed

    Marin-Burgin, Antonia; Eisenhart, F James; Kristan, William B; French, Kathleen A

    2006-02-01

    During development, many embryos show electrical coupling among neurons that is spatially and temporally regulated. For example, in vertebrate embryos extensive dye coupling is seen during the period of circuit formation, suggesting that electrical connections could pre-figure circuits, but it has been difficult to identify which neuronal types are coupled. We have used the leech Hirudo medicinalis to follow the development of electrical connections within the circuit that produces local bending. This circuit consists of three layers of neurons: four mechanosensory neurons (P cells), 17 identified interneurons, and approximately 24 excitatory and inhibitory motor neurons. These neurons can be identified in embryos, and we followed the spatial and temporal dynamics as specific connections developed. Injecting Neurobiotin into identified cells of the circuit revealed that electrical connections were established within this circuit in a precise manner from the beginning. Connections first appeared between motor neurons; mechanosensory neurons and interneurons started to connect at least a day later. This timing correlates with the development of behaviors, so the pattern of emerging connectivity could explain the appearance first of spontaneous behaviors (driven by a electrically coupled motor network) and then of evoked behaviors (when sensory neurons and interneurons are added to the circuit).

  5. From synapses to behavior: development of a sensory-motor circuit in the leech.

    PubMed

    Marin-Burgin, Antonia; Kristan, William B; French, Kathleen A

    2008-05-01

    The development of neuronal circuits has been advanced greatly by the use of imaging techniques that reveal the activity of neurons during the period when they are constructing synapses and forming circuits. This review focuses on experiments performed in leech embryos to characterize the development of a neuronal circuit that produces a simple segmental behavior called "local bending." The experiments combined electrophysiology, anatomy, and FRET-based voltage-sensitive dyes (VSDs). The VSDs offered two major advantages in these experiments: they allowed us to record simultaneously the activity of many neurons, and unlike other imaging techniques, they revealed inhibition as well as excitation. The results indicated that connections within the circuit are formed in a predictable sequence: initially neurons in the circuit are connected by electrical synapses, forming a network that itself generates an embryonic behavior and prefigures the adult circuit; later chemical synapses, including inhibitory connections, appear, "sculpting" the circuit to generate a different, mature behavior. In this developmental process, some of the electrical connections are completely replaced by chemical synapses, others are maintained into adulthood, and still others persist and share their targets with chemical synaptic connections.

  6. Intrinsic frequency response patterns in mechano-sensory neurons of the leech.

    PubMed

    Fischer, Linda; Scherbarth, Frank; Chagnaud, Boris; Felmy, Felix

    2017-07-15

    Animals employ mechano-sensory systems to detect and explore their environment. Mechano-sensation encompasses stimuli such as constant pressure, surface movement or vibrations at various intensities that need to be segregated in the central nervous system. Besides different receptor structures, sensory filtering via intrinsic response properties could provide a convenient way to solve this problem. In leech, three major mechano-sensory cell types can be distinguished, according to their stimulus sensitivity, as nociceptive, pressure and touch cells. Using intracellular recordings, we show that the different mechano-sensory neuron classes in Hirudo medicinalis differentially respond supra-threshold to distinct frequencies of sinusoidal current injections between 0.2 and 20 Hz. Nociceptive cells responded with a low-pass filter characteristic, pressure cells as high-pass filters and touch cells as an intermediate band-pass filter. Each class of mechano-sensory neurons is thus intrinsically tuned to a specific frequency range of voltage oscillation that could help segregate mechano-sensory information centrally. © 2017. Published by The Company of Biologists Ltd.

  7. Pleated septate junctions in leech photoreceptors: ultrastructure, arrangement of septa, gate and fence functions.

    PubMed

    Aschenbrenner, S; Walz, B

    1998-08-01

    The leech photoreceptor forms a unicellular epithelium: every cell surrounds an extracellular "vacuole" that is connected to the remaining extracellular space via narrow clefts containing pleated septate junctions. We analyzed the complete structural layout of all septa within the junctional complex in elastic brightfield stereo electron micrographs of semithin serial sections from photoreceptors infiltrated with colloidal lanthanum. The septa form tortuous interseptal corridors that are spatially continuous, and open ended basally and apically. Individual septa seem to be impermeable to lanthanum; interseptal corridors form the only diffusional pathway for this ion. The junctions form no diffusion barrier for the electron-dense tracer Ba2+, but they hinder the diffusion of various hydrophilic fluorescent dyes as demonstrated by confocal laser scanning microscopy (CLSM) of live cells. Even those dyes that penetrate gap junctions do not diffuse beyond the septate junctions. The aqueous diffusion pathway within the septal corridors is, therefore, less permeable than the gap-junctional pore. Our morphological results combined with published electrophysiological data suggest that the septa themselves are not completely tight for small physiologically relevant ions. We also examined, by CLSM, whether the septate junctions create a permeability barrier for the lateral diffusion of fluorescent lipophilic dyes incorporated into the peripheral membrane domain. AFC16, claimed to remain in the outer membrane leaflet, does not diffuse beyond the junctional region, whereas DiIC16, claimed to flip-flop, does. Thus, pleated septate junctions, like vertebrate tight junctions, contribute to the maintenance of cell polarity.

  8. Intrinsic frequency response patterns in mechano-sensory neurons of the leech

    PubMed Central

    Fischer, Linda; Scherbarth, Frank; Chagnaud, Boris

    2017-01-01

    ABSTRACT Animals employ mechano-sensory systems to detect and explore their environment. Mechano-sensation encompasses stimuli such as constant pressure, surface movement or vibrations at various intensities that need to be segregated in the central nervous system. Besides different receptor structures, sensory filtering via intrinsic response properties could provide a convenient way to solve this problem. In leech, three major mechano-sensory cell types can be distinguished, according to their stimulus sensitivity, as nociceptive, pressure and touch cells. Using intracellular recordings, we show that the different mechano-sensory neuron classes in Hirudo medicinalis differentially respond supra-threshold to distinct frequencies of sinusoidal current injections between 0.2 and 20 Hz. Nociceptive cells responded with a low-pass filter characteristic, pressure cells as high-pass filters and touch cells as an intermediate band-pass filter. Each class of mechano-sensory neurons is thus intrinsically tuned to a specific frequency range of voltage oscillation that could help segregate mechano-sensory information centrally. PMID:28546342

  9. Apparent intracellular Mg2+ buffering in neurons of the leech Hirudo medicinalis.

    PubMed Central

    Günzel, D; Zimmermann, F; Durry, S; Schlue, W R

    2001-01-01

    The apparent intracellular Mg2+ buffering, or muffling (sum of processes that damp changes in the free intracellular Mg2+ concentration, [Mg2+](i), e.g., buffering, extrusion, and sequestration), was investigated in Retzius neurons of the leech Hirudo medicinalis by iontophoretic injection of H+, OH-, or Mg2+. Simultaneously, changes in intracellular pH and the intracellular Mg2+, Na+, or K+ concentration were recorded with triple-barreled ion-selective microelectrodes. Cell volume changes were monitored measuring the tetramethylammonium (TMA) concentration in TMA-loaded neurons. Control measurements were carried out in electrolyte droplets (diameter 100-200 microm) placed on a silver wire under paraffin oil. Droplets with or without ATP, the presumed major intracellular Mg2+ buffer, were used to quantify the pH dependence of Mg2+ buffering and to determine the transport index of Mg2+ during iontophoretic injection. The observed pH dependence of [Mg2+](i) corresponded to what would be expected from Mg2+ buffering through ATP. The quantity of Mg2+ muffling, however, was considerably larger than what would be expected if ATP were the sole Mg2+ buffer. From the decrease in Mg2+ muffling in the nominal absence of extracellular Na+ it was estimated that almost 50% of the ATP-independent muffling is due to the action of Na+/Mg2+ antiport. PMID:11222292

  10. A stable prostacyclin-like substance produced by the medicinal leech Hirudo medicinalis.

    PubMed

    Nikonov, G I; Titova, E A; Seleznev, K G

    1999-08-01

    The medicinal leech Hirudo medicinalis produces a low-molecular mass compound with properties similar to those of prostacyclin. It extracted with organic solvent, had affinity to 6-keto-PGF1alpha antibodies, inhibited human platelet aggregation induced in vitro by thrombin (by 50% at 4 pg/ml), and caused hypotension and secretion of plasminogen (t-PA) into the blood stream of rats. A main distinction from prostacyclin is stability of the substance due to covalent binding with the polypeptide chain of destabilase. Because of the high aggregability of destabilase, the molecules of the protein-lipid complex are organized into micelles that can change their spatial orientation depending on the nature of the solvent. Incorporation of hirudin and blood plasma kallikrein inhibitor into the micelle structure causes the formation of liposomes (with a molecular mass of the structural monomer 25 kDa). This complex with polypeptides provides not only stability but also rapid transmembrane penetration. The pure prostacyclin-like substance has a molecular mass of 391 Da and can be produced on destruction of the destabilase polypeptide chain.

  11. Apparent intracellular Mg2+ buffering in neurons of the leech Hirudo medicinalis.

    PubMed

    Günzel, D; Zimmermann, F; Durry, S; Schlue, W R

    2001-03-01

    The apparent intracellular Mg2+ buffering, or muffling (sum of processes that damp changes in the free intracellular Mg2+ concentration, [Mg2+](i), e.g., buffering, extrusion, and sequestration), was investigated in Retzius neurons of the leech Hirudo medicinalis by iontophoretic injection of H+, OH-, or Mg2+. Simultaneously, changes in intracellular pH and the intracellular Mg2+, Na+, or K+ concentration were recorded with triple-barreled ion-selective microelectrodes. Cell volume changes were monitored measuring the tetramethylammonium (TMA) concentration in TMA-loaded neurons. Control measurements were carried out in electrolyte droplets (diameter 100-200 microm) placed on a silver wire under paraffin oil. Droplets with or without ATP, the presumed major intracellular Mg2+ buffer, were used to quantify the pH dependence of Mg2+ buffering and to determine the transport index of Mg2+ during iontophoretic injection. The observed pH dependence of [Mg2+](i) corresponded to what would be expected from Mg2+ buffering through ATP. The quantity of Mg2+ muffling, however, was considerably larger than what would be expected if ATP were the sole Mg2+ buffer. From the decrease in Mg2+ muffling in the nominal absence of extracellular Na+ it was estimated that almost 50% of the ATP-independent muffling is due to the action of Na+/Mg2+ antiport.

  12. A new genus and species of fish leeches Dolichobdella rubra, gen. n., sp. n., (Clitellata, Hirudinida, Piscicolidae) from the northern Sea ofJapan

    NASA Astrophysics Data System (ADS)

    Utevsky, Serge Y.; Chernyshev, Alexei V.

    2013-02-01

    A new species of fish leeches, Dolichobdella rubra gen. n., sp. n., was found in samples collected by RV Akademik M.A. Lavrentyev in the northern Sea of Japan from 470-528 m during the joint Russian-German expedition SoJaBio (Sea of Japan Biodiversity Studies) in August 2010. The leech does not exceed 13 mm in length and has the following morphological characteristics: body elongated, smooth, lacking gills and pulsatile vesicles; eyes and ocelli absent; coloration reddish; female gonopore larger than male one; 6pairs of testisacs; accessory glands, conductive tissue and copulatory area present; ovisacs short; bursa long; coelomic system reduced.

  13. Identification of iron and heme utilization genes in Aeromonas and their role in the colonization of the leech digestive tract

    PubMed Central

    Maltz, Michele; LeVarge, Barbara L.; Graf, Joerg

    2015-01-01

    It is known that many pathogens produce high-affinity iron uptake systems like siderophores and/or proteins for utilizing iron bound to heme-containing molecules, which facilitate iron-acquisition inside a host. In mutualistic digestive-tract associations, iron uptake systems have not been as well studied. We investigated the importance of two iron utilization systems within the beneficial digestive-tract association Aeromonas veronii and the medicinal leech, Hirudo verbana. Siderophores were detected in A. veronii using chrome azurol S. Using a mini Tn5, a transposon insertion in viuB generated a mutant unable to utilize iron using siderophores. The A. veronii genome was then searched for genes potentially involved in iron utilization bound to heme-containing molecules. A putative outer membrane heme receptor (hgpB) was identified with a transcriptional activator, termed hgpR, downstream. The hgpB gene was interrupted with an antibiotic resistance cassette in both the parent strain and the viuB mutant, yielding an hgpB mutant and a mutant with both iron uptake systems inactivated. In vitro assays indicated that hgpB is involved in utilizing iron bound to heme and that both iron utilization systems are important for A. veronii to grow in blood. In vivo colonization assays revealed that the ability to acquire iron from heme-containing molecules is critical for A. veronii to colonize the leech gut. Since iron and specifically heme utilization is important in this mutualistic relationship and has a potential role in virulence factor of other organisms, genomes from different Aeromonas strains (both clinical and environmental) were queried with iron utilization genes of A. veronii. This analysis revealed that in contrast to the siderophore utilization genes heme utilization genes are widely distributed among aeromonads. The importance of heme utilization in the colonization of the leech further confirms that symbiotic and pathogenic relationships possess similar

  14. Role of the tissue free amino acids in adaptation of medicinal leeches Hirudo medicinalis L., 1758 to extreme climatic conditions.

    PubMed

    Chernaya, L V; Kovalchuk, L A; Nokhrina, E S

    2016-01-01

    The first comparison of the spectra of free amino acids in tissues of the medicinal leeches H. medicinalis from different climatic and geographical Eurasian areas has been performed. Adaptation of H. medicinalis to extreme climatic conditions occurs via intensification of the amino acid metabolism resulting from a significant increase in the content of essential amino acids. Accumulation of arginine, histidine, and lysine (3.6-, 3.9-, and 2.0-fold increases, respectively) has proved to play a special protective role in adaptation of H. medicinalis to the low positive temperatures.

  15. Sodium-magnesium antiport in Retzius neurones of the leech Hirudo medicinalis.

    PubMed Central

    Günzel, D; Schlue, W R

    1996-01-01

    1. Intracellular free magnesium ([Mg2+]i) and sodium ([Na+]i) concentrations were measured in Retzius neurones of the leech Hirudo medicinalis using ion-sensitive microelectrodes. 2. The mean steady-state values for [Mg2+]i and [Na+]i were 0.46 mM (pMg, 3.34 +/- 0.23; range, 0.1-1.2 mM; n = 32) and 8.95 mM (pNa, 2.05 +/- 0.15; range, 5.1-15.5 mM, n = 21), respectively, at a mean membrane potential (Em) of -35.6 +/- 6.1 mV (n = 32). Thus, [Mg2+]i is far below the value calculated for a passive distribution (16.9 mM) but close to the equilibrium value calculated for a hypothetical 1 Na(+)-1 Mg2+ antiport (0.41 mM). 3. Simultaneous measurements of [Mg2+]i, [Na+]i and Em in Retzius neurones showed that an increase in the extracellular Mg2+ concentration ([Mg2+]o) resulted in an increase in [Mg2+]i, a parallel decrease in [Na+]i and a membrane depolarization, while a decrease in [Mg2+]o had opposite effects. These results are compatible with calculations based on a 1 Na(+)-1 Mg2+ antiport. 4. Na+ efflux at high [Mg2+]o still occurred when the Na(+)-K+ pump was inhibited by the application of ouabain or in K(+)-free solutions. This efflux was blocked by amiloride. 5. In the absence of extracellular Na+ ([Na+]o), no Mg2+ influx occurred. Mg2+ influx at high [Mg2+]o was even lower than in the presence of [Na+]o. Mg2+ efflux was blocked in the absence of [Na+]o. 6. The rate of Mg2+ extrusion was reduced by lowering [Na+]o, even if the Na+ gradient across the membrane remained almost unchanged. 7. Mg2+ efflux was blocked by amiloride (half-maximal effect at 0.25 mM amiloride; Hill coefficient, 1.3) but not by 5-(N-ethyl-N-isopropyl)-amiloride (EIPA). 8. No changes in intracellular Ca2+ and pH (pHi) could be detected when [Mg2+]o was varied between 1 and 30 mM. 9. Changing pHi by up to 0.4 pH units had no effect on [Mg2+]i. 10. The results suggest the presence of an electrogenic 1 Na(+)-1 Mg2+ antiport in leech Retzius neurones. This antiport can be reversed and is inhibited

  16. Deciphering the Structural Basis That Guides the Oxidative Folding of Leech-derived Tryptase Inhibitor*

    PubMed Central

    Pantoja-Uceda, David; Arolas, Joan L.; Aviles, Francesc X.; Santoro, Jorge; Ventura, Salvador; Sommerhoff, Christian P.

    2009-01-01

    Protein folding mechanisms have remained elusive mainly because of the transient nature of intermediates. Leech-derived tryptase inhibitor (LDTI) is a Kazal-type serine proteinase inhibitor that is emerging as an attractive model for folding studies. It comprises 46 amino acid residues with three disulfide bonds, with one located inside a small triple-stranded antiparallel β-sheet and with two involved in a cystine-stabilized α-helix, a motif that is widely distributed in bioactive peptides. Here, we analyzed the oxidative folding and reductive unfolding of LDTI by chromatographic and disulfide analyses of acid-trapped intermediates. It folds and unfolds, respectively, via sequential oxidation and reduction of the cysteine residues that give rise to a few 1- and 2-disulfide intermediates. Species containing two native disulfide bonds predominate during LDTI folding (IIa and IIc) and unfolding (IIa and IIb). Stop/go folding experiments demonstrate that only intermediate IIa is productive and oxidizes directly into the native form. The NMR structures of acid-trapped and further isolated IIa, IIb, and IIc reveal global folds similar to that of the native protein, including a native-like canonical inhibitory loop. Enzyme kinetics shows that both IIa and IIc are inhibitory-active, which may substantially reduce proteolysis of LDTI during its folding process. The results reported show that the kinetics of the folding reaction is modulated by the specific structural properties of the intermediates and together provide insights into the interdependence of conformational folding and the assembly of native disulfides during oxidative folding. PMID:19820233

  17. Dopamine activates the motor pattern for crawling in the medicinal leech.

    PubMed

    Puhl, Joshua G; Mesce, Karen A

    2008-04-16

    Locomotion in segmented animals is thought to be based on the coupling of "unit burst generators," but the biological nature of the unit burst generator has been revealed in only a few animal systems. We determined that dopamine (DA), a universal modulator of motor activity, is sufficient to activate fictive crawling in the medicinal leech, and can exert its actions within the smallest division of the animal's CNS, the segmental ganglion. In the entire isolated nerve cord or in the single ganglion, DA induced slow antiphasic bursting (approximately 15 s period) of motoneurons known to participate in the two-step elongation-contraction cycle underlying crawling behavior. During each cycle, the dorsal (DE-3) and ventral (VE-4) longitudinal excitor motoneurons fired approximately 180 degrees out of phase from the ventrolateral circular excitor motoneuron (CV), which marks the elongation phase. In many isolated whole nerve cords, DE-3 bursting progressed in an anterior to posterior direction with intersegmental phase delays appropriate for crawling. In the single ganglion, the dorsal (DI-1) and ventral (VI-2) inhibitory longitudinal motoneurons fired out of phase with each DE-3 burst, further confirming that the crawl unit burst generator exists in the single ganglion. All isolated ganglia of the CNS were competent to produce DA-induced robust fictive crawling, which typically lasted uninterrupted for 5-15 min. A quantitative analysis indicated that DA-induced crawling was not significantly different from electrically evoked or spontaneous crawling. We conclude that DA is sufficient to activate the full crawl motor program and that the kernel for crawling resides within each segmental ganglion.

  18. Cycling of Dense Core Vesicles Involved in Somatic Exocytosis of Serotonin by Leech Neurons

    PubMed Central

    Trueta, Citlali; Kuffler, Damien P.; De-Miguel, Francisco F.

    2012-01-01

    We studied the cycling of dense core vesicles producing somatic exocytosis of serotonin. Our experiments were made using electron microscopy and vesicle staining with fluorescent dye FM1-43 in Retzius neurons of the leech, which secrete serotonin from clusters of dense core vesicles in a frequency-dependent manner. Electron micrographs of neurons at rest or after 1 Hz stimulation showed two pools of dense core vesicles. A perinuclear pool near Golgi apparatuses, from which vesicles apparently form, and a peripheral pool with vesicle clusters at a distance from the plasma membrane. By contrast, after 20 Hz electrical stimulation 47% of the vesicle clusters were apposed to the plasma membrane, with some omega exocytosis structures. Dense core and small clear vesicles apparently originating from endocytosis were incorporated in multivesicular bodies. In another series of experiments, neurons were stimulated at 20 Hz while bathed in a solution containing peroxidase. Electron micrographs of these neurons contained gold particles coupled to anti-peroxidase antibodies in dense core vesicles and multivesicular bodies located near the plasma membrane. Cultured neurons depolarized with high potassium in the presence of FM1-43 displayed superficial fluorescent spots, each reflecting a vesicle cluster. A partial bleaching of the spots followed by another depolarization in the presence of FM1-43 produced restaining of some spots, other spots disappeared, some remained without restaining and new spots were formed. Several hours after electrical stimulation the FM1-43 spots accumulated at the center of the somata. This correlated with electron micrographs of multivesicular bodies releasing their contents near Golgi apparatuses. Our results suggest that dense core vesicle cycling related to somatic serotonin release involves two steps: the production of clear vesicles and multivesicular bodies after exocytosis, and the formation of new dense core vesicles in the perinuclear

  19. Systematics of the freshwater leech genus Hirudinaria Whitman, 1886 (Arhynchobdellida, Hirudinidae) from northeastern Thailand

    PubMed Central

    Tubtimon, Jaruwan; Jeratthitikul, Ekgachai; Sutcharit, Chirasak; Kongim, Bangon; Panha, Somsak

    2014-01-01

    Abstract In total, 435 specimens of the Southeast Asian freshwater leech species within the Hirudinidae family were collected from 17 locations of various types of aquatic habitats in northeastern Thailand. They were all morphologically placed within the genus Hirudinaria Whitman, 1886 and there were three distinct species: the common Hirudinaria manillensis, 78.2% of all collected specimens and at all 17 locations, Hirudinaria javanica at 20.3% of collected samples and from five locations and a rarer unidentified morphospecies (Hirudinaria sp.) with six samples from only two locations. The karyotypes of these three species were examined across their range in this study area for 38, 11 and 6 adult specimens of Hirudinaria manillensis, Hirudinaria javanica and Hirudinaria sp., respectively. This revealed different chromosome numbers among all three species, with Hirudinaria javanica having n = 13, 2n = 26, Hirudinaria manillensis lacked one small chromosome pair with n = 12, 2n = 24, and the unknown Hirudinaria sp. differed from any known Hirudinaria karyotypes in exhibiting a higher chromosome number (n = 14, 2n = 28) and a gradual change in size from large to small chromosomes. This suggests that the unknown Hirudinaria sp. is a new biological species. However, phylogenetic analysis based upon a 658 bp fragment of the cytochrome oxidase subunit I gene placed this unknown morphospecies within the Hirudinaria manillensis clade, perhaps then suggesting a recent sympatric speciation, although this requires further confirmation. Regardless, the chromosomes of all three species were asymmetric, most with telocentric elements. A distinct bi-armed chromosome marker was present on the first chromosome pair in Hirudinaria javanica, whilst it was on pairs 1, 2, 3 and 5 in Hirudinaria manillensis, and on pairs 3 and 5 for the unknown Hirudinaria sp. PMID:25493052

  20. Regional differences in BMP-dependence of dorsoventral patterning in the leech Helobdella

    PubMed Central

    Kuo, Dian-Han; Shankland, Marty; Weisblat, David A.

    2012-01-01

    In the leech Helobdella, the ectoderm exhibits a high degree of morphological homonomy between body segments, but pattern elements in lateral ectoderm arise via distinct cell lineages in the segments of the rostral and midbody regions. In each of the four rostral segments, a complete set of ventrolateral (O fate) and dorsolateral (P fate) ectodermal pattern elements arises from a single founder cell, op. In the 28 midbody and caudal segments, however, there are two initially indeterminate o/p founder cells; the more dorsal of these is induced to adopt the P fate by BMP5-8 emanating from the dorsalmost ectoderm, while the more ventral cell assumes the O fate. Previous work has suggested that the dorsoventral patterning of O and P fates differs in the rostral region, but the role of BMP signaling in those segments has not been investigated. We show here that suppression of dorsal BMP5-8 signaling (which effects a P-to-O fate change in the midbody) has no effect on the patterning of O and P fates in the rostral region. Furthermore, ectopic expression of BMP5-8 in the ventral ectoderm (which induces an O-to-P fate change in the midbody) has no effect in the rostral region. Finally, expression of a dominant-negative BMP receptor (which induces a P-to-O fate change in the midbody) fails to affect O/P patterning in the rostral region. Thus, the rostral segments appear to use some mechanism other than BMP signaling to pattern O and P cell fates along the dorsoventral axis. From a mechanistic standpoint, the OP lineage of the rostral segments and the O-P equivalence group of the midbody and caudal segments constitute distinct developmental modules that rely to differing degrees on positional cues from surrounding ectoderm in order to specify homonomous cell fates. PMID:22641012

  1. Phase relationships between segmentally organized oscillators in the leech heartbeat pattern generating network.

    PubMed

    Masino, Mark A; Calabrese, Ronald L

    2002-03-01

    Motor pattern generating networks that produce segmentally distributed motor outflow are often portrayed as a series of coupled segmental oscillators that produce a regular progression (constant phase differences) in their rhythmic activity. The leech heartbeat central pattern generator is paced by a core timing network, which consists of two coupled segmental oscillators in segmental ganglia 3 and 4. The segmental oscillators comprise paired mutually inhibitory oscillator interneurons and the processes of intersegmental coordinating interneurons. As a first step in understanding the coordination of segmental motor outflow by this pattern generator, we describe the functional synaptic interactions, and activity and phase relationships of the heart interneurons of the timing network, in isolated nerve cord preparations. In the timing network, most (approximately 75%) of the coordinating interneuron action potentials were generated at a primary spike initiation site located in ganglion 4 (G4). A secondary spike initiation site in ganglion 3 (G3) became active in the absence of activity at the primary site. Generally, the secondary site was characterized by a reluctance to burst and a lower spike frequency, when compared with the primary site. Oscillator interneurons in G3 inhibited spike activity at both initiation sites, whereas oscillator interneurons in G4 inhibited spike activity only at the primary initiation site. This asymmetry in the control of spike activity in the coordinating interneurons may account for the observation that the phase of the coordinating interneurons is more tightly linked to the G3 than G4 oscillator interneurons. The cycle period of the timing network and the phase difference between the ipsilateral G3 and G4 oscillator interneurons were regular within individual preparations, but varied among preparations. This variation in phase differences observed across preparations implies that modulated intrinsic membrane and synaptic properties

  2. Pseudarcicella hirudinis gen. nov., sp. nov., isolated from the skin of the medical leech Hirudo medicinalis.

    PubMed

    Kämpfer, Peter; Busse, Hans-Jürgen; Longaric, Igor; Rosselló-Móra, Ramon; Galatis, Harald; Lodders, Nicole

    2012-09-01

    A pinkish-pigmented, Gram-stain-negative, rod-shaped, non-spore-forming bacterium, strain E92(T), was isolated from the skin of the medical leech Hirudo medicinalis, on R2A agar. 16S rRNA gene sequence analysis of strain E92(T) showed a relatively low 16S rRNA gene sequence similarity (93.0-93.5 %) to representatives of the genus Arcicella and 91.5-92.0 % to members of the genus Flectobacillus. The polar lipid profile was composed of the major compounds phosphatidylethanolamine, an unidentified aminophospholipid, an unidentified aminolipid and an unidentified polar lipid; glycolipids were not detected. The major quinone was menquinone MK-7, and the major compound in the polyamine pattern was spermidine. The predominant fatty acids were C(16 : 1)ω5c and C(16 : 1)ω7c and/or iso-C(15 : 0) 2-OH (detected as summed feature 3). The isolate did not contain C(14 : 0) or the hydroxyl fatty acid iso-C(17 : 0) 3-OH found in all representatives of the genera Arcicella and Flectobacillus, but did produce C(18 : 1)ω7c and 11-methyl C(18 : 1)ω7c which are not found in these two genera. The DNA G+C content of strain E92(T) was 64.4 mol%. The unique 16S rRNA gene sequence, and specific chemotaxonomic and physiological data revealed that strain E92(T) represents a new genus in the family Cytophagaceae for which we propose the name Pseudarcicella hirudinis gen. nov., sp. nov., with the type strain of the type species as E92(T) (= LMG 26720(T) = CCM 7988(T)).

  3. Creation and reduction of a morphologically detailed model of a leech heart interneuron

    PubMed Central

    Tobin, Anne-Elise; Van Hooser, Stephen D.; Calabrese, Ronald L.

    2010-01-01

    Conductance-based neuron models aid in understanding the role intrinsic and synaptic currents play in producing neuronal activity. Incorporating morphological detail into a model allows for additional analysis of non-homogeneous distributions of active and synaptic conductances, as well as spatial segregation of electrical events. We developed a morphologically detailed “Full Model” of a leech heart interneuron that replicates reasonably well intracellular recordings from these interneurons. However, it comprises hundreds of compartments, each increasing parameter space and simulation time. To reduce the number of compartments of the Full Model, while preserving conductance densities and distributions, its compartments were grouped into functional groups that each share identical conductance densities. Each functional group was sequentially reduced to one or two compartments, preserving surface area, conductance densities and its contribution to input resistance. As a result, the input resistance and membrane time constant were preserved. The axial resistances of several compartments were rescaled to match the amplitude of synaptic currents and low-threshold calcium currents and the shape of action potentials to those in the Full Model. This reduced model, with intrinsic conductances, matched the activity of the Full Model for a variety of simulated current-clamp and voltage-clamp data. Because surface area and conductance distribution of the functional groups of the Full Model were maintained, parameter changes introduced into the reduced model can be directly translated to the Full Model. Thus, our computationally efficient reduced morphology model can be used as a tool for exploring the parameter space of the Full Model and in network simulations. PMID:16760352

  4. Endogenous and half-center bursting in morphologically-inspired models of leech heart interneurons

    PubMed Central

    Tobin, Anne-Elise; Calabrese, Ronald L.

    2010-01-01

    Based on a detailed morphology ‘Full Model’ of a leech heart interneuron, we previously developed a computationally-efficient, morphologically-inspired ‘Reduced Model’ to expedite tuning the model to produce endogenous bursting and alternating bursting when configured as a half-center oscillator (paired with reciprocally inhibitory synapses). To find conductance density distributions that produce endogenous bursting, we implemented a genetic algorithm automated parameter search. With multiple searches, we found eight parameter sets that produced endogenous bursting in the Reduced Model. When these parameter sets were applied to the Full Model, all produced endogenous bursting, although when the simulation time was extended from 80 s to 300 s, only four parameter sets produced sustained bursting in the Reduced Models. All parameter sets produced alternating half-center bursting in the Reduced and Full Models throughout 300 s. When conductance amplitudes were systematically varied for each of the four sustained burster sets, the effects on bursting activity differed, both for the same parameter set in the Reduced and Full Models and for different parameter sets with the same level of morphological detail. This implies that morphological detail can affect burst activity and that these parameter sets may represent different mechanisms for burst generation and/or regulation. We also tested the models with parameter variations that correspond to experimental manipulations. We conclude that whereas similar output can be achieved with multiple different parameter sets, perturbations such as conductance variations can highlight differences. Additionally, this work demonstrates both the utility and limitations of using simplified models to represent more morphologically-accurate models. PMID:16760353

  5. Microbial challenge promotes the regenerative process of the injured central nervous system of the medicinal leech by inducing the synthesis of antimicrobial peptides in neurons and microglia.

    PubMed

    Schikorski, David; Cuvillier-Hot, Virginie; Leippe, Matthias; Boidin-Wichlacz, Céline; Slomianny, Christian; Macagno, Eduardo; Salzet, Michel; Tasiemski, Aurélie

    2008-07-15

    Following trauma, the CNS of the medicinal leech, unlike the mammalian CNS, has a strong capacity to regenerate neurites and synaptic connections that restore normal function. In this study, we show that this regenerative process is enhanced by a controlled bacterial infection, suggesting that induction of regeneration of normal CNS function may depend critically upon the coinitiation of an immune response. We explore the interaction between the activation of a neuroimmune response and the process of regeneration by assaying the potential roles of two newly characterized antimicrobial peptides. Our data provide evidence that microbial components differentially induce the transcription, by microglial cells, of both antimicrobial peptide genes, the products of which accumulate rapidly at sites in the CNS undergoing regeneration following axotomy. Using a preparation of leech CNS depleted of microglial cells, we also demonstrate the production of antimicrobial peptides by neurons. Interestingly, in addition to exerting antibacterial properties, both peptides act as promoters of the regenerative process of axotomized leech CNS. These data are the first to report the neuronal synthesis of antimicrobial peptides and their participation in the immune response and the regeneration of the CNS. Thus, the leech CNS appears as an excellent model for studying the implication of immune molecules in neural repair.

  6. Hm-MyD88 and Hm-SARM: Two key regulators of the neuroimmune system and neural repair in the medicinal leech

    PubMed Central

    Rodet, F.; Tasiemski, A.; Boidin-Wichlacz, C.; Van Camp, C.; Vuillaume, C.; Slomianny, C.; Salzet, M.

    2015-01-01

    Unlike mammals, the CNS of the medicinal leech can regenerate damaged neurites, thus restoring neural functions after lesion. We previously demonstrated that the injured leech nerve cord is able to mount an immune response promoting the regenerative processes. Indeed neurons and microglia express sensing receptors like Hm-TLR1, a leech TLR ortholog, associated with chemokine release in response to a septic challenge or lesion. To gain insights into the TLR signaling pathways involved during these neuroimmune responses, members of the MyD88 family were investigated. In the present study, we report the characterization of Hm-MyD88 and Hm-SARM. The expression of their encoding gene was strongly regulated in leech CNS not only upon immune challenge but also during CNS repair, suggesting their involvement in both processes. This work also showed for the first time that differentiated neurons of the CNS could respond to LPS through a MyD88-dependent signalling pathway, while in mammals, studies describing the direct effect of LPS on neurons and the outcomes of such treatment are scarce and controversial. In the present study, we established that this PAMP induced the relocalization of Hm-MyD88 in isolated neurons. PMID:25880897

  7. Hm-MyD88 and Hm-SARM: two key regulators of the neuroimmune system and neural repair in the medicinal leech.

    PubMed

    Rodet, F; Tasiemski, A; Boidin-Wichlacz, C; Van Camp, C; Vuillaume, C; Slomianny, C; Salzet, M

    2015-04-16

    Unlike mammals, the CNS of the medicinal leech can regenerate damaged neurites, thus restoring neural functions after lesion. We previously demonstrated that the injured leech nerve cord is able to mount an immune response promoting the regenerative processes. Indeed neurons and microglia express sensing receptors like Hm-TLR1, a leech TLR ortholog, associated with chemokine release in response to a septic challenge or lesion. To gain insights into the TLR signaling pathways involved during these neuroimmune responses, members of the MyD88 family were investigated. In the present study, we report the characterization of Hm-MyD88 and Hm-SARM. The expression of their encoding gene was strongly regulated in leech CNS not only upon immune challenge but also during CNS repair, suggesting their involvement in both processes. This work also showed for the first time that differentiated neurons of the CNS could respond to LPS through a MyD88-dependent signalling pathway, while in mammals, studies describing the direct effect of LPS on neurons and the outcomes of such treatment are scarce and controversial. In the present study, we established that this PAMP induced the relocalization of Hm-MyD88 in isolated neurons.

  8. Population variation and individual maximum size in two leech populations: energy extraction from cannibalism or niche widening?

    PubMed

    Persson, Lennart; Elliott, J Malcolm

    2013-05-01

    The theory of cannibal dynamics predicts a link between population dynamics and individual life history. In particular, increased individual growth has, in both modeling and empirical studies, been shown to result from a destabilization of population dynamics. We used data from a long-term study of the dynamics of two leech (Erpobdella octoculata) populations to test the hypothesis that maximum size should be higher in a cycling population; one of the study populations exhibited a delayed feedback cycle while the other population showed no sign of cyclicity. A hump-shaped relationship between individual mass of 1-year-old leeches and offspring density the previous year was present in both populations. As predicted from the theory, the maximum mass of individuals was much larger in the fluctuating population. In contrast to predictions, the higher growth rate was not related to energy extraction from cannibalism. Instead, the higher individual mass is suggested to be due to increased availability of resources due to a niche widening with increased individual body mass. The larger individual mass in the fluctuating population was related to a stronger correlation between the densities of 1-year-old individuals and 2-year-old individuals the following year in this population. Although cannibalism was the major mechanism regulating population dynamics, its importance was negligible in terms of providing cannibalizing individuals with energy subsequently increasing their fecundity. Instead, the study identifies a need for theoretical and empirical studies on the largely unstudied interplay between ontogenetic niche shifts and cannibalistic population dynamics.

  9. The present state of the leech fauna (Annelida, Hirudinea) in the Upper Irtysh cascade of water reservoirs

    PubMed Central

    Fedorova, Lyudmila I.; Kaygorodova, Irina A.

    2016-01-01

    Abstract Hirudinea is a small and ecologically important group of aquatic organisms. However, up to date, the leech fauna of Kazakhstan is poorly studied. The presence of large under-collected areas, such as the Upper Irtysh basin, makes biodiversity studies concerning these invertebrates from Kazakhstan relevant. In this paper, the latest information on species diversity of the freshwater hirudofauna of the Upper Irtysh cascade of water reservoirs, the Kazakhstan part of Irtysh River, is presented. It includes 10 free-living and parasitic species, of which 7 and 9 inhabit the Shulbinsk and the Bukhtarma reservoirs, respectively. These species belong to 2 orders, 3 families and 6 genera. The faunal list highlights four potentially new morphological species (Alboglossiphonia sp., Erpobdella sp., Piscicola sp. 1 and Piscicola sp. 2). Besides them, another three species Erpobdella vilnensis, Helobdella stagnalis and Theromyzon tessulatum recorded for the first time in the area. The exact systematic position is stated for all leech taxa. Each species from the list accompanied with information on taxonomic synonymy, data on its geographic distribution, and brief summary of morphological and ecological characteristics. PMID:27408572

  10. Feeding-mediated distention inhibits swimming in the medicinal leech Abbreviated title: Inhibition of swimming by distention

    PubMed Central

    Gaudry, Quentin; Kristan, William B

    2010-01-01

    An animal’s response to a stimulus depends upon many factors such as age, hormonal state, experience, and its behavioral state. For example, an animal may suppress a behavior that is inappropriate or incompatible with its current state. In this study, we show that as a medicinal leech feeds, the distention that it incurs inhibits its expression of swimming. Distention slows the swimming pattern and decreases the number of swim cycles elicited by a test electrical stimulation; large distentions inhibit swimming altogether. We have previously shown that the ingestive phase of feeding inhibits behaviors by presynaptic inhibition of mechanosensory neurons. Distention has its effects downstream (e.g. gating and central pattern generating interneurons) from these sensory neurons and thus represents a novel mechanism for choosing between conflicting behaviors during feeding. Because removing the leech’s gut surgically did not eliminate the effects of body distention, we conclude that the receptors mediating the distention-induced suppression of swimming are likely to be located in the animal’s body wall. Together with previous findings, these new data show that leeches rely on two different decision-making networks to ensure that a biologically important behavior is not disrupted by other behaviors. PMID:20660257

  11. Comparison of the effects of cumene hydroperoxide and hydrogen peroxide on Retzius nerve cells of the leech Haemopis sanguisuga.

    PubMed

    Jovanovic, Zorica; Jovanovic, Svetlana

    2013-01-01

    Oxidative stress and the production of reactive oxygen species are known to play a major role in neuronal cell damage, but the exact mechanisms responsible for neuronal injury and death remain uncertain. In the present study, we examined the effects of oxidative stress on spontaneous spike activity and depolarizing outward potassium current by exposing the Retzius neurons of the leech to cumene hydroperoxide (CHP) and hydrogen peroxide (H(2)O(2)), the oxidants commonly used to examine oxidative mechanisms mediating cell death. We observed that relatively low concentrations of CHP (0.25, 1, and 1.5 mM) led to a marked prolongation of spontaneous repetitive activity. The prolonged action potentials showed an initial, spike-like depolarization followed by a plateau phase. In contrast, H(2)O(2) at the same and much higher concentrations (0.25 to 5 mM) did not significantly change the duration of spontaneous spike potentials of leech Retzius nerve cells (LRNCs). In the voltage clamp experiments, calcium-activated outward potassium currents, needed for the repolarization of the action potential, were suppressed with CHP, but not with H(2)O(2). The present findings indicate that CHP is a more potent oxidant and neurotoxin than H(2)O(2) and that the effect of CHP on the electrophysiological properties of LRNCs may be due to the inhibition of the potassium channels.

  12. Host specificity in Metamera sillasenorum, n. sp., a gregarine parasite of the leech Helobdella triserialis with notes on transmission dynamics.

    PubMed

    Wise, M R; Janovy, J; Wise, J C

    2000-06-01

    Eugregarines of the suborder Septatorina are apicomplexan parasites that are found mainly in arthropods. Some exceptions are species in the Metameridae that contains the only 5 septate gregarines recorded from annelids. The type genus is Metamera Duke, 1910 with 2 species, Metamera schubergi Duke, 1910, in European Glossiphonia complanata, and Metamera reynoldsi Jones, 1943, from North American G. complanata. Over the summers of 1995-1998, in Keith County, Nebraska, septate gregarines were found in the glossiphoniid leech Helobdella triserialis. The gregarines were determined to be a new species of Metamera, herein named Metamera sillasenorum. Measurements of size and body proportions of over 600 gregarines and 50 oocysts showed differences from measurements of M. schubergi and M. reynoldsi, and secondary septa in the deutomerite were rarely observed. Field observations indicated that M. sillasenorum is probably host specific. In the laboratory, leeches also exhibited strong feeding preferences; e.g., H. triserialis and G. complanata consumed only snails, whereas Helobdella stagnalis consumed only oligochaetes. Infection experiments demonstrated that freshwater snails ingest the oocysts and are required as mechanical vectors. Oocysts were passed unaltered through the snails' intestines. Glossiphonia complanata did not become infected regardless of heavy exposure to oocysts, although only 5 G. complanata were used in the experiments. The results show that host specificity of M. sillasenorum is most likely due to a combination of host-feeding habits and host-parasite compatibility.

  13. 9-Phenanthrol modulates postinhibitory rebound and afterhyperpolarizing potentials in an excitatory motor neuron of the medicinal leech.

    PubMed

    Angstadt, James D; Giordano, Joshua R; Goncalves, Alexander J

    2017-08-01

    Postinhibitory rebound (PIR) responses in leech dorsal excitatory motor neurons (cell DE-3) are eliminated by Ca(2+) channel blockers but also exhibit a strong dependence on extracellular Na(+). These features could be explained by a voltage-gated Ca(2+) current acting in concert with a Ca(2+)-activated nonspecific current (ICAN). In vertebrates, ICAN is associated with TRPM4 channels which are blocked selectively by 9-phenanthrol. Here, we show that 9-phenanthrol selectively inhibits a late phase of PIR and simultaneously enhances afterhyperpolarizing potentials (AHPs). Bath application of NNC 55-0396 or Cd(2+) combined with ion substitution experiments indicate that a low-voltage-activated Ca(2+) current plays a key role in generating PIR and that Ca(2+) influx through low- or high-voltage-activated Ca(2+) channels can trigger AHPs via activation of a Ca(2+)-dependent K(+) current. We also demonstrate modulation of rebound responses by other ICAN blockers such as gadolinium and flufenamic acid, as well as the calmodulin antagonist W-7. We discuss how these results provide additional insights into the specific types of ionic currents underlying rebound responses of motor neuron DE-3 in the medicinal leech.

  14. Functional screening of serine protease inhibitors in the medical leech Hirudo medicinalis monitored by intensity fading MALDI-TOF MS.

    PubMed

    Yanes, Oscar; Villanueva, Josep; Querol, Enrique; Aviles, Francesc X

    2005-10-01

    The blood-feeding invertebrates are a rich biological source of drugs and lead compounds to treat cardiovascular diseases because they have evolved highly efficient mechanisms to feed on their hosts by blocking blood coagulation. In this work, we focused our attention on the leech Hirudo medicinalis. We performed, by "intensity fading" MALDI-TOF mass spectrometry, a comprehensive detection and functional analysis of pre-existent peptides and small proteins with the capability of binding to trypsin-like proteases related to blood coagulation. Combining "intensity fading MS" and off-line LC prefractionation allowed us to detect more than 75 molecules present in the leech extract that interact specifically with a trypsin-like protease over a sample profile of nearly 2,000 different peptides/proteins in the 2-20-kDa range. Moreover we resolved 232 individual components from the complex mixture, 13 of which have high sequence homology with previously described serine protease inhibitors. Our findings indicate that such extracts are much more complex than expected. Additionally, intensity fading MS, when complemented with LC separation strategies, seems to be a useful tool to investigate complex biological samples, establishing a new bridge between profiling, functional peptidomics, and subsequent drug discovery.

  15. Coding and adaptation during mechanical stimulation in the leech nervous system.

    PubMed

    Pinato, G; Torre, V

    2000-12-15

    The experiments described here were designed to characterise sensory coding and adaptation during mechanical stimulation in the leech (Hirudo medicinalis). A chain of three ganglia and a segment of the body wall connected to the central ganglion were used. Eight extracellular suction pipettes and one or two intracellular electrodes were used to record action potentials from all mechanosensory neurones of the three ganglia. When the skin of the body wall was briefly touched with a filament exerting a force of about 2 mN, touch (T) cells in the central ganglion, but also those in adjacent ganglia (i.e. anterior and posterior), fired one or two action potentials. However, the threshold for action potential initiation was lower for T cells in the central ganglion than for those in adjacent ganglia. The timing of the first evoked action potential in a T cell was very reproducible with a jitter often lower than 100 us. Action potentials in T cells were not significantly correlated. When the force exerted by the filament was increased above 20 mN, pressure (P) cells in the central and neighbouring ganglia fired action potentials. Action potentials in P cells usually followed those evoked in T cells with a delay of about 20 ms and had a larger jitter of 0.5-10 ms. With stronger stimulations exceeding 50 mN, noxious (N) cells also fired action potentials. With such stimulations the majority of mechanosensory neurones in the three ganglia fired action potentials. The spatial properties of the whole receptive field of the mechanosensory neurones were explored by touching different parts of the skin. When the mechanical stimulation was applied for a longer time, i.e. 1 s, only P cells in the central ganglion continued to fire action potentials. P cells in neighbouring ganglia fully adapted after firing two or three action potentials.P cells in adjacent ganglia, having fully adapted to a steady mechanical stimulation of one part of the skin, fired action potentials following

  16. The LEECH Exoplanet Imaging Survey. Further constraints on the planet architecture of the HR 8799 system

    NASA Astrophysics Data System (ADS)

    Maire, A.-L.; Skemer, A. J.; Hinz, P. M.; Desidera, S.; Esposito, S.; Gratton, R.; Marzari, F.; Skrutskie, M. F.; Biller, B. A.; Defrère, D.; Bailey, V. P.; Leisenring, J. M.; Apai, D.; Bonnefoy, M.; Brandner, W.; Buenzli, E.; Claudi, R. U.; Close, L. M.; Crepp, J. R.; De Rosa, R. J.; Eisner, J. A.; Fortney, J. J.; Henning, T.; Hofmann, K.-H.; Kopytova, T. G.; Males, J. R.; Mesa, D.; Morzinski, K. M.; Oza, A.; Patience, J.; Pinna, E.; Rajan, A.; Schertl, D.; Schlieder, J. E.; Su, K. Y. L.; Vaz, A.; Ward-Duong, K.; Weigelt, G.; Woodward, C. E.

    2015-04-01

    Context. Astrometric monitoring of directly imaged exoplanets allows the study of their orbital parameters and system architectures. Because most directly imaged planets have long orbital periods (>20 AU), accurate astrometry is challenging when based on data acquired on timescales of a few years and usually with different instruments. The LMIRCam camera on the Large Binocular Telescope is being used for the LBT Exozodi Exoplanet Common Hunt (LEECH) survey to search for and characterize young and adolescent exoplanets in L' band (3.8 μm), including their system architectures. Aims: We first aim to provide a good astrometric calibration of LMIRCam. Then, we derive new astrometry, test the predictions of the orbital model of 8:4:2:1 mean motion resonance proposed for the system, and perform new orbital fitting of the HR 8799 bcde planets. We also present deep limits on a putative fifth planet inside the known planets. Methods: We use observations of HR 8799 and the Θ1 Ori C field obtained during the same run in October 2013. Results: We first characterize the distortion of LMIRCam. We determine a platescale and a true north orientation for the images of 10.707 ± 0.012 mas/pix and -0.430 ± 0.076°, respectively. The errors on the platescale and true north orientation translate into astrometric accuracies at a separation of 1'' of 1.1 mas and 1.3 mas, respectively. The measurements for all planets agree within 3σ with a predicted ephemeris. The orbital fitting based on the new astrometric measurements favors an architecture for the planetary system based on 8:4:2:1 mean motion resonance. The detection limits allow us to exclude a fifth planet slightly brighter or more massive than HR 8799 b at the location of the 2:1 resonance with HR 8799 e (~9.5 AU) and about twice as bright as HR 8799 cde at the location of the 3:1 resonance with HR 8799 e (~7.5 AU). The LBT is an international collaboration among institutions in the United States, Italy, and Germany. LBT

  17. Solubilization, molecular forms, purification and substrate specificity of two acetylcholinesterases in the medicinal leech (Hirudo medicinalis).

    PubMed Central

    Talesa, V; Grauso, M; Giovannini, E; Rosi, G; Toutant, J P

    1995-01-01

    Two acetylcholinesterases (AChE) differing in substrate and inhibitor specificities have been characterized in the medical leech (Hirudo medicinalis). A 'spontaneously-soluble' portion of AChE activity (SS-AChE) was recovered from haemolymph and from tissues dilacerated in low-salt buffer. A second portion of AChE activity was obtained after extraction of tissues in low-salt buffer alone or containing 1% Triton X-100 [detergent-soluble (DS-) AChE). Both enzymes were purified to homogeneity by affinity chromatography on edrophonium- and concanavalin A-Sepharose columns. Denaturing SDS/PAGE under reducing conditions gave one band at 30 kDa for purified SS-AChE and 66 kDa for DS-AChE. Sephadex G-200 chromatography indicated a molecular mass of 66 kDa for native SS-AChE and of 130 kDa for DS-AChE. SS-AChE showed a single peak sedimenting at 5.0 S in sucrose gradients with or without Triton X-100, suggesting that it was a hydrophylic monomer (G1). DS-AChE sedimented as a single 6.1-6.5 S peak in the presence of Triton X-100 and aggregated in the absence of detergent. A treatment with phosphatidylinositol-specific phospholipase C suppressed aggregation and gave a 7 S peak. DS-AChE was thus an amphiphilic glycolipid-anchored dimer. Substrate specificities were studied using p-nitrophenyl esters (acetate, propionate and butyrate) and corresponding thiocholine esters as substrates. SS-AChE displayed only limited variations in Km values with charged and uncharged substrates, suggesting a reduced influence of electrostatic interactions in the enzyme substrate affinity. By contrast, DS-AChE displayed higher Km values with uncharged than with charged substrates. SS-AChE was more sensitive to eserine and di-isopropyl fluorophosphate (IC50 5 x 10(-8) and 10(-8) M respectively) than DS-AChE (5 x 10(-7) and 5 x 10(-5) M. Images Figure 2 Figure 3 Figure 4 PMID:7702560

  18. Effect of conduction block at axon bifurcations on synaptic transmission to different postsynaptic neurones in the leech.

    PubMed Central

    Gu, X N

    1991-01-01

    1. The cutaneous receptive field of the medial pressure (mP) sensory neurone in the leech has been examined. The cell has one major receptive field and an anterior and a posterior minor receptive field, principally on lateral and dorsal skin. The two minor receptive fields are contiguous with the major receptive field and are innervated by fine anterior and posterior axons, but there is no overlap between major and minor receptive fields. 2. At low frequencies of stimulation of the minor receptive fields, conduction block takes place in the mP cell at the central branch point within the leech ganglion. 3. The mP cell synapses directly with many other cells in the leech ganglion, including the anterior pagoda (AP) cell, longitudinal (L) motoneurone and the annulus erector (AE) motoneurone, which were studied as a group of postsynaptic neurones. Conduction block in the mP cell affects its synaptic transmission to all three postsynaptic neurones, but the effect can be different in different postsynaptic neurones. Block at the central branch point for an impulse travelling along the anterior axon reduces transmission to the AE cell much more than to the AP or L cells, while block at the central branch for an impulse travelling along the posterior axon has the reverse effect. 4. The distribution of functional connections of the branches of the mP cell with each postsynaptic cell was studied. For this analysis, branches of the mP cell were selectively silenced either during conduction block or by laser microsurgery. Generally, nearly all of the functional connections with the L and AP cell are made by anterior branches of the mP cell while the connection with the AE cell was primarily made by posterior branches of the mP cell. 5. The possible sites of contact between the mP cell and postsynaptic cells were determined by injecting separate markers into the mP cell and a postsynaptic cell. In confirmation of physiology, the mP cell's posterior branches had few, if any

  19. Interaction of HmC1q with leech microglial cells: involvement of C1qBP-related molecule in the induction of cell chemotaxis

    PubMed Central

    2012-01-01

    Background In invertebrates, the medicinal leech is considered to be an interesting and appropriate model to study neuroimmune mechanisms. Indeed, this non-vertebrate animal can restore normal function of its central nervous system (CNS) after injury. Microglia accumulation at the damage site has been shown to be required for axon sprouting and for efficient regeneration. We characterized HmC1q as a novel chemotactic factor for leech microglial cell recruitment. In mammals, a C1q-binding protein (C1qBP alias gC1qR), which interacts with the globular head of C1q, has been reported to participate in C1q-mediated chemotaxis of blood immune cells. In this study, we evaluated the chemotactic activities of a recombinant form of HmC1q and its interaction with a newly characterized leech C1qBP that acts as its potential ligand. Methods Recombinant HmC1q (rHmC1q) was produced in the yeast Pichia pastoris. Chemotaxis assays were performed to investigate rHmC1q-dependent microglia migration. The involvement of a C1qBP-related molecule in this chemotaxis mechanism was assessed by flow cytometry and with affinity purification experiments. The cellular localization of C1qBP mRNA and protein in leech was investigated using immunohistochemistry and in situ hybridization techniques. Results rHmC1q-stimulated microglia migrate in a dose-dependent manner. This rHmC1q-induced chemotaxis was reduced when cells were preincubated with either anti-HmC1q or anti-human C1qBP antibodies. A C1qBP-related molecule was characterized in leech microglia. Conclusions A previous study showed that recruitment of microglia is observed after HmC1q release at the cut end of axons. Here, we demonstrate that rHmC1q-dependent chemotaxis might be driven via a HmC1q-binding protein located on the microglial cell surface. Taken together, these results highlight the importance of the interaction between C1q and C1qBP in microglial activation leading to nerve repair in the medicinal leech. PMID:22356764

  20. Interaction of HmC1q with leech microglial cells: involvement of C1qBP-related molecule in the induction of cell chemotaxis.

    PubMed

    Tahtouh, Muriel; Garçon-Bocquet, Annelise; Croq, Françoise; Vizioli, Jacopo; Sautière, Pierre-Eric; Van Camp, Christelle; Salzet, Michel; Nagnan-le Meillour, Patricia; Pestel, Joël; Lefebvre, Christophe

    2012-02-22

    In invertebrates, the medicinal leech is considered to be an interesting and appropriate model to study neuroimmune mechanisms. Indeed, this non-vertebrate animal can restore normal function of its central nervous system (CNS) after injury. Microglia accumulation at the damage site has been shown to be required for axon sprouting and for efficient regeneration. We characterized HmC1q as a novel chemotactic factor for leech microglial cell recruitment. In mammals, a C1q-binding protein (C1qBP alias gC1qR), which interacts with the globular head of C1q, has been reported to participate in C1q-mediated chemotaxis of blood immune cells. In this study, we evaluated the chemotactic activities of a recombinant form of HmC1q and its interaction with a newly characterized leech C1qBP that acts as its potential ligand. Recombinant HmC1q (rHmC1q) was produced in the yeast Pichia pastoris. Chemotaxis assays were performed to investigate rHmC1q-dependent microglia migration. The involvement of a C1qBP-related molecule in this chemotaxis mechanism was assessed by flow cytometry and with affinity purification experiments. The cellular localization of C1qBP mRNA and protein in leech was investigated using immunohistochemistry and in situ hybridization techniques. rHmC1q-stimulated microglia migrate in a dose-dependent manner. This rHmC1q-induced chemotaxis was reduced when cells were preincubated with either anti-HmC1q or anti-human C1qBP antibodies. A C1qBP-related molecule was characterized in leech microglia. A previous study showed that recruitment of microglia is observed after HmC1q release at the cut end of axons. Here, we demonstrate that rHmC1q-dependent chemotaxis might be driven via a HmC1q-binding protein located on the microglial cell surface. Taken together, these results highlight the importance of the interaction between C1q and C1qBP in microglial activation leading to nerve repair in the medicinal leech.

  1. Expression and characterization of the N-terminal half of antistasin, an anticoagulant protein derived from the leech Haementeria officinalis.

    PubMed

    Palladino, L O; Tung, J S; Dunwiddie, C; Alves, K; Lenny, A B; Przysiecki, C; Lehman, D; Nutt, E; Cuca, G C; Law, S W

    1991-02-01

    Antistasin, a 15-kDa anticoagulant protein isolated from the salivary glands of the Mexican leech Haementeria officinalis, has been shown to be a potent inhibitor of factor Xa in the blood coagulation cascade. Antistasin possesses a twofold internal homology between the N- and C-terminal halves of the molecule, suggesting a gene duplication event in the evolution of the antistasin gene. This structural feature also suggests that either or both halves of the protein may possess biological activity if expressed as separate domains. Because the N-terminal domain contains a factor Xa P1-reactive site, we chose to express this domain in an insect cell baculovirus expression system. Characterization of this recombinant half antistasin molecule reveals that the N-terminal domain inhibits factor Xa in vitro, with a K(i) of 1.7 nM.

  2. The action of 5-hydroxytryptamine and related compounds on the activity of retzius cells of the leech Hirudo medicinalis

    PubMed Central

    Smith, P.A.; Walker, R.J.

    1974-01-01

    1 The equipotent molar ratios of a range of tryptamine analogues, as compared with 5-hydroxytryptamine (5-HT), have been determined on the basis of their ability to hyperpolarize the membrane potential of the Retzius cell of the leech, Hirudo medicinalis. 2 The substitution of methyl, fluoro, chloro, methoxy or acetyl groups onto the 5-HT molecule progressively reduced the potency. 3 5-Methoxylation or terminal N-methylation of tryptamine considerably increased the potency of tryptamine but these compounds tended to depolarize cells rather than cause hyperpolarization. In some experiments they were ineffective on preparations pretreated with 5-HT. 4 It is suggested that these compounds may act by a different mechanism from the 5-hydroxylated indoles, perhaps involving a different receptor. PMID:4441793

  3. The neurotoxic effects of hydrogen peroxide and copper in Retzius nerve cells of the leech Haemopis sanguisuga

    PubMed Central

    Jovanovic, Zorica D.; Stanojevic, Marija B.; Nedeljkov, Vladimir B.

    2016-01-01

    ABSTRACT Oxidative stress and the generation of reactive oxygen species (ROS) play an important role in cellular damage. Electrophysiological analyses have shown that membrane transport proteins are susceptible to ROS. In the present study, oxidative stress was induced in Retzius nerve cells of the leech Haemopis sanguisuga by bath application of 1 mM of hydrogen peroxide (H2O2) and 0.02 mM of copper (Cu) for 20 min. The H2O2/Cu(II) produced considerable changes in the electrical properties of the Retzius nerve cells. Intracellular recording of the resting membrane potential revealed that the neuronal membrane was depolarized in the presence of H2O2/Cu(II). We found that the amplitude of action potentials decreased, while the duration augmented in a progressive way along the drug exposure time. The combined application of H2O2 and Cu(II) caused an initial excitation followed by depression of the spontaneous electrical activity. Voltage-clamp recordings revealed a second effect of the oxidant, a powerful inhibition of the outward potassium channels responsible for the repolarization of action potentials. The neurotoxic effect of H2O2/Cu(II) on the spontaneous spike electrogenesis and outward K+ current of Retzius nerve cells was reduced in the presence of hydroxyl radical scavengers, dimethylthiourea and dimethyl sulfoxide, but not mannitol. This study provides evidence for the oxidative modification of outward potassium channels in Retzius nerve cells. The oxidative mechanism of the H2O2/Cu(II) system action on the electrical properties of Retzius neurons proposed in this study might have a wider significance, referring not only to leeches but also to mammalian neurons. PMID:26935393

  4. A carboxypeptidase inhibitor from the medical leech Hirudo medicinalis. Isolation, sequence analysis, cDNA cloning, recombinant expression, and characterization.

    PubMed

    Reverter, D; Vendrell, J; Canals, F; Horstmann, J; Avilés, F X; Fritz, H; Sommerhoff, C P

    1998-12-04

    A novel metallocarboxypeptidase inhibitor was isolated from the medical leech Hirudo medicinalis. Amino acid sequence analysis provided a nearly complete primary structure. which was subsequently verified and completed by cDNA cloning using reverse transcriptase-polymerase chain reaction/rapid amplification of cDNA end techniques. The inhibitor, called LCI (leech carboxypeptidase inhibitor), is a cysteine-rich polypeptide composed of 66 amino acid residues. It does not show sequence similarity to any other protein except at its C-terminal end. In this region, the inhibitor shares the amino acid sequence -Thr-Cys-X-Pro-Tyr-Val-X with Solanacea carboxypeptidase inhibitors, suggesting a similar mechanism of inhibition where the C-terminal tail of the inhibitor interacts with the active center of metallocarboxypeptidases in a substrate-like manner. This hypothesis is supported by the hydrolytic release of the C-terminal glutamic acid residue of LCI after binding to the enzyme. Heterologous overexpression of LCI in Escherichia coli, either into the medium or as an intracellular thioredoxin fusion protein, yields a protein with full inhibitory activity. Both in the natural and recombinant forms, LCI is a tightly binding, competitive inhibitor of different types of pancreatic-like carboxypeptidases, with equilibrium dissociation constants Ki of 0.2-0.4 x 10(-9) M for the complexes with the pancreatic enzymes A1, A2, and B and plasma carboxypeptidase B. Circular dichroism and nuclear magnetic resonance spectroscopy analysis indicate that recombinant LCI is a compactly folded globular protein, stable to a wide range of pH and denaturing conditions.

  5. Symbiosis of Aeromonas veronii biovar sobria and Hirudo medicinalis, the medicinal leech: a novel model for digestive tract associations.

    PubMed

    Graf, J

    1999-01-01

    Hirudo medicinalis, the medicinal leech, is applied postoperatively in modern medicine. Infections by Aeromonas occur in up to 20% of patients unless a preemptive antibiotic treatment is administered. The associated infections demonstrate the need for a better understanding of the digestive tract flora of H. medicinalis. Early studies reported the presence of a single bacterial species in the digestive tract and suggested that these bacteria were endosymbionts contributing to the digestion of blood. In this study, we cultivated bacteria from the digestive tract and characterized them biochemically. The biochemical test results identified the isolates as Aeromonas veronii biovar sobria. This species identification was supported by sequence comparison of a variable region of the genes coding for 16S rRNA. In a colonization assay, a rifampin-resistant derivative of a symbiotic isolate was fed in a blood meal to H. medicinalis. The strain colonized the digestive tract rapidly and reached a concentration similar to that of the native bacterial flora. For the first 12 h, the in vivo doubling time was 1.2 h at 23 degreesC. After 12 h, at a density of 5 x 10(7) CFU/ml, the increase in viable counts ceased, suggesting a dramatic reduction in the bacterial growth rate. Two human fecal isolates, identified as Aeromonas hydrophila and A. veronii biovar sobria, were also able to colonize the digestive tract. These data demonstrate that the main culturable bacterium in the crop of H. medicinalis is A. veronii biovar sobria and that the medicinal leech can be used as a model for digestive tract association of Aeromonas species.

  6. Symbiosis of Aeromonas veronii Biovar sobria and Hirudo medicinalis, the Medicinal Leech: a Novel Model for Digestive Tract Associations

    PubMed Central

    Graf, Joerg

    1999-01-01

    Hirudo medicinalis, the medicinal leech, is applied postoperatively in modern medicine. Infections by Aeromonas occur in up to 20% of patients unless a preemptive antibiotic treatment is administered. The associated infections demonstrate the need for a better understanding of the digestive tract flora of H. medicinalis. Early studies reported the presence of a single bacterial species in the digestive tract and suggested that these bacteria were endosymbionts contributing to the digestion of blood. In this study, we cultivated bacteria from the digestive tract and characterized them biochemically. The biochemical test results identified the isolates as Aeromonas veronii biovar sobria. This species identification was supported by sequence comparison of a variable region of the genes coding for 16S rRNA. In a colonization assay, a rifampin-resistant derivative of a symbiotic isolate was fed in a blood meal to H. medicinalis. The strain colonized the digestive tract rapidly and reached a concentration similar to that of the native bacterial flora. For the first 12 h, the in vivo doubling time was 1.2 h at 23°C. After 12 h, at a density of 5 × 107 CFU/ml, the increase in viable counts ceased, suggesting a dramatic reduction in the bacterial growth rate. Two human fecal isolates, identified as Aeromonas hydrophila and A. veronii biovar sobria, were also able to colonize the digestive tract. These data demonstrate that the main culturable bacterium in the crop of H. medicinalis is A. veronii biovar sobria and that the medicinal leech can be used as a model for digestive tract association of Aeromonas species. PMID:9864188

  7. pH recovery from intracellular alkalinization in Retzius neurones of the leech central nervous system.

    PubMed Central

    Frey, G; Schlue, W R

    1993-01-01

    1. Neutral-carrier pH-sensitive microelectrodes were used to investigate intracellular pH (pHi) recovery from alkalinization in leech Retzius neurones in Hepes- and in CO2-HCO3(-)-buffered solution. The Retzius neurones were alkaline loaded by the addition and subsequent removal of 16 mM acetate, by changing from 5% CO2-27 mM HCO3- to 2% CO2-11 mM HCO3- or by changing from CO2-HCO3(-)- to Hepes-buffered solution. 2. In Hepes-buffered solution (pH 7.4) the mean pHi was 7.29 +/- 0.11 and the mean membrane potential -44.7 +/- 5.9 mV (mean +/- S.D.; n = 83). 3. The rate of pHi recovery from alkalinization increased with decreasing pH of the bathing medium (pHb). pHi changed about 0.30 pH units for a pHb unit change. 4. A decrease of extracellular buffer concentration (Hepes concentration lowered from 20 to 5 mM) caused an acidification of extracellular and intracellular pH and an acceleration of pHi recovery from alkalinization. 5. A depolarization of the Retzius cell membrane-induced by increasing the K+ concentration of the bathing medium from 4 to 20 mM (delta Em = 16.5 +/- 5.5 mV) or from 4 to 40 mM (delta Em = 24.8 +/- 3.5 mV)--evoked a decrease of pHi and an acceleration of pHi recovery from alkalinization. 6. The H+ current blocker Zn2+ (0.5 mM) inhibited pHi recovery from alkalinization at resting membrane potential as well as during depolarization. The inhibition was more pronounced during depolarization. 7. In Cl(-)-free, CO2-HCO3(-)-buffered solution pHi recovery from an alkaline load by changing from 5% CO2-27 mM HCO3- to 2% CO2-11 mM HCO3- was slowed by 48-71%. The rate of pHi recovery from an alkaline load induced by changing from CO2-HCO3- to Hepes buffer was reduced by 33-56% in Cl(-)-free solution. The removal of external Cl- did not affect pHi recovery in Hepes-buffered solution. 8. The pHi recovery from alkalinization was DIDS-insensitive in CO2-HCO3(-)- as in Hepes-buffered solutions and was not slowed in the absence of external Na+. 9. It is

  8. Mechanism of ammonia excretion in the freshwater leech Nephelopsis obscura: characterization of a primitive Rh protein and effects of high environmental ammonia

    PubMed Central

    Quijada-Rodriguez, Alex R.; Treberg, Jason R.

    2015-01-01

    Remarkably little is known about nitrogenous excretion in freshwater invertebrates. In the current study, the nitrogen excretion mechanism in the carnivorous ribbon leech, Nephelopsis obscura, was investigated. Excretion experiments showed that the ribbon leech is ammonotelic, excreting 166.0 ± 8.6 nmol·grams fresh weight (gFW)−1·h−1 ammonia and 14.7 ± 1.9 nmol·gFW−1·h−1 urea. Exposure to high and low pH hampered and enhanced, respectively, ammonia excretion rates, indicating an acid-linked ammonia trapping mechanism across the skin epithelia. Accordingly, compared with body tissues, the skin exhibited elevated mRNA expression levels of a newly identified Rhesus protein and at least in tendency the Na+/K+-ATPase. Pharmacological experiments and enzyme assays suggested an ammonia excretion mechanism that involves the V-ATPase, Na+/K+-ATPase, and carbonic anhydrase, but not necessarily a functional microtubule system. Most importantly, functional expression studies of the identified Rh protein cloned from leech skin tissue revealed an ammonia transport capability of this protein when expressed in yeast. The leech Rh-ammonia transporter (NoRhp) is a member of the primitive Rh protein family, which is a sister group to the common ancestor of vertebrate ammonia-transporting Rh proteins. Exposure to high environmental ammonia (HEA) caused a new adjustment of body ammonia, accompanied with a decrease in NoRhp and Na+/K+-ATPase mRNA levels, but unaltered ammonia excretion rates. To our knowledge, this is only the second comprehensive study regarding the ammonia excretion mechanisms in a freshwater invertebrate, but our results show that basic processes of ammonia excretion appear to also be comparable to those found in freshwater fish, suggesting an early evolution of ionoregulatory mechanisms in freshwater organisms. PMID:26180186

  9. Field Mapping, LiDAR Analysis and Shallow Geophysical Methods Define the Geometry and Kinematics of the Leech River Fault, an Active Forearc Structure in Northern Cascadia

    NASA Astrophysics Data System (ADS)

    Graham, A. C.; Morell, K. D.; Leonard, L. J.; Regalla, C.; Levson, V.

    2016-12-01

    We use LiDAR imagery, shallow geophysical methods, and structural/geomorphic mapping to constrain the geometry and kinematics of the Leech River fault, an active crustal structure in the northern Cascadia outer forearc on southern Vancouver Island, British Columbia. Field mapping along a 20 km section of the fault reveals a 100 - 500 m wide zone comprising multiple sub-vertical brittle faults cutting through units on either side of the main regional lithologic contact. Electrical resistivity and GPR surveys across scarps identified in LiDAR imagery likewise reveal steeply dipping structures that appear to deform late Quaternary units to within 2 m of the surface locally. A maximum age of the most recent faulting of 6430 ± 20 14C yr B.P. is constrained by a radiocarbon date from a deformed fluvial terrace. Holocene activity on the Leech River fault indicates that major active crustal faults of the Cascadia forearc extend further north than has been previously recognized, and suggests that the Leech River fault is likely an extension of the active fault network defined in western Washington. The complex geometry of the fault zone and sub-vertical orientation of individual strands are suggestive of a strike-slip fault within a transpressive regime. Further analyses of measurements of fault plane orientations and slip sense indicators will determine whether the sense of slip on the Leech River fault is dextral or sinistral. Determining the kinematics of this prominent forearc structure is a key component in elucidating the nature of forearc deformation in northern Cascadia in response to both the ongoing bending of the Olympic orocline and the overall dynamics of the Cascadia subduction zone.

  10. Austrobdella cairae n. sp., an Oioxenous Marine Leech (Clitellata: Piscicolidae) from the Banded Guitarfish, Zapteryx exasperata, in the Northeastern Pacific Ocean.

    PubMed

    Curran, Stephen S; Phillips, Anna J; Overstreet, Robin M; Benz, George W; Henningsen, Alan D

    2016-04-01

    A new marine leech is herein described from specimens infecting the external surfaces, including the mouth and cloaca, of the banded guitarfish, Zapteryx exasperate, captured in the Gulf of California and eastern Pacific Ocean off the coast of San Diego, California. The leech is assigned to Austrobdella by possessing continuous contractile coelomic channels that lie outside the somatic musculature along the lateral edges of the urosome (marginal lacunae), clitellar gland cells densely packed in the urosome, 5 pairs of testisacs, and 6-annulate mid-body somites. The new leech is distinguished from its 6 congeners on the basis of body size (maximum 10 mm long) and shape (sub-cylindrical trachelosome distinctly demarcated from wider urosome that is ventrally flattened, convex dorsally, and narrowing toward caudal sucker that is narrow, 20-25% of maximum body width), number of eyespots (2 pairs), shape and arrangement of the ovisacs (pyriform and limited to somites XII/XIII), and characteristics of the midgut (1 pair of mycetomes, 6 pairs of simple thin-walled crop ceca, ventral postceca wanting, and 2 pairs of dendritic diverticula emerging from anterior portion of thick-walled intestine). The new species occurs in the northeastern Pacific Ocean on a benthic elasmobranch. Examination of host specificity for each Austrobdella species using the quantitative Index of Phylogenetic Host Specificity revealed that the new species is 1 of 4 oioxenous specialists in the genus, and the remaining 3 congeners are relative generalists herein classified as euryxenous. This is the first time host specificity for members of the Piscicolidae has been quantitatively assessed. The analysis suggests that associations between marine leeches belonging in Austrobdella and their vertebrate hosts are driven by ecological influences rather than host taxonomic placement.

  11. Mechanism of ammonia excretion in the freshwater leech Nephelopsis obscura: characterization of a primitive Rh protein and effects of high environmental ammonia.

    PubMed

    Quijada-Rodriguez, Alex R; Treberg, Jason R; Weihrauch, Dirk

    2015-09-15

    Remarkably little is known about nitrogenous excretion in freshwater invertebrates. In the current study, the nitrogen excretion mechanism in the carnivorous ribbon leech, Nephelopsis obscura, was investigated. Excretion experiments showed that the ribbon leech is ammonotelic, excreting 166.0 ± 8.6 nmol·grams fresh weight (gFW)(-1)·h(-1) ammonia and 14.7 ± 1.9 nmol·gFW(-1)·h(-1) urea. Exposure to high and low pH hampered and enhanced, respectively, ammonia excretion rates, indicating an acid-linked ammonia trapping mechanism across the skin epithelia. Accordingly, compared with body tissues, the skin exhibited elevated mRNA expression levels of a newly identified Rhesus protein and at least in tendency the Na(+)/K(+)-ATPase. Pharmacological experiments and enzyme assays suggested an ammonia excretion mechanism that involves the V-ATPase, Na(+)/K(+)-ATPase, and carbonic anhydrase, but not necessarily a functional microtubule system. Most importantly, functional expression studies of the identified Rh protein cloned from leech skin tissue revealed an ammonia transport capability of this protein when expressed in yeast. The leech Rh-ammonia transporter (NoRhp) is a member of the primitive Rh protein family, which is a sister group to the common ancestor of vertebrate ammonia-transporting Rh proteins. Exposure to high environmental ammonia (HEA) caused a new adjustment of body ammonia, accompanied with a decrease in NoRhp and Na(+)/K(+)-ATPase mRNA levels, but unaltered ammonia excretion rates. To our knowledge, this is only the second comprehensive study regarding the ammonia excretion mechanisms in a freshwater invertebrate, but our results show that basic processes of ammonia excretion appear to also be comparable to those found in freshwater fish, suggesting an early evolution of ionoregulatory mechanisms in freshwater organisms.

  12. Field evaluation of deet, Repel Care, and three plant based essential oil repellents against mosquitoes, black flies (Diptera: Simuliidae) and land leeches (Arhynchobdellida: Haemadipsidae) in Thailand.

    PubMed

    Tawatsin, Apiwat; Thavara, Usavadee; Chansang, Uruyakorn; Chavalittumrong, Pranee; Boonruad, Thidarat; Wongsinkongman, Prapai; Bansidhi, Jaree; Mulla, Mir S

    2006-06-01

    Diethyl methyl benzamide, or deet, a commercial plant-based repellent (Repel Care), and essential ils from 3 species of plants (finger root rhizomes, guava leaves, and turmeric rhizomes), steam distillated and formulated as insect repellents, were evaluated in the field on human volunteers against hematophagous mosquitoes, black flies, and land leeches in Thailand. Field trials were conducted against wild mosquitoes in Bang Bua Thong District, Nonthaburi Province, and in the Thap Lan National Park Headquarters, Nadee District, Pranchinburi Province; anthroophilic black flies (Diptera: Simuliidae) at the Forestry Fire Control Station in Doi Inthanon National Park, Chomthong district, Chiang Mai Province; and land leeches (Arhynchobdellida: Haemadipsidae) in the Khao Yai National Park, Pak Chong District, Nakhon Ratchasima Province. The 3 experimental plant-based essential oil formulations as well as Repel Care and deet provided complete protection from mosquito landing and biting for up to 9 h (duration of the experiment). Similar results were obtained with the 5 products against black flies, providing 100% protection for 9 h but 96-82% protection after 10 and 11 h posttreatment. The 5 repellent products also provided 100% protection against land leeches for at least 8 h. Thi is the 1st report of repellency of plant-based repellents against black flies and land leeches in Thailand. The identification and availability of inexpensive sources of plant-based oils, i.e., finger root rhizomes, guava leaves, and turmeric rhizomes providing long-lasting repellency against blood-sucking organisms are promising leads into commercial production of relatively safe and effective repellents.

  13. Mg2+-malate co-transport, a mechanism for Na+-independent Mg2+ transport in neurons of the leech Hirudo medicinalis.

    PubMed

    Günzel, Dorothee; Hintz, Karin; Durry, Simone; Schlue, Wolf-Rüdiger

    2005-07-01

    Mg2+-extrusion from Mg2+-loaded neurons of the leech, Hirudo medicinalis, is mediated mainly by Na+/Mg2+ antiport. However, in a number of leech neurons, Mg2+ is extruded in the nominal absence of extracellular Na+, indicating the existence of an additional, Na+-independent Mg2+ transport mechanism. This mechanism was investigated using electrophysiological and microfluorimetrical techniques. The rate of Na+-independent Mg2+ extrusion from Mg2+-loaded leech neurons was found to be independent of extracellular Ca2+, K+, NO3-, HCO3-, SO4(2-), HPO4(2-), and of intra- and extracellular pH. Na+-independent Mg2+ extrusion was not inhibited by 4,4'-diisothiocyanatostilbene-2,2'-disulfonic acid (DIDS), furosemide, ouabain, vanadate, iodoacetate, 4-amino-hippurate, or alpha-cyano-4-hydroxycinnamate and was not influenced by changes in the membrane potential in voltage-clamp experiments. Na+-independent Mg2+ extrusion was, however, inhibited by the application of 2 mM probenecid, a blocker of organic anion transporters, suggesting that Mg2+ might be co-transported with organic anions. Extracellularly, of all organic anions tested (malate, citrate, lactate, alpha-ketoglutarate, and 4-amino-hippurate) only high, but physiological, concentrations of malate (30 mM) had a significant inhibitory effect on Na+-independent Mg2+ extrusion. Intracellularly, iontophoretically injected malate, citrate, or fura-2, but not Cl-, alpha-ketoglutarate, glutamate, succinate, or urate, were stimulating Na+-independent Mg2+ extrusion from those neurons that initially did not extrude Mg2+ in Na+-free solutions. Our data indicate that Mg2+ is co-transported with organic anions, preferably with malate, the predominant extracellular anion in the leech. The proposed model implies that, under experimental conditions, malate drives Mg2+ extrusion, whereas under physiological conditions, malate is actively taken up, driven by Mg2+, so that malate can be metabolized.

  14. The type II secretion system is essential for erythrocyte lysis and gut colonization by the leech digestive tract symbiont Aeromonas veronii.

    PubMed

    Maltz, Michele; Graf, Joerg

    2011-01-01

    Hemolysin and the type II secretion system (T2SS) have been shown to be important for virulence in many pathogens, but very few studies have shown their importance in beneficial microbes. Here, we investigated the importance of the type II secretion pathway in the beneficial digestive-tract association of Aeromonas veronii and the medicinal leech Hirudo verbana and revealed a critical role for the hemolysis of erythrocytes. A mutant with a miniTn5 insertion in exeM, which is involved in forming the inner membrane platform in the T2SS, was isolated by screening mutants for loss of hemolysis on blood agar plates. A hemolysis assay was used to quantify the mutant's deficiency in lysing sheep erythrocytes and revealed a 99.9% decrease compared to the parent strain. The importance of the T2SS in the colonization of the symbiotic host was assessed. Colonization assays revealed that the T2SS is critical for initial colonization of the leech gut. The defect was tied to the loss of hemolysin production by performing a colonization assay with blood containing lysed erythrocytes. This restored the colonization defect in the mutant. Complementation of the mutant using the promoter region and exeMN revealed that the T2SS is responsible for secreting hemolysin into the extracellular space and that both the T2SS and hemolysin export by the T2SS are critical for initial establishment of A. veronii in the leech gut.

  15. Phoretic interaction between the kangaroo leech Marsupiobdella africana (Hirudinea: Glossiphoniidae) and the cape river crab Potamonautes perlatus (Decapoda: Potamonautidae)☆

    PubMed Central

    Badets, Mathieu; Preez, Louis Du

    2013-01-01

    The South African leech Marsupiobdella africana is a temporary ectoparasite of the amphibian Xenopus laevis, has a phoretic association with a freshwater crab Potamonautes perlatus, and exhibits advanced parental care by incubating its offspring in a brood pouch. Because phoretic associations are usually regarded to favor the phoront’s dispersion, its occurrence within the biology of a parasitic species reflects an intimate context of interactions. In addition to phoresy, attachment to the crab may confer other advantages pertaining to offspring development and predator avoidance, dispersion and the parasitic life cycle. Two ponds where amphibian and crab hosts co-occur were sampled twice a month for a period of 1 year. The population dynamics of the leeches and their use of specific microhabitats as attachment sites on the crabs were also investigated. Results indicate a direct relationship between intra-specific variation in the sex ratio among captured crab hosts and the number of leeches recruited over time. The attachments to specific microhabitats on the hard surfaces of the host suggest a proximal proximate anti-predatory strategy. Finally, the importance of oxygen accessibility for the offspring development has been investigated experimentally. Results revealed a remarkable network of interactions linking all partners of this system raising the question as to whether the crabs merely act as a vehicle or play a role within the parasitic life cycle. PMID:24918071

  16. Genetic differentiation between two species of the medicinal leech, Hirudo medicinalis and the neglected H. verbana, based on random-amplified polymorphic DNA.

    PubMed

    Trontelj, Peter; Sotler, Marusa; Verovnik, Rudi

    2004-09-01

    The medicinal leech is one of the few parasitic invertebrates widely used in medicine and as a scientific model object. Because of a dramatic decline in its natural populations, it is subject to considerable conservation effort. Despite all attention, there is confusion regarding the taxonomic status of different morphological forms. The prevailing view is that all varieties of medicinal leech in Europe represent the same species, Hirudo medicinalis. However, the present study based on RAPD molecular markers demonstrates that a second European taxon, H. verbana, forms a distinct species. Phenetic clustering and principal coordinate analysis of eight populations revealed the same basic structure, reflecting taxonomic rather than geographic subdivision. Variation between species explained 60% of the total molecular variance (phiCT=0.60, P<0.001). Both taxa displayed a significant number of specific RAPD markers. Conversely, no specific fragment supporting the geographic association of both taxa was found. Since the stronghold of commercially exploited medicinal leech populations in southeastern Europe and Turkey belongs to H. verbana, most medicinal and scientific applications probably use this species, not H. medicinalis. Appropriate taxonomic correction of international conservation conventions and legislation is essential.

  17. Asymmetrization of first cleavage by transient disassembly of one spindle pole aster in the leech Helobdella robusta.

    PubMed

    Ren, Xiaoyun; Weisblat, David A

    2006-04-01

    Unequal first cleavage is characteristic of a diverse group of protostome animals. In the nematode Caenorhabditis elegans, unequal first cleavage is achieved through the interaction of an apparently symmetric mitotic spindle apparatus with a clearly polarized cell cortex. In the clitellate annelid Tubifex tubifex, by contrast, the spindle is monastral and contains only one gamma-tubulin-reactive centrosome; this monastral spindle is inherently asymmetric throughout mitosis. Here, we have used immunostaining for beta- and gamma-tubulin to follow spindle dynamics during the unequal first cleavage in another clitellate annelid, the leech Helobdella robusta. We find that the mitotic spindle is diastral and symmetric through early metaphase, then becomes asymmetric following the transient down-regulation of one centrosome, as judged by gamma-tubulin immunofluorescence. Low levels of drugs that affect microtubule dynamics can symmetrize the first cleavage without affecting the gamma-tubulin dynamics. Our results provide a striking example of the evolvability of cellular mechanisms underlying an unambiguously homologous developmental process.

  18. D quadrant specification in the leech Helobdella: actomyosin contractility controls the unequal cleavage of the CD blastomere

    PubMed Central

    Lyons, Deirdre C.; Weisblat, David A.

    2009-01-01

    The unequal division of the CD blastomere at second cleavage is critical in establishing the second embryonic axis in the leech Helobdella, as in other unequally cleaving spiralians. When CD divides, the larger D and smaller C blastomeres arise invariantly on the left and right sides of the embryo, respectively. Here we show that stereotyped cellular dynamics, including the formation of an intercellular blastocoel, culminate in a morphological left-right asymmetry in the 2-cell embryo, which precedes cytokinesis and predicts the chirality of the second cleavage. In contrast to the unequal first cleavage, the unequal second cleavage does not result from down-regulation of one centrosome, nor from an asymmetry within the spindle itself. Instead, the unequal cleavage of the CD cell entails a symmetric mitotic apparatus moving and anisotropically growing rightward in an actomyosin-dependent process. Our data reveal that mechanisms controlling the establishment of the D quadrant differ fundamentally even among the monophyletic clitellate annelids. Thus, while the homologous spiral cleavage pattern is highly conserved in this clade, it has diverged significantly at the level of cell biological mechanisms. This combination of operational conservation and mechanistic divergence begins to explain how the spiral cleavage program has remained so refractory to change while, paradoxically, accommodating numerous modifications throughout evolution. PMID:19607823

  19. Ionotropic and metabotropic activation of a neuronal chloride channel by serotonin and dopamine in the leech Hirudo medicinalis

    PubMed Central

    Ali, Declan W; Catarsi, Stefano; Drapeau, Pierre

    1998-01-01

    Cl− channels on the pressure-sensitive (P) neuron in the leech are directly activated by synaptic release of serotonin (5-HT) and are indirectly stimulated by the cAMP second messenger pathway, suggesting an unusual dual regulation of the channels. We have investigated the mode of action of 5-HT and dopamine (DA) on a Cl− channel in adult P cells in culture by recording from cell-attached patches.5-HT increased Cl− channel activity only when included in the recording pipette and not when applied in the bath.Pipette or, more effectively, bath application of DA led to an increase in Cl− channel activity. This effect was blocked by the potent and specific dopaminergic (DA1) receptor blocker, SCH-23390.The stimulation by DA, but not by 5-HT, was also blocked by the cAMP-dependent protein kinase A (PKA) inhibitor Rp-cAMP and was mimicked by the membrane-permeant cAMP analogue dibutyryl cAMP (db-cAMP).Our results show that 5-HT directly gates a Cl− channel that is also activated by DA via the cAMP pathway. This study demonstrates that a ligand-gated channel can be independently operated by another transmitter acting via a second messenger pathway. PMID:9547394

  20. Methylene blue blocks cGMP production and disrupts directed migration of microglia to nerve lesions in the leech CNS.

    PubMed

    Duan, Yuanli; Haugabook, Sharie J; Sahley, Christie L; Muller, Kenneth J

    2003-11-01

    Migration and accumulation of microglial cells at sites of injury are important for nerve repair. Recent studies on the leech central nervous system (CNS), in which synapse regeneration is successful, have shown that nitric oxide (NO) generated immediately after injury by endothelial nitric oxide synthase (eNOS) stops migrating microglia at the lesion. The present study obtained results indicating that NO may act earlier, on microglia migration, and aimed to determine mechanisms underlying NO's effects. Injury induced cGMP immunoreactivity at the lesion in a pattern similar to that of eNOS activity, immunoreactivity, and microglial cell accumulation, which were all focused there. The soluble guanylate cyclase (sGC) inhibitor methylene blue (MB) at 60 microM abolished cGMP immunoreactivity at lesions and blocked microglial cell migration and accumulation without interfering with axon conduction. Time-lapse video microscopy of microglia in living nerve cords showed MB did not reduce cell movement but reduced directed movement, with significantly more cells moving away from the lesion or reversing direction and fewer cells moving toward the lesion. The results indicate a new role for NO, directing the microglial cell migration as well as stopping it, and show that NO's action may be mediated by cGMP. Copyright 2003 Wiley Periodicals, Inc.

  1. Cathepsin L and cystatin B gene expression discriminates immune cœlomic cells in the leech Theromyzon tessulatum

    PubMed Central

    Lefebvre, Christophe; Vandenbulcke, Franck; Bocquet, Béatrice; Tasiemski, Aurélie; Desmons, Annie; Verstraete, Mathilde; Salzet, Michel; Cocquerelle, Claude

    2009-01-01

    Previous studies evidenced that cystatin B-like gene is specifically expressed and induced in large circulating cœlomic cells following bacterial challenge in the leech Theromyzon tessulatum. In order to understand the role of that cysteine proteinase inhibitor during immune response, we investigated the existence of members of cathepsin family. We cloned a cathepsin L-like gene and studied its tissue distribution. Immunohistochemical studies using anti-cathepsin L and anti-cystatin B antibodies and ultrastructural results demonstrated the presence of three distinct cœlomic cell populations, (1) the chloragocytes which were initially defined as large cœlomocytes, (2) the granular amœbocytes, and (3) small cœlomic cells. Among those cells, while chloragocytes contain cystatin B and cathepsin L, granular amœbocytes do only contain cathepsin L and third cell population contains neither cathepsin nor inhibitor. Finally, results evidenced that cathepsin L immunopositive granular amœbocytes are chemoattracted to the site of injury and phagocyte bacteria. PMID:18177937

  2. A dye mixture (Neurobiotin and Alexa 488) reveals extensive dye-coupling among neurons in leeches; physiology confirms the connections.

    PubMed

    Fan, Ruey-Jane; Marin-Burgin, Antonia; French, Kathleen A; Otto Friesen, W

    2005-12-01

    Although the neuronal circuits that generate leech movements have been studied for over 30 years, the list of interneurons (INs) in these circuits remains incomplete. Previous studies showed that some motor neurons (MNs) are electrically coupled to swim-related INs, e.g., rectifying junctions connect IN 28 to MN DI-1 (dorsal inhibitor), so we searched for additional neurons in these behavioral circuits by co-injecting Neurobiotin and Alexa Fluor 488 into segmental MNs DI-1, VI-2, DE-3 and VE-4. The high molecular weight Alexa dye is confined to the injected cell, whereas the smaller Neurobiotin molecules diffuse through gap junctions to reveal electrical coupling. We found that MNs were each dye-coupled to approximately 25 neurons, about half of which are likely to be INs. We also found that (1) dye-coupling was reliably correlated with physiologically confirmed electrical connections, (2) dye-coupling is unidirectional between MNs that are linked by rectifying connections, and (3) there are novel electrical connections between excitatory and inhibitory MNs, e.g. between excitatory MN VE-4 and inhibitory MN DI-1. The INs found in this study provide a pool of novel candidate neurons for future studies of behavioral circuits, including those underlying swimming, crawling, shortening, and bending movements.

  3. Infection, specificity and host manipulation of Australapatemon sp. (Trematoda, Strigeidae) in two sympatric species of leeches (Hirudinea).

    PubMed

    Karvonen, Anssi; Faltýnková, Anna; Choo, Jocelyn Mah; Valtonen, E Tellervo

    2017-09-01

    Factors that drive parasite specificity and differences in infection dynamics among alternative host species are important for ecology and evolution of host-parasite interactions, but still often poorly known in natural systems. Here, we investigated spatiotemporal dynamics of infection, host susceptibility and parasite-induced changes in host phenotype in a rarely explored host-parasite system, the Australapatemon sp. trematode infecting two sympatric species of freshwater leeches, Erpobdella octoculata and Helobdella stagnalis. We show significant variation in infection abundance between the host species in both space and time. Using experimental infections, we also show that most of this variation likely comes from interspecific differences in exposure rather than susceptibility. Moreover, we demonstrate that the hiding behaviour of E. octoculata, but not that of H. stagnalis, was impaired by the infection irrespective of the parasite abundance. This may increase susceptibility of E. octoculata to predation by the final avian host. We conclude that differences in patterns of infection and in behavioural alterations among alternative sympatric host species may arise in narrow spatial scales, which emphasises the importance of local infection and transmission dynamics for parasite life cycles.

  4. NMR structural characterization and computational predictions of the major intermediate in oxidative folding of leech carboxypeptidase inhibitor.

    PubMed

    Arolas, Joan L; D'Silva, Loyola; Popowicz, Grzegorz M; Aviles, Francesc X; Holak, Tad A; Ventura, Salvador

    2005-08-01

    The III-A intermediate constitutes the major rate-determining step in the oxidative folding of leech carboxypeptidase inhibitor (LCI). In this work, III-A has been directly purified from the folding reaction and structurally characterized by NMR spectroscopy. This species, containing three native disulfides, displays a highly native-like structure; however, it lacks some secondary structure elements, making it more flexible than native LCI. III-A represents a structurally determined example of a disulfide-insecure intermediate; direct oxidation of this species to the fully native protein seems to be restricted by the burial of its two free cysteine residues inside a native-like structure. We also show that theoretical approaches based on topological constraints predict with good accuracy the presence of this folding intermediate. Overall, the derived results suggest that, as it occurs with non-disulfide bonded proteins, native-like interactions between segments of secondary structure rather than the crosslinking of disulfide bonds direct the folding of LCI.

  5. Enhancing the secretory yields of leech carboxypeptidase inhibitor in Escherichia coli: influence of trigger factor and signal recognition particle.

    PubMed

    Puertas, Juan-Miguel; Nannenga, Brent L; Dornfeld, Kevin T; Betton, Jean-Michel; Baneyx, François

    2010-11-01

    The signal recognition particle (SRP) dependent secretion pathway is as an attractive alternative to Sec-dependent export for the production of disulfide-bonded and/or fast-folding recombinant proteins in the Escherichia coli periplasm. SRP, which shares a ribosomal attachment site with the molecular chaperone trigger factor (TF), recognizes highly hydrophobic signal sequence as they emerge from the ribosome and delivers ribosome nascent chain complexes to FtsY for subsequent cotranslational translocation of target proteins across the SecYEG pore. However, like in the case of Sec-dependent export, secretory yields can be limited by the accumulation of precursor proteins in the cytoplasm. Using leech carboxypeptidase inhibitor (LCI) fused to the SRP-dependent DsbA signal sequence as a model system, we show that a null mutation in the gene encoding TF (Deltatig) or SRP co-expression reduce pre-LCI accumulation by half, and that quantitative export can be achieved by combining the two strategies. Interestingly, enhanced precursor processing did not alter periplasmic LCI levels but increased the amount of protein excreted in the growth medium. All mature LCI was nearly fully active and an 80% increase in productivity was achieved in Deltatig cells alone due to their faster growth. Our results show that competition between SRP and TF can interfere with efficient export of recombinant proteins targeted to the SRP pathway and establish TF-deficient strains and SRP co-expression as a simple solution to improve yields.

  6. Sensitization and dishabituation of swim induction in the leech Hirudo medicinalis: role of serotonin and cyclic AMP.

    PubMed

    Zaccardi, Maria Luisa; Traina, Giovanna; Cataldo, Enrico; Brunelli, Marcello

    2004-08-31

    In this paper the role of serotonin (5HT) and cyclic AMP (cAMP) in sensitization and dishabituation of swim induction (SI) has been investigated in the leech Hirudo medicinalis. Electrical stimulation of the body wall evokes swimming activity with a constant latency. In animals with a disconnection between head ganglion and segmental ganglia, repetitive stimulation induces habituation of swimming whereas brushing on the dorsal skin provokes sensitization of a naïve response or dishabituation of a previously habituated response. Our findings indicate that 5HT is the neurotransmitter underlying both sensitization and dishabituation of SI. Injection of the 5HT receptor blocking agent methysergide impaires the onset of sensitization and dishabituation induced by brushing. Moreover, injection of 5HT mimics these forms of nonassociative learning, whereas injection of dopamine does not. Finally, the effects of 5HT are mediated by cAMP: (1) after injections of specific adenylate cyclase inhibitors such as MDL 12.330A or SQ22536, brushing becomes ineffective in facilitating the SI in either non-habituated or habituated animals. (2) 8Br-cAMP application mimics both sensitization and dishabituation of SI.

  7. Activity-dependent suppression of spontaneous spike generation in the Retzius neurons of the leech Hirudo medicinalis L.

    PubMed

    Rose, Tobias; Gras, Heribert; Hörner, Michael

    2006-12-01

    We report on factors affecting the spontaneous firing pattern of the identified serotonin-containing Retzius neurons of the medicinal leech. Increased firing activity induced by intracellular current injection is followed by a 'post-stimulus-depression' (PSD) without spiking for up to 23 s. PSD duration depends both on the duration and the amplitude of the injected current and correlates inversely with the spontaneous spiking activity. In contrast to serotonin-containing neurons in mammals, serotonin release from the Retzius cells presumably does not mediate the observed spike suppression in a self-inhibitory manner since robust PSD persists after synaptic isolation. Moreover, single additional spikes elicited at specific delays after spontaneously occurring action potentials are sufficient to significantly alter the firing pattern. Since sub-threshold current injections do not affect the ongoing spiking pattern and PSD persists in synaptically isolated preparations our data suggest that PSD reflects an endogenous and 'spike-dependent' mechanism controlling the spiking activity of Retzius cells in a use-dependent way.

  8. A 3-SYNAPSE POSITIVE FEEDBACK LOOP REGULATES THE EXCITABILITY OF AN INTERNEURON CRITICAL FOR SENSITIZATION IN THE LEECH

    PubMed Central

    Crisp, Kevin M.; Muller, Kenneth J.

    2007-01-01

    Sensitization of reflexive shortening in the leech has been linked to serotonin (5-HT)-induced changes in the excitability of a single interneuron, the S cell. This neuron is necessary for sensitization and complete dishabituation of reflexive shortening, during which it contributes to the sensory-motor reflex. The S cell does not contain 5-HT, which is released primarily from the Retzius (R) cells, whose firing enhances S-cell excitability. Here we show that the S cell excites the R cells, mainly via a fast disynaptic pathway in which the first synapse is the electrical junction between the S cell and the coupling interneurons, and the second synapse is a glutamatergic synapse of the coupling interneurons onto the R cells. The S cell-triggered excitatory postsynaptic potential in the R cell diminishes and nearly disappears in elevated concentrations of divalent cations because the coupling interneurons become inexcitable under these conditions. Serotonin released from the R cells feeds back upon the S cell and increases its excitability by activating a 5-HT7-like receptor; 5-methoxytryptamine (5-MeOT; 10 μM) mimics the effects of 5-HT on S cell excitability, and effects of both 5-HT and 5-MeOT are blocked by pimozide (10 μM) and SB-269970 (5 μM). This feedback loop may be critical for the full expression of sensitization of reflexive shortening. PMID:16571760

  9. Ultrastructural study of spermatogenesis and sperm in the African medicinal leech Hirudo troctina Johnson, 1816 (Annelida, Hirudinida).

    PubMed

    Ben Ahmed, Raja; Tekaya, Saïda; Urbisz, Anna Z; Świątek, Piotr

    2015-06-01

    This paper presents the process of spermatogenesis in the leech Hirudo troctina Johnson, 1816 using light, fluorescent and transmission electron microscopy. At the onset of spermatogenesis in testes, the pear-shaped spermatogonia divide mitotically without full cytokinesis and as a result isogenic groups are formed (clusters, clones) with 2, 4, 8, 16, 32, 64, 128 spermatogonia and, finally, 256 primary spermatocytes occur. The final meiotic divisions of spermatocytes give rise to clones with 1024 spermatids. There are hundreds of developing germ-line clones in each testis. In each clone, the male germ cells divide in full synchrony and they are in the same phase of spermatogenesis. During complex spermiogenesis each spermatid becomes a filiform spermatozoon with a helicoid nucleus, which is characterized by the presence of a long acrosome with two regions - anterior and posterior, which are followed by a helicoid nucleus, a midpiece with only one mitochondrion and a long flagellum. Our results were compared to those on other clitellate annelids that have been studied to date, especially to sperm formation in Hirudo medicinalis Linnaeus, 1785. Only minor differences were found in the length and the diameter of different organelles and the number of spermatids in germ-line clones.

  10. Effects of millimeter wave irradiation and equivalent thermal heating on the activity of individual neurons in the leech ganglion

    PubMed Central

    Romanenko, Sergii; Siegel, Peter H.; Wagenaar, Daniel A.

    2014-01-01

    Many of today's radiofrequency-emitting devices in telecommunication, telemedicine, transportation safety, and security/military applications use the millimeter wave (MMW) band (30–300 GHz). To evaluate the biological safety and possible applications of this radiofrequency band for neuroscience and neurology, we have investigated the physiological effects of low-intensity 60-GHz electromagnetic irradiation on individual neurons in the leech midbody ganglia. We applied incident power densities of 1, 2, and 4 mW/cm2 to the whole ganglion for a period of 1 min while recording the action potential with a standard sharp electrode electrophysiology setup. For comparison, the recognized U.S. safe exposure limit is 1 mW/cm2 for 6 min. During the exposure to MMWs and gradual bath heating at a rate of 0.04°C/s (2.4°C/min), the ganglionic neurons exhibited similar dose-dependent hyperpolarization of the plasma membrane and decrease in the action potential amplitude. However, narrowing of the action potential half-width during MMW irradiation at 4 mW/cm2 was 5 times more pronounced compared with that during equivalent bath heating of 0.6°C. Even more dramatic difference in the effects of MMW irradiation and bath heating was noted in the firing rate, which was suppressed at all applied MMW power densities and increased in a dose-dependent manner during gradual bath heating. The mechanism of enhanced narrowing of action potentials and suppressed firing by MMW irradiation, compared with that by gradual bath heating, is hypothesized to involve specific coupling of MMW energy with the neuronal plasma membrane. PMID:25122711

  11. Evidence for the uptake of neuronally derived choline by glial cells in the leech central nervous system.

    PubMed Central

    Wuttke, W A; Pentreath, V W

    1990-01-01

    1. With ion-sensitive microelectrodes based on the Corning exchanger 477317, the accumulation of an unidentified interfering substance was monitored in leech neuropile glial cells but not in neurons after a 10-fold increase in extracellular K+ concentration. Evidence is presented which shows that this substance may be choline. 2. The accumulation of interfering ions was not observed in Ca2(+)-free saline and was substantially reduced in the presence of eserine (a blocker of acetylcholinesterase). 3. In neuropile (and also packet) glial cells, extracellularly applied choline (10(-4) M) caused a steady increase in ion signal. This increase was not affected by removal of extracellular calcium, by hemicholinium-3 (a blocker of high-affinity choline uptake) or eserine. Shortly after the removal of choline from the saline the increase in ion signal stopped and the ion signal then decreased slowly to its original level. 4. Extracellular acetylcholine (10(-4) M) caused a similar increase in intracellular ion signal of neuropile glial cells to that caused by choline. This increase was blocked by eserine. 5. Extracellular choline caused a comparatively small increase in ion signal of Retzius neurones which was blocked by hemicholinium-3. In pressure neurones, choline or hemicholinium-3 had no effect on intracellular ion signal. 6. Autoradiographic analysis of [3H]choline uptake showed that most of the choline was taken up by glial cells in a time- and dose-dependent manner. Small but significant amounts of choline were taken up by neurones and connective tissue. 7. It is concluded that the neuropile and packet glial cells possess an effective choline uptake system which is activated by exogenous choline but also by choline that stems from enzymatic inactivation of acetylcholine released by neurones. Images Fig. 11 PMID:2324991

  12. Effects of millimeter wave irradiation and equivalent thermal heating on the activity of individual neurons in the leech ganglion.

    PubMed

    Romanenko, Sergii; Siegel, Peter H; Wagenaar, Daniel A; Pikov, Victor

    2014-11-15

    Many of today's radiofrequency-emitting devices in telecommunication, telemedicine, transportation safety, and security/military applications use the millimeter wave (MMW) band (30-300 GHz). To evaluate the biological safety and possible applications of this radiofrequency band for neuroscience and neurology, we have investigated the physiological effects of low-intensity 60-GHz electromagnetic irradiation on individual neurons in the leech midbody ganglia. We applied incident power densities of 1, 2, and 4 mW/cm(2) to the whole ganglion for a period of 1 min while recording the action potential with a standard sharp electrode electrophysiology setup. For comparison, the recognized U.S. safe exposure limit is 1 mW/cm(2) for 6 min. During the exposure to MMWs and gradual bath heating at a rate of 0.04°C/s (2.4°C/min), the ganglionic neurons exhibited similar dose-dependent hyperpolarization of the plasma membrane and decrease in the action potential amplitude. However, narrowing of the action potential half-width during MMW irradiation at 4 mW/cm(2) was 5 times more pronounced compared with that during equivalent bath heating of 0.6°C. Even more dramatic difference in the effects of MMW irradiation and bath heating was noted in the firing rate, which was suppressed at all applied MMW power densities and increased in a dose-dependent manner during gradual bath heating. The mechanism of enhanced narrowing of action potentials and suppressed firing by MMW irradiation, compared with that by gradual bath heating, is hypothesized to involve specific coupling of MMW energy with the neuronal plasma membrane. Copyright © 2014 the American Physiological Society.

  13. Ingested Blood Contributes to the Specificity of the Symbiosis of Aeromonas veronii Biovar Sobria and Hirudo medicinalis, the Medicinal Leech

    PubMed Central

    Indergand, Stefan; Graf, Joerg

    2000-01-01

    Hirudo medicinalis, the medicinal leech, usually carries in its digestive tract a pure culture of Aeromonas veronii bv. sobria. Such specificity is unusual for digestive tracts that are normally colonized by a complex microbial consortium. Important questions for the symbiotic interaction and for the medical application after microvascular surgery are whether other bacteria can proliferate or at least persist in the digestive tract of H. medicinalis and what factors contribute to the reported specificity. Using a colonization assay, we were able to compare experimentally the ability of clinical isolates and of a symbiotic strain to colonize H. medicinalis. The symbiotic A. veronii bv. sobria strain proliferated well and persisted for at least 7 days inside the digestive tract. In contrast, the proliferation of Pseudomonas aeruginosa and Staphylococcus aureus was inhibited inside the animal compared to growth in the in vitro control, indicating that the ingested blood was modified within the digestive tract. However, both strains were able to persist in the digestive tract for at least 7 days. For an Escherichia coli strain, the viable counts decreased approximately 1,000-fold within 42 h. The decrease of viable E. coli could be prevented by interfering with the activation of the membrane-attack complex of the complement system that is present in blood. This suggests that the membrane-attack complex remained active inside H. medicinalis and prevented the proliferation of sensitive bacteria. Thus, antimicrobial properties of the ingested vertebrate blood contribute to the specificity of the A. veronii-H. medicinalis symbiosis, in addition to modifications of the blood inside the digestive tract of H. medicinalis. PMID:11055917

  14. Ingested blood contributes to the specificity of the symbiosis of Aeromonas veronii biovar sobria and Hirudo medicinalis, the medicinal leech.

    PubMed

    Indergand, S; Graf, J

    2000-11-01

    Hirudo medicinalis, the medicinal leech, usually carries in its digestive tract a pure culture of Aeromonas veronii bv. sobria. Such specificity is unusual for digestive tracts that are normally colonized by a complex microbial consortium. Important questions for the symbiotic interaction and for the medical application after microvascular surgery are whether other bacteria can proliferate or at least persist in the digestive tract of H. medicinalis and what factors contribute to the reported specificity. Using a colonization assay, we were able to compare experimentally the ability of clinical isolates and of a symbiotic strain to colonize H. medicinalis. The symbiotic A. veronii bv. sobria strain proliferated well and persisted for at least 7 days inside the digestive tract. In contrast, the proliferation of Pseudomonas aeruginosa and Staphylococcus aureus was inhibited inside the animal compared to growth in the in vitro control, indicating that the ingested blood was modified within the digestive tract. However, both strains were able to persist in the digestive tract for at least 7 days. For an Escherichia coli strain, the viable counts decreased approximately 1, 000-fold within 42 h. The decrease of viable E. coli could be prevented by interfering with the activation of the membrane-attack complex of the complement system that is present in blood. This suggests that the membrane-attack complex remained active inside H. medicinalis and prevented the proliferation of sensitive bacteria. Thus, antimicrobial properties of the ingested vertebrate blood contribute to the specificity of the A. veronii-H. medicinalis symbiosis, in addition to modifications of the blood inside the digestive tract of H. medicinalis.

  15. Solving a Bloody Mess: B-Vitamin Independent Metabolic Convergence among Gammaproteobacterial Obligate Endosymbionts from Blood-Feeding Arthropods and the Leech Haementeria officinalis

    PubMed Central

    Manzano-Marín, Alejandro; Oceguera-Figueroa, Alejandro; Latorre, Amparo; Jiménez-García, Luis F.; Moya, Andres

    2015-01-01

    Endosymbiosis is a common phenomenon in nature, especially between bacteria and insects, whose typically unbalanced diets are usually complemented by their obligate endosymbionts. While much interest and focus has been directed toward phloem-feeders like aphids and mealybugs, blood-feeders such as the Lone star tick (Amblyomma americanum), Glossina flies, and the human body louse (Pediculus humanus corporis) depend on obligate endosymbionts which complement their B-vitamin-deficient diets, and thus are required for growth and survival. Glossiphoniid leeches have also been found to harbor distinct endosymbionts housed in specialized organs. Here, we present the genome of the bacterial endosymbiont from Haementeria officinalis, first of a glossiphoniid leech. This as-yet-unnamed endosymbiont belongs to the Gammaproteobacteria, has a pleomorphic shape and is restricted to bacteriocytes. For this bacterial endosymbiont, we propose the name Candidatus Providencia siddallii. This symbiont possesses a highly reduced genome with high A+T content and a reduced set of metabolic capabilities, all of which are common characteristics of ancient obligate endosymbionts of arthropods. Its genome has retained many pathways related to the biosynthesis of B-vitamins, pointing toward a role in supplementing the blood-restricted diet of its host. Through comparative genomics against the endosymbionts of A. americanum, Glossina flies, and P. humanus corporis, we were able to detect a high degree of metabolic convergence among these four very distantly related endosymbiotic bacteria. PMID:26454017

  16. Solving a Bloody Mess: B-Vitamin Independent Metabolic Convergence among Gammaproteobacterial Obligate Endosymbionts from Blood-Feeding Arthropods and the Leech Haementeria officinalis.

    PubMed

    Manzano-Marín, Alejandro; Oceguera-Figueroa, Alejandro; Latorre, Amparo; Jiménez-García, Luis F; Moya, Andres

    2015-10-09

    Endosymbiosis is a common phenomenon in nature, especially between bacteria and insects, whose typically unbalanced diets are usually complemented by their obligate endosymbionts. While much interest and focus has been directed toward phloem-feeders like aphids and mealybugs, blood-feeders such as the Lone star tick (Amblyomma americanum), Glossina flies, and the human body louse (Pediculus humanus corporis) depend on obligate endosymbionts which complement their B-vitamin-deficient diets, and thus are required for growth and survival. Glossiphoniid leeches have also been found to harbor distinct endosymbionts housed in specialized organs. Here, we present the genome of the bacterial endosymbiont from Haementeria officinalis, first of a glossiphoniid leech. This as-yet-unnamed endosymbiont belongs to the Gammaproteobacteria, has a pleomorphic shape and is restricted to bacteriocytes. For this bacterial endosymbiont, we propose the name Candidatus Providencia siddallii. This symbiont possesses a highly reduced genome with high A+T content and a reduced set of metabolic capabilities, all of which are common characteristics of ancient obligate endosymbionts of arthropods. Its genome has retained many pathways related to the biosynthesis of B-vitamins, pointing toward a role in supplementing the blood-restricted diet of its host. Through comparative genomics against the endosymbionts of A. americanum, Glossina flies, and P. humanus corporis, we were able to detect a high degree of metabolic convergence among these four very distantly related endosymbiotic bacteria.

  17. Transcription and protein synthesis inhibitors influence long-term effects of acetyl-l-carnitine on non-associative learning in the leech.

    PubMed

    Traina, Giovanna; Scuri, Rossana

    2015-01-01

    Acetyl-l-carnitine (ALC) is the principal acetyl ester of L-carnitine and it plays an essential role in intermediary metabolism. ALC affects several targets in the nervous system. Along this line of investigation, we analyzed the long-term effects of ALC on elementary nonassociative learning in the swimming induction model of the leech Hirudo medicinalis, in which nociceptive stimulation of the dorsal skin produces a more rapid swim response to a test stimulus (sensitization). In this simplified model a single ALC administration blocked the sensitizing effects of nociceptive stimulation in swim induction showing increasingly long lasting effects. Herein, we have analyzed the long-term effects of ALC on sensitization and dishabituation. Leeches were treated with inhibitors of either transcription or protein synthesis 30 min after the administration of ALC and, subsequently, subjected to noxious stimuli: the animals exhibited a sensitized swimming response 6 days after ALC treatment but not after 2 hours indicating that the long-term suppressive effects of ALC on sensitization/dishabituation needed mRNA and protein synthesis.

  18. Fine-scale fault structures revealed by earthquake relocations near the Leech River Fault, southern Vancouver Island

    NASA Astrophysics Data System (ADS)

    Li, G.; Liu, Y.; Regalla, C.; Morell, K. D.

    2016-12-01

    We present results of microseismic relocations that delineate potentially seismogenic structures along the Leech River Fault (LRF), southern Vancouver Island. The potential seismic activity of the LRF in particular has been the focus of many recent investigations, given its proximity to Victoria, the capitol of British Columbia. We investigate the seismic evidence of subsurface structures near the LRF by relocating 1528 earthquakes reported by Canadian National Seismograph Network (CNSN) catalog from 1985 to 2015 using the HypoDD [Waldhauser, F., 2001] method. This relocation procedure reveals two general northeast dipping structures representing the subsurface fractured deformation zone beneath the LRF and the San Juan Islands area, respectively. The subsurface structure underlying the LRF extends about 30 km along the LRF strike and has a dip of about 45o, which is generally consistent with fault dip inferred from seismic imaging [Green et al., 1987]. In addition, we use the k-mean++ clustering algorithm [Lloyd, 1982; Arthur et al., 2007] to select 5-subclusters for individual relocations in order to investigate possible finer faulting structures. The relocated sub-cluster seismicity, combined with earthquake focal mechanism solutions from previous studies, delineates the subsurface geometry of minor faults near the LRF. Relocation results also show the eastern segment is seismically active while the western segment lacks seismicity. We suggest two potential explanations for this marked variation in seismic behavior. First, seismicity might reflect variations in the regionals tress field [Balfour et al., 2011]. The orientation of the maximum horizontal compressive stress near the LRF is 10-30° clockwise from the strike of eastern segment, which would promote right lateral slip, but is nearly orthogonal to the strike of the western segment, which would inhibit slip. Second, foliations and mylonitic fabric are generally stronger, more developed, and exist over a

  19. THE LEECH EXOPLANET IMAGING SURVEY: CHARACTERIZATION OF THE COLDEST DIRECTLY IMAGED EXOPLANET, GJ 504 b, AND EVIDENCE FOR SUPERSTELLAR METALLICITY

    SciTech Connect

    Skemer, Andrew J.; Leisenring, Jarron; Bailey, Vanessa; Hinz, Philip; Defrére, Denis; Apai, Dániel; Close, Laird; Eisner, Josh; Morley, Caroline V.; Fortney, Jonathan; Zimmerman, Neil T.; Buenzli, Esther; Bonnefoy, Mickael; Biller, Beth; Brandner, Wolfgang; Skrutskie, Michael F.; Esposito, Simone; Crepp, Justin R.; De Rosa, Robert J.; Desidera, Silvano; and others

    2016-02-01

    As gas giant planets and brown dwarfs radiate away the residual heat from their formation, they cool through a spectral type transition from L to T, which encompasses the dissipation of cloud opacity and the appearance of strong methane absorption. While there are hundreds of known T-type brown dwarfs, the first generation of directly imaged exoplanets were all L type. Recently, Kuzuhara et al. announced the discovery of GJ 504 b, the first T dwarf exoplanet. GJ 504 b provides a unique opportunity to study the atmosphere of a new type of exoplanet with a ∼500 K temperature that bridges the gap between the first directly imaged planets (∼1000 K) and our own solar system's Jupiter (∼130 K). We observed GJ 504 b in three narrow L-band filters (3.71, 3.88, and 4.00 μm), spanning the red end of the broad methane fundamental absorption feature (3.3 μm) as part of the LBTI Exozodi Exoplanet Common Hunt (LEECH) exoplanet imaging survey. By comparing our new photometry and literature photometry with a grid of custom model atmospheres, we were able to fit GJ 504 b's unusual spectral energy distribution for the first time. We find that GJ 504 b is well fit by models with the following parameters: T{sub eff} = 544 ± 10 K, g < 600 m s{sup −2}, [M/H] = 0.60 ± 0.12, cloud opacity parameter of f{sub sed} = 2–5, R = 0.96 ± 0.07 R{sub Jup}, and log(L) = −6.13 ± 0.03 L{sub ⊙}, implying a hot start mass of 3–30 M{sub jup} for a conservative age range of 0.1–6.5 Gyr. Of particular interest, our model fits suggest that GJ 504 b has a superstellar metallicity. Since planet formation can create objects with nonstellar metallicities, while binary star formation cannot, this result suggests that GJ 504 b formed like a planet, not like a binary companion.

  20. The LEECH Exoplanet Imaging Survey: Characterization of the Coldest Directly Imaged Exoplanet, GJ 504 b, and Evidence for Superstellar Metallicity

    NASA Astrophysics Data System (ADS)

    Skemer, Andrew J.; Morley, Caroline V.; Zimmerman, Neil T.; Skrutskie, Michael F.; Leisenring, Jarron; Buenzli, Esther; Bonnefoy, Mickael; Bailey, Vanessa; Hinz, Philip; Defrére, Denis; Esposito, Simone; Apai, Dániel; Biller, Beth; Brandner, Wolfgang; Close, Laird; Crepp, Justin R.; De Rosa, Robert J.; Desidera, Silvano; Eisner, Josh; Fortney, Jonathan; Freedman, Richard; Henning, Thomas; Hofmann, Karl-Heinz; Kopytova, Taisiya; Lupu, Roxana; Maire, Anne-Lise; Males, Jared R.; Marley, Mark; Morzinski, Katie; Oza, Apurva; Patience, Jenny; Rajan, Abhijith; Rieke, George; Schertl, Dieter; Schlieder, Joshua; Stone, Jordan; Su, Kate; Vaz, Amali; Visscher, Channon; Ward-Duong, Kimberly; Weigelt, Gerd; Woodward, Charles E.

    2016-02-01

    As gas giant planets and brown dwarfs radiate away the residual heat from their formation, they cool through a spectral type transition from L to T, which encompasses the dissipation of cloud opacity and the appearance of strong methane absorption. While there are hundreds of known T-type brown dwarfs, the first generation of directly imaged exoplanets were all L type. Recently, Kuzuhara et al. announced the discovery of GJ 504 b, the first T dwarf exoplanet. GJ 504 b provides a unique opportunity to study the atmosphere of a new type of exoplanet with a ˜500 K temperature that bridges the gap between the first directly imaged planets (˜1000 K) and our own solar system's Jupiter (˜130 K). We observed GJ 504 b in three narrow L-band filters (3.71, 3.88, and 4.00 μm), spanning the red end of the broad methane fundamental absorption feature (3.3 μm) as part of the LBTI Exozodi Exoplanet Common Hunt (LEECH) exoplanet imaging survey. By comparing our new photometry and literature photometry with a grid of custom model atmospheres, we were able to fit GJ 504 b's unusual spectral energy distribution for the first time. We find that GJ 504 b is well fit by models with the following parameters: Teff = 544 ± 10 K, g < 600 m s-2, [M/H] = 0.60 ± 0.12, cloud opacity parameter of fsed = 2-5, R = 0.96 ± 0.07 RJup, and log(L) = -6.13 ± 0.03 L⊙, implying a hot start mass of 3-30 Mjup for a conservative age range of 0.1-6.5 Gyr. Of particular interest, our model fits suggest that GJ 504 b has a superstellar metallicity. Since planet formation can create objects with nonstellar metallicities, while binary star formation cannot, this result suggests that GJ 504 b formed like a planet, not like a binary companion. The LBT is an international collaboration among institutions in the United States, Italy, and Germany. LBT Corporation partners are the University of Arizona on behalf of the Arizona university system; Istituto Nazionale di Astrophisica, Italy; LBT

  1. Activation of AMPA/kainate receptors but not acetylcholine receptors causes Mg2+ influx into Retzius neurones of the leech Hirudo medicinalis.

    PubMed

    Muller, Anja; Gunzel, Dorothee; Schlue, Wolf-Rudiger

    2003-12-01

    In Retzius neurones of the medicinal leech, Hirudo medicinalis, kainate activates ionotropic glutamate receptors classified as AMPA/kainate receptors. Activation of the AMPA/kainate receptor-coupled cation channels evokes a marked depolarization, intracellular acidification, and increases in the intracellular concentrations of Na+ ([Na+]i) and Ca2+. Qualitatively similar changes are observed upon the application of carbachol, an activator of acetylcholine receptor-coupled cation channels. Using multibarrelled ion-selective microelectrodes it was demonstrated that kainate, but not carbachol, caused additional increases in the intracellular free Mg2+ concentration ([Mg2+]i). Experiments were designed to investigate whether this kainate-induced [Mg2+]i increase was due to a direct Mg2+ influx through the AMPA/kainate receptor-coupled cation channels or a secondary effect due to the depolarization or the ionic changes. It was found that: (a) Similar [Mg2+]i increases were evoked by the application of glutamate or aspartate. (b) All kainate-induced effects were inhibited by the glutamatergic antagonist DNQX. (c) The magnitude of the [Mg2+]i increases depended on the extracellular Mg2+ concentration. (d) A reduction of the extracellular Ca2+ concentration increased kainate-induced [Mg2+]i increases, excluding possible Ca2+ interference at the Mg2+-selective microelectrode or at intracellular buffer sites. (e) Neither depolarizations evoked by the application of 30 mM K+, nor [Na+]i increases induced by the inhibition of the Na+/K+ ATPase caused comparable [Mg2+]i increases. (f) Inhibitors of voltage-dependent Ca2+ channels did not affect the kainate-induced [Mg2+]i increases. Moreover, previous experiments had already shown that intracellular acidification evoked by the application of 20 mM propionate did not cause changes in [Mg2+]i. The results indicate that kainate-induced [Mg2+]i increases in leech Retzius neurones are due to an influx of extracellular Mg2+ through

  2. Activation of AMPA/Kainate Receptors but Not Acetylcholine Receptors Causes Mg2+ Influx into Retzius Neurones of the Leech Hirudo medicinalis

    PubMed Central

    Müller, Anja; Günzel, Dorothee; Schlue, Wolf-Rüdiger

    2003-01-01

    In Retzius neurones of the medicinal leech, Hirudo medicinalis, kainate activates ionotropic glutamate receptors classified as AMPA/kainate receptors. Activation of the AMPA/kainate receptor–coupled cation channels evokes a marked depolarization, intracellular acidification, and increases in the intracellular concentrations of Na+ ([Na+]i) and Ca2+. Qualitatively similar changes are observed upon the application of carbachol, an activator of acetylcholine receptor-coupled cation channels. Using multibarrelled ion-selective microelectrodes it was demonstrated that kainate, but not carbachol, caused additional increases in the intracellular free Mg2+ concentration ([Mg2+]i). Experiments were designed to investigate whether this kainate-induced [Mg2+]i increase was due to a direct Mg2+ influx through the AMPA/kainate receptor–coupled cation channels or a secondary effect due to the depolarization or the ionic changes. It was found that: (a) Similar [Mg2+]i increases were evoked by the application of glutamate or aspartate. (b) All kainate-induced effects were inhibited by the glutamatergic antagonist DNQX. (c) The magnitude of the [Mg2+]i increases depended on the extracellular Mg2+ concentration. (d) A reduction of the extracellular Ca2+ concentration increased kainate-induced [Mg2+]i increases, excluding possible Ca2+ interference at the Mg2+-selective microelectrode or at intracellular buffer sites. (e) Neither depolarizations evoked by the application of 30 mM K+, nor [Na+]i increases induced by the inhibition of the Na+/K+ ATPase caused comparable [Mg2+]i increases. (f) Inhibitors of voltage-dependent Ca2+ channels did not affect the kainate-induced [Mg2+]i increases. Moreover, previous experiments had already shown that intracellular acidification evoked by the application of 20 mM propionate did not cause changes in [Mg2+]i. The results indicate that kainate-induced [Mg2+]i increases in leech Retzius neurones are due to an influx of extracellular Mg2

  3. Na(+)/K(+) pump interacts with the h-current to control bursting activity in central pattern generator neurons of leeches.

    PubMed

    Kueh, Daniel; Barnett, William H; Cymbalyuk, Gennady S; Calabrese, Ronald L

    2016-09-02

    The dynamics of different ionic currents shape the bursting activity of neurons and networks that control motor output. Despite being ubiquitous in all animal cells, the contribution of the Na(+)/K(+) pump current to such bursting activity has not been well studied. We used monensin, a Na(+)/H(+) antiporter, to examine the role of the pump on the bursting activity of oscillator heart interneurons in leeches. When we stimulated the pump with monensin, the period of these neurons decreased significantly, an effect that was prevented or reversed when the h-current was blocked by Cs(+). The decreased period could also occur if the pump was inhibited with strophanthidin or K(+)-free saline. Our monensin results were reproduced in model, which explains the pump's contributions to bursting activity based on Na(+) dynamics. Our results indicate that a dynamically oscillating pump current that interacts with the h-current can regulate the bursting activity of neurons and networks.

  4. Ca(2+)-dependent and Ca2+ release-dependent excitation in leech photoreceptors: evidence from a novel "inside-out" cell model.

    PubMed

    Walz, Bernd; Liebherr, Helga; Ukhanov, Kyrill

    2003-07-01

    We have developed a novel, electrophysiologically intact and light-sensitive "inside-out" cell model (IOCM) of microvillar photoreceptors of the leech Hirudo medicinalis. Light responses recorded from the IOCM with sharp microelectrodes are depolarizations with amplitudes of up to 50-60 mV. In darkness, graded elevations of the free Ca(2+) concentration in the "intracellular medium" (ICM) reversibly increase the conductance of the microvillar membrane leading to Ca(2+)-induced graded voltage changes up to approximately 50 mV. The threshold for Ca(2+)-induced voltage changes is approximately 0.06 microM, EC(50) is approximately 1.2 microM, and saturation occurs at approximately 20 microM free Ca(2+). Small Ca(2+) elevations (<0.6 microM) produce discrete waves of depolarization resembling quantum bumps. Stimulating IOCMs with short (20-ms) and long (5-s) light stimuli produces transient light responses (repolarization within ca. 200 ms) in an ICM containing only 10nM free Ca(2+). At 0.44 microM free Ca(2+) in the ICM, the microvillar membrane depolarizes by 10-20 mV and responses to 5-s light steps have an initial transient component and a plateau component, similar to responses in intact cells. Generation of the plateau component in IOCMs is suppressed by heparin and cyclopiazonic acid (CPA), agents that block inositol 1,4,5-trisphosphate (Ins(1,4,5)P(3))-induced Ca(2+) release from and Ca(2+) uptake into the endoplasmic reticulum (ER). These results indicate that there is a Ca(2+)-dependent conductance in the microvillar membrane and that the light-induced Ins(1,4,5)P(3)- and Ca(2+) release-mediated intracellular Ca(2+) elevation in leech photoreceptors contributes to the generation of the receptor potential, particularly the plateau component of responses to long steps of light.

  5. A large fragment approach to DNA synthesis: total synthesis of a gene for the protease inhibitor eglin c from the leech Hirudo medicinalis and its expression in E. coli.

    PubMed Central

    Rink, H; Liersch, M; Sieber, P; Meyer, F

    1984-01-01

    A DNA containing the coding sequence for the proteinase inhibitor protein, eglin c, from the leech Hirudo medicinalis has been obtained by enzymatic assembly of chemically synthesized DNA fragments. The synthetic gene consists of a 232 base-pair fragment containing initiation and termination codon signals with restriction enzyme recognition sites conveniently placed for cloning into a plasmid vector. Only six oligonucleotides from 34 to 61 bases in length, sharing pairwise stretches of complementary regions at their 3'-termini, were prepared by phosphotriester solid-phase synthesis. The oligomers were annealed pairwise and converted into double stranded DNA fragments by DNA polymerase I mediated repair synthesis. The fragments were assembled by ligation, and the synthetic gene was expressed in high yield in E. coli under the transcriptional control of the E. coli tryptophan promoter. The expression product was purified to homogeneity and was shown to have similar physicochemical and identical biological properties as the authentic protein isolated from the leech. Images PMID:6382168

  6. Evolution of clitellate phaosomes from rhabdomeric photoreceptor cells of polychaetes – a study in the leech Helobdella robusta (Annelida, Sedentaria, Clitellata)

    PubMed Central

    2013-01-01

    Introduction In Annelida two types of photoreceptor cells (PRCs) are regarded as generally present, rhabdomeric and ciliary PRCs. In certain taxa, however, an additional type of PRC may occur, the so called phaosomal PRC. Whereas the former two types of PRCs are always organized as an epithelium with their sensory processes projecting into an extracellular cavity formed by the PRCs and (pigmented) supportive cells, phaosomes are seemingly intracellular vacuoles housing the sensory processes. Phaosomal PRCs are the only type of PRC found in one major annelid group, Clitellata. Several hypotheses have been put forward explaining the evolutionary origin of the clitellate phaosomes. To elucidate the evolution of clitellate PRC and eyes the leech Helobdella robusta, for which a sequenced genome is available, was chosen. Results TEM observations showed that extraocular and ocular PRCs are structurally identical. Bioinformatic analyses revealed predictions for four opsin genes, three of which could be amplified. All belong to the rhabdomeric opsin family and phylogenetic analyses showed them in a derived position within annelid opsins. Gene expression studies showed two of them expressed in the eye and in the extraocular PRCs. Polychaete eye-typic key enzymes for ommochromme and pterin shading pigments synthesis are not expressed in leech eyes. Conclusions By comparative gene-expression studies we herein provide strong evidence that the phaosomal PRCs typical of Clitellata are derived from the rhabdomeric PRCs characteristic for polychaete adult eyes. Thus, they represent a highly derived type of PRC that evolved in the stem lineage of Clitellata rather than another, primitive type of PRC in Metazoa. Evolution of these PRCs in Clitellata is related to a loss of the primary eyes and most of their photoreceptive elements except for the rhabdomeric PRCs. Most likely this happened while changing to an endobenthic mode of life. This hypothesis of PRC evolution is in accordance

  7. Description of a new leech species of Helobdella (Clitellata: Glossiphoniidae) from Mexico with a review of Mexican congeners and a taxonomic key.

    PubMed

    Salas-Montiel, Ricardo; Phillips, Anna J; De Leon, Gerardo Perez-Ponce; Oceguera-Figueroa, Alejandro

    2014-12-19

    To date, six species of the leech genus Helobdella have been recorded from Mexico: Helobdella atli, Helobdella elongata, Helobdella octatestisaca, Helobdella socimulcensis, Helobdella virginiae and Helobdella temiscoensis n. sp. This new species is characterized by a lanceolate body, the presence of a nuchal scute, uniform brown pigment on both dorsal and ventral surfaces, the absence of papillae, well-separated eyespots, six pairs of testisacs and five pairs of crop caeca, the last of which forms posterior caeca. In addition, we provide new geographic records for Helobdella species from Mexico resulting from our own collections, vouchers deposited at the Colección Nacional de Helmintos from the Instituto de Biología, UNAM, Mexico and vouchers at the Invertebrate Zoology Collection of the Smithsonian's National Museum of Natural History (USNM) Washington D.C., USA. We present a comprehensive review of Mexican Helobdella species, including the new species, with notes on the characteristic morphology and geographic distribution of each species with 91 new records from 20 states. In addition, we provide a taxonomic key for the identification of the Mexican species.

  8. Comparison of the effects of millimeter wave irradiation, general bath heating, and localized heating on neuronal activity in the leech ganglion

    NASA Astrophysics Data System (ADS)

    Romanenko, Sergii; Siegel, Peter H.; Wagenaar, Daniel A.; Pikov, Victor

    2013-02-01

    The use of electrically-induced neuromodulation has grown in importance in the treatment of multiple neurological disorders such as Parkinson's disease, dystonia, epilepsy, chronic pain, cluster headaches and others. While electrical current can be applied locally, it requires placing stimulation electrodes in direct contact with the neural tissue. Our goal is to develop a method for localized application of electromagnetic energy to the brain without direct tissue contact. Toward this goal, we are experimenting with the wireless transmission of millimeter wave (MMW) energy in the 10-100 GHz frequency range, where penetration and focusing can be traded off to provide non-contact irradiation of the cerebral cortex. Initial experiments have been conducted on freshly-isolated leech ganglia to evaluate the real-time changes in the activity of individual neurons upon exposure to the MMW radiation. The initial results indicate that low-intensity MMWs can partially suppress the neuronal activity. This is in contrast to general bath heating, which had an excitatory effect on the neuronal activity. Further studies are underway to determine the changes in the state of the membrane channels that might be responsible for the observed neuromodulatory effects.

  9. Lineage analysis of micromere 4d, a super-phylotypic cell for Lophotrochozoa, in the leech Helobdella and the sludgeworm Tubifex

    PubMed Central

    Gline, Stephanie E.; Nakamoto, Ayaki; Cho, Sung-Jin; Chi, Candace; Weisblat, David A.

    2011-01-01

    The super-phylum Lophotrochozoa contains the plurality of extant animal phyla and exhibits a corresponding diversity of adult body plans. Moreover, in contrast to Ecdysozoa and Deuterostomia, most lophotrochozoans exhibit a conserved pattern of stereotyped early divisions called spiral cleavage. In particular, bilateral mesoderm in most lophotrochozoan species arises from the progeny of micromere 4d, which is assumed to be homologous with a similar cell in the embryo of the ancestral lophotrochozoan, more than 650 million years ago. Thus, distinguishing the conserved and diversified features of cell fates in the 4d lineage among modern spiralians is required to understand how lophotrochozoan diversity has evolved by changes in developmental processes. Here we analyze cell fates for the early progeny of the bilateral daughters (M teloblasts) of micromere 4d in the leech Helobdella sp. Austin, a clitellate annelid. We show that the first six progeny of the M teloblasts (em1–em6) contribute five different sets of progeny to non-segmental mesoderm, mainly in the head and in the lining of the digestive tract. The latter feature, associated with cells em1 and em2 in Helobdella, is seen with the M teloblast lineage in a second clitellate species, the sludgeworm Tubifex tubifex and, on the basis of previously published work, in the initial progeny of the M teloblast homologs in molluscan species, suggesting that it may be an ancestral feature of lophotrochozoan development. PMID:21295566

  10. Na+/K+ pump interacts with the h-current to control bursting activity in central pattern generator neurons of leeches

    PubMed Central

    Kueh, Daniel; Barnett, William H; Cymbalyuk, Gennady S; Calabrese, Ronald L

    2016-01-01

    The dynamics of different ionic currents shape the bursting activity of neurons and networks that control motor output. Despite being ubiquitous in all animal cells, the contribution of the Na+/K+ pump current to such bursting activity has not been well studied. We used monensin, a Na+/H+ antiporter, to examine the role of the pump on the bursting activity of oscillator heart interneurons in leeches. When we stimulated the pump with monensin, the period of these neurons decreased significantly, an effect that was prevented or reversed when the h-current was blocked by Cs+. The decreased period could also occur if the pump was inhibited with strophanthidin or K+-free saline. Our monensin results were reproduced in model, which explains the pump’s contributions to bursting activity based on Na+ dynamics. Our results indicate that a dynamically oscillating pump current that interacts with the h-current can regulate the bursting activity of neurons and networks. DOI: http://dx.doi.org/10.7554/eLife.19322.001 PMID:27588351

  11. Differential Expression of Conserved Germ Line Markers and Delayed Segregation of Male and Female Primordial Germ Cells in a Hermaphrodite, the Leech Helobdella

    PubMed Central

    Cho, Sung-Jin; Vallès, Yvonne; Weisblat, David A.

    2014-01-01

    In sexually reproducing animals, primordial germ cells (PGCs) are often set aside early in embryogenesis, a strategy that minimizes the risk of genomic damage associated with replication and mitosis during the cell cycle. Here, we have used germ line markers (piwi, vasa, and nanos) and microinjected cell lineage tracers to show that PGC specification in the leech genus Helobdella follows a different scenario: in this hermaphrodite, the male and female PGCs segregate from somatic lineages only after more than 20 rounds of zygotic mitosis; the male and female PGCs share the same (mesodermal) cell lineage for 19 rounds of zygotic mitosis. Moreover, while all three markers are expressed in both male and female reproductive tissues of the adult, they are expressed differentially between the male and female PGCs of the developing embryo: piwi and vasa are expressed preferentially in female PGCs at a time when nanos is expressed preferentially in male PGCs. A priori, the delayed segregation of male and female PGCs from somatic tissues and from one another increases the probability of mutations affecting both male and female PGCs of a given individual. We speculate that this suite of features, combined with a capacity for self-fertilization, may contribute to the dramatically rearranged genome of Helobdella robusta relative to other animals. PMID:24217283

  12. Developmental transition to bilaterally symmetric cell divisions is regulated by Pax-mediated transcription in embryos of the leech Helobdella austinensis.

    PubMed

    Schmerer, Matthew W; Null, Ryan W; Shankland, Marty

    2013-10-01

    The leech embryo develops by spiral cleavage, and establishes the symmetry properties of its adult body plan through the bilaterally symmetric divisions of mesodermal proteloblast DM″ and ectodermal proteloblast DNOPQ‴. We here show that transcriptional inhibitors α-amanitin and actinomycin D specifically disrupt the symmetry and orientation of these two proteloblast cell divisions while having no apparent effect on the timing or geometry of other divisions. Transcriptional inhibition had a similar effect on both proteloblasts, i.e. cytokinesis was highly asymmetric and the cleavage plane roughly orthogonal to that seen during normal development. These findings suggest that zygotic gene product(s) are required, either directly or indirectly, for the correct placement of the proteloblast cleavage furrow. The same phenotypes were also observed following in vivo expression of dominant-negative Pax gene constructs. These dominant-negative phenotypes depended on protein/DNA interaction, and could be rescued by coexpression of full length Pax proteins. However, symmetric cleavage of the mesodermal proteloblast was rescued by full length constructs of either Hau-Paxβ1 or Hau-Pax2/5/8, while only Hau-Paxβ1 rescued the symmetry of ectodermal cleavage. We conclude that both proteloblasts need Pax-mediated transcription to adopt their normally symmetric cleavage patterns, but differ in terms of the specific Pax proteins required. The implication of these findings for the evolution of spiral cleavage is discussed. © 2013 Elsevier Inc. All rights reserved.

  13. The marine leech Stibarobdella loricata (Harding, 1924) (Hirudinea, Piscicolidae), parasitic on the angel shark Squatina spp. and sandtiger shark Carcharias taurus Rafinesque, 1810 (Chondrichthyes: Squatinidae, Carchariidae) in Southern Brazilian waters.

    PubMed

    Soto, J M R

    2003-11-01

    The presence of the marine leech, Stibarobdella loricata (Harding, 1924) (Hirudinea, Piscicolidae), is reported on the southern coast of Brazil, based on seven lots with 47 specimens, between 71 and 182 mm in total length, collected on the dorsal region of angel sharks, Squatina argentina (Marini, 1930); S. guggenheim Marini, 1936; S. punctata Marini, 1936 (Chondrichthyes, Squatinidae); and on the head of a sandtiger shark, Carcharias taurus Rafinesque, 1810 (Chondrichthyes, Carchariidae). This is the first record of S. loricata in the western Atlantic and of its parasitic association with S. argentina, S. guggenheim, S. punctata, and C. taurus.

  14. Mucinivorans hirudinis gen. nov., sp. nov., an anaerobic, mucin-degrading bacterium isolated from the digestive tract of the medicinal leech Hirudo verbana

    PubMed Central

    Nelson, Michael C.; Bomar, Lindsey; Maltz, Michele

    2015-01-01

    Three anaerobic bacterial strains were isolated from the digestive tract of the medicinal leech Hirudo verbana, using mucin as the primary carbon and energy source. These strains, designated M3T, M4 and M6, were Gram-stain-negative, non-spore-forming and non-motile. Cells were elongated bacilli approximately 2.4 µm long and 0.6 µm wide. Growth only occurred anaerobically under mesophilic and neutral pH conditions. All three strains could utilize multiple simple and complex sugars as carbon sources, with glucose fermented to acid by-products. The DNA G+C contents of strains M3T, M4 and M6 were 44.9, 44.8 and 44.8 mol%, respectively. The major cellular fatty acid of strain M3T was iso-C15 : 0. Phylogenetic analysis of full-length 16S rRNA gene sequences revealed that the three strains shared >99 % similarity with each other and represent a new lineage within the family Rikenellaceae of the order Bacteroidales, phylum Bacteroidetes. The most closely related bacteria to strain M3T based on 16S rRNA gene sequences were Rikenella microfusus DSM 15922T (87.3 % similarity) and Alistipes finegoldii AHN 2437T (87.4 %). On the basis of phenotypic, genotypic and physiological evidence, strains M3T, M4 and M6 are proposed as representing a novel species of a new genus within the family Rikenellaceae, for which the name Mucinivorans hirudinis gen. nov., sp. nov. is proposed. The type strain of Mucinivorans hirudinis is M3T ( = ATCC BAA-2553T = DSM 27344T). PMID:25563920

  15. Na+-dependent regulation of the free Mg2+ concentration in neuropile glial cells and P neurones of the leech Hirudo medicinalis.

    PubMed

    Hintz, K; Günzel, D; Schlue, W R

    1999-02-01

    To investigate the Mg2+ regulation in neuropile glial (NG) cells and pressure (P) neurones of the leech Hirudo medicinalis the intracellular free Mg2+ ([Mg2+]i) and Na+ ([Na+]i) concentrations, as well as the membrane potential (Em), were measured using Mg2+- and Na+-selective microelectrodes. The mean steady-state values of [Mg2+]i were found to be 0.91 mM (mean Em=-63.6 mV) in NG cells and 0.20 mM (mean Em=-40.6 mV) in P neurones with a [Na+]i of 6.92 mM (mean Em=-61.6 mV) and 7.76 mM (mean Em=-38.5 mV), respectively. When the extracellular Mg2+ concentration ([Mg2+]o) was elevated, [Mg2+]i in P neurones increased within 5-20 min whereas in NG cells a [Mg2+]i increase occurred only after long-term exposure (6 h). After [Mg2+]o was reduced back to 1 mM, a reduction of the extracellular Na+ concentration ([Na+]o) decreased the inwardly directed Na+ gradient and reduced the rate of Mg2+ extrusion considerably in both NG cells and P neurones. In P neurones Mg2+ extrusion was reduced to 15.4% in Na+-free solutions and to 6.0% in the presence of 2 mM amiloride. Mg2+ extrusion from NG cells was reduced to 6.2% in Na+-free solutions. The results suggest that the major [Mg2+]i-regulating mechanism in both cell types is Na+/ Mg2+ antiport. In P neurones a second, Na+-independent Mg2+ extrusion system may exist.

  16. Testing the spatial and temporal framework of speciation in an ancient lake species flock: the leech genus Dina (Hirudinea: Erpobdellidae) in Lake Ohrid

    NASA Astrophysics Data System (ADS)

    Trajanovski, S.; Albrecht, C.; Schreiber, K.; Schultheiß, R.; Stadler, T.; Benke, M.; Wilke, T.

    2010-07-01

    Ancient Lake Ohrid on the Balkan Peninsula is considered to be the oldest ancient lake in Europe with a suggested Plio-Pleistocene age. Its exact geological age, however, remains unknown. Therefore, molecular clock data of Lake Ohrid biota may serve as an independent constraint of available geological data, and may thus also help to refine age estimates. Such evolutionary data may also help unravel potential biotic and abiotic factors that promote speciation events. Here, mitochondrial sequencing data of one of the largest groups of endemic taxa in Lake Ohrid, the leech genus Dina, is used to test whether it represents an ancient lake species flock, to study the role of horizontal and vertical barriers in Lake Ohrid for evolutionary events, to estimate the onset of intralacustrine diversification in this group based on molecular clock analyses, and to compare this data with data from other endemic species for providing an approximate time frame for the origin of Lake Ohrid. Based on the criteria speciosity, monophyly and endemicity, it can be concluded that Lake Ohrid Dina, indeed, represents an ancient lake species flock. Lineage sorting of its species, however, does not seem to be complete. Analyses of population structures of Dina spp. in the Ohrid watershed indicate a horizontal zonation of haplotypes from spring and lake populations, corroborating the role of lake-side springs, particularly the southern feeder springs, for evolutionary processes in endemic Ohrid taxa. Vertical differentiation of lake taxa, however, appears to be limited, though differences between populations from the littoral and the profundal are apparent. Molecular clock analyses indicate that the most recent common ancestor of extant species of this flock is approximately 1.99±0.83 Ma old, whereas the split of the Lake Ohrid Dina flock from a potential sister taxon outside the lake is estimated at 8.30±3.60 Ma. Comparisons with other groups of endemic Ohrid species indicated that in all

  17. Testing the spatial and temporal framework of speciation in an ancient lake species flock: the leech genus Dina (Hirudinea: Erpobdellidae) in Lake Ohrid

    NASA Astrophysics Data System (ADS)

    Trajanovski, S.; Albrecht, C.; Schreiber, K.; Schultheiß, R.; Stadler, T.; Benke, M.; Wilke, T.

    2010-11-01

    Ancient Lake Ohrid on the Balkan Peninsula is considered to be the oldest ancient lake in Europe with a suggested Plio-/Pleistocene age. Its exact geological age, however, remains unknown. Therefore, molecular clock data of Lake Ohrid biota may serve as an independent constraint of available geological data, and may thus help to refine age estimates. Such evolutionary data may also help unravel potential biotic and abiotic factors that promote speciation events. Here, mitochondrial sequencing data of one of the largest groups of endemic taxa in the Ohrid watershed, the leech genus Dina, is used to test whether it represents an ancient lake species flock, to study the role of potential horizontal and vertical barriers in the watershed for evolutionary events, to estimate the onset of diversification in this group based on molecular clock analyses, and to compare this data with data from other endemic species for providing an approximate time frame for the origin of Lake Ohrid. Based on the criteria speciosity, monophyly and endemicity, it can be concluded that Dina spp. from the Ohrid watershed, indeed, represents an ancient lake species flock. Lineage sorting of its species, however, does not seem to be complete and/or hybridization may occur. Analyses of population structures of Dina spp. in the Ohrid watershed indicate a horizontal zonation of haplotypes from spring and lake populations, corroborating the role of lake-side springs, particularly the southern feeder springs, for evolutionary processes in endemic Ohrid taxa. Vertical differentiation of lake taxa, however, appears to be limited, though differences between populations from the littoral and the profundal are apparent. Molecular clock analyses indicate that the most recent common ancestor of extant species of this flock is approximately 1.99 ± 0.83 million years (Ma) old, whereas the split of the Ohrid Dina flock from a potential sister taxon outside the lake is estimated at 8.30 ± 3.60 Ma

  18. Using a model to assess the role of the spatiotemporal pattern of inhibitory input and intrasegmental electrical coupling in the intersegmental and side-to-side coordination of motor neurons by the leech heartbeat central pattern generator.

    PubMed

    García, Paul S; Wright, Terrence M; Cunningham, Ian R; Calabrese, Ronald L

    2008-09-01

    Previously we presented a quantitative description of the spatiotemporal pattern of inhibitory synaptic input from the heartbeat central pattern generator (CPG) to segmental motor neurons that drive heartbeat in the medicinal leech and the resultant coordination of CPG interneurons and motor neurons. To begin elucidating the mechanisms of coordination, we explore intersegmental and side-to-side coordination in an ensemble model of all heart motor neurons and their known synaptic inputs and electrical coupling. Model motor neuron intrinsic properties were kept simple, enabling us to determine the extent to which input and electrical coupling acting together can account for observed coordination in the living system in the absence of a substantive contribution from the motor neurons themselves. The living system produces an asymmetric motor pattern: motor neurons on one side fire nearly in synchrony (synchronous), whereas on the other they fire in a rear-to-front progression (peristaltic). The model reproduces the general trends of intersegmental and side-to-side phase relations among motor neurons, but the match with the living system is not quantitatively accurate. Thus realistic (experimentally determined) inputs do not produce similarly realistic output in our model, suggesting that motor neuron intrinsic properties may contribute to their coordination. By varying parameters that determine electrical coupling, conduction delays, intraburst synaptic plasticity, and motor neuron excitability, we show that the most important determinant of intersegmental and side-to-side phase relations in the model was the spatiotemporal pattern of synaptic inputs, although phasing was influenced significantly by electrical coupling.

  19. LEECH: LBTI Exozodi Exoplanet Common Hunt

    NASA Astrophysics Data System (ADS)

    Skemer, Andrew

    We propose to perform a direct exoplanet imaging survey around nearby (<25 pc), intermediate-aged (0.1-1.0 Gyr) stars using the LBT adaptive optics system (LBTAO) and the mid-infrared imager, LBTI. LBTAO is the first of the next generation extreme AO systems to come online and the only one that will operate in the mid-infrared (>3μm), where old/cool planets are brightest, and adaptive optics performance is superb (>95% Strehl ratio for LBTAO). The proposed survey will leverage 60 nights already-allocated to the NASA LBTI exozodiacal dust survey, which will target nearby stars with nulling interferometry to search for faint, warm debris disks. LBTI has a 3-5μm imager/spectrograph (LMIRCam) and an 8-13μm imager/spectrograph/nuller (NOMIC), which can be operated simultaneously using a beamsplitter, meaning that LMIRCam can search for gas-giant planets while NOMIC measures exozodiacal emission. Executing these two surveys simultaneously will greatly increase the scientific productivity of 60 already-allocated NASA nights by 1) creating exoplanet discovery opportunities and 2) providing a synergetic data set for studying debris disks (exozodis) as signposts of giant planets. The exozodi survey sample is older than the samples of other planned direct imaging surveys, which look at younger stars (<100 Myr) due to the fact that planets become fainter as they age. LBTI is still sensitive to planets around older stars because it operates at L (3.8 μm) where evolutionary models predict planets fade more slowly than at the wavelengths used by most direct-imaging surveys, (H; 1.65 μm). The ability to detect planets around nearby (<25 pc) intermediante-aged (0.1-1.0 Gyr) stars presents several scientific opportunities: 1) A variety of evolutionary models (hot-start, core-accretion/cold-start, warm-start) predict different cooling curves for extrasolar planets, based on different initial conditions. By imaging a population of old planets, we will determine how the planet population evolves by comparing our results to the young planet surveys (e.g. GPI and SPHERE). 2) Initial results show that young planets (such as HR 8799 bcde and 2M1207 b) have very different atmospheres from their older and more massive brown-dwarf analogs. Our sample is intermediate in age between the very young planets that will be discovered by other direct imaging surveys and the very old field brown- dwarf population. Multi-wavelength characterization of our old-planet sample will provide a key intermediate step for atmospheric models. 3) Our sample is composed of the nearest stars, which have been extensively studied by other types of surveys (Doppler RV planet searches, Spitzer and Herschel debris disk observations, and the concurrent LBTI exozodiacal dust survey). The combination of all of these data will reveal the overall architectures of planetary systems by constraining the presence of inner giant planets with RV data, outer giant planets with LBTI direct imaging, hot debris disks with LBTI exozodi data, and cold debris disks with Spitzer and Herschel. Based on planet population models and on-sky estimates of the LBTAO/LBTI instrument performance, we expect to discover ~2-7 planets by observing 117 nearby stars over the next 4 years. These planets will be older and nearer to the Sun than planets found by other surveys, and will benefit from synergetic data from several other surveys that target the nearest stars. Using these new discoveries, we will constrain the atmospheric and evolutionary models for intermediate-aged planets, and determine the extent to which debris disks can be used as signposts of extrasolar planets.

  20. Structural Stability Evaluation, Leech Lake Dam.

    DTIC Science & Technology

    1981-09-01

    81-22 _ 4. TITLE (a Pad Subtitle) 5. TYPE OF REPORT & PERIOD COVERED (j Final ect dQ TRUCTURAL STABILITY EVALUATION . PERFR ". W ar E. acRMeIN G...allowable values; consequently, post- tensioning of the monoliths to the foundation will be one solution to correct these deficiencies. Therefore, it is...Various types of pressuremeters are now being used in the United States; a self-boring pressuremeter shows great prom- ise for future use. 38. In

  1. Back to past leeches: repeated phlebotomies and cardiovascular risk

    PubMed Central

    2012-01-01

    In patients with metabolic syndrome, body iron overload exacerbates insulin resistance, impairment of glucose metabolism, endothelium dysfunction and coronary artery responses. Conversely, iron depletion is effective to ameliorate glucose metabolism and dysfunctional endothelium. Most of its effectiveness seems to occur through the amelioration of systemic and hepatic insulin resistance. In a study published by BMC Medicine, Michalsen et al. demonstrated a dramatic improvement of blood pressure, serum glucose and lipids after removing 550 to 800 ml of blood in subjects with metabolic syndrome. This effect was apparently independent of changes in insulin resistance, in contrast to previous cross-sectional and cohort studies investigating the association between iron overload, insulin resistance and cardiovascular disease. Despite drawbacks in the study design, its findings may lead the way to investigations aimed at exploring iron-dependent regulatory mechanisms of vascular tone in healthy individuals and patients with metabolic disease, thus providing a rationale for novel preventive and therapeutic strategies to counteract hypertension. Please see related article: http://www.biomedcentral.com/1741-7015/10/54 PMID:22647488

  2. Chinese Medicinal Leech: Ethnopharmacology, Phytochemistry, and Pharmacological Activities

    PubMed Central

    Dong, Han; Ren, Ji-Xiang; Wang, Jing-Jing; Ding, Li-Shuai; Zhao, Jian-Jun; Liu, Song-Yan; Gao, Hui-Min

    2016-01-01

    Hirudo (Shuizhi in Chinese) is an important Chinese medicine, which possesses many therapeutic properties for the treatment of the cerebral hemorrhage and other thrombosis-related diseases. The phytochemical investigation gave more than 51 compounds including pteridines, phosphatidylcholines, glycosphingolipids, and sterols, as well as some bioactive peptides from the Shuizhi derived from three animal species recorded in the current Chinese Pharmacopoeia. The pharmacological studies on the Shuizhi have revealed various activities such as anticoagulation, antithrombosis, antiatherosclerosis, antiplatelet aggregation, antitumor and anti-inflammatory as well as hemorheology improvement, and protective effects against cerebral ischemia-reperfusion injury. However, some important issues based on the traditional uses of Shuizhi are still not clear. The aim of the present review is to provide comprehensive knowledge on the ethnopharmacology, phytochemistry, and pharmacological activities of Shuizhi. It will provide a potential guidance in exploring main active compounds of Shuizhi and interpreting the action mechanism for the further research. PMID:27274755

  3. `Don't forget your leech socks'! Children's learning during an Eden Education Officer's workshop

    NASA Astrophysics Data System (ADS)

    Bowker, Rob; Jasper, Andy

    2007-04-01

    This study looked at 30 primary aged children between 10 and 11 years old who were visiting the Eden Project, Cornwall and participating in workshops led and designed by the Eden Education Officers. The study attempted to directly test the effects of the Education Officers' workshops on children's learning. Personal meaning mapping, a constructivist approach was used to measure change in the children's understanding along four semi-independent dimensions: extent, breadth, depth and mastery. The children's mastery of the subject regardless of ability was improved. Most of the workshop aims, such as ‘to recognise some plant adaptations in the humid tropics climate’ were shown to have been met. The research also highlighted areas to be reviewed, such as educating children about the way indigenous people use plants for their survival and also how children can transfer knowledge, so that they can understand the significance of plants in their own daily lives.

  4. ANATOMY, LIFE HISTORY AND DISTRIBUTION OF THE PARASITIC LEECH OLIGOBDELLA BIANNULATA (MOORE, 1900) (EUHIRUDINEA: GLOSSIPHONIIDAE)

    EPA Science Inventory

    Oligobdella biannulata (Moore, 1900) is a rare, endemic species originally described from a mountain stream near Blowing Rock, North Carolina. Specimens of this species were collected seasonally from fall 1999 to winter 2001 with four new county records in North Carolina (Avery,...

  5. The first data on species diversity of leeches (Hirudinea) in the Irtysh River Basin, East Kazakhstan.

    PubMed

    Kaygorodova, Irina A; Fedorova, Lyudmila I

    2016-07-27

    The Irtysh River is one of the major waterways of the Republic of Kazakhstan. The river originates in the territory of Mongolia and China, flows through the East Kazakhstan and falls into the Ob River in the Western Siberia of Russia. The Kazakhstan section of the river, with length of 1698 km and water resources about 43.8 km3 (Dyusebaeva 2012), is situated on the border of the West Siberian Plain, the Altai Mountains and the Kazakh Uplands. As the river crosses various natural zones, characteristics of its hydrological regime is very diverse. In the south, in the area of Ust-Kamenogorsk, the valley of the Irtysh extends. In the upper reaches up to the city of Semey (formerly Semipalatinsk), the river is surrounded by plain and mountain landscapes. Downstream from Semey, the Irtysh enters the West Siberian Plain and flows to north-west through dry steppes. The riverbed is unstable: sometimes it is very tortuous, sometimes it is broken into branches, and, there are a lot of sand spits. The hydrological level of the Kazakhstan part of the Irtysh River is regulated with the cascade of storage reservoirs near the towns Bukhtarma, Ust-Kamenogorsk and Shulbinsk.

  6. Characterization of a voltage-dependent conductance in the basolateral membrane of leech skin epithelium.

    PubMed

    Schnizler, M; Clauss, W

    1998-05-01

    Voltage clamp studies were performed on the dorsal integument of Hirudo medicinalis. Under apical calcium-free conditions an inward-directed component of transepithelial current was activated by changes of transepithelial voltage. Depolarization caused up to 50% increase of the transepithelial sodium current. Hyperpolarization had no comparable effects. With calcium (1.8 mM) or amiloride (100 microM) in the apical solution and in sodium-free solutions the inward-directed current failed to increase after depolarization. Activation also occurred under chloride-free conditions. Permeabilization of the apical membrane by nystatin (5 microM) increased the current activation significantly. After nystatin, calcium as well as amiloride lost their inhibitory effects. This indicates a basolateral localization of the voltage-dependent conductance. Vesicle insertion or cytoskeletal structures are probably not involved in regulation, as seen by the lack of effects of brefeldin A and the cytochalasins B and D. However, serosal hyposmolar solutions (170 mosmol.1(-1)) caused a reinforced activation of the current. Our results indicate a voltage-dependent conductance in a tight sodium-absorbing epithelium.

  7. "Don't Forget Your Leech Socks"! Children's Learning during an Eden Education Officer's Workshop

    ERIC Educational Resources Information Center

    Bowker, Rob; Jasper, Andy

    2007-01-01

    This study looked at 30 primary aged children between 10 and 11 years old who were visiting the Eden Project, Cornwall and participating in workshops led and designed by the Eden Education Officers. The study attempted to directly test the effects of the Education Officers' workshops on children's learning. Personal meaning mapping, a…

  8. [Leeching in the treatment of chronic inflammatory and dystrophic diseases of the salivary glands].

    PubMed

    Abal'masov, D V; Afanas'ev, V V; Pozharitskaia, M M

    2003-01-01

    Twenty patients with chronic inflammatory and dystrophic diseases of the salivary glands (sialadenitis, sialadenosis) were examined and treated using hirudotherapy. Positive clinical shifts were observed in 50% patients; the most pronounced therapeutic effect was observed in patients with sialadenosis. Hirudotherapy was ineffective in patients with chronic parenchymatous parotitis paralleled by Sjogren's syndrome.

  9. The complete mitochondrial genome of Sasakia funebris (Leech) (Lepidoptera: Nymphalidae) and comparison with other Apaturinae insects.

    PubMed

    Wang, Ju Ping; Cao, Tian Wen; Xuan, Shan Bin; Wang, Hui; Zhang, Min; Ma, En bo

    2013-09-10

    Sasakia funebris, a member of the lepidopteran family, Nymphalidae (superfamily Papilionoidea) is a rare species and is found only in some areas of South China. In this study, the 15,233 bp long complete mitochondrial genome of S. funebris was determined, and harbors the gene arrangement identical to all other sequenced lepidopteran insects. The nucleotide composition of the genome is highly A+T biased, accounting for 81.2%. All protein-coding genes (PCGs) start with typical ATN codons, except for COI which begins with the CGA codon. All tRNAs have a typical clover-leaf secondary structure, except for tRNASer(AGN), the dihydrouridine (DHU) arm of which forms a simple loop. The S. funebris A+T-rich region of 370 bp contains several features common to the Lepidoptera insects, including the motif ATAGA followed by a 19 bp poly-T stretch, and two tandem repeats consisting of 18 bp repeat units and 14 bp repeat units. The phylogenetic analyses of Apaturinae based on mitogenome sequences showed: (S. funebris+Sasakia charonda)+(Apatura metis+Apatura ilia). This result is consistent with the morphological classification.

  10. Predatory leeches (Hirudinida) may contribute to amphibian declines in the Lassen, California.

    Treesearch

    Stead Jonathan; Karen Pope

    2010-01-01

    Researchers have documented precipitous declines in Cascades Frog (Rana cascadae) populations in the southern portion of the species' range, in the Lassen region of California. Reasons for the declines, however, have not been elucidated. In addition to common, widespread causes, an understanding of local community interactions may be necessary...

  11. From leeches to personalized medicine: evolving concepts in the management of polycythemia vera

    PubMed Central

    Vannucchi, Alessandro M.

    2017-01-01

    Polycythemia vera is a clonal disorder of hematopoietic stem/progenitor cells. It manifests as an expansion of red cell mass. It is the most common chronic myeloproliferative neoplasm. In virtually all cases, it is characterized by a V617F point mutation in JAK2 exon 14 or less common mutations in exon 12. The landmark discovery of the autonomously activated JAK/STAT signaling pathway paved the way for the clinical development of the first target drug, the JAK1 and JAK2 inhibitor ruxolitinib. This is now approved for patients with resistance or intolerance to hydroxyurea. Phlebotomies and hydroxyurea are still the cornerstone of treatment, and aim to prevent the first appearance or recurrence of cardiovascular events that, together with progression to post-polycythemia vera myelofibrosis and leukemia, represent the main causes of death. Interferon-α is an alternative drug and has been shown to induce molecular remissions. It is currently undergoing phase III trials that might eventually lead to its approval for clinical use. The last few years have witnessed important advances towards an accurate early diagnosis of polycythemia vera, greater understanding of its pathogenesis, and improved patient management. This review will focus on the most recent achievements and will aim to unify the different concepts involved in a personalized approach to the patient with polycythemia vera. In spite of many recent advances in the understanding of its pathogenesis and improved disease management, polycythemia vera remains a life-threatening myeloproliferative neoplasm for which there is no cure. This review will present a critical overview of evolving concepts in diagnosis and treatment of this disease. PMID:27884974

  12. Land Leeches of the g.Haemadipsa (Haemadipsoidea:Haemadipsidae). II. Bibliography.

    DTIC Science & Technology

    1982-05-01

    Sangsues Terrestres). Bull Soc Pathol Exot 10:640-675, 1917 10. DEQUAL, L. Nuovi Irudinei esotici del Museo Zoologico di Torino. Boll Mus Zool Anat Comp...A. Forest pests man should avoid. Philipp J For 3:191-201, 1940 DEQUAL, L. Nuovi Irudinei esotici del Museo Zoologico di Torino. BtI)i Mus Zool Anat

  13. "Don't Forget Your Leech Socks"! Children's Learning during an Eden Education Officer's Workshop

    ERIC Educational Resources Information Center

    Bowker, Rob; Jasper, Andy

    2007-01-01

    This study looked at 30 primary aged children between 10 and 11 years old who were visiting the Eden Project, Cornwall and participating in workshops led and designed by the Eden Education Officers. The study attempted to directly test the effects of the Education Officers' workshops on children's learning. Personal meaning mapping, a…

  14. ANATOMY, LIFE HISTORY AND DISTRIBUTION OF THE PARASITIC LEECH OLIGOBDELLA BIANNULATA (MOORE, 1900) (EUHIRUDINEA: GLOSSIPHONIIDAE)

    EPA Science Inventory

    Oligobdella biannulata (Moore, 1900) is a rare, endemic species originally described from a mountain stream near Blowing Rock, North Carolina. Specimens of this species were collected seasonally from fall 1999 to winter 2001 with four new county records in North Carolina (Avery,...

  15. [Scheme for dispersion analysis for evaluating heritability of reproductive traits in medicinal leech Hirudo medicinalis L].

    PubMed

    Utevskaia, O M; Atramentova, L A

    2003-10-01

    ANOVA designs for estimating heritabilities of fecundity traits in Hirudo medicinalis including batch size, the number of juveniles per cocoon, and juvenile weight. Accounting for reproduction mode of this species, different types of kinship were identified, which were taken into account in the ANOVA designs: juveniles from one batch and one cocoon were considered respectively full sibs and polyzygotic twins. Variation components were analyzed in the following kinship groups: for batch size, in full sibs; for the number of juveniles per cocoon, between sibships, full sins, and replicates; and for juvenile weight, in full sibs and polyzygotic twins. Using these designs, heritabilities of basic reproductive traits of H. medicinalis were obtained: for batch size, h2 = 0.35-0.40, H2 = 0.40-0.45; for the number of juveniles per cocoon, h2 = 0.33-0.36, H2 = 0.36-0.39; and for juvenile weight, h2 = 0.40-0.44, H2 = 0.44-0.48.

  16. The Iberian Leech: Napoleon’s Counterinsurgency Operations in the Peninsula, 1807-1810

    DTIC Science & Technology

    2005-06-17

    1807 Napoleon’s armies were not the same well trained, veteran armies that had marched so swiftly from their training camps in Boulogne to defeat the...the village of Carabanchel Alto murdered a French captain.5 In the same month, General Lejuene reported from the city of Burgos: I was just going to...thirst after money overcame all fear of punishment.46 The looting and pillaging got so out-of-hand that one of Marshal Ney’s aides-de- camp remarked

  17. THE LEECH EXOPLANET IMAGING SURVEY: ORBIT AND COMPONENT MASSES OF THE INTERMEDIATE-AGE, LATE-TYPE BINARY NO UMa

    SciTech Connect

    Schlieder, Joshua E.; Skemer, Andrew J.; Hinz, Philip; Leisenring, Jarron; Defrère, Denis; Close, Laird M.; Eisner, Josh A.; Maire, Anne-Lise; Desidera, Silvano; Skrutskie, Michael F.; Bailey, Vanessa; Esposito, Simone; Strassmeier, Klaus G.; Weber, Michael; Biller, Beth A.; Bonnefoy, Mickaël; Buenzli, Esther; Henning, Thomas; Crepp, Justin R.; Hofmann, Karl-Heinz; and others

    2016-02-10

    We present high-resolution Large Binocular Telescope LBTI/LMIRcam images of the spectroscopic and astrometric binary NO UMa obtained as part of the LBT Interferometer Exozodi Exoplanet Common Hunt exoplanet imaging survey. Our H-, K{sub s}-, and L′-band observations resolve the system at angular separations <0.″09. The components exhibit significant orbital motion over a span of ∼7 months. We combine our imaging data with archival images, published speckle interferometry measurements, and existing spectroscopic velocity data to solve the full orbital solution and estimate component masses. The masses of the K2.0 ± 0.5 primary and K6.5 ± 0.5 secondary are 0.83 ± 0.02 M{sub ⊙} and 0.64 ± 0.02 M{sub ⊙}, respectively. We also derive a system distance of d = 25.87 ± 0.02 pc and revise the Galactic kinematics of NO UMa. Our revised Galactic kinematics confirm NO UMa as a nuclear member of the ∼500 Myr old Ursa Major moving group, and it is thus a mass and age benchmark. We compare the masses of the NO UMa binary components to those predicted by five sets of stellar evolution models at the age of the Ursa Major group. We find excellent agreement between our measured masses and model predictions with little systematic scatter between the models. NO UMa joins the short list of nearby, bright, late-type binaries having known ages and fully characterized orbits.

  18. A new and alien species of ``oyster leech'' (Platyhelminthes, Polycladida, Stylochidae) from the brackish North Sea Canal, The Netherlands

    NASA Astrophysics Data System (ADS)

    Sluys, Ronald; Faubel, Anno; Rajagopal, Sanjeevi; Velde, Gerard Van Der

    2005-11-01

    A new species of polyclad flatworm, Imogine necopinata Sluys, sp. nov., is described from a brackish habitat in The Netherlands. Taxonomic affinities with Asian species and the ecology of the animals suggest that the species is an introduced, exotic component of the Dutch fauna. The new species belongs to a group of worms with species that are known to predate on oysters.

  19. Transcriptome Sequence of the Bloodstream Form of Trypanoplasma borreli, a Hematozoic Parasite of Fish Transmitted by Leeches

    PubMed Central

    Carrington, Mark; Dóró, Eva; Wiegertjes, Geert F.

    2017-01-01

    ABSTRACT Here, we report a transcriptome sequence of Trypanoplasma borreli isolated from its natural host, the common carp, Cyprinus carpio. The transcriptome allows an analysis of abundant cell surface proteins and acts as a comparator for understanding the evolution and pathogenicity of other Kinetoplastida, including several that infect humans. PMID:28254980

  20. History of evidence-based medicine. Oranges, chloride of lime and leeches: barriers to teaching old dogs new tricks.

    PubMed

    Doherty, Steven

    2005-08-01

    Knowledge translation is the process of taking evidence from research and applying it in clinical practice. In this article I will cite some pivotal moments in the history of medicine to highlight the difficulties and delays associated with getting evidence into practice. These historical examples have much in common with modern medical trials and quality improvement processes. I will also review the reasons why evidence is not used and consider what factors facilitate the uptake of evidence. Understanding these concepts will make it easier for individual clinicians and institutions to change clinical behaviour and provide a starting point for those looking at implementing 'new' practices, new therapies and clinical guidelines. Finally, I will offer a list of criteria that clinicians might choose to consider when deciding on whether or not to adopt a new practice, treatment or concept.

  1. Hydrogeology and ground-water quality of glacial-drift aquifers, Leech Lake Indian Reservation, north-central Minnesota

    USGS Publications Warehouse

    Lindgren, R.J.

    1996-01-01

    Water collected from wells completed in the unconfined aquifer in residential and recreational land-use areas had concentrations of arsenic, cadmium, chromium, copper, lead, mercury, and cyanide equal to or less than 6 micrograms per liter. Concentrations of organic-acid herbicides in water from three wells screened in the unconfined aquifer in managed-forest land-use areas were all below detection levels. Concentrations of U.S. Environmental Protection Agency priority pollutants in water from three wells screened in the unconfined aquifer and from one well screened in the uppermost confined aquifer were also all below detection levels.

  2. New Host and Distribution Records of the Leech Placobdella sophieaeOceguera-Figueroa et al., 2010 (Hirudinida: Glossiphoniidae)

    USGS Publications Warehouse

    Moser, William E.; Bowerman, Jay; Hovingh, Peter; Pearl, Christopher A.; Oceguera-Figueroa, Alajandro

    2014-01-01

    Placobdella sophieae Oceguera-Figueroa et al., 2010 (Hirudinida: Glossiphoniidae) is reported from Oregon, California, and British Columbia for the first time. New hosts reported for P. sophieae include Taricha granulosa (rough-skinned newt), Rana pretiosa (Oregon spotted frog), and Anaxyrus boreas (western toad). Placobdella sophieae exhibits relatively low host specificity and all amphibians occurring in the Pacific Northwest are potential hosts.

  3. Cultural Resources Survey Subterranean Cable Right-of-Way Adjacent to Leech Lake River, Cass County, Minnesota.

    DTIC Science & Technology

    1980-09-01

    of his youthful enthusiasm and his considerable muscle ; the latter provided his expertise in soils and vegetation both in the field and by preparing...requested, the Contractin . Officer will provide thle Contractor w-ith a letter of introduction signed by the St. Paul District Engineer explaining the

  4. The LEECH Exoplanet Imaging Survey: Orbit and Component Masses of the Intermediate-Age, Late-Type Binary NO UMa

    NASA Astrophysics Data System (ADS)

    Schlieder, Joshua E.; Skemer, Andrew J.; Maire, Anne-Lise; Desidera, Silvano; Hinz, Philip; Skrutskie, Michael F.; Leisenring, Jarron; Bailey, Vanessa; Defrère, Denis; Esposito, Simone; Strassmeier, Klaus G.; Weber, Michael; Biller, Beth A.; Bonnefoy, Mickaël; Buenzli, Esther; Close, Laird M.; Crepp, Justin R.; Eisner, Josh A.; Hofmann, Karl-Heinz; Henning, Thomas; Morzinski, Katie M.; Schertl, Dieter; Weigelt, Gerd; Woodward, Charles E.

    2016-02-01

    We present high-resolution Large Binocular Telescope LBTI/LMIRcam images of the spectroscopic and astrometric binary NO UMa obtained as part of the LBT Interferometer Exozodi Exoplanet Common Hunt exoplanet imaging survey. Our H-, Ks-, and L‧-band observations resolve the system at angular separations <0.″09. The components exhibit significant orbital motion over a span of ∼7 months. We combine our imaging data with archival images, published speckle interferometry measurements, and existing spectroscopic velocity data to solve the full orbital solution and estimate component masses. The masses of the K2.0 ± 0.5 primary and K6.5 ± 0.5 secondary are 0.83 ± 0.02 M⊙ and 0.64 ± 0.02 M⊙, respectively. We also derive a system distance of d = 25.87 ± 0.02 pc and revise the Galactic kinematics of NO UMa. Our revised Galactic kinematics confirm NO UMa as a nuclear member of the ∼500 Myr old Ursa Major moving group, and it is thus a mass and age benchmark. We compare the masses of the NO UMa binary components to those predicted by five sets of stellar evolution models at the age of the Ursa Major group. We find excellent agreement between our measured masses and model predictions with little systematic scatter between the models. NO UMa joins the short list of nearby, bright, late-type binaries having known ages and fully characterized orbits. Based on data obtained with the STELLA robotic telescope in Tenerife, an AIP facility jointly operated by AIP and IAC.

  5. Lox6, a leech Dfd ortholog, is expressed in the central nervous system and in peripheral sensory structures.

    PubMed

    Wong, V Y; Macagno, E R

    1998-03-01

    Sequence analysis of a newly isolated Hirudo medicinalis cDNA containing an Antennapedia (Antp)-class homeobox suggests that the corresponding gene, Lox6, is an ortholog of the Drosophila Deformed (Dfd) gene. In situ hybridization of whole-mounted preparations shows that the major sites of Lox6 expression during embryogenesis are the central nervous system (CNS) and the peripheral sensory system. Lox6 mRNA can be detected in a subset of neurons in each ganglion from the subesophageal ganglion (RG2) to the most posterior ganglion, with the highest level of expression seen in RG3. Peripherally, Lox6 is expressed principally in the primordia of the sensillae and in the eyes. This pattern of expression of Lox6 suggests that one of its functions may be to contribute to the diversification of neuronal phenotypes.

  6. Oogenesis in the leech Glossiphonia heteroclita (Annelida, Hirudinea, Glossiphoniidae) II. Vitellogenesis, follicle cell structure and egg shell formation.

    PubMed

    Swiatek, Piotr

    2006-08-01

    By the end of previtellogenesis, the oocytes of Glossiphonia heteroclita gradually protrude into the ovary cavity. As a result they lose contact with the ovary cord (which begins to degenerate) and float freely within the hemocoelomic fluid. The oocyte's ooplasm is rich in numerous well-developed Golgi complexes showing high secretory activity, normal and transforming mitochondria, cisternae of rER and vast amounts of ribosomes. The transforming mitochondria become small lipid droplets as vitellogenesis progresses. The oolemma forms microvilli, numerous coated pits and vesicles occur at the base of the microvilli, and the first yolk spheres appear in the peripheral ooplasm. A mixed mechanism of vitellogenesis is suggested. The eggs are covered by a thin vitelline envelope with microvilli projecting through it. The envelope is formed by the oocyte. The vitelline envelope is produced by exocytosis of vesicles containing two kinds of material, one of which is electron-dense and seems not to participate in envelope formation. The cortical ooplasm of fully grown oocytes contains many cytoskeletal elements (F-actin) and numerous membrane-bound vesicles filled with stratified content. Those vesicles probably are cortical granules. The follicle cells surrounding growing oocytes have the following features: (1) they do not lie on a basal lamina; (2) their plasma membrane folds deeply, forming invaginations which eventually seem to form channels throughout their cytoplasm; (3) the plasma membrane facing the ovary lumen is lined with a layer of dense material; and (4) the plasma membrane facing the oocyte forms thin projections which intermingle with the oocyte microvilli. In late oogenesis, the follicle cells detach from the oocytes and degenerate in the ovary lumen.

  7. New records of the chaetiferous leech-like annelid Paracanthobdella livanowi (Epshtein, 1966) (Annelida: Clitellata: Acanthobdellida) from Kamchatka, Russia.

    PubMed

    Utevsky, Serge Y; Sokolov, Sergei G; Shedko, Marina B

    2013-01-01

    Acanthobdellidans are unique in their organisation and phylogenetic relationships due to having transitional characters that combine features of oligochaetous and achaetous annelids. Alongside the relatively well-studied Acanthobdella peledina Grube, 1851, there is another member of the group, Paracanthobdella livanowi (Epshtein, 1966), with five rows of chaetae and an anterior sucker. It appears that the anterior sucker is weakly developed in small juveniles but acquires a deep cavity in adults. Smaller individuals of P. livanowi can be distinguished from A. peledina, which does not possess an anterior sucker, by the varying breadth of their chaetae. The mid-body segment consists of two doubled annuli in juveniles and is quadri-annulate in large individuals. In Kamchatka freshwaters, hosts of P. livanowi mostly include Salvelinus spp. and more rarely Gasterosteus aculeatus, Oncorhynchus mykiss and O. kisutch. New information on the distribution and the biology of P. livanowi is presented.

  8. Actinobdella inequinnulata (Annelida: Hirudinida:Rhynchobdellida:Glossiphoniidae) from White Crappie, Pomoxis annularis (Perciformes: Centrarchidae), in Arkansas, USA

    EPA Science Inventory

    One of 4 (25%) white crappie, Pomoxis annularis from the Ouachita River, Dallas County, Arkansas, was found to be infested with 8 glossiphoniid leeches, Actinobdella inequiannulata Moore, 1901. Leeches were removed from within the oeprculum on gills and gill arches. This leech i...

  9. Actinobdella inequinnulata (Annelida: Hirudinida:Rhynchobdellida:Glossiphoniidae) from White Crappie, Pomoxis annularis (Perciformes: Centrarchidae), in Arkansas, USA

    EPA Science Inventory

    One of 4 (25%) white crappie, Pomoxis annularis from the Ouachita River, Dallas County, Arkansas, was found to be infested with 8 glossiphoniid leeches, Actinobdella inequiannulata Moore, 1901. Leeches were removed from within the oeprculum on gills and gill arches. This leech i...

  10. Cultural Resources Investigation of the Reservoir Shorelines: Gull Lake, Leech Lake, Pine River and Lake Pokegama. Volume 2. Site Location Data.

    DTIC Science & Technology

    1979-06-01

    affiliation Malmo (Middle Woodland ); Blackduck 4. Location: Sec. 4 Twp. 135N R. 29W Verbal description E NWk north end Upper Gull; just east of bridge over...affiliation Woodland 4. Location: Sec. 25 Twp. 134N R. 30W Verbal description NWk SEk on the southeast arm of the southwest part of Wilson Bay, Gull...mound, village, etc.) mound? burial? 2. Map reference USGS Gull Lake 3. Cultural affiliation Woodland and Historic 4. Location: Sec. 21 Tp. 135N R. 29W

  11. Upper Mississippi River Headwaters Reservoirs Damsites Cultural Resources Investigation. Lake Winnibigoshish, Leech Lake, Pokegema Lake, Sandy Lake, Pine River and Gull Lake Reservoirs, Minnesota

    DTIC Science & Technology

    1988-02-01

    Wanzer also reported on an outbreak of small- pox among the Chippewa laborers at the dam, and on the Chippewa’s attitudes toward the construction of the...dwelling, laborers’ quarters, engineers’ quarters, a dining hall, an office building, an officer’s ho.se, a wood shed, a chicken coop, a barn, a...a barn, carpenter and blacksmith shops, two warehouses, a wood shed and a chicken coop (Figure 42). The "new" dwelling was destroyed by another fire

  12. [Infections Associated with Treatment with Hirudo medicinalis - Report of Two Cases and Review of the Literature].

    PubMed

    Reese, K; Gümbel, D; Seifert, J; Daeschlein, G; Napp, M; Ekkernkamp, A

    2015-06-01

    Medicinal leeches are well-established for promoting venous drainage in transplants/flaps and analgesia in osteoarthritis. Although medicinal leeches are bred and kept under controlled conditions, they are colonised by a genuine species-specific bacterial flora. Therapeutic application of leeches implies skin penetration carrying an a priori risk of infection. We report 2 cases with different indications for medicinal leech therapy. In both cases wound infection occurred in close temporal and spatial correlation or with evidence of a leech-associated germ that could be treated successfully. An unclarified complication rate warrants strict indications for the application of medicinal leeches. Preventive measures are currently tested. © Georg Thieme Verlag KG Stuttgart · New York.

  13. Hirudotherapy in Medicine and Dentistry

    PubMed Central

    Garg, Aarti; Narang, Ridhi; Das, Sunanda

    2015-01-01

    The concept of Unani medicine is based on balancing body humours, the imbalance of which causes diseases. The application of leech therapy in medical and dental science is well recognized. Although easy and non-invasive, complications also exist. The article aims to presents a brief review on the applications of leech therapy. The physiological effect, along with its therapeutic role in cancer, diabetes and dentistry have been underlined. Complications of leech therapy have also been dealt with. PMID:26817000

  14. Molecular Identification of Haemadipsa rjukjuana (Hirudiniformes: Haemadipsidae) in Gageo Island, Korea.

    PubMed

    Won, Sohyun; Park, Bae Keun; Kim, Baek Jun; Kim, Hye Won; Kang, Jun Gu; Park, Tae Seo; Seo, Hong Yul; Eun, Ye; Kim, Ki Gyoung; Chae, Joon Seok

    2014-04-01

    There are 60 species of blood-feeding land leeches, 50 species belonging to the family Haemadipsidae and 10 species belonging to the family Xerobdellidae. Despite recent papers on the land leeches, their taxonomic identification is not fully understood, especially at a species level. In Korea, there have been no historical records of the terrestrial leeches, but recently an unrecorded blood-feeding land leech was discovered at Gageo-do (Island), Korea. Molecular analysis was used to identify the species of 29 leeches collected from Mt. Dock-Sil in Gageo-do. Conventional PCR was conducted using nuclear 18S rRNA and mitochondrial cytochrome c oxidase subunit 1 (CO1) genetic marker. The 18S rRNA sequences revealed that the leeches share 99.9% identity with Haemadipsa rjukjuana (inhabiting Taiwan), and the CO1 sequences revealed that the leeches are very close to H. rjukjuana (inhabiting Taiwan). The CO1 sequences were separated into 2 categories, 1 with 94.6% and the other with 94.3% similarity to the H. rjukjuana L00115A (inhabiting Taiwan). This new finding of the land leech is the first record in Korea. In addition, the north range of the distribution of the blood-feeding leech (Hirudiniformes: Haemadipisidae) should be reconsidered including Korea.

  15. BIOLOGY OF THE LEECH ACTINOBDELLA INEQUIANNULATA MOORE, 1901 (ANNELIDA: HIRUDINEA: RHYNCHOBDELLIDA: GLOSSIPHONIIDAE), PARASITIC ON THE WHITE SUCKER, CATOSTOMUS COMMERSONI LACEPEDE, 1803 AND THE LONGNOSE SUCKER, CATOSTOMUS CATOSTOMUS FORSTER, 1773, IN ALGONQUIN PROVINCIAL PARK, ONTARIO, CANADA

    EPA Science Inventory

    Actinobdella inequiannulata was found on the white sucker, Catostomus commersoni, and less frequently on the longnose sucker, Catostomus catostomus, in Algonquin Provincial Park, Ontario, Canada. Catostomus commersoni parasitized with Act. inequiannulata was collected from July ...

  16. BIOLOGY OF THE LEECH ACTINOBDELLA INEQUIANNULATA MOORE, 1901 (ANNELIDA: HIRUDINEA: RHYNCHOBDELLIDA: GLOSSIPHONIIDAE), PARASITIC ON THE WHITE SUCKER, CATOSTOMUS COMMERSONI LACEPEDE, 1803 AND THE LONGNOSE SUCKER, CATOSTOMUS CATOSTOMUS FORSTER, 1773, IN ALGONQUIN PROVINCIAL PARK, ONTARIO, CANADA

    EPA Science Inventory

    Actinobdella inequiannulata was found on the white sucker, Catostomus commersoni, and less frequently on the longnose sucker, Catostomus catostomus, in Algonquin Provincial Park, Ontario, Canada. Catostomus commersoni parasitized with Act. inequiannulata was collected from July ...

  17. Mississippi Headwaters Reservoirs Oral History Interviews, Series 1 and 2

    DTIC Science & Technology

    1988-06-01

    up. Boat and livery from Wooded country Detroit)Ashelmans Resort, Pelican Lake. Lakes Leech, Sand. (52 lier day and ua) V-) (2) Pettibone Log,&o... Pettibone , Prop. River Leech. (610 per week ad up) (1𔃺 Livery John Rahm’s Auto and Horse Livery, Hotels Stanton, 82 per dlay, capacity 50. Detroit, Minn

  18. Oral hirudiniasis in a stray dog, first report in Italy

    PubMed Central

    RAELE, Donato Antonio; GALANTE, Domenico; CAFIERO, Maria Assunta

    2015-01-01

    In June 2014, a male stray dog was recovered at Ente Nazionale di Protezione Animali (ENPA) kennel of Manfredonia, Apulia region, showing oral bleeding and physical prostration. The dog fell in a water canal and was trapped. During the clinical examination, a specimen of leech was revealed into its oral cavity. The parasite, probably entered by drinking unfiltered and contaminated water, has been identified as an adult of aquatic leech Limnatis nilotica. Leeches could overrun wide variety of animals, and few reports about blood sucking leech infestations in mammals are available in literature. This paper describes here the first oral hirudiniasis in a dog in Italy and highlights the possibility of human nasopharyngeal leech-related infection in Apulia region. PMID:26004432

  19. High prevalence of buccal ulcerations in largemouth bass, Micropterus salmoides (Centrarchidae) from Michigan inland lakes associated with Myzobdella lugubris Leidy 1851 (Annelida: Hirudinea)

    PubMed Central

    Faisal, M.; Schulz, C.; Eissa, A.; Whelan, G.

    2011-01-01

    Widespread mouth ulcerations were observed in largemouth bass collected from eight inland lakes in the Lower Peninsula of Michigan during the summer months of 2002 and 2003. These ulcerations were associated with, and most likely caused by, leech parasitism. Through the use of morphological dichotomous keys, it was determined that all leeches collected are of one species: Myzobdella lugubris. Among the eight lakes examined, Lake Orion and Devils Lake had the highest prevalence of leech parasitism (34% and 29%, respectively) and mouth ulcerations (53% and 68%, respectively). Statistical analyses demonstrated that leech and ulcer prevalence varied significantly from one lake to the other. Additionally, it was determined that the relationship between the prevalence of ulcers and the prevalence of leech attachment is significant, indicating that leech parasitism is most likely the cause of ulceration. The ulcers exhibited deep hemorrhagic centers and raised irregular edges. Affected areas lost their epithelial lining and submucosa, with masses of bacteria colonizing the damaged tissues. Since largemouth bass is a popular global sportfish and critical to the food web of inland lakes, there are concerns that the presence of leeches, damaged buccal mucosa, and general unsightliness may negatively affect this important sportfishery. PMID:21395209

  20. Bleeding and cupping.

    PubMed Central

    Turk, J. L.; Allen, E.

    1983-01-01

    Bleeding and cupping have been used in medicine since ancient times in the treatment of fevers and local inflammatory disorders. Local bleeding, by 'wet cupping', was effected by a scarificator or by leeches. John Hunter recommended venesection in moderation but preferred leeches for local bleeding. Bleeding as an accepted therapeutic practice went out of vogue in the middle of the nineteenth century as a result of the introduction of modern scientific methods. Dry cupping and the use of leeches, as counter irritants, persisted until the middle of this century. Images Fig. 1 Fig. 2 Fig. 3 Fig. 4 Fig. 5 Fig. 6 PMID:6338802

  1. Head lice

    MedlinePlus

    ... PA: Elsevier Saunders; 2015:chap 294. Elston DM. Arthropods and leeches. In: Goldman L, Schafer AI, eds. ... 15. Kliegman RM, Stanton BF, St Geme JW. Arthropods bites and infestations. In: Kliegman RM, Stanton BF, ...

  2. Trypanosoma humboldti n. sp. from the Chilean catshark, Schroederichthys chilensis (Guichenot, 1848).

    PubMed

    Morillas, J; George-Nascimento, M; Valeria, H; Khan, R A

    1987-08-01

    The morphology of Trypanosoma humboldti n. sp. is described from living and stained specimens obtained from the blood of a catshark, Schroederichthys chilensis. This represents the first report of a trypanosome in fish from the eastern Pacific Ocean. It is distinguished by its size and apparent lack of pleomorphism. The presence of a leech, Branchellion ravenellii, attached to the catshark, raises the possibility that it can act as a vector. Additionally, this leech is recorded for the first time from the Pacific Ocean.

  3. Multiple pseudolymphomas caused by Hirudo medicinalis therapy.

    PubMed

    Smolle, J; Cerroni, L; Kerl, H

    2000-11-01

    Therapy with medicinal leeches (Hirudo medicinalis) is now frequently applied in plastic surgery and in the management of chronic venous insufficiency. We observed a patient in whom firm, brown-red, pea-sized papules developed at each site where leeches had been applied on the lower legs. Histology, immunohistology, and molecular analysis of T-cell receptor and immunoglobulin heavy chain gene rearrangement proved these lesions to be follicular pseudolymphomas.

  4. Hirudo Medicinalis and the plastic surgeon.

    PubMed

    Whitaker, I S; Izadi, D; Oliver, D W; Monteath, G; Butler, P E

    2004-06-01

    Medicinal leech therapy is an ancient craft that dates back to ancient Egypt and the beginnings of civilisation. The popularity of Hirudo Medicinalis has varied throughout history, reaching such a peak in Europe in the early 19th century that supplies were exhausted. During the latter half of the 19th century, their use fell out of favour, as they did not fit in with the emerging concepts of modern medicine. Leeches have enjoyed a renaissance in the world of reconstructive microsurgery during recent years, and their first reported use in alleviating venous engorgement following flap surgery was reported in this journal [M Derganc, F Zdravic, Venous congestion of flaps treated by application of leeches, Br J Plast Surg 13 (1960) 187]. Contemporary plastic and reconstructive surgeons in units throughout the United Kingdom and Ireland continue to use leeches to aid salvage of failing flaps. We carried out a survey of all 62 plastic surgery units in the United Kingdom and the Republic of Ireland to assess the current extent of use, and to investigate current practice. We have shown that the majority of plastic surgery units in the UK and Ireland use leeches post-operatively and that the average number of patients requiring leech therapy was 10 cases per unit per year. Almost all units use antibiotic prophylaxis, but the type of antibiotic and combination used is variable. We outline current practice and suggest a protocol for the use of leeches. Whilst the use of leeches is widespread, the plastic surgery community has progressed little in defining indications for their use or in achieving an accepted protocol for their application in units throughout the UK and Ireland.

  5. Hirudotherapy in veterinary medicine.

    PubMed

    Sobczak, Natalia; Kantyka, Magdalena

    2014-01-01

    The saliva of medicinal leeches, e.g., Hirudo medicinalis and Hirudo verbana commonly used in hirudotherapy, contains more than 100 bioactive substances with various therapeutic effects, including anticoagulant, vasodilator, thrombolytic, anti-inflammatory and anaesthetic properties. Recently, leeches have been used very successfully in veterinary medicine to treat many diseases of animals, especially dogs, cats and horses. The most common indications for the use of leeches are hip and elbow dysplasia, acute and chronic arthritis, diseases associated with inflammation of tendons, ligaments, and fascia, diseases of the vertebrae and the treatment of scars. Leech therapy is a painless procedure which takes an average of 30 to 120 minutes, the time being dependent on the size of the animal. All leeches used in medical procedures should originate only from certified biofarms. The maintenance of sterile conditions for the culture, transport and storage of medical leeches is very important to protect animals from microbial infections. Hirudotherapy is successfully used in veterinary medicine, especially when traditional treatment is not effective, the effects of treatment are too slow, or after surgery, when the tissues may be threatened by venous congestion.

  6. Which way is up? Asymmetric spectral input along the dorsal-ventral axis influences postural responses in an amphibious annelid.

    PubMed

    Jellies, John

    2014-11-01

    Medicinal leeches are predatory annelids that exhibit countershading and reside in aquatic environments where light levels might be variable. They also leave the water and must contend with terrestrial environments. Yet, leeches generally maintain a dorsal upward position despite lacking statocysts. Leeches respond visually to both green and near-ultraviolet (UV) light. I used LEDs to test the hypothesis that ventral, but not dorsal UV would evoke compensatory movements to orient the body. Untethered leeches were tested using LEDs emitting at red (632 nm), green (513 nm), blue (455 nm) and UV (372 nm). UV light evoked responses in 100 % of trials and the leeches often rotated the ventral surface away from it. Visible light evoked no or modest responses (12-15 % of trials) and no body rotation. Electrophysiological recordings showed that ventral sensilla responded best to UV, dorsal sensilla to green. Additionally, a higher order interneuron that is engaged in a variety of parallel networks responded vigorously to UV presented ventrally, and both the visible and UV responses exhibited pronounced light adaptation. These results strongly support the suggestion that a dorsal light reflex in the leech uses spectral comparisons across the dorsal-ventral axis rather than, or in addition to, luminance.

  7. Evidence of ectoparasite-induced endocrine disruption in an imperiled giant salamander, the eastern hellbender (Cryptobranchus alleganiensis).

    PubMed

    DuRant, Sarah E; Hopkins, William A; Davis, Andrew K; Romero, L Michael

    2015-07-01

    Parasitic leeches and trypanosomes release chemical signals into their hosts to evade immuno-detection, but it is unknown whether these compounds manipulate host behavior or endocrine physiology. We determined whether parasitic infections with leeches and/or trypanosomes affected the immune and stress response of an imperiled giant species of amphibian, the eastern hellbender (Cryptobranchus alleganiensis Daudin). We monitored corticosterone and white blood cell counts in response to restraint and injection with adrenocorticotropin hormone (ACTH) or saline for up to 50 h. The presence of leeches dampened hellbender corticosterone responses to restraint and reduced diel patterns of plasma corticosterone. Injection with ACTH restored the normal inter-renal responses of hellbenders, suggesting that leeches, possibly through neurotransmitters in leech saliva, cause down-regulation of corticosterone release at the level of the pituitary or hypothalamus. Infection with leeches also increased the relative abundance of eosinophils, white blood cells often recruited into circulation in response to parasitic infection. Lastly, neutrophil to lymphocyte (N:L) ratios increased in all animals after 24 h of capture and remained elevated for up to 50 h, but these temporal dynamics did not differ with parasite infection. Trypanosome infection did not affect any aspect of hellbender physiology that we measured. Our findings reveal a previously undocumented host-parasite dynamic. While the functional significance to the parasite is unclear, the physiological and behavioral implications for the host are great, given the important role of glucocorticoids in regulating physiology and behavior.

  8. Bacterial flora of Hirudo medicinalis and their antibiotic sensitivities in the Middle Black Sea Region, Turkey.

    PubMed

    Eroglu, C; Hokelek, M; Guneren, E; Esen, S; Pekbay, A; Uysal, O A

    2001-07-01

    The rate of infectious complications of leech therapy is almost 20% because Hirudo medicinalis has endosymbiotic bacteria. The aim of this study was to determine the bacterial flora of H. medicinalis and their antibiotic sensitivities in a region in Turkey. Sixteen adult leeches were collected in Middle Black Sea region, Turkey. They were rubbed onto blood agar plates directly under ether anesthesia to obtain surface cultures. They were then killed to obtain mouth and gut cultures. Culture swabs were applied to blood agar, eosin methylene blue agar, and ampicillin blood agar plates. Gut contents were applied to blood culture medium as well. Bacteria were isolated in 15 of 16 leech surfaces, in 7 of 16 mouths, and in 15 of 16 guts. Isolated bacteria were identified with Analytical Profile Index 32 E and Analytical Profile Index 20 NE (fermentative and nonfermentative respectively). Most common types of cultured bacteria were Aeromonas hydrophila (N = 25), Ochrobacter anthropi (N = 23), nonfermenting Gram-negative rods (N = 12), Acinetobacter lwoffi (N = 3), and A. sobria (N = 2) in 73 isolates. A standard disk diffusion test was performed on isolated bacteria. All isolates were 100% susceptible to ciprofloxacin, cefotaxime, ceftazidime, gentamicin, and trimethoprim/sulfamethoxazole. Because leeches are carriers of Aeromonas and other bacteria, appropriate antibiotic prophylaxis should be administrated to the patient who needs leech therapy. Antibacterial agents can be determined by the resistance pattern of the bacterial flora of regional H. medicinalis.

  9. The Hirudo medicinalis species complex

    NASA Astrophysics Data System (ADS)

    Kutschera, U.

    2012-05-01

    Recently, Hildebrandt and Lemke (Naturwissenschaften 98:995-1008, 2011) argued that the taxonomic status of the three European medicinal leeches, Hirudo medicinalis Linnaeus 1758, Hirudo verbana Carena 1820, and Hirudo orientalis Utevsky and Trontelj (Parasitol Res 98:61-66, 2005) is "questionable" since "all three species interbreed in the laboratory". This statement is in conflict with data published by Elliott and Kutschera (Freshwater Reviews 4:21-41, 2011), indicating that these leeches, which are reciprocally copulating hermaphrodites, represent reproductively isolated biospecies. Here, I summarize evidence indicating that these three European taxa, plus the North African "dragon leech" ( Hirudo troctina Johnson 1816), must be interpreted as a complex of closely related species, and that the economically most important taxon H. verbana is polymorphic.

  10. The Hirudo medicinalis species complex.

    PubMed

    Kutschera, U

    2012-05-01

    Recently, Hildebrandt and Lemke (Naturwissenschaften 98:995-1008, 2011) argued that the taxonomic status of the three European medicinal leeches, Hirudo medicinalis Linnaeus 1758, Hirudo verbana Carena 1820, and Hirudo orientalis Utevsky and Trontelj (Parasitol Res 98:61-66, 2005) is "questionable" since "all three species interbreed in the laboratory". This statement is in conflict with data published by Elliott and Kutschera (Freshwater Reviews 4:21-41, 2011), indicating that these leeches, which are reciprocally copulating hermaphrodites, represent reproductively isolated biospecies. Here, I summarize evidence indicating that these three European taxa, plus the North African "dragon leech" (Hirudo troctina Johnson 1816), must be interpreted as a complex of closely related species, and that the economically most important taxon H. verbana is polymorphic.

  11. Partitioned Analysis for Multidisciplinary Problems Involving Structures.

    DTIC Science & Technology

    1986-04-30

    ATTN: R FRANK JOINT STRAT TGT PLANNING STAFF BDM CORP ATTN: JK (ATTN: DNA REP) ATTN: F LEECH LAWRENCE LIVERMORE NATIONAL LABORATORY BDM CORPORATION...KAMAN SCIENCES CORPATTN: LIBRARY/B. KINSLOW AIR FORCE INSTITUTE OF TECHNOLOGY/EN KAMAN SCIENCES CORPORATION ATTN: LIBRARY/AFIT/LDEE ATTN: DASIAC AIR

  12. Laboratory-Scale Airblast Precursor Experiments. Volume 3. HOB (Height of Burst) Studies Micro-Mach Experiments

    DTIC Science & Technology

    1989-03-31

    HEADQUARTERS USAF/IN ATTN: IN RM 4A932 APPLIED RESEARCH ASSOCIATES, INCATTN: R FRANK SECRETARY OF AF/AQQS ATTN: AF/RDQI BDM CORPATTN: F LEECH STRATEGIC...SOUTHWEST RESEARCH INSTITUTE ATTN: B KINSLOW ATTN: A WENZEL KAMAN SCIENCES CORP SRI INTERNATIONAL ATTN: E CONRAD ATTN: J COLTON KAMAN SCIENCES CORPORATION

  13. High Speed Imaging using Nanoprobe Arrays

    DTIC Science & Technology

    2010-06-23

    6774 2006. 13 B. Gotsmann, U. Dürig, J. Frommer , and C. J. Hawker, Adv. Funct. Mater. 16, 1499 2006. 14 S. Bakbak, P. J. Leech, B. E. Carson, S...2006. [36] B. Gotsmann, U. Dürig, J. Frommer , and C. J. Hawker, “Exploiting chemical switching in a Diels-Alder polymer for nanoscale probe litho

  14. Compare and Contrast Inductive and Deductive Research Approaches

    ERIC Educational Resources Information Center

    Soiferman, L. Karen

    2010-01-01

    This discussion paper compares and contrasts "inductive" and "deductive" research approaches as described by Trochim (2006) and Plano Clark and Creswell (2007). It also examines the "exploratory" and "confirmatory" approaches by Onwueghuzie and Leech (2005) with respect to the assumption each holds about the nature of knowledge. The paper starts…

  15. Predicting High School Completion Using Student Performance in High School Algebra: A Mixed Methods Research Study

    ERIC Educational Resources Information Center

    Chiado, Wendy S.

    2012-01-01

    Too many of our nation's youth have failed to complete high school. Determining why so many of our nation's students fail to graduate is a complex, multi-faceted problem and beyond the scope of any one study. The study presented herein utilized a thirteen-step mixed methods model developed by Leech and Onwuegbuzie (2007) to demonstrate within a…

  16. Qualitative Analysis Techniques for the Review of the Literature

    ERIC Educational Resources Information Center

    Onwuegbuzie, Anthony J.; Leech, Nancy L.; Collins, Kathleen M. T.

    2012-01-01

    In this article, we provide a framework for analyzing and interpreting sources that inform a literature review or, as it is more aptly called, a research synthesis. Specifically, using Leech and Onwuegbuzie's (2007, 2008) frameworks, we delineate how the following four major source types inform research syntheses: talk, observations,…

  17. 76 FR 11494 - List of Recipients of Indian Health Scholarships Under the Indian Health Scholarship Program

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-03-02

    ... Rocky Boy's Reservation, Montana Bernard, Kenneth Richard Lee, Harvard Medical School, Turtle Mountain... Minnesota, Leech Lake Band, Minnesota Chippewa Tribe, Minnesota Heth (Taylor), Talara K., Connors State... Charles, University of Buffalo, St. Regis Band of Mohawk Indians of New York Taylor, Tara Lynn, Lewis and...

  18. Silicon-Neuron Junction: Capacitive Stimulation of an Individual Neuron on a Silicon Chip

    NASA Astrophysics Data System (ADS)

    Fromherz, Peter; Stett, Alfred

    1995-08-01

    An identified nerve cell of the leech is attached to a planar silicon microstructure of p-doped silicon covered by a thin layer of insulating silicon oxide. A voltage step, applied between silicon and electrolyte, induces a capacitive transient in the cell which elicits an action potential. The capacitive extracellular stimulation is described by an equivalent electrical four-pole.

  19. Hitchhiking of host biology by beneficial symbionts enhances transmission

    PubMed Central

    Ott, Brittany M.; Cruciger, Michael; Dacks, Andrew M.; Rio, Rita V. M.

    2014-01-01

    Transmission plays a key role in the evolution of symbiosis. Mixed mode transmission combines horizontal and vertical mechanisms for symbiont acquisition. However, features that enable mixed transmission are poorly understood. Here, we determine the mechanistic basis for the recruitment of the beneficial bacterium, Aeromonas veronii by the leech, Hirudo verbana. We demonstrate that host mucosal secretions complement imperfect symbiont vertical transmission. First, we show that the A. veronii population within secretions originates from the host digestive tract and proliferates synchronously with shedding frequency, demonstrating the coupling of partner biology. Furthermore, leeches are attracted to these castings with oral contact proving sufficient for symbiont transmission. Leech attraction to mucus is not affected by the symbiont state of either the host or mucus, suggesting that A. veronii exploits preexisting host behavior and physiological traits. A dual transmission mode, integrating multiple layers of host contributions, may prove evolutionarily advantageous for a wide range of symbioses. Using such a strategy, host infection is ensured, while also providing access to a higher genetic diversity of symbionts. Countless host-associated microbes exhibit mixed mode transmission, supporting the use of the leech symbiosis as a model for enhancing our understanding of the specificity, establishment and persistence of microbiotas. PMID:25059557

  20. 77 FR 2085 - Notice of Inventory Completion: Minnesota Indian Affairs Council, Bemidji, MN

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-01-13

    ... Leech Lake Band of the Minnesota Chippewa Tribe, Minnesota; the Minnesota Chippewa Tribe, Minnesota; and... removed from the Rock Lake area, Becker County, MN, by unknown person(s) and donated to the Becker County... from the north shore of Detroit Lake (Steffen Lot), Becker County, MN, by unknown person(s)...

  1. The Most Frequently Used English Phrasal Verbs in American and British English: A Multicorpus Examination

    ERIC Educational Resources Information Center

    Liu, Dilin

    2011-01-01

    This study uses the Corpus of Contemporary American English and the British National Corpus as data and Biber, Johansson, Leech, Conrad, and Finegan's (1999) and Gardner and Davies' (2007) informative studies as a starting point and reference. The study offers a cross-English variety and cross-register examination of the use of English phrasal…

  2. Politeness Principle in Cross-Culture Communication

    ERIC Educational Resources Information Center

    Huang, Yongliang

    2008-01-01

    As we all know, different people hold different views about politeness. To be polite, Leech thinks you should follow "Politeness Principle" while Levinson suggests paying attention to others' "Face Wants". Sometimes what the Chinese people considered to be polite may not be true according to western culture. In order to…

  3. 77 FR 2083 - Notice of Inventory Completion: Minnesota Indian Affairs Council, Bemidji, MN

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-01-13

    ..., one individual were recovered near Ice Cracking Lake during road construction and donated to the... professional staff in consultation with representatives of the Leech Lake Band of the Minnesota Chippewa Tribe... individual were recovered from the north side of Height of Land Lake, site 21BK48, by Mr. William Krause...

  4. Three-dimensional visualisation of developmental stages of an apicomplexan fish blood parasite in its invertebrate host

    PubMed Central

    2011-01-01

    Background Although widely used in medicine, the application of three-dimensional (3D) imaging to parasitology appears limited to date. In this study, developmental stages of a marine fish haemogregarine, Haemogregarina curvata (Apicomplexa: Adeleorina), were investigated in their leech vector, Zeylanicobdella arugamensis; this involved 3D visualisation of brightfield and confocal microscopy images of histological sections through infected leech salivary gland cells. Findings 3D assessment demonstrated the morphology of the haemogregarine stages, their spatial layout, and their relationship with enlarged host cells showing reduced cellular content. Haemogregarine meronts, located marginally within leech salivary gland cells, had small tail-like connections to the host cell limiting membrane; this parasite-host cell interface was not visible in two-dimensional (2D) light micrographs and no records of a similar connection in apicomplexan development have been traced. Conclusions This is likely the first account of the use of 3D visualisation to study developmental stages of an apicomplexan parasite in its invertebrate vector. Elucidation of the extent of development of the haemogregarine within the leech salivary cells, together with the unusual connections between meronts and the host cell membrane, illustrates the future potential of 3D visualisation in parasite-vector biology. PMID:22107751

  5. The Paradox of Fight or Flight - A Leadership Guide to Understanding and Mitigating Operational Stress Injuries

    DTIC Science & Technology

    2006-04-01

    reproduction , both parents transfer sequences or copied portions of their DNA to their offspring. A particular type of DNA - called mitochondrial DNA - is...strength, willpower and self-control.59 To remedy a nostalgic patient, doctors prescribed purging, opium, leeches, and hypnotic emulsions. Less

  6. The Most Frequently Used English Phrasal Verbs in American and British English: A Multicorpus Examination

    ERIC Educational Resources Information Center

    Liu, Dilin

    2011-01-01

    This study uses the Corpus of Contemporary American English and the British National Corpus as data and Biber, Johansson, Leech, Conrad, and Finegan's (1999) and Gardner and Davies' (2007) informative studies as a starting point and reference. The study offers a cross-English variety and cross-register examination of the use of English phrasal…

  7. GABA-ergic neurons in the leach central nervous system

    SciTech Connect

    Cline, H.T.

    1985-01-01

    GABA is a candidate for an inhibitory neurotransmitter in the leech central nervous system because of the well-documented inhibitory action of GABA in other invertebrates. To demonstrate that GABA meets the criteria used to identify a substance as a neurotransmitter, the author examined GABA metabolism and synaptic interactions of inhibitory motor neurons in two leech species, Hirudo medicinalis and Haementeria ghilianii. Segmental ganglia of the leech ventral nerve cord and identified inhibitors have the capacity to synthesize GABA when incubated in the presence of the precursor glutamate. Application of GABA to cell bodies of excitatory motor neurons or muscle fibers innervated by the inhibitors hyperpolarizes the membrane potential of the target cell and activates a chloride ion conductance channel, similar to the inhibitory membrane response following intracellular stimulation of the inhibitor. Bicuculline methiodide (5 x 10/sup -5/M), GABA receptor antagonist, blocks reversibly the response to applied GABA and the inhibitory synaptic inputs onto the postsynaptic neurons or muscle fibers without interfering with their excitatory inputs. Furthermore, the inhibitors are included among approximately 25 neurons per segmental ganglion that take up GABA by a high affinity uptake system, as revealed by /sup 3/H-GABA-autoradiography. The development of the capacities to synthesize and to take up GABA were examined in leech embryos. The embryos are able to synthesize GABA at early stages of the development of the nervous system, before any neurons have extended neutrites.

  8. Seventh Annual Distinguished Lecture Series in Special Education and Rehabilitation; Summer Session 1968.

    ERIC Educational Resources Information Center

    MacLeech, Bert, Ed.; Schrader, Donald R., Ed.

    Lectures are concerned with the nurturing of intellect by Maurice Freehill, a new concept in rehabilitation by Bert MacLeech, principles of residential therapy as a tool of rehabilitation by Edward L. French, the progress of special education by Romaine P. Mackie, and behavior principles by Norris G. Haring. The priorities and territories of…

  9. Characterizations of Hirudo medicinalis DNA promoters for targeted gene expression.

    PubMed

    Baker, Michael W; Macagno, Eduardo R

    2006-09-30

    The expression of exogenous genes in neurons and other cells has become a powerful means for studying the function of encoded proteins. We report here on the isolation and functional analysis of three Hirudo medicinalis actin gene promoters and the 5' UTR of a leech elongation factor-1alpha (HmEF-1alpha) gene. In situ hybridization labeling revealed that the EF-1alpha gene and one of the actins had pan-neuronal expression, whereas, the other two actin genes were expressed by the embryo's body wall musculature. Comparative analysis shows that they all display many features typical of actin and EF-1alpha promoters from other species, including canonical TATA box sequences and predicted general transcription factor binding sites (such as CCATT, CarB boxes and CG-rich regions). The ability of these 5' UTR sequences to drive expression of the enhanced green fluorescent protein (EGFP), leech cytoplasmic actin and leech synaptobrevin was examined. Direct intracellular nuclear, but not cytoplasmic, microinjection of each of the promoter sequences was found to produce reliably cellular expression of the reporter construct in both neuronal and muscle cells. These results introduce reliable and effective methods to selectively express genes in individual cells of the leech in vivo during embryonic development.

  10. Antimicrobial prophylaxis during Hirudo medicinalis therapy: a multicenter study.

    PubMed

    Kruer, Rachel M; Barton, Cassie A; Roberti, Gregory; Gilbert, Brian; McMillian, Wesley D

    2015-03-01

    Medicinal leeches (Hirudo medicinalis) are indicated for salvage of tissue flaps, grafts, or replants when venous congestion threatens tissue viability. The purpose of this study was to evaluate the efficacy of prophylactic antimicrobial agents in patients who received medicinal leech therapy. A multicenter retrospective cohort study of all adult patients between January 1, 2010, and February 28, 2013, who received medicinal leech therapy was conducted. Antimicrobial prophylaxis was documented in 54 (91.5%) of the included patients, ciprofloxacin, trimethoprim-sulfamethoxazole, piperacillin-tazobactam, and ceftriaxone in 33 (61.1%), 18 (33.3%), 2 (3.7%), and 2 (3.7%) patients, respectively. Surgical site infection (SSI) was found in seven (11.9%) patients, all of whom received antimicrobial prophylaxis. Aeromonas spp. was isolated in four infections, and all isolates were resistant to the chosen prophylactic agent. The SSI incidence was similar between antimicrobial prophylaxis agents. Trimethoprim-sulfamethoxazole and ciprofloxacin appear equally effective at preventing leech-associated infections. Thieme Medical Publishers 333 Seventh Avenue, New York, NY 10001, USA.

  11. Pragmatic Transfer.

    ERIC Educational Resources Information Center

    Kasper, Gabriele

    1992-01-01

    Attempting to clarify the concept of pragmatic transfer, this article proposes as a basic distinction Leech/Thomas' dichotomy of sociopragmatics versus pragmalinguistics, presenting evidence for transfer at both levels. Issues discussed include pragmatic universals in speech act realization, conditions for pragmatic transfer, communicative…

  12. Doctoral Students' Reasons for Reading Empirical Research Articles: A Mixed Analysis

    ERIC Educational Resources Information Center

    Burgess, Melissa L.; Benge, Cindy; Onwuegbuzie, Anthony J.; Mallette, Marla H.

    2012-01-01

    Little is known about reading ability among doctoral students. Thus, we used a fully mixed concurrent equal status design (Leech & Onwuegbuzie, 2009) to examine 205 doctoral students in the College of Education and their reasons for reading research articles. A thematic analysis revealed 5 themes (subsumed by 2 meta-themes) explaining reasons…

  13. Study on the Effects of Diverting Water into Upper Burnt Pocket, Navigation Pool Number 18, Illinois and A Field Test of the Regression Simulation Model Previously Developed on Navigation Pool Number 8.

    DTIC Science & Technology

    1981-03-01

    earthworms ( Annelida : Oligochaeta). Pages 143-156 in C. W. Hart, Jr. and S. L. H. Fuller, eds. Pollution ecology of freshwater invertebrates...Site, Navigation Pool No. 18, Upper Mississippi River, between 1978 and 1980. NEMATODA round worms ANNELIDA Oligochaeta earth worms Hirudinea leeches

  14. Cell dialysis by sharp electrodes can cause nonphysiological changes in neuron properties.

    PubMed

    Hooper, Scott L; Thuma, Jeffrey B; Guschlbauer, Christoph; Schmidt, Joachim; Büschges, Ansgar

    2015-08-01

    We recorded from lobster and leech neurons with two sharp electrodes filled with solutions often used with these preparations (lobster: 0.6 M K2SO4 or 2.5 M KAc; leech: 4 M KAc), with solutions approximately matching neuron cytoplasm ion concentrations, and with 6.5 M KAc (lobster, leech) and 0.6 M KAc (lobster). We measured membrane potential, input resistance, and transient and sustained depolarization-activated outward current amplitudes in leech and these neuron properties and hyperpolarization-activated current time constant in lobster, every 10 min for 60 min after electrode penetration. Neuron properties varied with electrode fill. For fills with molarities ≥2.5 M, neuron properties also varied strongly with time after electrode penetration. Depending on the property being examined, these variations could be large. In leech, cell size also increased with noncytoplasmic fills. The changes in neuron properties could be due to the ions being injected from the electrodes during current injection. We tested this possibility in lobster with the 2.5 M KAc electrode fill by making measurements only 10 and 60 min after penetration. Neuron properties still changed, although the changes were less extreme. Making measurements every 2 min showed that the time-dependent variations in neuron properties occurred in concert with each other. Neuron property changes with high molarity electrode-fill solutions were great enough to decrease neuron firing strongly. An experiment with (14)C-glucose electrode fill confirmed earlier work showing substantial leak from sharp electrodes. Sharp electrode work should thus be performed with cytoplasm-matched electrode fills. Copyright © 2015 the American Physiological Society.

  15. Haematological and immunological characteristics of eastern hellbenders (Cryptobranchus alleganiensis alleganiensis) infected and co-infected with endo- and ectoparasites

    PubMed Central

    Hopkins, William A.; Fallon, Jesse A.; Beck, Michelle L.; Coe, Brittney H.; Jachowski, Catherine M. B.

    2016-01-01

    Disease is among the leading causes of the global decline in amphibian populations. In North America, parasites and pathogens are among the factors implicated in precipitous population declines of the giant hellbender salamander (Cryptobranchus alleganiensis), but the incidence of infections and the responses of hellbenders to infections remain poorly studied. Here, we document the prevalence of leech and trypanosome infections in a wild population of eastern hellbenders (Cryptobranchus alleganiensis alleganiensis) and describe haematological and immunological characteristics of hellbenders harbouring these infections. We hypothesized that hellbenders parasitized by trypanosomes would be anaemic, that individuals infected with either or both parasites would exhibit shifts in white blood cell counts and that hellbenders infected with leeches would exhibit altered plasma bactericidal capacity. We found that 24 and 68% of hellbenders in our sample population were infected with leeches and trypanosomes, respectively, and 20% were co-infected with both parasites. We found no evidence suggestive of anaemia among infected individuals. However, hellbenders infected with either or both parasites exhibited marked shifts in circulating white blood cells that were consistent with predictable responses to parasitic infection. Additionally, we found that hellbenders harbouring leeches had much higher plasma bactericidal capacity than individuals without leeches, and we offer multiple potential mechanistic explanations for this observation. We also found evidence that cellular and serological immune responses to parasites were less robust in juvenile than adult hellbenders. This finding warrants further investigation in light of the demographic characteristics, specifically the scarcity of juvenile age classes, of hellbender populations where disease is a possible contributor to declines. Finally, we describe two methodological advances that will improve future studies seeking to

  16. Haematological and immunological characteristics of eastern hellbenders (Cryptobranchus alleganiensis alleganiensis) infected and co-infected with endo- and ectoparasites.

    PubMed

    Hopkins, William A; Fallon, Jesse A; Beck, Michelle L; Coe, Brittney H; Jachowski, Catherine M B

    2016-01-01

    Disease is among the leading causes of the global decline in amphibian populations. In North America, parasites and pathogens are among the factors implicated in precipitous population declines of the giant hellbender salamander (Cryptobranchus alleganiensis), but the incidence of infections and the responses of hellbenders to infections remain poorly studied. Here, we document the prevalence of leech and trypanosome infections in a wild population of eastern hellbenders (Cryptobranchus alleganiensis alleganiensis) and describe haematological and immunological characteristics of hellbenders harbouring these infections. We hypothesized that hellbenders parasitized by trypanosomes would be anaemic, that individuals infected with either or both parasites would exhibit shifts in white blood cell counts and that hellbenders infected with leeches would exhibit altered plasma bactericidal capacity. We found that 24 and 68% of hellbenders in our sample population were infected with leeches and trypanosomes, respectively, and 20% were co-infected with both parasites. We found no evidence suggestive of anaemia among infected individuals. However, hellbenders infected with either or both parasites exhibited marked shifts in circulating white blood cells that were consistent with predictable responses to parasitic infection. Additionally, we found that hellbenders harbouring leeches had much higher plasma bactericidal capacity than individuals without leeches, and we offer multiple potential mechanistic explanations for this observation. We also found evidence that cellular and serological immune responses to parasites were less robust in juvenile than adult hellbenders. This finding warrants further investigation in light of the demographic characteristics, specifically the scarcity of juvenile age classes, of hellbender populations where disease is a possible contributor to declines. Finally, we describe two methodological advances that will improve future studies seeking to

  17. Cell dialysis by sharp electrodes can cause nonphysiological changes in neuron properties

    PubMed Central

    Hooper, Scott L.; Guschlbauer, Christoph; Schmidt, Joachim; Büschges, Ansgar

    2015-01-01

    We recorded from lobster and leech neurons with two sharp electrodes filled with solutions often used with these preparations (lobster: 0.6 M K2SO4 or 2.5 M KAc; leech: 4 M KAc), with solutions approximately matching neuron cytoplasm ion concentrations, and with 6.5 M KAc (lobster, leech) and 0.6 M KAc (lobster). We measured membrane potential, input resistance, and transient and sustained depolarization-activated outward current amplitudes in leech and these neuron properties and hyperpolarization-activated current time constant in lobster, every 10 min for 60 min after electrode penetration. Neuron properties varied with electrode fill. For fills with molarities ≥2.5 M, neuron properties also varied strongly with time after electrode penetration. Depending on the property being examined, these variations could be large. In leech, cell size also increased with noncytoplasmic fills. The changes in neuron properties could be due to the ions being injected from the electrodes during current injection. We tested this possibility in lobster with the 2.5 M KAc electrode fill by making measurements only 10 and 60 min after penetration. Neuron properties still changed, although the changes were less extreme. Making measurements every 2 min showed that the time-dependent variations in neuron properties occurred in concert with each other. Neuron property changes with high molarity electrode-fill solutions were great enough to decrease neuron firing strongly. An experiment with 14C-glucose electrode fill confirmed earlier work showing substantial leak from sharp electrodes. Sharp electrode work should thus be performed with cytoplasm-matched electrode fills. PMID:26063785

  18. Hirudo medicinalis: a platform for investigating genes in neural repair.

    PubMed

    Wang, W Z; Emes, R D; Christoffers, K; Verrall, J; Blackshaw, S E

    2005-03-01

    We have used the nervous system of the medicinal leech as a preparation to study the molecular basis of neural repair. The leech central nervous system, unlike mammalian CNS, can regenerate to restore function, and contains identified nerve cells of known function and connectivity. We have constructed subtractive cDNA probes from whole and regenerating ganglia of the ventral nerve cord and have used these to screen a serotonergic Retzius neuron library. This identifies genes that are regulated as a result of axotomy, and are expressed by the Retzius cell. This approach identifies many genes, both novel and known. Many of the known genes identified have homologues in vertebrates, including man. For example, genes encoding thioredoxin (TRX), Rough Endoplasmic Reticulum Protein 1 (RER-1) and ATP synthase are upregulated at 24 h postinjury in leech nerve cord. To investigate the functional role of regulated genes in neuron regrowth we are using microinjection of antisense oligonucleotides in combination with horseradish peroxidase to knock down expression of a chosen gene and to assess regeneration in single neurons in 3-D ganglion culture. As an example of this approach we describe experiments to microinject antisense oligonucleotide to a leech isoform of the structural protein, Protein 4.1. Our approach thus identifies genes regulated at different times after injury that may underpin the intrinsic ability of leech neurons to survive damage, to initiate regrowth programs and to remake functional connections. It enables us to determine the time course of gene expression in the regenerating nerve cord, and to study the effects of gene knockdown in identified neurons regenerating in defined conditions in culture.

  19. Genetic variation in Whitmania pigra, Hirudo nipponica and Poecilobdella manillensis, three endemic and endangered species in China using SSR and TRAP markers.

    PubMed

    Liu, Fei; Guo, Qiao-Sheng; Shi, Hong-Zhuan; Cheng, Bo-Xing; Lu, Yu-Xi; Gou, Ling; Wang, Jia; Shen, Wen-Biao; Yan, Shi-Meng; Wu, Man-Jun

    2016-04-01

    Leeches are not only important medicinal animals worldwide but also are endangered. We aimed to (i) explore the level of genetic diversity within/among populations of three leeches, (ii) assess genetic differentiation among these three leeches, and (iii) discuss an appropriate strategy for conserving leech germplasm. A total of 315 individuals of Whitmania pigra, Hirudo nipponica and Poecilobdella manillensis from 21 populations were collected in China and Vietnam. The genetic structure and genetic diversity among and within the 21 populations were evaluated using target region amplified polymorphism (TRAP) and simple sequence repeat (SSR) markers. Sixteen pairs of TRAP primers generated a total of 398 fragments, of which 396 (99.50%) were polymorphic; fourteen pairs of SSR primers generated a total of 60 fragments, of which 59 (98.33%) were polymorphic. Shannon's index (I) and Nei's gene diversity index (H) for the three leeches were high at the species level (I=0.4980 and H=0.3323 for TRAPs, I=0.4487 and H=0.2969 for SSRs in W. pigra; I=0.4147/0.3769, H=0.2788/0.2566 for H. nipponica; and I=0.4616/0.4717, H=0.3099/0.3203 for P. manillensis). However, low genetic diversity was determined at the population level; the average genetic diversity measures within populations were H=0.1767/0.1376, I=0.2589/0.2043 for W. pigra, H=0.2149/0.2021, I=0.3184/0.3000 for H. nipponica and H=0.2850/0.2724, I=0.4152/0.3967 for P. manillensis. We conclude that there was limited gene exchange within/among populations and species, as the gene flow number (Nm) was 0.5493/0.5807. However, for all three species, the genetic diversity was different at the population level. Gene differentiation (Gst) and Nm were 0.4682 /0.5364 and 0.5678/0.4321 for W. pigra, 0.2294/0.2127 and 1.6797/1.8512 for H. nipponica and 0.1214/0.1496 and 3.6202/2.8412 for P. manillensis. STRUCTURE analysis, Unweighted Pair-Group Method with Arithmetic means (UPGMA) cluster analysis and Principal Coordinates Analysis

  20. Evolutionary Origin of Body Axis Segmentation in Annelids and Arthropods

    NASA Technical Reports Server (NTRS)

    Shankland, S. Martin

    2003-01-01

    During the period of this report, we have made a number of important discoveries. To date this work has led to 4 peer-reviewed publications in primary research journals plus 1 minireview and 1 chapter in the proceedings of a meeting. Publications resulting from this grant support are enumerated at the end of the report. Two additional, on-going studies also described. 1. Using laser cell ablation, we have obtained evidence that an annelid - the leech Helobdella robusta - patterns the anteroposterior (AP) polarity of its nascent segment primordia independent of cell interactions oriented along the AP axis. 2. We cloned a Helobdella homologue (hro-hh) of the Drosophila segment polarity gene hedgehog, and used in situ hybridization and northern blots to characterize its expression in the embryo. 3. We have used laser cell ablations to examine the possible role of cell interactions during the developmental patterning of the 4 rostralmost "head" segments of the leech Helobdella robusta.

  1. Fossilized spermatozoa preserved in a 50-Myr-old annelid cocoon from Antarctica

    PubMed Central

    Bomfleur, Benjamin; Mörs, Thomas; Ferraguti, Marco; Reguero, Marcelo A.; McLoughlin, Stephen

    2015-01-01

    The origin and evolution of clitellate annelids—earthworms, leeches and their relatives—is poorly understood, partly because body fossils of these delicate organisms are exceedingly rare. The distinctive egg cases (cocoons) of Clitellata, however, are relatively common in the fossil record, although their potential for phylogenetic studies has remained largely unexplored. Here, we report the remarkable discovery of fossilized spermatozoa preserved within the secreted wall layers of a 50-Myr-old clitellate cocoon from Antarctica, representing the oldest fossil animal sperm yet known. Sperm characters are highly informative for the classification of extant Annelida. The Antarctic fossil spermatozoa have several features that point to affinities with the peculiar, leech-like ‘crayfish worms' (Branchiobdellida). We anticipate that systematic surveys of cocoon fossils coupled with advances in non-destructive analytical methods may open a new window into the evolution of minute, soft-bodied life forms that are otherwise only rarely observed in the fossil record. PMID:26179804

  2. Evolutionary Origin of Body Axis Segmentation in Annelids and Arthropods

    NASA Technical Reports Server (NTRS)

    Shankland, S. Martin

    2003-01-01

    During the period of this report, we have made a number of important discoveries. To date this work has led to 4 peer-reviewed publications in primary research journals plus 1 minireview and 1 chapter in the proceedings of a meeting. Publications resulting from this grant support are enumerated at the end of the report. Two additional, on-going studies also described. 1. Using laser cell ablation, we have obtained evidence that an annelid - the leech Helobdella robusta - patterns the anteroposterior (AP) polarity of its nascent segment primordia independent of cell interactions oriented along the AP axis. 2. We cloned a Helobdella homologue (hro-hh) of the Drosophila segment polarity gene hedgehog, and used in situ hybridization and northern blots to characterize its expression in the embryo. 3. We have used laser cell ablations to examine the possible role of cell interactions during the developmental patterning of the 4 rostralmost "head" segments of the leech Helobdella robusta.

  3. A review of Biston Leach, 1815 (Lepidoptera, Geometridae, Ennominae) from China, with description of one new species

    PubMed Central

    Jiang, Nan; Xue, Dayong; Han, Hongxiang

    2011-01-01

    Abstract The genus Biston Leach, 1815 is reviewed for China. Seventeen species are recognized, of which Biston mediolata sp. n. is described. Biston pustulata (Warren, 1896) and Biston panterinaria exanthemata (Moore, 1888) are newly recorded for China. The following new synonyms are established: Biston suppressaria suppressaria (Guenée, 1858) (= Biston suppressaria benescripta (Prout, 1915), syn. n. = Biston luculentus Inoue, 1992 syn. n.); Biston falcata (Warren, 1893) (= Amphidasis erilda Oberthür, 1910, syn. n. = Amphidasis clorinda Oberthür, 1910, syn. n. = Biston emarginaria Leech, 1897, syn. n.); Biston panterinaria panterinaria (Bremer & Grey, 1853) (= Biston panterinaria abraxata (Leech, 1889), syn. n. = Biston panterinaria lienpingensis (Wehrli, 1939), syn. n. = B. panterinaria szechuanensis (Wehrli, 1939), syn. n.). Biston falcata satura (Wehrli,1941), comb. n. is proposed. A key to Chinese Biston and diagnoses for Chinese species are provided. Illustrations of external features and genitalia are presented. PMID:22259309

  4. Protein and cholesterol electrophoresis of plasma samples from captive cownose ray (Rhinoptera bonasus).

    PubMed

    Cray, Carolyn; Rodriguez, Marilyn; Field, Cara; McDermott, Alexa; Leppert, Lynda; Clauss, Tonya; Bossart, Gregory D

    2015-11-01

    Our study was undertaken to assess the application of semiautomated methods available at the reference laboratory level for the evaluation of plasma protein and cholesterol via electrophoresis in samples from cownose rays (Rhinoptera bonasus). Three groups of animals were assessed: clinically normal, clinically abnormal, and parasitized with leeches. As reported previously, the albumin band was negligible; the protein electrophoretograms were dominated by a large beta-globulin fraction. While the group of samples from the leech-parasitized rays did not show any large differences, the abnormal group exhibited significantly elevated total solids and cholesterol levels. The latter was related to a significant increase in very low density lipoprotein levels. The results demonstrate the potential application of these laboratory methods in quantitation of plasma proteins and cholesterol fractions in subclass Elasmobranchii.

  5. Biological clockwork underlying adaptive rhythmic movements

    PubMed Central

    Iwasaki, Tetsuya; Chen, Jun; Friesen, W. Otto

    2014-01-01

    Owing to the complexity of neuronal circuits, precise mathematical descriptions of brain functions remain an elusive ambition. A more modest focus of many neuroscientists, central pattern generators, are more tractable neuronal circuits specialized to generate rhythmic movements, including locomotion. The relative simplicity and well-defined motor functions of these circuits provide an opportunity for uncovering fundamental principles of neuronal information processing. Here we present the culmination of mathematical analysis that captures the adaptive behaviors emerging from interactions between a central pattern generator, the body, and the physical environment during locomotion. The biologically realistic model describes the undulatory motions of swimming leeches with quantitative accuracy and, without further parameter tuning, predicts the sweeping changes in oscillation patterns of leeches undulating in air or swimming in high-viscosity fluid. The study demonstrates that central pattern generators are capable of adapting oscillations to the environment through sensory feedback, but without guidance from the brain. PMID:24395788

  6. Fossilized spermatozoa preserved in a 50-Myr-old annelid cocoon from Antarctica.

    PubMed

    Bomfleur, Benjamin; Mörs, Thomas; Ferraguti, Marco; Reguero, Marcelo A; McLoughlin, Stephen

    2015-07-01

    The origin and evolution of clitellate annelids--earthworms, leeches and their relatives--is poorly understood, partly because body fossils of these delicate organisms are exceedingly rare. The distinctive egg cases (cocoons) of Clitellata, however, are relatively common in the fossil record, although their potential for phylogenetic studies has remained largely unexplored. Here, we report the remarkable discovery of fossilized spermatozoa preserved within the secreted wall layers of a 50-Myr-old clitellate cocoon from Antarctica, representing the oldest fossil animal sperm yet known. Sperm characters are highly informative for the classification of extant Annelida. The Antarctic fossil spermatozoa have several features that point to affinities with the peculiar, leech-like 'crayfish worms' (Branchiobdellida). We anticipate that systematic surveys of cocoon fossils coupled with advances in non-destructive analytical methods may open a new window into the evolution of minute, soft-bodied life forms that are otherwise only rarely observed in the fossil record.

  7. Acute toxic effects of two lampricides on twenty-one freshwater invertebrates

    USGS Publications Warehouse

    Rye, Robert P.; King, Everett Louis

    1976-01-01

    We conducted laboratory static bioassays to determine acute toxicity of two lampricides -- a 70% 2-aminoethanol salt of 5,2'dichloro-4'-nitrosalicylanilide (Bayer 73) and a mixture containing 98% 3-trifluoromethyl-4-nitrophenol (TFM) and 2% Bayer 73 (TFM-2B) -- to 21 freshwater invertebrates. LC50 values were determined for 24-h exposure periods at 12.8 C. Organisms relatively sensitive to Bayer 73 were a turbellarian (Dugesia tigrina), aquatic earthworms (Tubifex tubifex and Lumbriculus inconstans), snails (Physa sp.) and (Pleurocera sp.), a clam (Eliptio dilatatus), blackflies (Simulium sp.), leeches (Erpobdellidae), and a daphnid (Daphnia pulex). The invertebrates most sensitive to TFM-2B were turbellarians, aquatic earthworms (Tubifex), snails (Physa), blackflies, leeches, and burrowing mayflies (Hexagenia sp.). Bayer 73 was generally much more toxic to the test organisms than TFM-2B. At lampricidal concentrations, TFM-2B was more highly selective than Bayer 73 against larval sea lampreys (Petromyzon marinus).

  8. A new species of Oceanobdella (Hirudinida: Piscicolidae) from the plain sculpin, Myoxocephalus jaok, from Bristol Bay, Alaska.

    PubMed

    Burreson, Eugene M; Williams, Julianne I

    2004-08-01

    Oceanobdella khani n. sp. is described from the plain sculpin, Myoxocephalus jaok, from Bristol Bay, Alaska, in the eastern Bering Sea. Prevalence was never greater than 10% in any single collection; maximum intensity was 12 leeches per host, but most fish had 5 or fewer leeches. Oceanobdella khani possesses generic characters of small oral sucker, 5 pairs of testisacs, unremarkable terminal male reproductive organs, coelomic system lacking pulsatile vesicles, and 3 pairs of eyes on the oral sucker-trachelosome. Oceanobdella khani is distinguished from other species in the genus by solid black pigmentation on the urosome, clitellum, trachelosome, and most of the oral sucker except for an unpigmented margin. The pigmentation of the caudal sucker is highly variable, ranging from overall mottled blackish gray to completely unpigmented. The caudal sucker lacks ocelli. Intestinal ceca are large, crop ceca are directed anteriorly, and postceca are separate for their entire length.

  9. A review of Biston Leach, 1815 (Lepidoptera, Geometridae, Ennominae) from China, with description of one new species.

    PubMed

    Jiang, Nan; Xue, Dayong; Han, Hongxiang

    2011-01-01

    The genus Biston Leach, 1815 is reviewed for China. Seventeen species are recognized, of which Biston mediolatasp. n. is described. Biston pustulata (Warren, 1896) and Biston panterinariaexanthemata (Moore, 1888) are newly recorded for China. The following new synonyms are established: Biston suppressaria suppressaria (Guenée, 1858) (= Biston suppressaria benescripta (Prout, 1915), syn. n. = Biston luculentus Inoue, 1992 syn. n.); Biston falcata (Warren, 1893) (= Amphidasiserilda Oberthür, 1910, syn. n. = Amphidasis clorinda Oberthür, 1910, syn. n. = Biston emarginaria Leech, 1897, syn. n.); Biston panterinaria panterinaria (Bremer & Grey, 1853) (= Biston panterinaria abraxata (Leech, 1889), syn. n. = Biston panterinaria lienpingensis (Wehrli, 1939), syn. n. = B. panterinaria szechuanensis (Wehrli, 1939), syn. n.). Biston falcata satura (Wehrli,1941), comb. n. is proposed. A key to Chinese Biston and diagnoses for Chinese species are provided. Illustrations of external features and genitalia are presented.

  10. Vascular endothelial growth factor is involved in neoangiogenesis in Hirudo medicinalis (Annelida, Hirudinea).

    PubMed

    Tettamanti, G; Grimaldi, A; Valvassori, R; Rinaldi, L; de Eguileor, M

    2003-06-21

    Vascular endothelial growth factor (VEGF) is fundamental in vertebrates for correct development of blood vessels. However, there are only few data about the presence of VEGF in invertebrates. In this study the role of VEGF in neovessel formation is investigated in Hirudo medicinalis. The leech is able to respond to administration of human VEGF by formation of new vessels. The response of H. medicinalis to this growth factor is explained by the presence of two specific VEGF-like receptors (Flt-1/VEGFR-1 and Flk-1/VEGFR-2) as demonstrated by immunohistochemistry and biochemical analysis. The VEGF-like produced by this annelid following surgical stimulation determines not only blood vessel formation, proliferation of vascular endothelial cells but also an increase of cytoplasmic calcium levels. The administration of specific VEGF receptor antibodies can inhibit angiogenesis in leeches previously stimulated with VEGF.

  11. [Taxonomic diversity of parasites in agnathans and fishes from the Volga River basin. VI. Acanthocephala, Hirudinea and Bivalvia].

    PubMed

    Molodozhnikova, N M; Zhokhov, A E

    2008-01-01

    The checklist of Acanthocephala, Hirudinea, and Bivalvia parasitizing agnathans and fishes in the Volga River basin is presented. Hosts and areas of distribution are indicated for each parasites species. The checklist includes 10 species of acanthocephalans, 7 species of leeches, and 9 species of Bivalvia (at the glochidium stage) from 45 fish species. None of the given parasite species is alien for the Volga River basin.

  12. Sediment Management at the Watershed Level

    DTIC Science & Technology

    2012-08-01

    James R. Leech and David Biedenharn PURPOSE: This Coastal and Hydraulics Engineering Technical Note (CHETN) is an approach to analyze and manage...States (U.S.). The magnitude of this problem is illustrated in the Corps of Engineers ’ final report to Congress for the Streambank Erosion Control...of riverine habitats and ecosystems. This complexity has led some engineers to conclude that treatment of sediment sources at the watershed level

  13. On the taxonomy of the genus Acronicta (Lepidoptera, Noctuidae). III. Acronicta lutea species group.

    PubMed

    Volynkin, Anton V; Stüning, Dieter; Matov, Alexej Yu

    2016-11-17

    The present paper is the third in a series of articles on the genus Acronicta Ochsenheimer, 1816 (Volynkin et al. 2015; Volynkin 2015). Here we focus on the taxonomy of the Acronicta lutea species group, designating the lectotype of Acronicta lutea (Bremer & Grey, 1852), revising synonym status of Acronycta metaxantha funesta Draudt, 1950 and Pharetra leucoptera Butler, 1881, transferring Acronicta nigricans (Leech, 1900) to the subgenus Viminia Chapman, 1890 and introducing the new synonymy A. nigricans = Acronicta regifica Draudt, 1937 syn. n.

  14. Family Practice History: Bloodletting

    PubMed Central

    Curtis, Peter

    1981-01-01

    For 2000 years bloodletting was an accepted form of treatment. During this time, the indications and philosophical basis for lancing, cupping, and the application of leeches evolved in conjunction with advances in anatomy and physiology. In the late 19th century discoveries by tissue pathologists using new diagnostic techniques undermined earlier dogma and bloodletting quickly became a highly suspect form of treatment. ImagesFig. 1 PMID:21289758

  15. Ectoparasites and classification.

    PubMed

    Hopla, C E; Durden, L A; Keirans, J E

    1994-12-01

    The authors present an introductory overview of the principal groups of ectoparasites (flukes, leeches, crustaceans, insects, arachnids, lampreys and vampire bats) associated with domestic animals. Currently-accepted higher-level classifications are outlined for these parasites. Almost all significant ectoparasites of domestic animals are invertebrates, the majority being arthropods (crustaceans, insects and arachnids). Some of these ectoparasites are of particular importance as vectors of pathogens. Many ectoparasite species are host-specific, and vector species typically transmit characteristic pathogens.

  16. Larval stages of Neoplagioporus elongatus (Goto and Ozaki, 1930) (Opecoelidae: Plagioporinae), with notes on potential second intermediate hosts.

    PubMed

    Yano, Ayaka; Urabe, Misako

    2017-04-01

    The morphology of sporocysts and cercariae of Neoplagioporus elongatus (Goto and Ozaki, 1930) is described for the first time. A cotylomicrocercous cercaria obtained from the sorbeoconch snail Semisulcospira nakasekoae was confirmed to be the cercaria of N. elongatus, based on the degree of sequence identity of the COI gene to that of adult worms. Freshwater annelids (oligochaetes and leeches) and some aquatic insects (odonates) were demonstrated experimentally to be potential second intermediate hosts.

  17. Biological Monitoring of Pesticides, Heavy Metals and Other Contaminants at Rocky Mountain Arsenal. Phase 1

    DTIC Science & Technology

    1979-07-01

    herbivorous , while the ground beetle is carnivorous , and both serve as major prey items for insectivorous vertebrates. Leeches and snails are representative...most conspicuous small mammal on RMA and is a strict herbivore that feeds on both above-ground plant parts and roots. The deer mouse is the most abundant...mammal inhabiting RMA. It is widely distributed throughout the Arsenal, but occupies a small home range. It is omnivorous in its food habits, 6 2-4

  18. Effects of extracellular purines on ion transport across the integument of Hirudo medicinalis.

    PubMed

    Schnizler, Mikael; Buss, Mirjam; Clauss, Wolfgang

    2002-09-01

    Little is known about the long-term regulation of epithelial ion transport in invertebrates and the specific mediators involved. For some years, we have been investigating the short-term regulation of transepithelial ion transport across the dorsal integument of the leech Hirudo medicinalis, and we have established a model of Na+ uptake. In the present study, we investigated the effect of long-term acclimation on transintegumental ion transport by adapting leeches to high-salinity conditions. We dissected segments of dorsal integument and measured ion currents in Ussing chamber experiments. Electrophysiological variables, such as transepithelial potential (V(T)) and short-circuit-current (I(sc)), were profoundly affected by adaptation to high-salinity conditions. The total transepithelial Na+ current (I(Na)) decreased from 7.66+/-0.82 to 4.6+/-0.54 microA cm(-2) in preparations adapted to high salinity. The involvement of epithelial Na+ channels was determined as current inhibition (I(ami)) by apical application of amiloride; Na+ channels were equally active in control epithelia and epithelia from leeches adapted to high salinity. Removal of Ca2+ from the apical solutions, which is believed to reduce intracellular Ca2+ concentrations, equalized transepithelial variables between high-salt-adapted integuments and control integuments. Extracellular purines regulate transepithelial Cl- secretion and Na+ absorption. In a variety of tissues we tested ATP and adenosine for their effects on epithelial transport. Examination of integuments from pondwater- and high-salinity-adapted leeches revealed different sensitivities for these purines. Apical and basolateral application of ATP both stimulated transepithelial Na+ uptake and I(ami). Adenosine upregulated non-Na+ currents and acted from the basolateral side only. Apical Ca2+-free conditions attenuated these effects of purines on transepithelial currents. Extracellular UTP had no effect on ion transport.

  19. Complement resistance is essential for colonization of the digestive tract of Hirudo medicinalis by Aeromonas strains.

    PubMed

    Braschler, Thomas R; Merino, Susana; Tomás, Juan M; Graf, Joerg

    2003-07-01

    From the crop of the medicinal leech, Hirudo medicinalis, only Aeromonas veronii bv. sobria can be cultured consistently. Serum-sensitive A. veronii mutants were unable to colonize H. medicinalis, indicating the importance of the mammalian complement system for this unusual simplicity. Complementation of one selected mutant restored its ability to colonize. Serum-sensitive mutants are the first mutant class with a colonization defect for this symbiosis.

  20. Complement Resistance Is Essential for Colonization of the Digestive Tract of Hirudo medicinalis by Aeromonas Strains

    PubMed Central

    Braschler, Thomas R.; Merino, Susana; Tomás, Juan M.; Graf, Joerg

    2003-01-01

    From the crop of the medicinal leech, Hirudo medicinalis, only Aeromonas veronii bv. sobria can be cultured consistently. Serum-sensitive A. veronii mutants were unable to colonize H. medicinalis, indicating the importance of the mammalian complement system for this unusual simplicity. Complementation of one selected mutant restored its ability to colonize. Serum-sensitive mutants are the first mutant class with a colonization defect for this symbiosis. PMID:12839811

  1. Challenge to Change: Developing Leaders for the Nonkinetic Fight

    DTIC Science & Technology

    2009-06-15

    more of discourse on history. Later, several recognized the end of the presentation was approaching as Goodfriend’s tone shifted and his pace...brigadier general representing the Field Artillery, interjected . “Sir, there are still comments about how IO leeches into public affairs and undermines our...it is coming together. I am most encouraged. The strong network of our communicators and their abilities as a team to shape a public discourse

  2. Timing and Scope of Genomic Expansion within Annelida: Evidence from Homeoboxes in the Genome of the Earthworm Eisenia fetida

    PubMed Central

    Zwarycz, Allison S.; Nossa, Carlos W.; Putnam, Nicholas H.; Ryan, Joseph F.

    2016-01-01

    Annelida represents a large and morphologically diverse group of bilaterian organisms. The recently published polychaete and leech genome sequences revealed an equally dynamic range of diversity at the genomic level. The availability of more annelid genomes will allow for the identification of evolutionary genomic events that helped shape the annelid lineage and better understand the diversity within the group. We sequenced and assembled the genome of the common earthworm, Eisenia fetida. As a first pass at understanding the diversity within the group, we classified 363 earthworm homeoboxes and compared them with those of the leech Helobdella robusta and the polychaete Capitella teleta. We inferred many gene expansions occurring in the lineage connecting the most recent common ancestor (MRCA) of Capitella and Eisenia to the Eisenia/Helobdella MRCA. Likewise, the lineage leading from the Eisenia/Helobdella MRCA to the leech H. robusta has experienced substantial gains and losses. However, the lineage leading from Eisenia/Helobdella MRCA to E. fetida is characterized by extraordinary levels of homeobox gain. The evolutionary dynamics observed in the homeoboxes of these lineages are very likely to be generalizable to all genes. These genome expansions and losses have likely contributed to the remarkable biology exhibited in this group. These results provide a new perspective from which to understand the diversity within these lineages, show the utility of sub-draft genome assemblies for understanding genomic evolution, and provide a critical resource from which the biology of these animals can be studied. PMID:26659921

  3. Artery-Only Ear Replantation in a Child: A Case Report With Daily Photographic Documentation

    PubMed Central

    Mendenhall, Shaun D.; Sawyer, Justin D.

    2016-01-01

    Objective: Ear replantation poses a significant technical challenge even for the skilled microsurgeon. Many ear amputations result from avulsion and thus have damaged and often diminutive vessels with a paucity of veins. Artery-only replantation is an option for ear salvage, but little is published on the clinical course and appearance after this procedure. Methods: A subtotal ear replantation was performed on a 10-year-old boy without a venous anastomosis. Leech therapy was used to manage venous congestion postoperatively, and daily photography was performed to document the clinical course. Results: Postoperative venous congestion was successfully managed with leech therapy. Four days after the replantation, arterial thrombosis occurred that required a take back and salvage with an interposition vein graft for arterial repair. Native venous drainage and arterial revascularization from skin edges were evident by postoperative day 12, and leeches were discontinued on day 14. The patient required debridement of the posterior ear and superior helix necrotic skin, with burying of the upper portion of the ear in a superior auricular skin flap. The ear was subsequently released from the head, and the exposed portions were covered successfully with a full-thickness skin graft. Conclusions: While arterial and venous anastomoses should always be attempted, arterial-only ear replantation can provide excellent results when venous congestion is properly managed. Daily photography can be a useful tool to monitor subtle skin color changes that may indicate native venous drainage and arterial revascularization. PMID:28101290

  4. Compensatory plasticity restores locomotion after chronic removal of descending projections

    PubMed Central

    Harley, Cynthia M.; Reilly, Melissa G.; Stewart, Christopher; Schlegel, Chantel; Morley, Emma; Puhl, Joshua G.; Nagel, Christian; Crisp, Kevin M.

    2015-01-01

    Homeostatic plasticity is an important attribute of neurons and their networks, enabling functional recovery after perturbation. Furthermore, the directed nature of this plasticity may hold a key to the restoration of locomotion after spinal cord injury. Here we studied the recovery of crawling in the leech Hirudo verbana after descending cephalic fibers were surgically separated from crawl central pattern generators shown previously to be regulated by dopamine. We observed that immediately after nerve cord transection leeches were unable to crawl, but remarkably, after a day to weeks, animals began to show elements of crawling and intersegmental coordination. Over a similar time course, excessive swimming due to the loss of descending inhibition returned to control levels. Additionally, removal of the brain did not prevent crawl recovery, indicating that connectivity of severed descending neurons was not essential. After crawl recovery, a subset of animals received a second transection immediately below the anterior-most ganglion remaining. Similar to their initial transection, a loss of crawling with subsequent recovery was observed. These data, in recovered individuals, support the idea that compensatory plasticity directly below the site of injury is essential for the initiation and coordination of crawling. We maintain that the leech provides a valuable model to understand the neural mechanisms underlying locomotor recovery after injury because of its experimental accessibility, segmental organization, and dependence on higher-order control involved in the initiation, modulation, and coordination of locomotor behavior. PMID:25787951

  5. Artery-Only Ear Replantation in a Child: A Case Report With Daily Photographic Documentation.

    PubMed

    Mendenhall, Shaun D; Sawyer, Justin D; Adkinson, Joshua M

    2016-01-01

    Objective: Ear replantation poses a significant technical challenge even for the skilled microsurgeon. Many ear amputations result from avulsion and thus have damaged and often diminutive vessels with a paucity of veins. Artery-only replantation is an option for ear salvage, but little is published on the clinical course and appearance after this procedure. Methods: A subtotal ear replantation was performed on a 10-year-old boy without a venous anastomosis. Leech therapy was used to manage venous congestion postoperatively, and daily photography was performed to document the clinical course. Results: Postoperative venous congestion was successfully managed with leech therapy. Four days after the replantation, arterial thrombosis occurred that required a take back and salvage with an interposition vein graft for arterial repair. Native venous drainage and arterial revascularization from skin edges were evident by postoperative day 12, and leeches were discontinued on day 14. The patient required debridement of the posterior ear and superior helix necrotic skin, with burying of the upper portion of the ear in a superior auricular skin flap. The ear was subsequently released from the head, and the exposed portions were covered successfully with a full-thickness skin graft. Conclusions: While arterial and venous anastomoses should always be attempted, arterial-only ear replantation can provide excellent results when venous congestion is properly managed. Daily photography can be a useful tool to monitor subtle skin color changes that may indicate native venous drainage and arterial revascularization.

  6. [Does garlic protect against vampires? An experimental study].

    PubMed

    Sandvik, H; Baerheim, A

    1994-12-10

    Vampires are feared everywhere, but the Balkan region has been especially haunted. Garlic has been regarded as an effective prophylactic against vampires. We wanted to explore this alleged effect experimentally. Owing to the lack of vampires, we used leeches instead. In strictly standardized research surroundings, the leeches were to attach themselves to either a hand smeared with garlic or to a clean hand. The garlic-smeared hand was preferred in two out of three cases (95% confidence interval 50.4% to 80.4%). When they preferred the garlic the leeches used only 14.9 seconds to attach themselves, compared with 44.9 seconds when going to the non-garlic hand (p < 0.05). The traditional belief that garlic has prophylactic properties is probably wrong. The reverse may in fact be true. This study indicates that garlic possibly attracts vampires. Therefore to avoid a Balkan-like development in Norway, restrictions on the use of garlic should be considered.

  7. [Contact dermatitis with Hirudo medicinalis].

    PubMed

    Blaise, S; Le Brun, V; Sparsa, A; Delrous, J-L; Bonnetblanc, J-M

    2002-12-01

    Hirudine is the first natural anti-coagulant ever described. It is used for its anti-coagulant properties in plastic surgery or for the treatment of post-phlebitic diseases. Natural hirudine is extracted from the saliva of the Hirudo medicinalis leech, but it can also be found in crushed leech and included in a cream (Hirucrème). Side effects to hirudine are considered to be rare. We report a contact eczema caused by an extract from the medicinal Hirudo medicinalis leech. This was confirmed by the patch tests. However, we noticed a negativity of these tests with two analogs of the recombinant hirudine. Several cases of contact dermatitis with Hirucreme have been described. The analogs of recombinant hirudine, which share similar biological activity, have a very close molecular structure. They are indicated via the systemic route for thrombopenia related to heparin for the prevention of severe thromboses. The negative patch tests does not allow definite conclusion, but they prove that these molecules do not always lead to cross-allergies.

  8. Differentially expressed genes in Hirudo medicinalis ganglia after acetyl-L-carnitine treatment.

    PubMed

    Federighi, Giuseppe; Macchi, Monica; Bernardi, Rodolfo; Scuri, Rossana; Brunelli, Marcello; Durante, Mauro; Traina, Giovanna

    2013-01-01

    Acetyl-L-carnitine (ALC) is a naturally occurring substance that, when administered at supra-physiological concentration, is neuroprotective. It is involved in membrane stabilization and in enhancement of mitochondrial functions. It is a molecule of considerable interest for its clinical application in various neural disorders, including Alzheimer's disease and painful neuropathies. ALC is known to improve the cognitive capability of aged animals chronically treated with the drug and, recently, it has been reported that it impairs forms of non-associative learning in the leech. In the present study the effects of ALC on gene expression have been analyzed in the leech Hirudo medicinalis. The suppression subtractive hybridisation methodology was used for the generation of subtracted cDNA libraries and the subsequent identification of differentially expressed transcripts in the leech nervous system after ALC treatment. The method detects differentially but also little expressed transcripts of genes whose sequence or identity is still unknown. We report that a single administration of ALC is able to modulate positively the expression of genes coding for functions that reveal a lasting effect of ALC on the invertebrate, and confirm the neuroprotective and neuromodulative role of the substance. In addition an important finding is the modulation of genes of vegetal origin. This might be considered an instance of ectosymbiotic mutualism.

  9. Differentially Expressed Genes in Hirudo medicinalis Ganglia after Acetyl-L-Carnitine Treatment

    PubMed Central

    Federighi, Giuseppe; Macchi, Monica; Bernardi, Rodolfo; Scuri, Rossana; Brunelli, Marcello; Durante, Mauro; Traina, Giovanna

    2013-01-01

    Acetyl-l-carnitine (ALC) is a naturally occurring substance that, when administered at supra-physiological concentration, is neuroprotective. It is involved in membrane stabilization and in enhancement of mitochondrial functions. It is a molecule of considerable interest for its clinical application in various neural disorders, including Alzheimer’s disease and painful neuropathies. ALC is known to improve the cognitive capability of aged animals chronically treated with the drug and, recently, it has been reported that it impairs forms of non-associative learning in the leech. In the present study the effects of ALC on gene expression have been analyzed in the leech Hirudo medicinalis. The suppression subtractive hybridisation methodology was used for the generation of subtracted cDNA libraries and the subsequent identification of differentially expressed transcripts in the leech nervous system after ALC treatment. The method detects differentially but also little expressed transcripts of genes whose sequence or identity is still unknown. We report that a single administration of ALC is able to modulate positively the expression of genes coding for functions that reveal a lasting effect of ALC on the invertebrate, and confirm the neuroprotective and neuromodulative role of the substance. In addition an important finding is the modulation of genes of vegetal origin. This might be considered an instance of ectosymbiotic mutualism. PMID:23308261

  10. Association of LAR-like receptor protein tyrosine phosphatases with an enabled homolog in Hirudo medicinalis.

    PubMed

    Biswas, Subhas C; Dutt, Anindita; Baker, Michael W; Macagno, Eduardo R

    2002-12-01

    Receptor protein tyrosine phosphatases (RPTPs) are thought to play important functions in pathfinding and target recognition by growing neuronal processes. The leech RPTPs HmLAR1 and HmLAR2 are expressed selectively by central neurons, Comb cells, and peripheral muscle tissues in the Hirudo medicinalis embryo. To explore the functions of HmLARs, we have sought to determine their physiological substrates. We report here the cloning and embryonic expression of Lena, the leech homolog of Enabled, a cytosolic protein implicated in actin-based cell motility. Lena is expressed in embryonic central neurons and in the Comb cell. We present experimental evidences indicating that Lena associates selectively with the intracellular domain of HmLAR1 and HmLAR2. Additionally, RNA interference (RNAi) of HmLAR1 in intact leech embryos leads to the hyperphosphorylation of Lena. We propose, therefore, that Lena is an in vivo substrate of HmLAR1 in neurons and perhaps of HmLAR2 in the Comb cells.

  11. Timing and Scope of Genomic Expansion within Annelida: Evidence from Homeoboxes in the Genome of the Earthworm Eisenia fetida.

    PubMed

    Zwarycz, Allison S; Nossa, Carlos W; Putnam, Nicholas H; Ryan, Joseph F

    2015-12-10

    Annelida represents a large and morphologically diverse group of bilaterian organisms. The recently published polychaete and leech genome sequences revealed an equally dynamic range of diversity at the genomic level. The availability of more annelid genomes will allow for the identification of evolutionary genomic events that helped shape the annelid lineage and better understand the diversity within the group. We sequenced and assembled the genome of the common earthworm, Eisenia fetida. As a first pass at understanding the diversity within the group, we classified 363 earthworm homeoboxes and compared them with those of the leech Helobdella robusta and the polychaete Capitella teleta. We inferred many gene expansions occurring in the lineage connecting the most recent common ancestor (MRCA) of Capitella and Eisenia to the Eisenia/Helobdella MRCA. Likewise, the lineage leading from the Eisenia/Helobdella MRCA to the leech H. robusta has experienced substantial gains and losses. However, the lineage leading from Eisenia/Helobdella MRCA to E. fetida is characterized by extraordinary levels of homeobox gain. The evolutionary dynamics observed in the homeoboxes of these lineages are very likely to be generalizable to all genes. These genome expansions and losses have likely contributed to the remarkable biology exhibited in this group. These results provide a new perspective from which to understand the diversity within these lineages, show the utility of sub-draft genome assemblies for understanding genomic evolution, and provide a critical resource from which the biology of these animals can be studied.

  12. Real-time biomimetic Central Pattern Generators in an FPGA for hybrid experiments.

    PubMed

    Ambroise, Matthieu; Levi, Timothée; Joucla, Sébastien; Yvert, Blaise; Saïghi, Sylvain

    2013-01-01

    This investigation of the leech heartbeat neural network system led to the development of a low resources, real-time, biomimetic digital hardware for use in hybrid experiments. The leech heartbeat neural network is one of the simplest central pattern generators (CPG). In biology, CPG provide the rhythmic bursts of spikes that form the basis for all muscle contraction orders (heartbeat) and locomotion (walking, running, etc.). The leech neural network system was previously investigated and this CPG formalized in the Hodgkin-Huxley neural model (HH), the most complex devised to date. However, the resources required for a neural model are proportional to its complexity. In response to this issue, this article describes a biomimetic implementation of a network of 240 CPGs in an FPGA (Field Programmable Gate Array), using a simple model (Izhikevich) and proposes a new synapse model: activity-dependent depression synapse. The network implementation architecture operates on a single computation core. This digital system works in real-time, requires few resources, and has the same bursting activity behavior as the complex model. The implementation of this CPG was initially validated by comparing it with a simulation of the complex model. Its activity was then matched with pharmacological data from the rat spinal cord activity. This digital system opens the way for future hybrid experiments and represents an important step toward hybridization of biological tissue and artificial neural networks. This CPG network is also likely to be useful for mimicking the locomotion activity of various animals and developing hybrid experiments for neuroprosthesis development.

  13. Whitmania Pigra Whitman Extracts Inhibit Lipopolysaccharide Induced Rat Vascular Smooth Muscle Cells Migration and their Adhesion Ability to THP-1 and RAW 264.7 Cells.

    PubMed

    Li, Shuaishuai; Cheng, Long; An, Dengkun; Song, Shuliang; Liang, Hao; Chu, Fulong; Ji, Aiguo

    2017-03-01

    Atherosclerosis is a kind of chronic inflammatory disease. A crucial pathology change of atherosclerosis is the migration of activated VSMCs to the intima where they interact with leukocytes by expressing adhesion molecules, including intercellular cell adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1). Moreover, monocyte chemoattractant protein-1 (MCP-1) expressed by VSMCs plays an important role in recruiting monocytes and macrophages. Leech (Whitmania pigra Whitman) is a traditional Chinese medicine to treat cardiovascular diseases including atherosclerosis, however previous research has rarely reported the molecular mechanism for its curative effect. Thus, our study focuses on the effects of leech extracts on the expression of inflammatory factors, adhesion molecules and MCP-1 in rat VSMCs. In our present study, wound-healing assay and Boyden chamber model were applied to evaluate the anti-migration effect of LEE (Leech Enzyme Extracts) on LPS induced VSMCs. The anti-adhesion effect was assessed using DiI-labeled THP-1 and RAW264.7. LEE suppressed LPS-induced VSMCs migration and decreased the chemotaxis and adhesive capacity of THP-1 and RAW264.7 to LPS-stimulated VSMCs. LEE also attenuated the upregulation of a variety of pro-atherosclerotic factors by inhibiting the phosphorylation of p38 MAPK. LEE was also observed to prevent NF-κB p65 nuclear localization using immune-fluorescent staining. In conclusion, LEE suppresses LPS-induced upregulation of inflammatory factors, adhesion molecules and MCP-1 in rat VSMCs mainly via inhibiting the p38 MAPK/NF-κB pathways, thus partly uncovered LEE's molecular mechanisms for its therapeutic effect on atherosclerosis.

  14. Real-time biomimetic Central Pattern Generators in an FPGA for hybrid experiments

    PubMed Central

    Ambroise, Matthieu; Levi, Timothée; Joucla, Sébastien; Yvert, Blaise; Saïghi, Sylvain

    2013-01-01

    This investigation of the leech heartbeat neural network system led to the development of a low resources, real-time, biomimetic digital hardware for use in hybrid experiments. The leech heartbeat neural network is one of the simplest central pattern generators (CPG). In biology, CPG provide the rhythmic bursts of spikes that form the basis for all muscle contraction orders (heartbeat) and locomotion (walking, running, etc.). The leech neural network system was previously investigated and this CPG formalized in the Hodgkin–Huxley neural model (HH), the most complex devised to date. However, the resources required for a neural model are proportional to its complexity. In response to this issue, this article describes a biomimetic implementation of a network of 240 CPGs in an FPGA (Field Programmable Gate Array), using a simple model (Izhikevich) and proposes a new synapse model: activity-dependent depression synapse. The network implementation architecture operates on a single computation core. This digital system works in real-time, requires few resources, and has the same bursting activity behavior as the complex model. The implementation of this CPG was initially validated by comparing it with a simulation of the complex model. Its activity was then matched with pharmacological data from the rat spinal cord activity. This digital system opens the way for future hybrid experiments and represents an important step toward hybridization of biological tissue and artificial neural networks. This CPG network is also likely to be useful for mimicking the locomotion activity of various animals and developing hybrid experiments for neuroprosthesis development. PMID:24319408

  15. Complex evolutionary history of the Aeromonas veronii group revealed by host interaction and DNA sequence data.

    PubMed

    Silver, Adam C; Williams, David; Faucher, Joshua; Horneman, Amy J; Gogarten, J Peter; Graf, Joerg

    2011-02-16

    Aeromonas veronii biovar sobria, Aeromonas veronii biovar veronii, and Aeromonas allosaccharophila are a closely related group of organisms, the Aeromonas veronii Group, that inhabit a wide range of host animals as a symbiont or pathogen. In this study, the ability of various strains to colonize the medicinal leech as a model for beneficial symbiosis and to kill wax worm larvae as a model for virulence was determined. Isolates cultured from the leech out-competed other strains in the leech model, while most strains were virulent in the wax worms. Three housekeeping genes, recA, dnaJ and gyrB, the gene encoding chitinase, chiA, and four loci associated with the type three secretion system, ascV, ascFG, aexT, and aexU were sequenced. The phylogenetic reconstruction failed to produce one consensus tree that was compatible with most of the individual genes. The Approximately Unbiased test and the Genetic Algorithm for Recombination Detection both provided further support for differing evolutionary histories among this group of genes. Two contrasting tests detected recombination within aexU, ascFG, ascV, dnaJ, and gyrB but not in aexT or chiA. Quartet decomposition analysis indicated a complex recent evolutionary history for these strains with a high frequency of horizontal gene transfer between several but not among all strains. In this study we demonstrate that at least for some strains, horizontal gene transfer occurs at a sufficient frequency to blur the signal from vertically inherited genes, despite strains being adapted to distinct niches. Simply increasing the number of genes included in the analysis is unlikely to overcome this challenge in organisms that occupy multiple niches and can exchange DNA between strains specialized to different niches. Instead, the detection of genes critical in the adaptation to specific niches may help to reveal the physiological specialization of these strains.

  16. Complex Evolutionary History of the Aeromonas veronii Group Revealed by Host Interaction and DNA Sequence Data

    PubMed Central

    Faucher, Joshua; Horneman, Amy J.; Gogarten, J. Peter; Graf, Joerg

    2011-01-01

    Aeromonas veronii biovar sobria, Aeromonas veronii biovar veronii, and Aeromonas allosaccharophila are a closely related group of organisms, the Aeromonas veronii Group, that inhabit a wide range of host animals as a symbiont or pathogen. In this study, the ability of various strains to colonize the medicinal leech as a model for beneficial symbiosis and to kill wax worm larvae as a model for virulence was determined. Isolates cultured from the leech out-competed other strains in the leech model, while most strains were virulent in the wax worms. Three housekeeping genes, recA, dnaJ and gyrB, the gene encoding chitinase, chiA, and four loci associated with the type three secretion system, ascV, ascFG, aexT, and aexU were sequenced. The phylogenetic reconstruction failed to produce one consensus tree that was compatible with most of the individual genes. The Approximately Unbiased test and the Genetic Algorithm for Recombination Detection both provided further support for differing evolutionary histories among this group of genes. Two contrasting tests detected recombination within aexU, ascFG, ascV, dnaJ, and gyrB but not in aexT or chiA. Quartet decomposition analysis indicated a complex recent evolutionary history for these strains with a high frequency of horizontal gene transfer between several but not among all strains. In this study we demonstrate that at least for some strains, horizontal gene transfer occurs at a sufficient frequency to blur the signal from vertically inherited genes, despite strains being adapted to distinct niches. Simply increasing the number of genes included in the analysis is unlikely to overcome this challenge in organisms that occupy multiple niches and can exchange DNA between strains specialized to different niches. Instead, the detection of genes critical in the adaptation to specific niches may help to reveal the physiological specialization of these strains. PMID:21359176

  17. Botanical repellents and pesticides traditionally used against hematophagous invertebrates in Lao People's Democratic Republic: a comparative study of plants used in 66 villages.

    PubMed

    de Boer, Hugo; Vongsombath, Chanda; Pålsson, Katinka; Björk, Lars; Jaenson, Thomas G T

    2010-05-01

    Hematophagous parasites such as leeches, ticks, mites, lice, bedbugs, mosquitoes, and myiasis-producing fly larvae are common health problems in Lao People's Democratic Republic. Several arthropod-borne infections, e.g., malaria, dengue fever, and Japanese encephalitis, are endemic there. Effective vector control methods including the use of pesticides, insecticide-treated bed nets, and synthetic and plant-based repellents are important means of control against such invertebrates and the pathogens they may transmit or directly cause. In this study, we documented traditional knowledge on plants that are used to repel or kill hematophagous arthropods, including mosquitoes, bedbugs, human lice, mites and ticks, fly larvae, and blood-sucking leeches. Structured interviews were carried out in 66 villages comprising 17 ethnic groups, covering a range of cultures, throughout Lao People's Democratic Republic. A total of 92 plant species was recorded as traditional repellents (including plants for pesticidal usages) in 123 different plant-ectoparasite combinations. The number and species of plants, and animal taxa repelled (or killed) per plant species differed per region, village, and ethnic group. Traditional use was confirmed in the scientific literature for 74 of these plant species, and for an additional 13 species using literature on closely related species. The use of botanical repellents and pesticides from many plant species is common and widespread in the Lao countryside. In the future, the identification of the active components in certain plants to develop more optimal, inexpensive repellents, insecticides, acaricides, or antileech compounds as alternatives to synthetic repellents/pesticides against blood-feeding insects, ticks, mites, and leeches is warranted.

  18. Ex-vivo detection of neural events using THz BioMEMS.

    PubMed

    Abbas, Abdennour; Dargent, Thomas; Croix, Dominique; Salzet, Michel; Bocquet, Bertrand

    2009-09-01

    Electromagnetic frequencies up to a few terahertz (THz) can yield real-time and noninvasive measurements on biological matter. Unfortunately, strong absorption in aqueous solutions and low spatial resolution return difficult free-space investigations. A new approach based on integrated THz circuits was used. The authors designed and fabricated a BioMEMS (Biological MicroElectro-Mechanical System) compatible with microfluidic circulation and electromagnetic propagation. It is dedicated to the ex vivo detection of nitric oxide synthase (NOS) activity, which is involved in neurodegenerative phenomena. The biological model was a leech's central nervous system. After its injury, the production of NO was observed and measured in the far-THz spectral domain. The nerve cord was put inside a BioMEMS realized in polydimethylsiloxane (PDMS) sealed on a glass wafer. Glass is a good material for supporting high-frequency integrated waveguides such as coplanar waveguides (CPWs). Measurements were performed with vectorial network analyser (VNA). The transmission parameter in the frequency range of 0.14-0.22 THz was measured through CPWs located just below the microchannel containing the injured leech nerve cord. The lesion caused a decreased transmission coefficient due to the NOS activity. L-NAME was injected inside the microchannel and it was verified that it inhibits this activity. It was demonstrated that THz spectroscopy can detect a biochemical event, such as NOS activity around an injured leech's nerve cord, in real time. Future studies will be dedicated to quantitative measurements of the reaction products using the sophisticated management of several drugs allowed with microfluidic microsystems.

  19. Patterns of presynaptic activity and synaptic strength interact to produce motor output.

    PubMed

    Wright, Terrence Michael; Calabrese, Ronald L

    2011-11-30

    Motor neuron activity is coordinated by premotor networks into a functional motor pattern by complex patterns of synaptic drive. These patterns combine both the temporal pattern of spikes of the premotor network and the profiles of synaptic strengths (i.e., conductances). Given the complexity of premotor networks in vertebrates, it has been difficult to ascertain the relative contributions of temporal patterns and synaptic strength profiles to the motor patterns observed in these animals. Here, we use the leech (Hirudo sp.) heartbeat central pattern generator (CPG), in which we can measure both the temporal pattern and the synaptic strength profiles of the entire premotor network and the motor outflow in individual animals. In this system, a series of motor neurons all receive input from the same premotor interneurons of the CPG but must be coordinated differentially to produce a functional pattern. These properties allow a theoretical and experimental dissection of the rules that govern how temporal patterns and synaptic strength profiles are combined in motor neurons so that functional motor patterns emerge, including an analysis of the impact of animal-to-animal variation in input to such variation in output. In the leech, segmental heart motor neurons are coordinated alternately in a synchronous and peristaltic pattern. We show that synchronous motor patterns result from a nearly synchronous premotor temporal pattern produced by the leech heartbeat CPG. For peristaltic motor patterns, the staggered premotor temporal pattern determines the phase range over which segmental motor neurons can fire while synaptic strength profiles define the intersegmental motor phase progression realized.

  20. Patterns of Presynaptic Activity and Synaptic Strength Interact to Produce Motor Output

    PubMed Central

    Wright, Terrence Michael; Calabrese, Ronald L.

    2014-01-01

    Motor neuron activity is coordinated by premotor networks into a functional motor pattern by complex patterns of synaptic drive. These patterns combine both the temporal pattern of spikes of the premotor network and the profiles of synaptic strengths (i.e., conductances). Given the complexity of premotor networks in vertebrates, it has been difficult to ascertain the relative contributions of temporal patterns and synaptic strength profiles to the motor patterns observed in these animals. Here, we use the leech (Hirudo sp.) heartbeat central pattern generator (CPG), in which we can measure both the temporal pattern and the synaptic strength profiles of the entire premotor network and the motor outflow in individual animals. In this system, a series of motor neurons all receive input from the same premotor interneurons of the CPG but must be coordinated differentially to produce a functional pattern. These properties allow a theoretical and experimental dissection of the rules that govern how temporal patterns and synaptic strength profiles are combined in motor neurons so that functional motor patterns emerge, including an analysis of the impact of animal-to-animal variation in input to such variation in output. In the leech, segmental heart motor neurons are coordinated alternately in a synchronous and peristaltic pattern. We show that synchronous motor patterns result from a nearly synchronous premotor temporal pattern produced by the leech heartbeat CPG. For peristaltic motor patterns, the staggered premotor temporal pattern determines the phase range over which segmental motor neurons can fire while synaptic strength profiles define the intersegmental motor phase progression realized. PMID:22131417

  1. In Vivo Isolation and Characterization of Stem Cells with Diverse Phenotypes Using Growth Factor Impregnated Biomatrices

    PubMed Central

    Grimaldi, Annalisa; Bianchi, Cristiano; Greco, Gabriella; Tettamanti, Gianluca; Noonan, Douglas M.; Valvassori, Roberto; de Eguileor, Magda

    2008-01-01

    Background The stimulation to differentiate into specific cell types for somatic stem cells is largely due to a series of internal and external signals coming from the microenvironment that surrounds the stem cell. Even though intensive research has been made, the basic mechanisms of plasticity and/or the molecules regulating stem cells proliferation and differentiation are not completely determined. Potential answers concerning the problems could be derived from the studies of stem cells in culture. Methodology/Principle Findings We combine a new procedure (using the matrigel biopolymer supplemented with a selected cytokine/growth factor) with classic techniques such as light, confocal and electron microscopy, immunohistochemistry and cell culture, to perform an analysis on stem cells involved in the leech (Hirudo medicinalis) repair tissues. The leech has a relative anatomical simplicity and is a reliable model for studying a variety of basic events, such as tissue repair, which has a striking similarity with vertebrate responses. Our data demonstrate that the injection of an appropriate combination of the matrigel biopolymer supplemented with a selected cytokine/growth factor in the leech Hirudo medicinalis is a remarkably effective tool for isolating a specific cell population in vivo. A comparative analysis of biopolymer in vivo sorted stem cells indicates that VEGF recruited cells of a hematopoietic/endothelial phenotype whereas MCP-1/CCL2 isolated cells that were of an early myeloid lineage. Conclusion Our paper describes, for the first time, a method allowing not only the isolation of a specific cell population in relation to the cytokine utilized but also the possibility to culture a precise cell type whose isolation is otherwise quite difficult. This approach could be broadly applied to isolate stem cells of diverse origins based on the recruitment stimuli employed. PMID:18382683

  2. Examination of interaction of trypanosome infection and crude oil exposure on hematology of the longhorn sculpin (Myoxocephalus octodecemspinosus)

    SciTech Connect

    Kiceniuk, J.W.; Khan, R.A.; Dawe, M.; Williams, U.

    1982-04-01

    Adult longhorn sculpins were collected from Conception Bay, Newfoundland, and infected with trypanosomes via leeches. One group of these fish were exposed to water-accommodated Venezuelan crude oil, while the other served as controls. The results of the study suggests that water soluble fractions of Venezuelan crude oil have a minor effect on the hematology of sculpins in the concentration range 150-300 ppb, but that trypanosomes appear to potentiate the effect of oil on blood hemoglobin content of trypanosome infected fish. If this is found to be the case in active species of fish, the decrease in oxygen carrying capacity would likely limit the aerobic capacity of fish. (JMT)

  3. HmCRIP, a cysteine-rich intestinal protein, is expressed by an identified regenerating nerve cell.

    PubMed

    Emes, R D; Wang, W-Z; Lanary, K; Blackshaw, S E

    2003-01-02

    A Hirudo medicinalis cDNA isolated from regenerating CNS tissue at 24 h post-axotomy was identified as a leech homologue of the mammalian cysteine-rich intestinal proteins (CRIPs) and named HmCRIP. HmCRIP is up-regulated within 6 h of axotomy, peaking at 24 h. This is the first demonstration of a CRIP homologue in regenerating CNS and in a serotonergic neurone. In rodents CRIP is an important factor in the regulation of the inflammatory immune response through control of Th1/Th2 differentiation. The role of HmCRIP in the regeneration competent environment of the annelid central nervous system is discussed.

  4. Synchronization Patterns and Related Problems in Combinatorial Analysis and Graph Theory

    DTIC Science & Technology

    1981-06-01

    APPENDIX 86 REFERENCES 90 Li ~1 ivF LIST OF FIGURES Figure Page 1-1 The Leech Tree 6 2-1 The Graph L and the Graph R 9 2-2 The Dodecahedron and Two...forest F with 4 nodes, but in this case Ithe direct count to find the number takes more work. D Figure 2-2. The Dodecahedron and Two Copies of the...with n - 2m nodes, girth > 4, and d - 3, then G m - 2 Om-4) T -(Zm 3 24m + 100m 147). The dodecahedron , D, and one graph, P, consisting of two copies

  5. First record of a digenean from invasive lionfish, Pterois cf. volitans, (Scorpaeniformes: Scorpaenidae) in the northwestern Atlantic Ocean.

    PubMed

    Bullard, S A; Barse, A M; Curran, S S; Morris, J A

    2011-10-01

    Adults of Lecithochirium floridense (Digenea: Hemiuridae) parasitized the stomach in each of 22 necropsied lionfish, Pterois cf. volitans (Scorpaeniformes: Scorpaenidae) (prevalence  =  100%, mean intensity  =  11), captured in the northwestern Atlantic Ocean off Beaufort, North Carolina (34°14.83'N, 76°35.25'W). This is the first report of a digenean from the invasive lionfish and that of L. floridense from a species of Pterois. The leech specimen previously identified as Myzobdella lugubris from P. volitans in the northwestern Atlantic Ocean is re-identified as Trachelobdella lubrica based on a study of the original voucher specimen.

  6. Corrections to the theory and the optimal line in the swimming diagram of Taylor (1952).

    PubMed

    Humphrey, Joseph A C; Chen, Jun; Iwasaki, Tetsuya; Friesen, W Otto

    2010-08-06

    The analysis of undulatory swimming gaits requires knowledge of the fluid forces acting on the animal body during swimming. In his classical 1952 paper, Taylor analysed this problem using a 'resistive-force' theory. The theory was used to characterize the undulatory gaits that result in the smallest energy dissipation to the fluid for a given swim velocity. The optimal gaits thus found were compared with data recorded from movies of a snake and a leech swimming. This report identifies and corrects a mathematical error in Taylor's paper, showing that his theory applies even better to animals of circular cross section.

  7. The biology of some intraerythrocytic parasites of fishes, amphibia and reptiles.

    PubMed

    Davies, A J; Johnston, M R

    2000-01-01

    Fishes, amphibia and reptiles, the ectothermic vertebrates, are hosts for a variety of intraerythrocytic parasites including protists, prokaryotes, viruses and structures of uncertain status. These parasites may experience host temperature fluctuations, host reproductive strategies, population genetics, host habitat and migratory behaviour quite unlike those of endothermic hosts. Few blood infections of fishes, amphibia and reptiles have proven pathogenicity, in contrast to the many intraerythrocytic parasites of mammals and some birds which harm their hosts. Although not given the attention afforded to intraerythrocytic parasites of endotherms, those of ectotherms have been studied for more than a century. This review reports on the diversity, general biology and phylogeny of intraerythrocytic parasites of ectotherms. The existence of taxonomic confusion is emphasized and the main taxonomic features of most of the 23 better characterized genera, particularly the kinetoplastid and apicomplexan protists, are summarized. Transmission of protistan infections of aquatic ectotherms is also discussed. Leeches can transfer sporozoties or merozoites to the vertebrate host during feeding. Dormant sporozoites of Lankesterella may permit transmission of species of this genus between vertebrates by predation. The fish haemogregarine, Haemogregarina bigemina, probably has gnathiid isopods, rather than leeches, as its definitive hosts. Hepatozoon spp. in aquatic hosts, and Progarnia of caiman, may also use invertebrate hosts other than leeches. Protistan infections of terrestrial or semi-terrestrial hosts are transmitted by a variety of arthropods, or, in some cases, leeches, contaminated paratenic hosts, or sporocysts free in water. Transfer of protists between vertebrates by predation and congenitally may also occur. The biology of the host cells of these infections, the red blood cells of ectotherm vertebrates, is summarized and compared with that of mammalian erythrocytes

  8. [Dental aspects of general symptoms in the 18th century].

    PubMed

    Forrai, Judit

    2009-05-24

    In the 18th century, numerous diseases with symptoms of oral cavity were cured by chirurgien-dentist, barber-surgeons, or tooth drawer. The so called "dentitio difficilis" was blamed for the high children mortality, therefore gum cut or use of leeches was advised as a treatment. Both acute and chronic type of gum inflammation was called scurvy. It seems that the mechanical removal of plaque was enough to cure the scurvy as it was written in advertisements from that time. Syphilis was present in the everyday life throughout centuries, and assumed to cause different stigmas in the oral cavity. Today we consider theses stigmas as the toxic signs of mercury treatment.

  9. Behavioral studies of contaminant effects on aquatic invertebrates: A review of Russian investigations

    USGS Publications Warehouse

    Nepomnyashchikh, V. A.; DeLonay, V. A.; Little, E.E.; Henshel, D. S.; Bengston, David A.

    1996-01-01

    Studies by Russian scientists have documented significant alterations and impairment of critical behavioral functions in aquatic organisms following exposure to environmental contaminants. Behavioral responses disrupted by sublethal exposure to toxicants are intimately involved in habitat selection, foraging, competition, predator-prey relationships, and reproduction, and are essential to survival. Behavioral responses of benthic invertebrates have received considerable study in Russia. A range of invertebrate taxa have been studied, including leeches, insects, molluscs, plankton, and crustaceans. In addition, aquatic invertebrates exhibit a large number of behavioral responses which are sensitive to contaminant exposure and are easily quantified. Standardized behavioral methodologies for measuring contaminant effects are being developed.

  10. Interpretative Analysis of Surficial Sediments as an Aid in Transport Studies of Dredged Materials Cape Canaveral, Florida.

    DTIC Science & Technology

    1984-03-01

    of Cocoa leech, nornda, to 40-55 ft of uster, on the inner continental shelf. Drefte isposal material io composed of clay, silt, ad fine send remved...of Cape Canaveral and 4.5 miles east of Cocoa Beach, Florida (Site B, Figure 1), has been in use since 1974. During the past 9 years, approximately...and direction diagrams, calculated for Cocoa Beach for the 10-year period (1945-1947 and 1950-1958), were located on project plansheets (SM 1979). A

  11. Beauty and the beast: Superconformal symmetry in a monster module

    NASA Astrophysics Data System (ADS)

    Dixon, L.; Ginsparg, P.; Harvey, J.

    1988-06-01

    Frenkel, Lepowsky, and Meurman have constructed a representation of the largest sporadic simple finite group, the Fischer-Griess monster, as the automorphism group of the operator product algebra of a conformal field theory with central charge c=24. In string terminology, their construction corresponds to compactification on a Z 2 asymmetric orbifold constructed from the torus R 24/∧, where ∧ is the Leech lattice. In this note we point out that their construction naturally embodies as well a larger algebraic structure, namely a super-Virasoro algebra with central charge ĉ=16, with the supersymmetry generator constructed in terms of bosonic twist fields.

  12. NASA Education Stakeholder's Summit

    NASA Image and Video Library

    2010-09-12

    NASA Student Ambassadors and Facilitator are seen on a panel at the NASA Education Stakeholders’ Summit One Stop Shopping Initiative (OSSI), Monday, Sep. 13, 2010, at the Westfields Marriott Conference Center in Chantilly, VA. From left to right are: Quenton Bonds, University of South Florida; Geoffrey Wawrzyniak, Purdue University; Heriberto Reynoso, University of Texas at Brownsville; Marie Kingbird-Lowry, Leech Lake Tribal College; Kareen Borders, University of Washington; Katelyn Doran, University of North Carolina at Charlotte and Ashanti Johnson, PhD, Executive Director, Institute for Broadening Participation. (Photo Credit: NASA/Carla Cioffi)

  13. Clifford Algebras and Their Decomposition into Conjugate Fermionic Heisenberg Algebras

    NASA Astrophysics Data System (ADS)

    Catto, Sultan; Gürcan, Yasemin; Khalfan, Amish; Kurt, Levent; Kato La, V.

    2016-10-01

    We discuss a construction scheme for Clifford numbers of arbitrary dimension. The scheme is based upon performing direct products of the Pauli spin and identity matrices. Conjugate fermionic algebras can then be formed by considering linear combinations of the Clifford numbers and the Hermitian conjugates of such combinations. Fermionic algebras are important in investigating systems that follow Fermi-Dirac statistics. We will further comment on the applications of Clifford algebras to Fueter analyticity, twistors, color algebras, M-theory and Leech lattice as well as unification of ancient and modern geometries through them.

  14. Bioactive proteins and peptides isolated from Chinese medicines with pharmaceutical potential.

    PubMed

    Wong, Kam Lok; Wong, Ricky Ngok Shun; Zhang, Liang; Liu, Wing Keung; Ng, Tzi Bun; Shaw, Pang Chui; Kwok, Philip Chi Lip; Lai, Yau Ming; Zhang, Zhang Jin; Zhang, Yanbo; Tong, Yao; Cheung, Ho-Pan; Lu, Jia; Sze, Stephen Cho Wing

    2014-01-01

    Some protein pharmaceuticals from Chinese medicine have been developed to treat cardiovascular diseases, genetic diseases, and cancer. Bioactive proteins with various pharmacological properties have been successfully isolated from animals such as Hirudo medicinalis (medicinal leech), Eisenia fetida (earthworm), and Mesobuthus martensii (Chinese scorpion), and from herbal medicines derived from species such as Cordyceps militaris, Ganoderma, Momordica cochinchinensis, Viscum album, Poria cocos, Senna obtusifolia, Panax notoginseng, Smilax glabra, Ginkgo biloba, Dioscorea batatas, and Trichosanthes kirilowii. This article reviews the isolation methods, molecular characteristics, bioactivities, pharmacological properties, and potential uses of bioactive proteins originating from these Chinese medicines.

  15. Bioactive proteins and peptides isolated from Chinese medicines with pharmaceutical potential

    PubMed Central

    2014-01-01

    Some protein pharmaceuticals from Chinese medicine have been developed to treat cardiovascular diseases, genetic diseases, and cancer. Bioactive proteins with various pharmacological properties have been successfully isolated from animals such as Hirudo medicinalis (medicinal leech), Eisenia fetida (earthworm), and Mesobuthus martensii (Chinese scorpion), and from herbal medicines derived from species such as Cordyceps militaris, Ganoderma, Momordica cochinchinensis, Viscum album, Poria cocos, Senna obtusifolia, Panax notoginseng, Smilax glabra, Ginkgo biloba, Dioscorea batatas, and Trichosanthes kirilowii. This article reviews the isolation methods, molecular characteristics, bioactivities, pharmacological properties, and potential uses of bioactive proteins originating from these Chinese medicines. PMID:25067942

  16. Unusual foreign body in the larynx: a bee.

    PubMed

    İlhan, Ethem; Yaman, Handan; Dost, Burhan; Köse, Gökçe Akman; Yaman, Hüseyin

    2015-01-01

    Foreign body lodgement in the larynx is a rare situation. Our review of the literature revealed no living foreign body in larynx except for laryngeal leeches and anisakiasis. In this article, we report a patient with unusual laryngeal foreign body lodgement: a bee which presented with sudden odynophagia and stinging sensation in throat. The bee was detected on the laryngeal mucosa in indirect laryngoscopic examination and removed immediately under general anesthesia in apneic period. In this case report, we describe the importance of detailed anamnesis and laryngeal examination even if the patient has no severe symptoms.

  17. Genetic Regulatory Networks in Embryogenesis and Evolution

    NASA Technical Reports Server (NTRS)

    1998-01-01

    The article introduces a series of papers that were originally presented at a workshop titled Genetic Regulatory Network in Embryogenesis and Evaluation. Contents include the following: evolution of cleavage programs in relationship to axial specification and body plan evolution, changes in cell lineage specification elucidate evolutionary relations in spiralia, axial patterning in the leech: developmental mechanisms and evolutionary implications, hox genes in arthropod development and evolution, heterochronic genes in development and evolution, a common theme for LIM homeobox gene function across phylogeny, and mechanisms of specification in ascidian embryos.

  18. Neuroelectronic device based on nanocoax arrays

    NASA Astrophysics Data System (ADS)

    Naughton, Jeffrey R.; Lundberg, Jaclyn N.; Varela, Juan A.; Burns, Michael J.; Chiles, Thomas C.; Christianson, John P.; Naughton, Michael J.

    2015-03-01

    We report on development of a nanocoax-based neuroelectronic array. The device has been used in real time to noninvasively couple to a ganglion sac located along the main nerve cord of the leech Hirudo medicinalis. This allowed for extracellular recording of synaptic activity in the form of spontaneous synapse firing in pre- and post-synaptic somata, with the next target being recording of local field potentials from rat hippocampal cells. We also discuss an alteration of the architecture to facilitate optical integration of the nanoarray, toward utilizing the so-modified device to elicit / inhibit action potentials in optogenetically-modified cells.

  19. [Neurite-stimulating activity of secretions from the Hirudo medicinalis salivary gland in cultured sensory neurons].

    PubMed

    Chalisova, N I; Pennijajnen, V P; Baskova, I P; Zavalova, L L; Bazanova, A V

    2001-12-01

    Components of leech (Hirudo medicinalis) salivary gland secretion (high molecular mass bdelin B, bdellastasin, eglin C, destabilize) were used to investigate their neurotrophic effect on organotypic culture of dorsal root ganglia of the 10-11-day old chicken embryos. Destabilize and high molecular mass bdelin B in concentrations of 0.01, 0.02, 0.05 and 0.1 ng/ml, bdellastasin in concentrations of 0.02 and 0.05 ng/ml, eglin C in concentrations of 0.1 ng/ml exhibit neurite-stimulating effect as compared to the control.

  20. Toll-like receptor genes (TLRs) from Capitella capitata and Helobdella robusta (Annelida).

    PubMed

    Davidson, Charis R; Best, Natalie M; Francis, Joseph W; Cooper, Edwin L; Wood, Todd Charles

    2008-01-01

    Toll-like receptors (TLRs) are an important part of the innate immunity system and are found throughout the animal kingdom, but have not yet been reported in annelids. We searched shotgun reads of the genomes of the leech Helobdella and polychaete Capitella for TLR homologs. We found 105 TLR homologs in Capitella and 16 in Helobdella. The deduced phylogeny of these sequences, together with TLRs from other animal phyla, reveals three major clades. One clade consists of a mixture of both vertebrates and invertebrates, including sequences from Capitella and Helobdella, while the other two clades contain only invertebrate TLRs.

  1. Minnesota Tribal Coalition - Final Report

    SciTech Connect

    Michael Triplett

    2006-12-19

    The project helped tribal leaders, staff and community members on the Grand Portage, Leech Lake, and White Earth Reservations better understand their community's energy usage, assess local resources that might be utilized to reduce energy consumption and begin to formulate appropriate development strategies. The principal guiding interest was to assess energy usage and the potential for wind resource development on each of the three reservations. Key tribal staff became familiar with wind energy technology and assessment methodologies that will be of continued use as each tribe moves forward with development projects. The findings were that wind resources are available at each reservation with varying degrees of potential for development. At White Earth moderate to excellent resources are present at White Earth village and along the U.S. 59 corridor sufficient to be tapped to serve several scattered tribal complexes. At Grand Portage a former community television repeater tower site provides a viable elevated location for a wind turbine to serve the tribal community settlement. At Leech Lake, while most constrained by tree cover, a site adjacent to a casino holds promise for the newer taller wind turbines now coming to market at ever-increasing taller rotor heights. The project developed considerable data of importance regarding the potential for wind development on and near each reservation.

  2. Effects of high-rate electrical stimulation upon firing in modelled and real neurons.

    PubMed

    Krauthamer, V; Crosheck, T

    2002-05-01

    Many medical devices use high-rate, low-amplitude currents to affect neural function. This study examined the effect of stimulation rate upon action potential threshold and sustained firing rate for two model neurons, the rabbit myelinated fibre and the unmyelinated leech touch sensory cell. These model neurons were constructed with the NEURON simulator from electrophysiological data. Alternating-phase current pulses (0-1250 Hz), of fixed phase duration (0.2 ms), were used to stimulate the neurons, and propagation success or failure was measured. One effect of the high pulse rates was to cause a net depolarisation, and this was verified by the relief of action potential conduction block by 500 Hz extracellular stimulation in leech neurons. The models also predicted that the neurons would maintain maximum sustained firing at a number of different stimulation rates. For example, at twice threshold, the myelinated model followed the stimulus up to 500 Hz stimulation, half the stimulus rate up to 850 Hz stimulation, and it did not fire at 1250 Hz stimulation. By contrast, the unmyelinated neuron model had a lower maximum firing rate of 190 Hz, and this rate was obtained at a number of stimulation rates, up to 1250 Hz. The myelinated model also predicted sustained firing with 1240 Hz stimulation at threshold current, but no firing when the current level was doubled. Most of these effects are explained by the interaction of stimulus pulses with the cell's refractory period.

  3. [Parasites-invaders of the Volga river basin: history of invasion, perspective of dispersion, possibility of epizootic].

    PubMed

    Zhokhov, A E; Pugacheva, M N

    2001-01-01

    The fauna of fish parasites in the Volga river increased by 15 species in last time. These parasites have invaded together with introduced hosts or dispersed after destruction of some physical and ecological barriers. The infusoria Ambiphrya ameuri, cestode Cestobothrium acheilognathi, trematodes Amurotrema dombrovskajae and Sanguinicola skrabini have been introduced together with their hosts. A creation of water reservoirs destroyed ecological barriers and created favorable conditions for the migration and dispersion of parasites. The cestodes Eubothrium rugosum, Proteocephalus longicollis, and nematode Cystidicola farionis migrated from the North, Aspidogaster limacoides migrated from the South. The leech Caspiobdella fadejevi, trematodes Rossicotrema donicus, Apophallus muehlingi, Niccola skrjabini, Plagioporus skrjabini migrated through the Volga-Don channel. Some invader have already finished their dispersion in the water reservoirs of Volga river, other parasites still continue this process. South border of the E. rugosum range is in the Kujbyshev water basin, the leech C. fadejevi is distributed in all water basins, trematodes R. donicus, A. muehlingi and P. skrjabini are found in the Volga delta, while N. skrjabini has already reached the Saratov water basin. Perspectives of new invaders and epizootic significance of invaders is discussed.

  4. Recombinant destabilase-lysozyme: synthesis de novo in E. coli and action mechanism of the enzyme expressed in Spodoptera frugiperda.

    PubMed

    Zavalova, L L; Baskova, I P; Barsova, E V; Snezhkov, E V; Akopov, S B; Lopatin, S A

    2004-07-01

    Destabilase-lysozyme (DL) from salivary gland secretion of the medicinal leech (Hirudo medicinalis) is as a member of the invertebrate lysozyme family, which sharply differs from other lysozyme families. In this study, DL lysozyme function was confirmed during expression of a gene encoding DL in Escherichia coli. Several constructs of the expression vectors pKK OmpA and pET-3A with or without bacterial, leech, or yeast signal peptides (SP) were used. The use of a construct without signal peptide genes resulted in normal growth of the transformed cells. Transformation of E. coli cells with the constructs containing SP was accompanied by the disruption of the forming cells. The use of the expression vector pET-32 LTC-System for production of DL as a fusion protein with thioredoxin also resulted in normal cell growth. However, specific activity of DL isolated from such cells was significantly lower than that of enzyme purified from extracts of Spodoptera frugiperda cells, which were infected with the baculovirus vector carrying DL cDNA. It is shown that the action mechanism of invertebrate lysozyme does not differ from that of other families: recombinant DL from S. frugiperda extracts catalyzed cleavage of synthetic substrate, hexamer of N-acetylglucosamine, to di- and tetramers, which is typical for enzymatic function of other lysozyme families.

  5. [Studies on effects of water temperature, stocking density and feeding cycle on growth and feeding in Hirudo nipponica].

    PubMed

    Lu, Yu-Xi; Cheng, Bo-Xing; Guo, Qiao-Sheng; Liu, Fei; Shi, Hong-Zhuan; Guo, Ling

    2017-07-01

    Effects of different water temperature, stocking density and feeding cycle on growth, feeding and survival of Hirudo nipponica have been studied, six temperature gradients were set: 18, 22, 26, 30, 34 and 38 ℃, five stocking density gradients were set: 30, 60, 120, 180 and 240 leech/L, four feeding cycle gradients were set: 2, 5, 10 and 20 d, respectively. The results showed that there exists a significant regression relationship between water temperature and specific growth rate: y=-0.016 5x²+0.836 9x-6.847 5(R²=0.990 8)(P<0.05), a regression analysis indicated that specific growth rate reached the maximum (3.76) at 25.36 ℃. When water temperature was beyond 30 ℃, the survival rate significantly decreased as water temperature increased (P<0.05). The specific growth rate and survival rate decreased as stocking density increased. A linear relationship exists between the feeding cycle and the SGR: y=-0.094 1x+3.832 9(R²=0.992 7). From this study, it can be concluded that the optimal water temperature and stocking density for the growth of H. nipponica is 22-26 ℃ and 30-120 leech/L, respectively. Copyright© by the Chinese Pharmaceutical Association.

  6. Emergent Space-Time

    NASA Astrophysics Data System (ADS)

    Chapline, George

    It has been shown that a nonlinear Schrödinger equation in 2+1 dimensions equipped with an SU(N) Chern-Simons gauge field can provide an exact description of certain self-dual Einstein spaces in the limit N-=∞. Ricci flat Einstein spaces can then be viewed as arising from a quantum pairing of the classical self-dual and anti-self-dual solutions. In this chapter, we will outline how this theory of empty space-time might be generalized to include matter and vacuum energy by transplanting the nonlinear Schrödinger equation used to construct Einstein spaces to the 25+1-dimensional Lorentzian Leech lattice. If the distinguished 2 spatial dimensions underlying the construction of Einstein spaces are identified with a hexagonal lattice section of the Leech lattice, the wave-function becomes an 11 × 11 matrix that can represent fermion and boson degrees of freedom (DOF) associated with 2-form and Yang-Mills gauge symmetries. The resulting theory of gravity and matter in 3+1 dimensions is not supersymmetric, which provides an entry for a vacuum energy. Indeed, in the case of a Lemaitre cosmological model, the emergent space-time will naturally have a vacuum energy on the order of the observed cosmological constant.

  7. Control of traveling-wave oscillations and bifurcation behavior in central pattern generators

    NASA Astrophysics Data System (ADS)

    Landsman, Alexandra S.; Slotine, Jean-Jacques

    2012-10-01

    Understanding synchronous and traveling-wave oscillations, particularly as they relate to transitions between different types of behavior, is a central problem in modeling biological systems. Here, we address this problem in the context of central pattern generators (CPGs). We use contraction theory to establish the global stability of a traveling-wave or synchronous oscillation, determined by the type of coupling. This opens the door to better design of coupling architectures to create the desired type of stable oscillations. We then use coupling that is both amplitude and phase dependent to create either globally stable synchronous or traveling-wave solutions. Using the CPG motor neuron network of a leech as an example, we show that while both traveling and synchronous oscillations can be achieved by several types of coupling, the transition between different types of behavior is dictated by a specific coupling architecture. In particular, it is only the “repulsive” but not the commonly used phase or rotational coupling that can explain the transition to high-frequency synchronous oscillations that have been observed in the heartbeat pattern generator of a leech. This shows that the overall dynamics of a CPG can be highly sensitive to the type of coupling used, even for coupling architectures that are widely believed to produce the same qualitative behavior.

  8. Control of traveling-wave oscillations and bifurcation behavior in central pattern generators.

    PubMed

    Landsman, Alexandra S; Slotine, Jean-Jacques

    2012-10-01

    Understanding synchronous and traveling-wave oscillations, particularly as they relate to transitions between different types of behavior, is a central problem in modeling biological systems. Here, we address this problem in the context of central pattern generators (CPGs). We use contraction theory to establish the global stability of a traveling-wave or synchronous oscillation, determined by the type of coupling. This opens the door to better design of coupling architectures to create the desired type of stable oscillations. We then use coupling that is both amplitude and phase dependent to create either globally stable synchronous or traveling-wave solutions. Using the CPG motor neuron network of a leech as an example, we show that while both traveling and synchronous oscillations can be achieved by several types of coupling, the transition between different types of behavior is dictated by a specific coupling architecture. In particular, it is only the "repulsive" but not the commonly used phase or rotational coupling that can explain the transition to high-frequency synchronous oscillations that have been observed in the heartbeat pattern generator of a leech. This shows that the overall dynamics of a CPG can be highly sensitive to the type of coupling used, even for coupling architectures that are widely believed to produce the same qualitative behavior.

  9. Therapeutic Response for Functional Abdominal Pain in Children with Occult Constipation: Laxatives versus Prokinetic Drugs

    PubMed Central

    2017-01-01

    The relationship between functional abdominal pain (FAP) and occult constipation (OC) in children who did not meet the Rome III criteria for constipation has rarely been reported. This study aimed to estimate the prevalence of OC in patients with FAP and to compare the effectiveness of prokinetic drugs and laxatives for FAP and OC. Pediatric outpatients (n = 212; aged 4–15 years) who satisfied the Rome III criteria for childhood FAP were divided into 2 groups based on Leech scores: group 1 < 8; group 2 ≥ 8. Group 2 received either prokinetic drugs or laxatives and pain severity was assessed after 2 weeks, 1 month, and 3 months. A total 52.4% (111/212) of patients had OC in this study. More patients who received laxatives had reduced pain scores compared with those who received prokinetic drugs. Those treated with laxatives in group 2 had a better response than those treated with prokinetic drugs throughout the study period (P < 0.001, P < 0.001, and P = 0.002 after 2 weeks, 1 month, and 3 months, respectively). OC was frequently encountered in children with FAP. Laxatives can be more effective than prokinetic drugs for relieving symptoms of FAP in children with a Leech score ≥ 8 and suspected OC. PMID:27914138

  10. Tag jumps illuminated--reducing sequence-to-sample misidentifications in metabarcoding studies.

    PubMed

    Schnell, Ida Baerholm; Bohmann, Kristine; Gilbert, M Thomas P

    2015-11-01

    Metabarcoding of environmental samples on second-generation sequencing platforms has rapidly become a valuable tool for ecological studies. A fundamental assumption of this approach is the reliance on being able to track tagged amplicons back to the samples from which they originated. In this study, we address the problem of sequences in metabarcoding sequencing outputs with false combinations of used tags (tag jumps). Unless these sequences can be identified and excluded from downstream analyses, tag jumps creating sequences with false, but already used tag combinations, can cause incorrect assignment of sequences to samples and artificially inflate diversity. In this study, we document and investigate tag jumping in metabarcoding studies on Illumina sequencing platforms by amplifying mixed-template extracts obtained from bat droppings and leech gut contents with tagged generic arthropod and mammal primers, respectively. We found that an average of 2.6% and 2.1% of sequences had tag combinations, which could be explained by tag jumping in the leech and bat diet study, respectively. We suggest that tag jumping can happen during blunt-ending of pools of tagged amplicons during library build and as a consequence of chimera formation during bulk amplification of tagged amplicons during library index PCR. We argue that tag jumping and contamination between libraries represents a considerable challenge for Illumina-based metabarcoding studies, and suggest measures to avoid false assignment of tag jumping-derived sequences to samples. © 2015 John Wiley & Sons Ltd.

  11. Recombinant decorsin: dynamics of the RGD recognition site.

    PubMed Central

    Krezel, A. M.; Ulmer, J. S.; Wagner, G.; Lazarus, R. A.

    2000-01-01

    Decorsin is an antagonist of integrin alphaIIbbeta3 and a potent platelet aggregation inhibitor. A synthetic gene encoding decorsin, originally isolated from the leech Macrobdella decora, was designed, constructed, and expressed in Escherichia coli. The synthetic gene was fused to the stII signal sequence and expressed under the transcriptional control of the E. coli alkaline phosphatase promoter. The protein was purified by size-exclusion filtration of the periplasmic contents followed by reversed-phase high-performance liquid chromatography. Purified recombinant decorsin was found to be indistinguishable from leech-derived decorsin based on amino acid composition, mass spectral analysis, and biological activity assays. Complete sequential assignments of 1H and proton bound 13C resonances were established. Stereospecific assignments of 21 of 25 nondegenerate b-methylene groups were determined. The RGD adhesion site recognized by integrin receptors was found at the apex of a most exposed hairpin loop. The dynamic behavior of decorsin was analyzed using several independent NMR parameters. Although the loop containing the RGD sequence is the most flexible one in decorsin, the conformation of the RGD site itself is more restricted than in other proteins with similar activities. PMID:10975565

  12. Automated video analysis of animal movements using Gabor orientation filters.

    PubMed

    Wagenaar, Daniel A; Kristan, Wiliam B

    2010-03-01

    To quantify locomotory behavior, tools for determining the location and shape of an animal's body are a first requirement. Video recording is a convenient technology to store raw movement data, but extracting body coordinates from video recordings is a nontrivial task. The algorithm described in this paper solves this task for videos of leeches or other quasi-linear animals in a manner inspired by the mammalian visual processing system: the video frames are fed through a bank of Gabor filters, which locally detect segments of the animal at a particular orientation. The algorithm assumes that the image location with maximal filter output lies on the animal's body and traces its shape out in both directions from there. The algorithm successfully extracted location and shape information from video clips of swimming leeches, as well as from still photographs of swimming and crawling snakes. A Matlab implementation with a graphical user interface is available online, and should make this algorithm conveniently usable in many other contexts.

  13. Phenotypically plastic responses of green frog embryos to conflicting predation risk.

    PubMed

    Ireland, D H; Wirsing, A J; Murray, D L

    2007-05-01

    Predators have been shown to alter the timing of switch points between life history stages, but few studies have addressed switch point plasticity in prey exposed simultaneously to conflicting predation pressure. We tested hatching responses of green frog (Rana clamitans) embryos subject to perceived predation risk from chemical cues released by two stage-specific predators, predicting that these predators would elicit: (1) directional hatching responses when presented independently, and (2) intermediate phenotypic responses when presented simultaneously. R. clamitans embryos in outdoor exclosures were exposed to cues from an egg predator (freshwater leeches; Nephelopsis obscura), a larval predator (dragonfly nymphs, Aeschna canadensis), and both predators in a 2 x 2 factorial experiment, and changes in hatchling size, hatchling developmental stage, and hatching time were compared to those for control embryos. Leeches alone induced embryos to hatch at a smaller size and an earlier developmental stage than controls, while dragonfly nymphs elicited a delay in egg hatching time that was associated with larger size and later developmental stage at hatching. Embryos failed to respond to simultaneous exposure to both predators, implying that responses to each occurred concurrently and were therefore dampened. Our results indicate that prey under threat from conflicting predators may manifest intermediate defensive phenotypes. Such intermediate responses may result in elevated rates of prey mortality with possible consequences at the population level.

  14. Rhopalobdella japonica n. gen., n. sp. (Hirudinea, Piscicolidae) from Dasyatis akajei (Chondrichthyes: Dasyatididae) in the northwestern Pacific.

    PubMed

    Burreson, E M; Kearn, G C

    2000-08-01

    A new genus and species of piscicolid leech in the Platybdellinae inhabits the oral cavity of Dasyatis akajei in the northwestern Pacific Ocean near Tanabe, Japan. The genus Rhopalobdella n. gen. is characterized externally by very small oral and caudal suckers and a smooth body that is widest just posterior to the clitellum. Eyespots and ocelli are lacking. The coelom is spacious with large segmental connecting sinuses between dorsal and ventral sinuses. There are 5 pairs of testisacs, an unusually extensive epididymis, and a very large bursa. Conducting tissue is absent. There are 2 pairs of esophageal diverticula and very well developed nephridia. Rhopalobdella japonica n. gen. n. sp. is characterized by a urosome that tapers strongly to the caudal sucker and by a single gonopore; the common oviduct opens into the posterior portion of the bursa. The coelomic and excretory systems resemble Aestabdella, but in other respects the genera are quite different. This is the first marine leech reported from rays in the northwestern Pacific.

  15. Identifying genes for neuron survival and axon outgrowth in Hirudo medicinalis.

    PubMed

    Blackshaw, S E; Babington, E J; Emes, R D; Malek, J; Wang, W-Z

    2004-01-01

    We have studied the molecular basis of nervous system repair in invertebrate (Hirudo medicinalis) nerve cells. Unlike in mammals, neurons in invertebrates survive injury and regrow processes to restore the connections that they held before the damage occurred. To identify genes whose expression is regulated after injury, we have used subtractive probes, constructed from regenerating and non-regenerating ganglia from the leech Hirudo medicinalis, to screen cDNA libraries made from whole leech CNS or from identified microdissected neurons. We have identified genes of known or predicted function as well as novel genes. Known genes up-regulated within hours of injury and that are widely expressed in invertebrate and mammalian cells include thioredoxin and tubulin. Other known genes, e.g. Cysteine Rich Intestinal Protein (CRIP), have previously been identified in mammalian cells though not in regenerating adult neurons. Two regulated genes identified, myohemerythrin and the novel protein ReN3 are exclusively expressed in invertebrates. Thus our approach has enabled us to identify genes, present in a neuron of known function, that are up- and down-regulated within hours of axotomy, and that may underpin the intrinsic ability of invertebrate neurons to survive damage and initiate regrowth programmes.

  16. The Lan3-2 glycoepitope of Hirudo medicinalis consists of beta-(1,4)-linked mannopyranose: cell type-specific glycans of H. medicinalis.

    PubMed

    Huang, Linjuan; Hollingsworth, Rawle I; Haslam, Stuart M; Morris, Howard R; Dell, Anne; Zipser, Birgit

    2008-12-01

    While glycosyltransferases are restrictively expressed in invertebrate model organisms, little is known of their glycan end products. One such restrictively expressed glycoepitope was localized to sensory and epithelial cells of leech and Caenorhabditis elegans using the Lan3-2 monoclonal antibody. A biological function for the neural Lan3-2 epitope was previously determined in the leech. Here we report on the chemical structure of this mannosidic epitope harvested from whole Hirudo medicinalis. Crude glycans were liberated from glycoproteins by hydrazinolysis. Re-N-acetylated glycans were subjected to immunoaffinity purification. The affinity-purified glycans were fractioned by size chromatography into oligosaccharides and polysaccharides. Lan3-2 oligosaccharide structure was characterized by gas chromatography of alditol acetates, methylation analysis, 500 MHz 1H NMR spectroscopy, matrix-assisted laser desorption/ionization mass spectrometry, and electrospray ionization tandem MS-MS of permethylated derivatives. The predominant components of the Lan3-2 oligosaccharide fraction were a series of linear beta-(1,4)-linked mannose polymers. The homologous expression of the Lan3-2 epitope in C. elegans will facilitate the exploration of its glycosylation pathway. Other invertebrates expressing the Lan3-2 epitope are Planaria dugesia, Capitella sp. I and Lumbriculus variegatus. The glycoepitope was not detected in the diploblastic animals Hydra littoralis and Aptaisia sp. or in deuterostomes.

  17. Identifying genes for neuron survival and axon outgrowth in Hirudo medicinalis

    PubMed Central

    Blackshaw, SE; Babington, EJ; Emes, RD; Malek, J; Wang, W-Z

    2004-01-01

    We have studied the molecular basis of nervous system repair in invertebrate (Hirudo medicinalis) nerve cells. Unlike in mammals, neurons in invertebrates survive injury and regrow processes to restore the connections that they held before the damage occurred. To identify genes whose expression is regulated after injury, we have used subtractive probes, constructed from regenerating and non-regenerating ganglia from the leech Hirudo medicinalis, to screen cDNA libraries made from whole leech CNS or from identified microdissected neurons. We have identified genes of known or predicted function as well as novel genes. Known genes up-regulated within hours of injury and that are widely expressed in invertebrate and mammalian cells include thioredoxin and tubulin. Other known genes, e.g. Cysteine Rich Intestinal Protein (CRIP), have previously been identified in mammalian cells though not in regenerating adult neurons. Two regulated genes identified, myohemerythrin and the novel protein ReN3 are exclusively expressed in invertebrates. Thus our approach has enabled us to identify genes, present in a neuron of known function, that are up- and down-regulated within hours of axotomy, and that may underpin the intrinsic ability of invertebrate neurons to survive damage and initiate regrowth programmes. PMID:14690474

  18. Diagnostic value of plain abdominal radiography in stroke patients with bowel dysfunction.

    PubMed

    Moon, Hyo Jeong; Noh, Se Eung; Kim, Ji Hee; Joo, Min Cheol

    2015-04-01

    To evaluate the diagnostic value of plain abdominal radiography in stroke patients with bowel dysfunction. A total of 59 stroke patients were recruited and assigned into constipation or non-constipation group. Patients were interviewed to obtain clinical information, constipation score, and Bristol stool form scale. The total and segmental colon transit time (CTT) was measured using radio-opaque markers (Kolomark). The degree of stool retention was evaluated by plain abdominal radiography and scored by two different methods (Starreveld score and Leech score). The relationship between the clinical aspects, CTT, and stool retention score using plain abdominal radiography was determined. Average constipation score was 4.59±2.16. Average Bristol stool form scale was 3.86±1.13. The total and segmental CTTs showed significant differences between the constipation and non-constipation groups. There was statistically significant (p<0.05) correlation between the total CTT and constipation score or between Starreveld score and Leech score. Each segmental CTT showed significant correlation (p<0.05) between segmental stool retention scores. The stool retention score showed significant correlation with constipation score as well as total and segmental CTT. Thus, plain abdominal radiography is a simple and convenient method for the evaluation of bowel dysfunction in stroke patients.

  19. Feedback Signal from Motoneurons Influences a Rhythmic Pattern Generator.

    PubMed

    Rotstein, Horacio G; Schneider, Elisa; Szczupak, Lidia

    2017-09-20

    Motoneurons are not mere output units of neuronal circuits that control motor behavior but participate in pattern generation. Research on the circuit that controls the crawling motor behavior in leeches indicated that motoneurons participate as modulators of this rhythmic motor pattern. Crawling results from successive bouts of elongation and contraction of the whole leech body. In the isolated segmental ganglia, dopamine can induce a rhythmic antiphasic activity of the motoneurons that control contraction (DE-3 motoneurons) and elongation (CV motoneurons). The study was performed in isolated ganglia where manipulation of the activity of specific motoneurons was performed in the course of fictive crawling (crawling). In this study, the membrane potential of CV was manipulated while crawling was monitored through the rhythmic activity of DE-3. Matching behavioral observations that show that elongation dominates the rhythmic pattern, the electrophysiological activity of CV motoneurons dominates the cycle. Brief excitation of CV motoneurons during crawling episodes resets the rhythmic activity of DE-3, indicating that CV feeds back to the rhythmic pattern generator. CV hyperpolarization accelerated the rhythm to an extent that depended on the magnitude of the cycle period, suggesting that CV exerted a positive feedback on the unit(s) of the pattern generator that controls the elongation phase. A simple computational model was implemented to test the consequences of such feedback. The simulations indicate that the duty cycle of CV depended on the strength of the positive feedback between CV and the pattern generator circuit.SIGNIFICANCE STATEMENT Rhythmic movements of animals are controlled by neuronal networks that have been conceived as hierarchical structures. At the basis of this hierarchy, we find the motoneurons, few neurons at the top control global aspects of the behavior (e.g., onset, duration); and within these two ends, specific neuronal circuits control the

  20. Ab-initio study of polypyrrole as a pervaporation membrane

    NASA Astrophysics Data System (ADS)

    Botelho, Andre; Lin, Xi

    2010-03-01

    The affinities between polypyrrole, water, ethanol and a sulfonate-carrying ion were calculated from first principles. All interactions were demonstrably hydrogen bonds between the oxygen from the sulfonate groups and the hydrogen in the hydroxyl (for water and ethanol) and amine groups (for polypyrrole). Each sulfonate group was shown to form three hydrogen bonds, with any of the three other types of molecules, allowing the ion complex to bind to multiple polypyrrole chains, water, and ethanol molecules simultaneously. The energies indicated a higher affinity between the ion and poypyrrole, second highest between water and the ion, and the lowest for ethanol and the ion. A high affinity of the ion to the polymer backbone is desirable to prevent leeching. The ion was found to have a higher affinity to water molecules than to ethanol, confirming the system's selectivity in separating water from ethanol.