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Bacterial symbiont and salivary peptide evolution in the context of leech phylogeny

  • DOI:
  • Published online: 10 May 2011

The evolutionary history of leeches is employed as a general framework for understanding more than merely the systematics of this charismatic group of annelid worms, and serves as a basis for understanding blood-feeding related correlates ranging from the specifics of gut-associated bacterial symbionts to salivary anticoagulant peptides. A variety of medicinal leech families were examined for intraluminal crop bacterial symbionts. Species of Aeromonas and Bacteroidetes were characterized with DNA gyrase B and 16S rDNA. Bacteroidetes isolates were found to be much more phylogenetically diverse and suggested stronger evidence of phylogenetic correlation than the gammaproteobacteria. Patterns that look like co-speciation with limited taxon sampling do not in the full context of phylogeny. Bioactive compounds that are expressed as gene products, like those in leech salivary glands, have ‘passed the test’ of evolutionary selection. We produced and bioinformatically mined salivary gland EST libraries across medicinal leech lineages to experimentally and statistically evaluate whether evolutionary selection on peptides can identify structure-function activities of known therapeutically relevant bioactive compounds like antithrombin, hirudin and antistasin. The combined information content of a well corroborated leech phylogeny and broad taxonomic coverage of expressed proteins leads to a rich understanding of evolution and function in leech history.

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*Corresponding author: Mark E Siddall. E-mail:
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This list contains references from the content that can be linked to their source. For a full set of references and notes please see the PDF or HTML where available.

K. Apakupakul , M. E. Siddall and E. M. Burreson (1999). Higher level relationships of leeches (Annelida: Clitellata: Euhirudinea) based on morphology and gene sequences. Molecular Phylogenetics and Evolution 12, 350359. doi:10.1006/mpev.1999.0639.

D. A. Bader , U. Roshan and A. Stamatakis (2006). Computational grand challenges in assembling the tree of life: problems and solutions. Advances in Computers 68, 127176. doi:10.1016/S0065-2458(06)68004-2.

I. P. Baskova and L. L. Zavalova (2001). Proteinase inhibitors from the medicinal leech Hirudo medicinalis. Biochemistry Moscow 66, 703717. doi:10.1023/A:1010223325313.

A. G. G. Batchelor , P. Davison and L. Sully (1984). The salvage of congested skin flaps by the application of leeches. British Journal of Plastic Surgery 37, 358360. doi:10.1016/0007-1226(84)90079-1.

E. Borda , A. Oceguera-Figueroa and M. E. Siddall (2008). On the evolution, classification and biogeography of some haemadipsoid leeches (Hirudinida: Arhynchobdellida: Hirudiniformes). Molecular Phylogenetics and Evolution 46, 142–54. doi:10.1016/j.ympev.2007.09.006.

E. Borda and M. E. Siddall (2004). Arhynchobdellida (Annelida: Oligochaeta: Hirudinida): phylogenetic relationships and evolution. Molecular Phylogenetics and Evolution 30, 213225. doi:10.1016/j.ympev.2003.09.002.

E. Borda and M. E. Siddall (2011). Insights into the evolutionary history of IndoPacific bloodfeeding terrestrial leeches (Hirudinida: Arhynchobdellida: Haemadipisdae). Invertebrate Systematics. 24, 456472. doi:10.1071/IS10013.

K. Bourtzis and T. A. Miller (2006). Insect Symbiosis, Volume 2. CRC Press/Taylor and Frances, Boca Raton, FL, London.

M. Derganc and F. Zdravic (1960). Venous congestion of flaps treated by application of leeches. British Journal of Plastic Surgery 13, 187192.

P. V. Dunlap , J. C. Ast , S. Kimura , A. Fukui , T. Yoshino and H. Endo (2007). Phylogenetic analysis of host-symbiont specificity and codivergence in bioluminescent symbioses. Cladistics 23, 507532. doi:10.1111/j.1096-0031.2007.00157.x.

F. Faria , L. Junqueira-de-Azevedo Ide , P. L. Ho , M. U. Sampaio and A. M. Chudzinski-Tavassi (2005). Gene expression in the salivary complexes from Haementeria depressa leech through the generation of expressed sequence tags. Gene 349, 173185. doi:10.1016/j.gene.2004. 12.022.

J. S. Farris , V. A. Albert , M. Källersjö , D. Lipscomb and A. G. Kluge (1996). Parsimony jackknifing outperforms neighbor-joining. Cladistics 12, 99124. doi:10.1111/j.1096-0031.1996.tb00196.x.

G. Giribet (2008). Assembling the lophotrochozoan (=spiralian) tree of life. Philosophical Transactions of the Royal Society B 363, 15131522. doi:10.1098/rstb.2007.2241.

P. A. Goloboff , J. S. Farris and K. C. Nixon (2008). TNT, a free program for phylogenetic analysis. Cladistics 24, 774786. doi:10.1111/j.1096-0031.2008.00217.x.

J. Graf (2002). The effect of symbionts on the physiology of Hirudo medicinalis, the medicinal leech. Invertebrate Reproduction and Development 41, 269275.

A. Greinacher , H. Völpel , U. Janssens , V. Hach-Wunderle , B. Kemkes-Matthes , P. Eichler , H. G. Mueller-Velten and B. Pötzsch (1999). Recombinant hirudin (Lepirudin) provides safe and effective anticoagulation in patients with heparin-induced thrombocytopenia. Circulation 99, 7380.

G. Haas (1924). Versuche der Blutauswaschung am Lebenden mit Hilfe der Dialyse. Wiener Klinische Wochenschrift 4, 1314. doi:10.1007/BF01745400.

C. T. Hittinger , M. Johnston , J. T. Tossberg and A. Rokas (2010). Leveraging skewed transcript abundance by RNA-Seq to increase the genomic depth of the tree of life. Proceedings of the National Academy of Sciences, USA 107, 14761481. doi: 10.1073/pnas.0910449107.

W. A. Jackson (2001). A short guide to humoral medicine. TRENDS in Pharmacological Sciences 22, 487489.

A. J. Keading , J. C. Ast , M. M. Pearce , H. Urbanczyk , S. Kimura , H. Endo , M. Nakamura and P. V. Dunlap (2007). Phylogenetic diversity and cosymbiosis in the bioluminescent symbioses of “Photobacterium mandapamensis. Applied and Environmental Microbiology 73, 31733182. doi:10.1128/AEM.02212-06.

Y. Kiskuchi and T. Fukatsu (2002). Endosymbiotic bacteria in the esophageal organ of glossiphoniid leeches. Applied and Environmental Microbiology 68, 46374641. doi:10.1128/AEM.68.9.4637-4641.2002.

Y. Kikuchi and J. Graf (2007). Spatial and temporal population dynamics of a naturally occurring two-species microbial community inside the digestive tract of the medicinal leech. Applied and Environmental Microbiology 73, 19841991. doi:10.1128/AEM.01833-06.

J. R. Kimbell and M. J. McFall-Ngai (2003). The squid-Vibrio symbioses: from demes to genes. Integrative and Comparative Biology 43, 254260. doi: 10.1093/icb/43.2.254.

J. R. Kimbell , M. J. McFall-Ngai and G. K. Roderick (2002). Two genetically distinct populations of bobtail squid, Euprymna scolopes, exist on the island of Oahu. Pacific Science 56, 347355.

S. L. Kosakovsky Pond , S. D. W. Frost and S. V. Muse. (2005). HyPhy: hypothesis testing using phylogenies. Bioinformatics 21, 676679. doi:10.1093/bioinformatics/bti079.

C. Kosiol , T. Vinar , R. R. da Fonseca , M. J. Hubisz , C. D. Bustamante , R. Nielsen and A. Siepel (2008). Patterns of positive selection in six mammalian genomes. PLoS Genetics 4, e1000144. doi:10.1371/journal.pgen.1000144.

D. M. Kristensen , L. Kannan , M. K. Coleman , Y. I. Wolf , A. Sorokin , E. V. Koonin and A. Mushegian (2010). A low-polynomial algorithm for assembling clusters of orthologous groups from intergenomic symmetric best matches. Bioinformatics 26, 14811487.

S. Kvist , S. A. Montanari , H. Yi , B. Fuks and M. E. Siddall (2011). Teaching Biodiversity & Evolutionary Biology in a North American Marine Coastal Environment. The American Biology Teacher 73, 7277. doi:10.1525/abt.2011.73.2.4.

I. Lakatos (1971). History of science and its rational reconstruction. Proceedings of the Biennial Meeting of the Philosophy of Science Association 1970, 91136

R. Lapatto , U. Krengel , H. A. Schreuder , A. Arkema , B. de Boer , K. H. Kalk , W. G. Hol , P. D. Grootenhuis , J. W. Mulders , R. Dijkema , H. J. Theunissen and B. W. Dijkstra (1997). X-ray structure of antistasin at 1·9 A resolution and its modelled complex with blood coagulation factor Xa. EMBO Journal 16, 51515161. doi:10.1093/emboj/16.17.5151.

A. S. Laufer , M. E. Siddall and J. Graf (2008). Characterization of the digestive-tract microbiota of Hirudo orientalis, a European medicinal leech. Applied and Environmental Microbiology 74, 61516154. doi:10.1128/AEM.00795-08.

T. A. Mason , P. J. McIlroy and D. H. Shain (2004). A cysteine-rich protein in the Theromyzon (Annelida: Hirudinea) cocoon membrane. Federation of European Biochemical Societies Letters 561, 167172. doi:10.1016/S0014-5793(04)00167-X.

F. Markwardt (1992). Hirudin: The promising antithrombotic. Cardiovascular Drug Reviews 10, 211232. doi:10.1111/j.1527-3466.1992.tb00247.x.

D. J. McLaughlin , D. S. Hibbett , F. Lutzoni , J. W. Spatafora’ and R. Vilgalys (2009). The search for the fungal tree of life. Trends in Microbiology 17, 488497. doi:10.1016/j.tim.2009.08.001.

G-S. Min , I. N. Sarkar , and M. E. Siddall (2010). Salivary transcriptome of the North American medicinal leech, Macrobdella decora. Journal of Parasitology 96, 12111221. doi:10.1645/GE-2496.1.

A. Morabia (1996). P. C. A. Louis and the birth of clinical epidemiology. Journal of Clinical Epidemiology 49, 13271333. doi:10.1016/S0895-4356(96)00294-6.

N. A. Moran (2001). The coevolution of bacterial endosymbionts and phloem-feeding insects. Annals of the Missouri Botanical Garden 88, 3544.

R. Munro , M. Siddall , S. S. Desser , and R. T. Sawyer (1992). The leech as a tool for studying comparative haematology. Comparative Haematology International 2, 7578. doi:10.1007/BF00186263.

M. K. Nishiguchi (2002). Host-symbiont recognition in the environmentally transmitted sepiolid squid-Vibrio mutualism. Microbial Ecology 44, 1018. doi:10.1007/BF03036870.

M. K. Nishiguchi , J. E. Lopez and S. V. Boletzky (2004). Enlightenment of old ideas from new investigations: more questions regarding the evolution of bacteriogenic light organs in squids. Evolution and Development 6, 4149. doi:10.1111/j.1525-142X.2004.04009.x.

K. C. Nixon (1999). The parsimony ratchet, a new method for rapid parsimony analysis. Cladistics 15, 407414. doi:10.1111/j.1096-0031.1999.tb00277.x.

J. A. A. Nylander , U. Olsson , P. Alström and I. Sanmartín (2008). Accounting for phylogenetic uncertainty in biogeography: a Bayesian approach to Dispersal-vicariance Analysis of the thrushes (Aves: Turdus). Systematic Biology 57, 257268. doi: 10.1080/10635150802044003.

R. D. M. Page (1990). Component analysis: a valiant failure? Cladistics 6, 119136. doi:10.1111/j.1096-0031.1990.tb00532.x.

S. L. Perkins , R. B. Budinoff and M. E. Siddall (2005). New gammaproteobacteria associated with blood-feeding leeches and a broad phylogenetic analysis of leech endosymbionts. Applied and Environmental Microbiology 71, 52195224. doi:10.1128/AEM.71.9.5219-5224.2005.

A. J. Phillips , R. Arauco-Brown , A. Oceguera-Figueroa , G. P. Gomez , M. Beltran , Y.-T. Lai and M. E. Siddall (2010). Tyrannobdella rex n. gen. n. sp. and the evolutionary origins of mucosal leech infestations. PLoS ONE 5, e10057. doi:10.1371/journal.pone.0010057.

A. J. Phillips and M. E. Siddall (2005). Phylogeny of the New World medicinal leech family Macrobdellidae (Oligochaeta: Hirudinida: Arhynchobdellida). Zoologica Scripta 34, 559564. doi:10.1111/j.1463-6409.2005.00210.x.

A. J. Phillips and M. E. Siddall (2009). Poly-paraphyly of Hirudinidae: Many lineages of medicinal leeches. BMC Evolutionary Biology 9, 246256. doi:10.1186/1471-2148-9-246.

S. L. K. Pond and S. D. W. Frost (2005). Datamonkey: rapid detection of selective pressure on individual sites of codon alignments. Bioinformatics 21, 25312533. doi: 10.1093/bioinformatics/bti320.

A. F. Y. Poon , F. I. Lewis , S. L. Kosakovsky Pond and S. D. W. Frost (2007). An evolutionary-network model reveals stratified interactions in the V3 loop of the HIV-1 envelope. PLoS Computational Biology 3, e231. doi:10.1371/journal.pcbi.0030231.

R. H. Ree and S. A. Smith (2008). Maximum likelihood inference of geographic range evolution by dispersal, local extinction, and cladogenesis. Systematic Biology 57, 414. doi: 10.1080/10635150701883881.

R. V. M. Rio , M. Anderegg and J. Graf (2007). Characterization of a catalase gene from Aeromonas veronii, the digestive-tract symbiont of the medicinal leech. Microbiology 153, 18971906. doi:10.1099/mic.0.2006/003020-0

R. V. M. Rio , M. Maltz , B. McCormick , A. Reiss and J. Graf (2009). Symbiont succession during embryonic development of the European medicinal leech, Hirudo verbana. Applied and Environmental Microbiology 75, 68906895. doi:10.1128/AEM.01129-09.

E. P. Rocha , J. M. Smith , L. D. Hurst , M. T. Holden , J. E. Cooper , N. H. Smith and E. J. Feil (2005). Comparisons of dN/dS are time dependent for closely related bacterial genomes. Journal of Theoretical Biology 239, 226–35.

F. Ronquist (1997). Dispersal-vicariance analysis: a new approach to the quantification of historical biogeography. Systematic Biology 46, 195203. doi: 10.1093/sysbio/46.1.195.

K. Scheffler , D. P. Martin and C. Seoighe (2006). Robust inference of positive selection from recombining coding sequences. Bioinformatics 22, 24932499. doi: 10.1093/bioinformatics/btl427.

M. E. Siddall (2002). Phylogeny and revision of the leech family Erpobdellidae (Hirudinida: Oligochaeta). Invertebrate Systematics 16, 16. doi:10.1071/IT01011.

M. E. Siddall , K. Apakupakul , E. M. Burreson , K. A. Coates , C. Erséus , S. R. Gelder , M. Källersjö and H. Trapido-Rosenthal (2001). Validating Livanow: Molecular data agree that leeches, Branchiobdellidans and Acanthobdella peledina form a monophyletic group of Oligochaetes. Molecular Phylogenetics and Evolution 21, 346351. doi:10.1006/mpev.2001.1021.

M. E. Siddall , R. B. Budinoff , and E. Borda (2005). Phylogenetic evaluation of systematics and biogeography of the leech family Glossphoniidae. Invertebrate Systematics 19, 105112. doi:10.1071/IS04034.

M. E. Siddall and E. M. Burreson (1998). Phylogeny of leeches (Hirudinea) based on mitochondrial cytochrome c Oxidase Subunit I. Molecular Phylogenetics and Evolution 9, 156162. doi:10.1006/mpev.1997.0455.

M. E. Siddall and E. M. Burreson (1995). Phylogeny of the Euhirudinea: Independent evolution of blood feeding by leeches? Canadian Journal of Zoology 73, 10481064. doi:10.1139/z95-125.

M. E. Siddall , S. L. Perkins and S. S. Desser (2004). Leech mycetome endosymbionts are a new lineage of alphaproteobacteria related to the Rhizobiaceae. Molecular Phylogenetics and Evolution 30, 178186. doi:10.1016/S1055-7903(03)00184-2.

M. E. Siddall , P. Trontelj , S. Y. Utevsky , M. Nkamany and K. S. Macdonald (2007 a). Diverse molecular data demonstrate that commercially available medicinal leeches are not Hirudo medicinalis. Proceedings of the Royal Society B 274, 14811487. doi: 10.1098/rspb.2007.0248.

M. E. Siddall , P. L. Worthen , M. Johnson and J. Graf (2007 b). Novel role for Aeromonas jandaei as a digestive tract symbiont of the North American medicinal leech. Applied and Environmental Microbiology 73, 655658. doi: 10.1128/AEM.01282-06.

A. Silver , N. M. Rabinowitz , S. Küffer and J. Graf (2007). Identification of Aeromonas veronii genes required for colonization of the medicinal leech, Hirudo verbana. Journal of Bacteriology 189, 67636772. doi:10.1128/JB.00685-07.

W. Soto , J. Gutierrez , M. D. Remmenga and M. K. Nishiguchi (2009). Salinity and temperature effects of physiological responses of Vibrio fischeri from diverse ecological niches. Microbial Ecology 57, 140150. doi:10.1007/s00248-008-9412-9.

S. Y. Utevsky and P. Trontelj (2004). Phylogenetic relationships of fish leeches (Hirudinea, Piscicolidae) based on mitochondrial DNA sequences and morphological data. Zoologica Scripta 33, 375385. doi:10.1111/j.0300-3256.2004.00156.x.

R. L. Warren , G. G. Sutton , S. M. J. Jones and R. A. Holt (2007). Assembling millions of short DNA sequences using SSAKE. Bioinformatics 23, 500501. doi:10.1093/bioinformatics/btl629.

I. S. Whitaker , C. Kamya , E. A. Azzopardi , J. Graf , M. Kon and W. C. Lineaweaver (2009). Preventing infective complications following leech therapy: Is practice keeping pace with current research? Microsurgery 29, 619625. doi:10.1002/micr.20666.

J. I. Williams and E. M. Burreson (2006). Phylogeny of the fish leeches (Oligochaeta, Hirudinida, Piscicolidae) based on nuclear and mitochondrial genes and morphology. Zoologica Scripta 35, 627639. doi:10.1111/j.1463-6409.2006.00246.x.

P. L. Worthen , C. J. Gode and J. Graf (2006). Culture-independent characterization of the digestive-tract microbiota of the medicinal leech reveals a tripartite symbiosis. Applied and Environmental Microbiology 72, 47754781. doi:10.1128/AEM.00356-06.

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