Get 20M+ Full-Text Papers For Less Than $1.50/day. Start a 14-Day Trial for You or Your Team.

Learn More →

Evolution of salivary secretions in haematophagous animals

Evolution of salivary secretions in haematophagous animals BioscienceHorizons Volume 10 2017 10.1093/biohorizons/hzw015 .............................................. .................................................. .................................................. ............... Review article Evolution of salivary secretions in haematophagous animals Francesca L. Ware and Martin R. Luck School of Biosciences, University of Nottingham, Sutton Bonington Campus, Loughborough, Leicester LE12 5RD, UK *Corresponding author: 27 Braishfield Gardens, Bournemouth, Dorset BH8 0QA, UK. Email: francesca.ware@yahoo.co.uk Supervisor: Prof. Martin Luck, School of Biosciences, University of Nottingham, Sutton Bonington Campus, Loughborough, Leicester LE12 5RD, UK. .............................................. .................................................. .................................................. ............... Haemostasis is the prevention of blood fluidity in vertebrates and is the first stage of wound healing. Haematophagous animals use the blood of vertebrates as their sole source of nutrition and have evolved many salivary constituents to counteract the haemostatic response of their prey. These animals and their saliva have been studied for many years, with some applications in medicine. The purpose of this study is to compare the salivary constituents of leeches (Hirudinae), ticks (Argasidae and Ixodidae) and vampire bats (Desmodontinae) and to consider their evolutionary origin. Salivary constituents include plasminogen activa- tors (PAs), anticoagulants (activated factor X, FXa; inhibitors), vasodilators, platelet aggregation inhibitors (PAgI) and thrombin inhibitors. The animals studied all tend to possess an anticoagulant and a form of apyrase (PAgI) to assist with blood feeding. Ticks and vampire bats have a form of PA but the leech does not. The vampire bat has a PAgI but no vasodilator. The animals studied are from taxonomically unrelated groups but exploit similar mechanisms of action to facilitate their haematophagy. Given that the haematophagous lifestyle of these animals developed much later than their common ancestors, we conclude that their mechanisms for haematophagy have arisen by convergent evolution. Some molecules, e.g. serine proteases found in invertebrate saliva, are probably derived from a common ancestral gene. The possible paths that have led to evolution of vam- pire bat salivary components are considered. Further research into the homology of these salivary constituents is required to give insight into how these animals adapted to haematophagy and their further therapeutic potential. Key words: haematophagous, saliva, evolution, leech, tick, vampire bat Submitted on 21 September 2015; editorial decision on 28 November 2016 .............................................. .................................................. .................................................. ............... number of species, we have chosen to examine examples Introduction representing distinctly different phyla. We describe and com- pare the range of active components in the saliva of haema- Haematophagous animals are those which rely on blood from tophagous animals in the following three groups: leeches other animals as their only source of nutrition. These parasitic (Hirudinae), ticks (Ixodidae and Argasidae) and vampire bats creatures have evolved highly specific salivary molecules that (Desmodontinae). We then consider whether these components counteract the haemostatic response of the host and also exert may have arisen through convergent or divergent evolution. limited behavioural control. Whilst considerable research has been carried out on the salivary components and their poten- Haemostasis (Fig. 1) is the process by which blood flow stops tial for the treatment of human disease, few papers have at the site of an injury. It is one of the first responses to vascular explored their evolution. Because this unusual feeding method damage and initiates further processes including wound repair. is displayed across the animal kingdom but in a limited Briefly, coagulation (reviewed by Palta, Saroa and Palta, 2014) ............................................................................................... .................................................................. © The Author 2017. Published by Oxford University Press. This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/ licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com Review article Bioscience Horizons � Volume 10 2017 ............................................................................................... .................................................................. serine proteases). Plasmin is the active component of the plas- minogen (fibrinolytic) system (reviewed by Chapin and Hajjar, 2015), responsible for the eventual breakdown of fibrin fibres, fibrinogen, FV, FVIII, FXII and prothrombin at the start of the wound repair process (Tellgren-Roth et al., 2009; Hall, 2011; Barrett et al., 2012). Leeches Leeches are invertebrate, segmented worms from the phylum Annelida, class Clitellata, subclass Hirudinae (reviewed by Abdualkader et al., 2013). Leeches live in slow flowing streams and fresh water ponds (reviewed by Hildebrandt and Lemke, 2011). Young leeches, having left the safety of the cocoon, feed on amphibians. When their mouth parts have matured, they move on to feed on animals with thicker skin and more nutritious blood, including birds and mammals (Hildebrandt and Lemke, 2011). Leeches, specifically the medicinal leech Hirudo medicina- lis, have been used by physicians as a medicinal therapy for various diseases since early civilization (reviewed by Munshi et al., 2008). Haycraft (1884) reported that the H. medicinalis produced a substance with anticoagulant properties. In fact, Figure 1. Haemostasis in vertebrates. A simplified account of the leech produces several anticoagulants and thrombolytics, haemostasis in vertebrates, including mammals, identifying possible stored in the salivary glands (Chopin et al., 2000). targets for the salivary molecules of haematophagous animals. Adapted from Law, Ribeiro and Wells (1992, review article) and Barrett The leech attaches to its prey using the anterior portion of et al. (2012). Dashed arrows indicate inhibition. 5-HT, serotonin; TXA , its sucker. It begins periodic tilting movements of its three jaws, thromboxane A2. in order to slice open the skin. The pumping action of the pha- ryngeal muscles sucks the blood from the destroyed blood ves- sels and lymph of the host into the leech’scrop (Lent et al., involves the formation of a loose haemostatic plug, which is 1988). These jaw movements also initiate the secretion of saliva then converted into a definitive clot (thrombus) containing from its unicellular salivary gland cells, located anteriorly in fibrin. The formation of fibrin is initiated by a cascade of segments three and nine (Hildebrandt and Lemke, 2011). The enzymic reactions (clotting factors of the intrinsic and extrinsic mechanism of protein release from gland cells and the biochem- pathways), the last element of which is a complex of activated ical events needed to synthesize salivary proteins are unknown substances, collectively termed the prothrombin activator. This (Hildebrandt and Lemke, 2011). Serotonin may stimulate sal- complex catalyses the conversion of prothrombin to thrombin iva excretion (Marshall and Lent, 1988; Hildebrandt and and is considered to be the rate limiting step in blood coagula- Lemke, 2011) and pharyngeal peristalsis (Lent et al.,1988). tion (Hall, 2011; Barrett et al., 2012). The ingestion of blood lasts roughly 25 min (Lent et al., Thrombin is a serine protease, which activates and poly- 1988). Only the red blood cells and plasma proteins are of merizes fibrinogen to form fibrin. Fibrin stabilizing factor nutritious value to the leech. Plasma and haem derivatives are (Factor XIII or FXIII; we use this system of abbreviation excreted over the following 4–6days, ensuring efficient diges- throughout), released from platelets within the developing tion. Table 1 shows the haematophagy-relevant components of thrombus, is activated by thrombin and produces a stable saliva in various leech species and their function in the host. cross-linked fibrin meshwork containing trapped blood cells, platelets and plasma. After the clot has formed (3–6 min), it begins to contract due to the release of pro-coagulant (clot- Anticoagulants encouraging) substances, which cross-link more adjacent The majority of the anticoagulants found in leeches are inhibi- fibrin fibres. The retracting fibrin meshwork pulls the edges of tors of FXa preventing the conversion of prothrombin to the damaged blood vessel together, assisting haemostasis thrombin of the vertebrate coagulation cascade (common path- (Hall, 2011; Barrett et al., 2012). way). Examples are Therostatin (isolated from Theromyzon A euglobulin called plasminogen (profibrinolysin) is found tessulatum; Chopin et al., 2000) and Antistatin. Antistatin within the plasma proteins trapped in the clot. Plasminogen is has also been proposed to have anti-metastatic properties converted into plasmin (fibrinolysin) by tissue-type (t-PA) or (Tuszynski, Gasic and Gasic, 1987). Ghilanten is an anticoagu- urokinase-type (u-PA) plasminogen activators (PAs; both are lant (FXa inhibitor) obtained from the leech Haementeria ............................................................................................... .................................................................. 2 Bioscience Horizons � Volume 10 2017 Review article ............................................................................................... .................................................................. ............................................................................................... .................................................................. Table 1. Haematophagy-related salivary constituents of different leech species and their functions in the host Leech species Molecule Function in host Reference Hirudo medicinalis Hirudin Thrombin inhibitor Jacobi (1904), Markwardt (1957), Nawarskas and (European medicinal Anderson (2001), Coppens et al. (2012), Jiang et al. leech) (2013) Bufrudin Thrombin inhibitor Electricwala et al. (1991), Abdualkader et al. (2013) Apyrase Agonist of platelets Rigbi, Orevi and Eldor (1996), Hildebrandt and Lemke (2011) Theromin Thrombin inhibitor Salzet et al. (2000) Mammalian-type Reduce platelet adherence Rigbi et al. (1987) collagenase Calin Platelet adhesion and activation inhibition Munro, Jones and Sawyer (1991), Abdualkader et al. (2013) Haementeria vizottoi Vizottin Anticoagulant (FXa inhibitor) Oliveira et al. (2012) Hyaluronidase Digests hyaluronic acid present in the ECM Linker, Hoffman and Meyer (1957), Hovingh and Linker (1999), Hildebrandt and Lemke (2011) Inhibitor of C1-complement Anti-inflammatory Baskova et al. (1988), Boskova and Zavalova (2001) system component LCI Regulator of fibrinolysis rate or inhibits carboxypeptidase Reverter et al. (1998), Hildebrandt and Lemke (2011) Eglin-c Impairment of neutrophils (alpha-chymotrypsin, subtilisin, Seemüller et al. (1977), Snider et al. (1985), Braun et al. chymosin, granulocyte proteinases, elastase and (1987), Zaidi et al. (2011) cathepsin G inhibitor) LDTI Possible role in supressing cell-mediated inflammatory Mϋhlhahn et al. (1994), Sommerhoff et al. (1994), reactions Stubbs et al. (1997); Hildebrandt and Lemke (2011) Hirustasin Antistatin-type serine protease inhibitor. Also trypsin, Söllner et al. (1994), Boskova and Zavalova (2001) alpha-chymotrypsin and neutrophil cathepsin G inhibitor Haementeria officinalis Antistatin Anticoagulant (FXa inhibitor) and anti-metastasis Tuszynski, Gasic and Gasic (1987) (Mexican Leech) Haementeria ghilianii Ghilanten Anticoagulant (FXa inhibitor) and anti-metastasis Brankamp et al. (1990), Blankenship et al. (1990) (Great/Giant Amazon Saratin PAgI Barnes et al. (2001), Abdualkader et al. (2013) Leech) Hementin Fibrinogenolytic metalloproteinase Swadesh, Huang and Budzynski (1990) Continued Review article Bioscience Horizons � Volume 10 2017 ............................................................................................... .................................................................. ghilianii (Blankenship et al.,1990; Brankamp et al.,1990), and is also believed to have anti-metastasic properties. Lefaxin is another anticoagulant (FXa inhibitor) from Haementeria depressa (Faria et al.,1999). The N-terminal chain of lefaxin shows no homology to antistatin or ghilianten. Another FXa inhibitor isolated from Haementeria vizottoi, called Vizottin, displays a different effect compared with other leech antistatin-like inhibitors (Oliveira et al., 2012). Vizottin prevents the formation of FXa by the extrinsic tenase complex (FVIIa: tissue factor) and also inhibits free and bound FXa, probably by interacting with the active site of FXa (Oliveira et al., 2012). Thrombin inhibitors A direct thrombin inhibitor is an anticoagulant, which binds to thrombin directly and blocks its activity (reviewed by Coppens et al.,2012). Hirudin (Jacobi, 1904) was isolated from H. med- icinalis by Markwardt (1957; reviewed by Zaidi et al.,2011). Hirudin is the most potent naturally occurring thrombin inhibi- tor (Zaidi et al.,2011; Jiang et al., 2013) and many of its syn- thetic derivatives are used in clinics on a daily basis (reviewed by Nawarskas and Anderson, 2001; Zaidi et al.,2011). Subsequently, a similar anti-thrombin called Bufrudin has also been isolated (Electricwala et al.,1991; Abdualkader et al., 2013). Hirudin and bufrudin have slightly different structural (N-terminal amino acid sequence) and immunological proper- ties (Electricwala et al.,1991; Zaidi et al.,2011). Hirustasin is an antistatin-type serine protease inhibitor and the first tissue kallikrein inhibitor identified in the leech (Söllner et al.,1994; Kallikrein-kinin system discovered by Abelous and Bardier, 1909, as cited by review Su, 2014 and Kraut, Frey and Werle, 1930). Despite similarities to antistatin (FXa inhibitor; differing reactive site sequence and proteinase activity), hirusta- sin does not inhibit blood coagulation in vitro, nor is it amido- lytic of isolated FXa. This suggests that the specificity of antistatin-type proteinases influences coagulation (reviewed by Söllner et al.,1994; Boskova and Zavalova, 2001). A further thrombin inhibitor, Theromin, has been isolated from the Duck Leech Theromyzon tessulatum. It has no sequence homology with any other thrombin inhibitor (Salzet et al., 2000). Analgesia Some kinins are potent activators of nociceptive nerve cells, which induce or enhance pain sensations (Steranka et al., 1988; Hildebrandt and Lemke, 2011). Tissue kallikreins are proteases that cleave inactive kininogens to become active kinins (Hildebrandt and Lemke, 2011). The presence of antistatin-type substances (e.g. Hirustasin) in leech saliva may be an indication that it could reduce local tissue kallikrein activity in host tissue around the feeding site, therefore pre- venting the production of pain-inducing kinins. It may have an analgesic effect on the host (Hildebrandt and Lemke, 2011). An alternative explanation for kinin suppression is the ............................................................................................... .................................................................. Table 1. Continued Leech species Molecule Function in host Reference Haementeria depressa Lefaxin Anticoagulant (FXa inhibitor) Faria et al. (1999), Abdualkader et al. (2013) Hementerin Fibrinogenolytic metalloproteinase Chudzinski-Tavassi et al. (1998) Theromyson tessulatum Therostatin Anticoagulant (FXa inhibitor) Chopin et al. (2000) (Duck leech) Theromacin Antimicrobial Tasiemski et al. (2004), Abdualkader et al. (2013) Theromyzin Antimicrobial Tasiemski et al. (2004), Abdualkader et al. (2013) ECM, extracellular matrix; LCI, leech carboxypeptidase inhibitor; LDTI, leech derived tryptase inhibitor; PAgI, platelet aggregation inhibitor. Bioscience Horizons � Volume 10 2017 Review article ............................................................................................... .................................................................. release of kininases into the wound to inactivate kinins by an adenosine 5’-diphosphate (ADP) diphosphohydrolase. The proteolytic cleavage. This removal of pain-inducing or sensi- latter removes the terminal phosphate group from ADP, gen- tizing agents from the feeding site would explain the analgesic erating AMP that does not bind to purinergic receptors, effect of leeches. thereby suppressing platelet adhesion (Rigbi, Orevi and Eldor, 1996; Hildebrandt and Lemke, 2011). Fibrinolysis/increasing blood flow Leech carboxypeptidase inhibitor (LCI; Reverter et al.,1998), Ticks inhibits a metalloproteinase (Carboxypeptidase B) responsible for the cleavage of kinin and therefore its inactivation in blood Ticks are ectoparasites of the order Parasitiformes, sub-order plasma. By inhibiting carboxypeptidase B, this increases kinin Ixodida (Black and Piesman, 1994), which have an obligate presence, which may result in increased pain and blood flow haematophagous lifestyle (reviewed by Kazimírová and to the feeding site. Whilst the increased blood flow would be Štibrániová, 2013). There are two major families, the beneficial to the leech, the increase in pain (kinin) would not. Ixodidae (hard; Black and Piesman, 1994; reviewed by Therefore, the LCI effect may be more relevant to inhibiting Francischetti et al., 2009) and Argasidae (soft). Ixodidae ticks carboxypeptidase B’ effect on fibrinolysis (Hildebrandt and are further partitioned into the metastriate (Dermacentor or Lemke, 2011). Rhipicephalusgenera spp.), which have short mouth parts and secrete a cement for attachment to the host (Francischetti Inhibitors of host immune system et al., 2009; Kazimírová and Štibrániová, 2013; reviewed by Sauer, 1977), and prostriate ticks, which use long, barbed The complement system is part of the innate immune system of mouth parts to stay attached to their host (Francischetti et al., vertebrates and is the first defence against invaders (e.g. bac- 2009; Kazimírová and Štibrániová, 2013). teria; Buchner, 1891). An inhibitor of complement component C1 (60–70 kDa), from leech saliva, blocks both the classic and Adult Ixodidae ticks prefer large mammals like humans alternative pathways of the complement system (Baskova et al., and especially, ruminant livestock (Maina et al., 2014). 1988; as cited and by Boskova and Zavalova, 2001)and is Immature Ixodidae ticks also feed on smaller mammals, birds anti-inflammatory (Hildebrandt and Lemke, 2011). and reptiles (Maina et al., 2014). Soft ticks (Argasidae) feed on a wide range of vertebrates including amphibians, reptiles Leech saliva contains a specific inhibitor of mast cells, leech- and birds (Kazimírová and Štibrániová, 2013). derived tryptase inhibitor (LDTI; Mϋhlhahn et al., 1994; Sommerhoff et al., 1994; Stubbs et al.,1997; Hildebrandt and The constituents of tick saliva diversify as they mature, Lemke, 2011). It exists in three isoforms with differing C- and quantities of various substances vary between each moult termini. Its biological function is not yet characterized, but it (instar) until sexual maturity is reached (Lloyd and Walker, may supress cell-mediated inflammation in host tissues at the 1995; Due et al., 2013; Juckett, 2013). feeding site (Hildebrandt and Lemke, 2011). The active constituents of tick saliva are shown in Table 2. A secretory protein found in leeches named Eglin-c Ticks form a haemorrhagic pool within the tissues of the host (Seemüller et al., 1977) binds to human neutrophils (Snider from which they feed (reviewed by Ribeiro, 1995). When a et al.,1985; Braun et al., 1987). Neutrophils are the most abun- tick is not attached to a host, salivary gland lobes produce dant white blood cell in mammals and part of the innate hygroscopic saliva. This helps the tick remain hydrated whilst immune system (Murphy, 2012). Eglin-c stops neutrophils at it waits for a host, sometimes for years (reviewed by Bowman the feeding site from entering the surrounding tissue, preventing and Sauer, 2004; Francischetti et al., 2009). Salivation is inflammation. It is therefore proposed as an anti-inflammatory believed to be under nervous control, involving cAMP and agent, which protects host tissues from destruction by endogen- calcium (Sauer, 1977). Ticks alternate in cycles of feeding and ous neutrophils (Hildebrandt and Lemke, 2011). salivation, each lasting 5–20 min (Gregson, 1967). After feed- ing, they fall off their hosts and become inactive and unwilling Hyaluronidase (Linker, Hoffman and Meyer, 1957; to reattach (Bowman and Sauer, 2004). It was long believed, Hovingh and Linker, 1999) digests hyaluronic acid present in as early as 27–79 AD, that ticks would die after gorging on the extracellular matrix of host tissues. Its secretion in leech blood due to lack of an anus. However, ticks do possess an saliva facilitates the distribution of other salivary molecules. anus and excrete mainly guanine and other nitrogenous waste These interfere with immune cell function in the host by in small amounts (Bowman and Sauer, 2004). mobilizing water molecules from proteoglycans and destabil- izing the matrix (Hildebrandt and Lemke, 2011). Vasodilators and inhibitors of immune Inhibitors of platelet activation and system function adherence A vasodilator assists the tick in its blood feeding by increasing Leech saliva contains two substances, which reduce host blood flow to feeding site (Ribeiro, Makoul and Robinson, platelet adherence to vessel walls at the feeding site: a 1988). The salivary vasodilators from Ixodes scapularis/dam- mammalian-type collagenase (Rigbi et al., 1987) and apyrase, mini are the arachidonic acid derivatives prostacyclin (PGI ; ............................................................................................... .................................................................. 5 Review article Bioscience Horizons � Volume 10 2017 ............................................................................................... .................................................................. Table 2. Haematophagy-related substances found in saliva of hard (Ixodidae) and soft (Argasidae) ticks and their functions in the host Tick species Molecule Function in host Reference Hard ticks (Ixodidae) Ixodes scapularis or Ixodes dammini (Deer/ PGI Platelet inhibitor Ribeiro, Makoul and Robinson (1988), Blacklegged tick) Kazimírová and Štibrániová (2013) PGE Vasodilator Ribeiro et al. (1985), Law, Ribeiro and Wells (1992) Metalloprotease Inhibits angiogenesis Valenzuela et al. (2002), Kazimírová and Štibrániová (2013) Ixolaris TFPI Lai et al. (2004), Francischetti et al. (2002b) - Kininase enzyme Ribeiro et al. (1985), Ribeiro and Mather (analgesic effect) (1998), Francischetti et al. (2009) Amblyomma americanum (Lone Star tick) PGE Vasodilator Ribeiro et al. (1992), Law, Ribeiro and Wells (1992), Ribeiro (1995) PGF Vasodilator Ribeiro et al. (1992) 2α Amblyomma hebraeum (South African Bont Tick) Amblin Thrombin inhibitor Lai et al. (2004) Amblyomma variegatum (Tropical Bont tick) Variegin Thrombin inhibitor Koh et al. (2007), Koh and Kini (2008) Peptide AP18 Enhances thrombin Koh et al. (2007) amidolytic activity - Anti-IL-8 Hajnická et al. (2001) Rhipicephalus/Boophilus microplus (Cattle/ PGE Vasodilator Dickinson et al. (1976), Higgs et al. Southern cattle tick) (1976), Law, Ribeiro and Wells (1992), Tatchell and Binnington (1973) Boophilin Thrombin inhibitor; Macedo-Ribeiro et al. (2008); Liao et al. serine protease (2009) inhibitor; potential anticoagulant Ixodidin Antimicrobial single- Fogaça et al. (2006) domain inhibitor/ protease inhibitor Rhipicephalus evertsi evertsi (Red-legged tick) Neurotoxin Paralysis of host Viljoen et al. (1986), Lloyd and Walker (1995) Hyalomma dromedarii (Camel tick) NTI-1 Thrombin inhibitor Ibrahim et al. (2001) NTI-2 Haemaphysalis Longicornis (Cattle/Bush/Shrub Longistatin PA and anticoagulant Anisuzzaman et al. (2010, 2011) tick) properties. (isolated from midgut) Hemalin Thrombin inhibitor Liao et al. (2009) Dermacentor variabilis (American dog/Wood Variabilin Platelet inhibitor Wang et al. (1996), Francischetti et al. tick) (Disintegrin) (2009) Dermacentor andersoni (Rocky Mountain - Anti-TNFα Ramachandra and Wikel (1995) wood tick) Continued ............................................................................................... .................................................................. 6 Bioscience Horizons � Volume 10 2017 Review article ............................................................................................... .................................................................. Table 2. Continued Tick species Molecule Function in host Reference Dermacentor reticulatus (Marsh/Ornate Cow/ - Anti-IL-8 Hajnická et al. (2001) American Levi tick), Rhipicephalus appendiculatus (Brown/Brown Ear tick), Haemaphysalis inermis (Winter tick), Ixodes ricinus (Sheep/Castor Bean/Deer tick) Soft ticks (Argasidae) Ornithodoros moubata (Soft/Argasid/African Ornithodorin Thrombin inhibitor van de Locht, Stubbs and Bode (1996), relapsing fever/Tampan tick/Eyeless Kazimírová and Štibrániová (2013) tampan) Moubatin PAgI Waxman and Connolly (1993), Basanova, Baskova and Zavalova (2002) TAP Anticoagulant (FXa Waxman et al. (1990) inhibitor) Ornithodoros savignyi (Sand tampan) Savignin Inhibits thrombin- Nienaber, Gaspar and Neitz (1999) induced platelet aggregation Apyrase Platelet inhibitor Mans et al. (1998a, 1998b), Kazimírová and Štibrániová (2013) Table includes all known putative secreted proteins, excluding housekeeping proteins or transcripts. PGI , prostaglandin I or prostacyclin; PGE , prostaglandin E ; ‘-’, name not specified; PGF , prostaglandin F ; NTI, non-competitive thrombin inhibitor; TFPI, 2 2 2 2 2α 2alpha tissue factor pathway inhibitor; TAP, tick anticoagulant peptide; PAgI, platelet aggregation inhibitor; PA, plasminogen activator. Ribeiro, Makoul and Robinson, 1988) and PGE (Ribeiro Anticoagulant and PA et al.,1985, reviewed by Valenzuela, 2004). PGE is also Haemaphysalis longicornis secretes a PA, characterized and foundinthe saliva of Amblyomma americanum (Ribeiro named Longistatin (Anisuzzaman et al.,2010, 2011). This has et al.,1992)and Boophilus microplus (Dickinson et al., high specificity to fibrin clot-bound plasminogen and has been 1976; Higgs et al., 1976; reviewed by Tatchell and proposed to have anticoagulant function also (Anisuzzaman Binnington, 1973; Law, Ribeiro and Wells, 1992). These et al.,2011). This combination of functions suggests that it is a may be involved in the generalized lymphocyte suppression critical component in the maintenance of blood pools in the of tick-infested species (Ribeiro et al., 1985; Law, Ribeiro feeding process of Ixodid ticks (Anisuzzaman et al., 2011). and Wells, 1992). Tick histamine-binding proteins (HBPs) are lipocalins, Platelet aggregation inhibitors which trap cationic, hydrophilic molecules. This is in contrast Tick saliva contains apyrase, a platelet agonist (Kazimírová and to most lipocalins that bind lipophilic compounds. This is pre- Štibrániová, 2013) and platelet inhibitors such as disintegrins sumably an adaptation to haematophagy; the salivary glands and prostacyclin. These inhibitors, for example Variabilin, con- of most if not all species of tick contain highly histamine- tain the arginine-glycine-aspartic acid (RGD) motif, which pre- specific binding proteins (Paesen et al., 1999, 2000). Evidence vents the binding of fibrinogen to platelets (Wang et al., 1996; suggests that expression of histamine is linked to an acquired Francischetti et al., 2009). A further platelet inhibitor called tick resistance (D. andersoni; reviewed by Wikel, 1982; Moubatin is capable of weakly inhibiting collagen-induced pla- Brossard and Wikel, 2004). telet aggregation (reviewed by Waxman and Connolly, 1993; The metalloprotease found in I. scapularis inhibits angio- Basanova, Baskova and Zavalova, 2002). genesis, the process of forming new blood capillaries (Valenzuela et al., 2002; Kazimírová and Štibrániová, Thrombin inhibitors 2013). Cytokines, including chemokines, interferons and tumour necrosis factor (TNF), are important in vertebrate Two non-competitive thrombin inhibitors, NTI-1 (3.2 kDa) and host immune responses, its development and expression. NTI-2 (14.9 kDa), have been isolated from Hyalomma drome- Tick saliva also contains anti-TNF (Ramachandra and darii (Ibrahim et al., 2001). A potent thrombin inhibitor found Wikel, 1995), anti-IL-8 (interleukin 8; Hajnická et al., 2001) in the tropical bont tick (Amblyomma variegatum), called and blocks other cytokine binding activities (Brossard and Variegin is a 32 residue polypeptide and one of the smallest Wikel, 2004). thrombin inhibitors discovered in nature (Koh et al., 2007, ............................................................................................... .................................................................. 7 Review article Bioscience Horizons � Volume 10 2017 ............................................................................................... .................................................................. reviewed by Koh and Kini, 2008). It has no structural similar- individuals (Greenhall and Schutt, 1996; Altringham, 2011; ities with thrombin inhibitors found in other haematophagous reviewed by Wimsatt, 1959). animals, including hirudin, Rhodniin and Theromin, even Vampire bats often roost in a group of 8–12 females; these though they have the same binding site on thrombin (Koh et al., are related and unrelated individuals with whom they per- 2007). Peptide AP18 (a synthetic peptide of variegin) has been form reciprocal altruism, the reduction of one animal’s fitness reportedto bindtoexosite-I, slightly enhancing thrombin ami- to increase that of another with the expectation that this dolytic activity (Koh et al., 2007). behaviour will be returned (Altringham, 2011). The fact that Boophilius are one-host ticks, which feed on cattle vampire bats perform this with those to whom they are not (Jongejan and Uilenberg, 2004). Boophilin, a proteinase closely related suggests a highly stable, social composition inhibitor, inhibits thrombin and interferes with serine pro- within the colony (Altringham, 2011). teases: trypsin and plasmin (Macedo-Ribeiro et al., 2008). Once the bat has located its prey (using long-range vision, Hemalin (Liao et al., 2009), a thrombin inhibitor of the olfaction, acute hearing and echolocation), it uses close-range Kunitz-type family, has high homology with boophilin thermal and mechanical sensitivity strategies to locate blood (Macedo-Ribeiro et al., 2008). The thrombin inhibitor capillaries below the skin surface (Jones, Teeling and Amblin (Lai et al., 2004) has been isolated from the haemo- Rossiter, 2013; Low et al., 2013). They attack only resting or lymph of ixodid tick Amblyomma hebraeum and displays sleeping prey and generally feed without disturbance sequence similarities to boophilin (Macedo-Ribeiro et al., (Wimsatt, 1959). The process must therefore be painless 2008) and Ixolaris (a tissue factor pathway inhibitor, TFPI; (Wimsatt, 1959), suggesting an analgesic in the saliva. Each Francischetti et al., 2002b). Inhibitors of the Kunitz family species possesses a characteristic lower lip median groove, with bovine-pancreatic-trypsin inhibitor domains are also which facilitates feeding (Wimsatt, 1959) but with small dif- common in ticks (reviewed by Corral-Rodríguez et al., 2009). ferences in anatomy (Greenhall and Schutt, 1996). Savignin is a thrombin inhibitor found in Ornithodoros savignyi, which inhibits thrombin-induced platelet aggrega- Desmodus rotundus possess upper and lower incisors. The tion (Nienaber, Gaspar and Neitz, 1999). It is a competitive, upper incisors diverge to create an upside-down ‘V’ shape. slow, tight-binding inhibitor that interacts with thrombin This creates a crater-like ‘divot’ 1–2 mm deep in the host’s tis- exosite-I and blocks its catalytic site (as does leech hirudin; sue. The bat laps up the blood via two straw-like ducts on the Nienaber, Gaspar and Neitz, 1999). ventral side of the tongue (Wimsatt, 1959), whilst saliva is released from the dorsal side of the tongue from the sub- maxillary gland (Low et al., 2013). It takes 20–30 min for a Other bat to take in ~25 ml, which is approximately 60% of their Neurotoxins found in the saliva of Rhipicephalus evertsi ever- bodyweight (Altringham, 2011). To prevent themselves from tsi cause host paralysis (Viljoen et al., 1986; Lloyd and being grounded by the weight and open to potential preda- Walker, 1995) and have presumably evolved to prevent the tors, their kidneys excrete excess plasma from the already host interrupting feeding sessions. Tick anticoagulant peptide ingested blood before they finish a feed (Altringham, 2011). (TAP), isolated from Ornithodoros moubata, functions as an The concoction of anticoagulants and other biochemicals inhibitor of FXa (Waxman et al., 1990). Ixodes scapularis in the bat’s saliva causes the normal response to this type of contains a kininase enzyme, which may contribute to the injury in the host to be delayed from minutes to hours (Low analgesic effect of bites (Ribeiro et al., 1985; Ribeiro and et al., 2013). The overall anticoagulant activity of the saliva Mather, 1998; Francischetti et al., 2009). decreases progressively after daily salivation and is restored after 4 days (Fernandez et al., 1998). Multiple feedings from an individual prey animal may increase its resistance to the Vampire bats anticoagulants, indicating that an immune response can be Three species of bat have evolved a haematophagous lifestyle: acquired (Delpieto and Russo, 2009). Diphylla ecaudata, Desmodus rotundus and Diaemus youngi. Anticoagulants These ‘vampire’ bats are mammals of the order Chiroptera, sub-order Microchiroptera, sub-family Desmodontinae Draculin is an anticoagulant found in vampire bat saliva (other members of the sub-order are insectivores). Avian (Apitz-Castro et al.,1995; Fernandez et al.,1998, 1999). It hosts are the only prey of the hairy-legged vampire bat (D. requires glycosylation of the native molecule to become a bio- ecaudata; Tellgren-Roth et al., 2009; Low et al., 2013). The logically active, non-competitive, tight-binding inhibitor of common vampire bat (D. rotundus) feeds substantially only FXa (Fernandez et al.,1998, 1999). It is also an inhibitor of on mammalian hosts, including livestock and humans (Low FIXa of the intrinsic pathway of blood coagulation (Fernandez et al., 2013). The white-winged vampire bat (D. youngi) et al., 1998; Basanova, Baskova and Zavalova, 2002; Oliveira feeds on both mammalian and avian blood (Low et al., et al.,2012). 2013). They are distributed widely over central and South America, typically living in caves, tree hollows and aban- Recent transcriptome and proteome studies of D. rotundus doned mines (Low et al., 2013)incoloniesof30–100 glands (Ma et al., 2013) revealed a 2-Kunitz-domain type ............................................................................................... .................................................................. 8 Bioscience Horizons � Volume 10 2017 Review article ............................................................................................... .................................................................. inhibitor, named Desmolaris (Ma et al., 2013). Desmolaris is This raises the question of whether divergent or convergent evo- a naturally deleted form of Kunitz-I-domainless TFPI and has lutionary mechanisms may have been in play. similar actions: it tightly binds FXIa of the extrinsic pathway We take divergent evolution to be the emergence in a spe- of the coagulation cascade (Ma et al., 2013). cies of an apomorphy not present in an ancestor (or the reten- tion of a characteristic in one descendant and its loss in Platelet aggregation inhibition and another descendant). This mechanism can be reliably invoked fibrinolytic activity when unique characteristics and mechanisms occur in a spe- cies or higher taxonomic group. We take convergent evolu- A platelet aggregation inhibitor (PAgI) (ADP-induced and tion to be evidenced by the presence of an apomorphy in thrombin-induced) in vampire bat saliva exerts its effects by more than one species when it is unlikely to have been present blocking ADP-binding sites or by specifically acting on the in their closest common ancestor. plasma cofactor involved in the platelet aggregation reaction (Hawkey, 1967). This molecule could be the apyrase or phos- The three animal groups represented here (mammals, phatase recently found in the saliva of D. rotundus arthropods and annelids) apparently diverged early in animal (Francischetti et al., 2013). evolution (reviewed by Sanetra et al., 2005). Mammals are deuterostomes while arthropods and annelids are proto- PA/fibrinolytic activity stomes (diverged from bilateria); arthropods are ecdysozoa and annelids are lophotrochozoa (diverged from protosto- The fibrinolytic activity of vampire bat saliva has been known mia). These major divergences probably occurred during the since 1932 (Bier, 1932). A PA (a serine protease) has been pre-Cambrian, Ediacaran era around 600 million years ago found and named D. rotundus salivary plasminogen activator (m.y.a.; Erwin and Davidson 2002; Peterson et al., 2008). In (DSPA), Desmokinase (Cartwright, 1974; Low et al.,2013)or addition, all three animals prey on amniote homeotherms— Desmoteplase (reviewed by Paciaroni, Medeiros and mammals and birds—which emerged considerably later than Bogousslavsky, 2009). It has been characterized (Hawkey, the parasite clades: 300–250 m.y.a. (Hellenius and Ruben 1966; Gardell et al.,1989) and cloned (Krätzschmar et al., 2004; Laurin and Reisz, 2011). 1991; Gulba, Praus and Witt, 1995; Francischetti et al.,2013). Four DSPAs (α1, α2, β and γ) have been isolated and character- Given that haematophagy is a rare phenomenon in each of ized (reviewed by Piechowski-Jozwiak and Bogousslavsky, the three groups, the likelihood that it is a characteristic which 2013); they are encoded by four different highly conserved has been retained from nearest common ancestors is extremely genes, each with distinct structure and properties (Krätzschmar small. Furthermore, assuming that the vertebrate hosts for the et al., 1991). All are homologous to human t-PA, except that haematophagous leeches and ticks emerged much later than the DSPAs are single-chain molecules and are dependent on the common ancestor of these parasites and that the mamma- fibrin as a cofactor (Liberatore et al.,2003). lian parasite–host relationships of the vampire bats must also have emerged later, we take the haematophagy of our three DSPA-α1 has been investigated for its pharmaceutical animals groups to be a convergently evolved lifestyle. potential and was in phase III clinical trials (Piechowski- Jozwiak and Bogousslavsky, 2013). Several substances found Given this conclusion, it would appear that any similarities in D. rotundus saliva (Table 3) could be of potential use in between the species in the mechanisms they employ to achieve thrombolytic therapy. haematophagy (preventing blood coagulation, nociception, would healing, etc.) must also have arisen convergently. Note, however, that the fundamental processes of coagulation, noci- Discussion ception and wound healing, against which haematophagous animals work, are largely common to mammals and birds. Haematophagous animals have evolved mechanisms, which They therefore define and limit the biochemical mechanisms, counteract the haemostatic response of their hosts. The which must have evolved for the haematophagous lifestyle to Triatominae sub-family of haematophagous bugs possesses be successful. A further possibility is that the mechanisms many beneficial compounds in their saliva to counteract haematophagous animals use are co-opted from, and adapta- haemostasis. Lipocalins, for example, also found in many other tions of, commonly existing biochemical processes. To this species of blood-feeding arthropods (Santos et al.,2007). The extent, they represent divergent evolution from their non- variety of mechanisms is extensive and several have been haematophagous cousins as well as convergent evolution exploited in medical applications (Abdualkader et al.,2013; towards unrelated similarity. reviewed by Cherniack, 2011; Piechowski-Jozwiak and Bogousslavsky, 2013). The three types of animal discussed in Most haematophagous species possesses at least one anti- this review represent taxonomically distinct groups and it is coagulant, which inhibits one or multiple factors involved clear that they exhibit similarities and differences in the in the mammalian coagulation cascade (Tables 1–4). mechanisms they employ. Other haematophagous parasites not Anticoagulants, most of which are FXa inhibitors, are the discussed (Table 4)such as sandflies (Lutzomyia longipalpis) dominant constituents of leech saliva. Although the N- also possess anti-coagulation mechanisms (Collin et al.,2012). terminal sequence of lefaxin showed no homology with other ............................................................................................... .................................................................. 9 Review article Bioscience Horizons � Volume 10 2017 ............................................................................................... .................................................................. Table 3. Haematophagy-related molecules found in the sub-maxillary and accessory glands of the vampire bat (D. rotundus) Molecule Function in host Reference Draculin Anticoagulant (FXa and FIXa Apitz-Castro et al. (1995), Fernandez et al. (1998, 1999), Basanova, inhibitor) Baskova and Zavalova (2002) Desmolaris Anticoagulant (FXIa Ma et al. (2013) inhibitor) Desmoteplase (DSPA-alpha-1) PA (fibrinolytic/ Bier (1932), Gardell et al. (1989), Krätzschmar et al. (1991), thrombolytic) Schleuning et al. (1992), Gulba, Praus and Witt (1995), Paciaroni, Merdeiros and Bogoussalvsky (2009); Tellgren-Roth et al. (2009); Francischetti et al. (2013), Piechowski-Jozwiak and Bogousslavsky (2013), reviewed by Patel, Ispoglou and Apostolakis (2014) Phosphatase Anti-platelet Hawkey (1967) Apyrase Anti-platelet Hawkey (1967) Pituitary adenylate cyclase Vasodilator activating peptide (PACAP) C-type natriuretic peptide (CNP) Vasodilator TNFα-stimulated gene 6 (TSG-6) Anti-inflammatory Lipophilin/secretoglobin precursors Anti-inflammatory (several) Lipocalin and other lipid carriers Anti-inflammatory Cystatin Anti-inflammatory (cycteine- type inhibitor) Chemokine CCL28 Antimicrobial (broad spectrum) β-Defensin, BPI/LBP/CETP family Antimicrobial lymphotoxin Kunitz domain (protease inhibitor Kunitz inhibitor domain) Neuroserpins Modulates activation of fibrinolysis triggered by DSPA Chymase Serine protease Clotting pathways serine proteases Protease DNAse Effects neutrophil function Source: Francischetti et al. (2013) unless otherwise stated specified. Further research is needed to confirm the function of some of these molecules in the host. Serine proteases are included because PAs belong to this family of enzymes. FIXa, activated coagulation factor IX; FXIa, activated coagulation factor XI; CCL28, (C-C motif) ligand 28; BPI/LBP/CETP family, bactericidal permeability-increasing protein/lipopolysaccharide-binding protein/cholesteryl ester transfer protein family; DNAse, deoxyribonuclease; PA, plasminogen activator; DSPA, D rotundus salivary plasminogen activator. leech anticoagulants it does show homology with Prolixin S Whilst evidence of analgesia is currently not available for derived from Rhodnius prolixus (Kissing bug; Hellmann and leeches, it is for Ixodes ticks (Ribeiro et al., 1985; Hawkins, 1965). The anticoagulant and anti-metastatic Hildebrandt and Lemke, 2011). Similar feeding physiology of agents antistatin and ghilanten are found in different species leeches has been observed in R. prolixus, suggesting that simi- of leech so it is most likely that these properties have devel- lar processes may be required for efficient blood feeding, also oped separately to facilitate blood feeding in these species. acquired convergently (Lent et al., 1988). ............................................................................................... .................................................................. 10 Bioscience Horizons � Volume 10 2017 Review article ............................................................................................... .................................................................. Table 4. Molecules in the saliva of haematophagous and non-haematophagous animals other than the three groups considered in detail in this review Species Molecule Function in host Reference Haematophagous animals Lutzomyia longipalpis (Sand fly) Maxadilan Vasodilator Ribeiro et al. (1989b), Law, Ribeiro and Wells (1992), Lerner et al. (1991). Lufaxin Anticoagulant (FXa Collin et al. (2012). inhibitor) - Anti-complement Cavalcante, Pereira and Gontijo (2003). Apyrase PAgI Ribeiro, Rossignol and Spielman (1986), Ribeiro (1995). Phlebotomus papatasi (Sand fly) Apyrase PAgI Ribeiro et al. (1989a), Ribeiro (1995). Phlebotomus argentipes Apyrase PAgI Ribeiro et al. (1989a), Ribeiro (1995). Phlebotomus perniciosus Apyrase PAgI Ribeiro et al. (1989a), Ribeiro (1995). Glossina morsitans (Savannah Tsetse fly) 5′nucleotidase-related PAgI Mant and Parker (1981), Caljon et al. Apyrase (2010). - Thrombin inhibitor Parker and Mant (1979), Ribeiro (1995). Glossina austeni (Savannah Tsetse fly) - PA Hawkins (1966), Hawkey (1967). Simulium vittatum (Black fly/Buffalo gnat) - Thrombin inhibitor Jacobs et al. (1990), Ribeiro (1995). - Anticoagulant (FVII Makonnen et al., unpublished inhibitor) observations as cited by Ribeiro (1995) - Anticoagulant (FXa Jacobs et al. (1990) inhibitor) - Anticoagulant (FVa Abebe et al. (1996), Basanova, Baskova inhibitor) and Zavalova (2002) Aedes aegypti (Yellow-fever mosquito) - Anticoagulant Stark and James (1995) (serine protease inhibitor of FXa) Apyrase PAgI Ribeiro et al. (1984), Vachereau and Ribeiro (1989) Oropsylla bacchii (Flea) Apyrase PAgI Ribeiro, Vaughan and Azad (1990b), Ribeiro (1995) Orchopea howardii (Squirrel Flea) Apyrase PAgI Ribeiro, Vaughan and Azad (1990b), Ribeiro (1995) Xenopsylla cheopis (Oriental Rat Flea) Apyrase PAgI Ribeiro, Vaughan and Azad (1990b), Ribeiro (1995) Rhodniini/Triatomini (Kissing/ Apyrase PAgI Ribeiro, Marinotti and Gonzales (1990a), Traitomines/Assassin/Conenose bug) Law, Ribeiro and Wells (1992), Santos et al. (2007) Dimiconin Anticoagulant (FXII Ishimaru et al. (2012) inhibitor) Continued ............................................................................................... .................................................................. 11 Review article Bioscience Horizons � Volume 10 2017 ............................................................................................... .................................................................. Table 4. Continued Species Molecule Function in host Reference Prolixin S (NP2) Anticoagulant Hellmann and Hawkins (1965), Ribeiro, Schneider and Guimarães (1995) Triabin Thrombin inhibitor Noeske-Jungblut et al. (1995), Ishimaru et al. (2012) Rhodniin Thrombin Inhibitor Friedrich et al. (1993) Pallidipin 1 and 2 Collagen-induced Noeske-Jungblut et al. (1994), Basanova, PAgI Baskova and Zavalova (2002), Ishimaru et al. (2012) R. prolixus aggregation PAgI Francischetti et al. (2000), Francischetti, inhibitor-1 (RPAI-1) Andersen and Ribeiro (2002a), Francischetti et al. (2009), Ishimaru et al. (2012) Triafestin-1 and -2 Plasma kallikrein- Isawa et al. (2007), Ishimaru et al. (2012). kinin system inhibitors (FXII and kininogen inhibitor) NP1-4 Vasodilator Ribeiro, Marinotti and Gonzales (1990a), Champagne, Nussenzveig and Ribeiro (1995), Basanova, Baskova and Zavalova (2002), reviewed by Champagne (2005) Lipocalins Transports small Santos et al. (2007) hydrophobic molecules Serine protease Protease Santos et al. (2007) -PA Hellmann and Hawkins (1964), Hawkins and Hellman (1966), Hawkey (1967) - Anticoagulant (FVIII Hellmann and Hawkins (1965, 1966) inhibitor) Eutriatoma maculatus (Assasin bug) Maculatin Thrombin Inhibitor Hellmann and Hawkins (1966) -PA Hellmann and Hawkins (1966) Culicoides sonorensis (Biting midge) TFPI1 and TFPI2 Protease inhibitors Campbell et al. (2005) - Hyaluronidase Campbell et al. (2005) Non-haematophagous animals Bombus terrestris (Buff-tailed/Large earth Bt-KTI Serine protease Qiu et al. (2013) Bumblebee) inhibitor (acts as a plasmin inhibitor) Scolopendra subspinipes mutilans - Anticoagulant (FXa Kong et al. (2013) (Chinese red-head/Chinese inhibitor) red-headed centipede) Scolonase Serine protease You et al. (2004) Continued ............................................................................................... .................................................................. 12 Bioscience Horizons � Volume 10 2017 Review article ............................................................................................... .................................................................. Table 4. Continued Species Molecule Function in host Reference Eisenia fetida (Redworm/Brandling/ Earthworm Fibrinolytic Thrombolytic/ Park et al. (1989) as cited by Sharma Panfish/Trout/Tiger/Red Wiggler/Red enzyme (EFE)/ Fibrinolytic et al. (2011), Mihara et al. (1991), Californian Earthworm), Lumbricus Lumbrokinase Sharma et al. (2011), Fu et al. (2013), rubellus, Eudrilus eugeniae (various Li et al. (2012) Earthworms) Antheraea pernyi (Chinese oak silkworm) Cocoonase Thrombolytic Geng et al. (2014) The table excludes putatively secreted proteins, housekeeping proteins/transcripts, substances with unknown function and substances not related to wound repair, thrombolysis or blood feeding. Most potent known to man (greater than calcitonin gene-related peptide). Serine proteases are included because PAs belong to this family of enzymes. ‘-’, name not specified; NP, nitroporins; TFPI1 and TFPI2, tissue factor pathway inhibitor 1 and 2; FVa, FVIIa, FXa, FXIIa, FXIIIa, activated coagulation factors V, VII, X, XII, XIII; PAgI, platelet aggregation inhibitor; PA, plasminogen activator. The PAs in the vampire bats are highly conserved, espe- Both haematophagous animals and non-haematophagous cially between the two species which feed upon mammals animals possess some sort of serine protease and serine prote- (D. rotundus and D. youngi; Tellgren-Roth et al., 2009). ase inhibitor (Qiu et al., 2013). The presence of serine prote- Evolution of the plasminogen gene may have led to the wide ase inhibitors in diverse groups of insects (honey bee, utilization of mammalian livestock by these two species and mosquito; Table 4; Qiu et al., 2013) suggests that endogenous consequently led to the rise of D. rotundus as the common suppressors of serine proteases, of which PA is a family mem- vampire bat (Tellgren-Roth et al., 2009). Absence of a PA in ber, are required at some stage of insect life and have been leeches suggests that the PA activity in the saliva of H. longi- conserved for reasons unconnected with blood feeding. cornis, Eutriatoma maculatus, R. prolixus (Hellmann and Insects heal wounds using cross-linking enzymes including Hawkins, 1964; Hawkins and Hellman, 1966) and D. rotun- transglutaminase, phenoloxidase (reviewed by Theopold dus is the result of convergent evolution. If it originated from et al., 2004) and lipophorin (Li et al., 2002) to form haemo- a common ancestral gene, it is unclear why the leech would lymph clots. No true orthologues of vertebrate blood clotting lose a potentially beneficial feature during its evolution. factors have been found in insects but several proteins with similar functional domains have been detected (Theopold The dual function of tick Longistatin, as a PA and anticoagu- et al., 2004). lant, indicates the importance of these components to species survival. It also suggests that other PAs are the result of replica- In line with the similarities between serine proteases and tion and division over the course of evolution. The evolutionary domains of coagulation factors, some serine proteases in the split of the PAs and anticoagulants might have led to the now protochordate Botryllus schosseri (star ascidian/golden star separate entities which can be observed in other invertebrates. tunicate) are homologous to vertebrate blood-coagulation Further investigation into the homology between Longistatin proteases (Ponczek, Bijak and Nowak, 2012). They par- and other PAs and anticoagulants may provide further insight ticipate in reactions involving the provoked aggregation (Bowman and Sauer 2004; Anisuzzaman et al., 2011). The of different cell-type colonies leading to cell clumping at thrombin inhibitor variegin, from the tick A. variegatum,has the site of contact. This suggests that a cascade of acti- no homology with other haematophagous animal thrombin vated serine proteases was initially a defence mechanism inhibitors and presumably evolved independently. Peptide (not associated with vascular injury; Wan et al.,2013), AP18 binding to exosite-I also in A. variegatum ticks is compar- which evolved into the vertebrate complement system and able to hirudin C terminus behaviour of leeches (Maraganore haemostatic response (Oren et al., 2008; Ponczek, Bijak et al., 1989; Naski et al.,1990), suggesting that these two and Nowak, 2012). sequences have a similar mechanism of action even though the Although the evidence suggests that PAs in invertebrates two species are phylogenetically distant (Koh et al., 2007). were derived from a common ancestor, this does not seem The vampire bat uses a distinct PAgI to block ADP- likely for the vampire bats. One hypothesis for the transition induced platelet aggregation. The PAgI of other haema- of vampire bats to haematophagy is arboreal feeding (Schutt, tophagous animals uses different biochemical mechanisms. 2008): they may have evolved from carnivorous bats whose Apyrase, which hydrolyses ATP and ADP, is commonly prey had grown too large for them to hunt by their usual found in haematophagous insects including the tsetse fly methods. A behavioural shift allowed the proto-vampires to Glossina morsitans (Mant and Parker, 1981; Caljon et al., utilize these larger animals as a food resource. 2010). Amounts of apyrase activity are elevated in blood- We hypothesize two alternative mechanisms for the devel- feeding animals compared with non-blood feeding insects, opment of the necessary salivary biochemistry in vampire suggesting exploitation of a pre-existing mechanism bats to support this transition: (Francischetti et al., 2009). ............................................................................................... .................................................................. 13 Review article Bioscience Horizons � Volume 10 2017 ............................................................................................... .................................................................. (1) Adaptation of blood haemostatic molecules into anti- completing a PhD, to ultimately become a lecturer in immun- haemostatic salivary molecules. Because PAs have been ology. F.L.W. designed the study, carried out the research and found in all vertebrates investigated so far (reviewed by wrote up the paper. She has primary responsibility for the Schleuning, 2001), it is possible that molecules present in paper. M.R.L. assisted in writing up the paper. a common ancestor could have evolved separate but par- allel functions. The mechanism for this would be gene duplication followed by mutation and selection. Since References other haematophagous animals possess different salivary components, the convergence to haematophagy, facili- Abdualkader, A. M., Ghawi, A. M., Alaama, M. et al. (2013) Leech thera- tated by duplication, must have resulted from selection peutic applications. Indian Journal of Pharmaceutical Sciences,75 according to different mammalian lifestyles or habitat. (2), 127–137. Whether this would have been possible in the relatively short evolutionary time available (between 6000 and Abebe, M., Ribeiro, J. M., Cupp, M. S. et al. (1996) Novel anticoagulant 2 000 000 years, with the Phyllostomidae family diver- from salivary glands of Simulium vittatum (Diptera: Simuliidae) inhi- ging at 35 m.y.a.; Schutt, 2008; Jones, Teeling and bits activity of coagulation factor V. Journal of Medical Entomology, Rossiter, 2013) is not clear. 33 (1), 173–176. (2) Exploitation of host blood proteins. Anti-haemostatic Abelous, J. E. and Bardier, E. (1909) ‘Les substances hypotensives de proteins could be obtained directly from the blood of the l’urine humaine normale’. CR Société de Biologie (in French), 66, hosts and exploited adaptively. For this to occur, bats 511–520. would need to absorb the materials intact and without destruction by their immune system, and be able to Altringham, J. D. (2011) Bats: From Evolution to Conservation, 2nd edn, exploit them in subsequent feeding. The blood coagula- Oxford University Press Inc, New York, USA, pp. 29–30 , 160–161. tion factors FVII, FX and FIX have similar sequence and Anisuzzaman, Islam, M. K., Alim, M. A. et al. (2011) Longistatin, a plas- domain arrangements in all vertebrates (Ponczek, Bijak minogen activator, is key to the availability of blood-meals for ixo- and Nowak, 2012) so it is possible that those derived did ticks. PLoS Pathogens, 7 (3), e1001312. from the host were well tolerated. Anisuzzaman, Islam, M. K., Miyoshi, T. et al. (2010) Longistatin, a novel EF-hand protein from the ixodid tick Haemaphysalis longicornis,is Conclusion required for acquisition of host blood-meals. International Journal for Parasitology, 40 (6), 721–729. Haematophagous animals possess many salivary constituents, which are capable of preventing haemostasis in their host, Apitz-Castro, R., Béguin, S., Tablante, A. et al. (1995) Purification and exploiting a range of different mechanisms. Given that hae- partial characterization of draculin, the anticoagulant factor present matophagy is considered to have evolved independently sev- in the saliva of vampire bats (Desmodus rotundus). Thrombosis and eral times, the active salivary components presumably Haemostasis, 73 (1), 94–100. emerged by convergent evolution. The derivation of these Barnes, C. S., Krafft, B., Frech, M. et al. (2001) Production and character- molecules has yet to be defined, but the salivary components ization of saratin, an inhibitor of von Willebrand factor-dependent of leeches and ticks at least could be adaptations of common platelet adhesion to collagen. Seminars in Thrombosis and ancestral genes. Hemostasis, 27 (4), 337–348. Recent advances in large-scale transcriptome analysis and Barrett, K. E., Barman, S. M., Boitano, S. et al. (2012) Ganong’s Review of structural proteomic analysis have made the identification of Medical Physiology, 24th edn, McGraw-Hill Companies Inc, China, such molecules easier. Identification of new molecules and pp. 564–569. comparative homology of current proteins, especially those with anti-metastasis and analgesic properties, may help Basanova, A. V., Baskova, I. P. and Zavalova, L. L. (2002) Vascular- explain their origin as well as the evolution to haematophagy. platelet and plasma hemostasis regulators from bloodsucking ani- mals. Biochemistry (Moscow), 67 (1), 143–150. Baskova, I. P., Nikonov, G. I., Mirkamali, E. G. et al. (1988) Influence of a Author biography preparation from the medicinal leech on phagocytosis and comple- ment system. Kazansky Medicinsky Zhurnal, 5, 334–336. Francesca graduated with a 2:1 BSc (Hons) Animal Science degree from the University of Nottingham in 2014. It was Bier, O. G. (1932) Action anticoagulante et fibrinolytique de l’extract during this time that she developed a passionate interest in des glands salivaires d’une chauve-souris hèmatophage (Desmodus immunology, infection and wound healing. Francesca was rufus). Comptes Rendus Hebdomadaires des Séances – Société de awarded one of fifteen School of Life Sciences Taught MSc Biologie, 110, 129–131. Scholarship and has now completed her MSc in Immunology and Allergy at the University of Nottingham. She hopes to Black, W. C. 4th and Piesman, J. (1994) Phylogeny of hard- and soft-tick continue in the field of immunology research, by one day taxa (Acari: Ixodida) based on mitochondrial 16S rDNA sequences. ............................................................................................... .................................................................. 14 Bioscience Horizons � Volume 10 2017 Review article ............................................................................................... .................................................................. Proceedings of the National Academy of Sciences of the United States Chudzinski-Tavassi, A. M., Kelen, E. M., de Paula Rosa, A. P. et al. (1998) of America, 91 (21), 10034–10038. Fibrino(geno)lytic properties of purified hementerin, a metallopro- teinase from the leech Haementeria depressa. Thrombosis and Blankenship, D. T., Brankamp, R. G., Manley, G. D. et al. (1990) Amino Hemostasis, 80 (1), 155–160. acid sequence of ghilanten: anticoagulant-antimetastatic principle of the South American leech, Haementeria ghilianii. Biochemical and Collin, N., Assumpção, T. C. F., Mizurini, D. M. et al. (2012) Lufaxin, a Biophysical Research Communications, 166 (3), 1384–1389. novel factor Xa inhibitor from the salivary gland of the sand fly Lutzomyia longipalpis blocks protease-activated receptor 2 activa- Boskova, I. P. and Zavalova, L. L. (2001) Proteinase inhibitors from the medi- tion and inhibits inflammation and thrombosis in vivo. cinal leech Hirudo medicinalis. Biochemistry (Moscow), 66 (7), 703–714. Arteriosclerosis, Thrombosis and Vascular Biology, 32 (9), 2185–2198. Bowman, A. S. and Sauer, J. R. (2004) Tick salivary glands: function, Coppens, M., Eikelboom, J. W., Gustafsson, D. et al. (2012) Translational physiology and future. Parasitology, 129 (Suppl), S67–S81. success stories: development of direct thrombin inhibitors. Brankamp, R. G., Blankenship, D. T., Sunkara, P. S. et al. (1990) Circulation Research, 111, 920–929. Ghilantens: anticoagulant-antimetastatic proteins from the South Corral-Rodríguez, M. A., Macedo-Ribeiro, S., Barbosa Pereira, P. J. B. American leech, Haementeria ghilianii. The Journal of Laboratory et al. (2009) Tick-derived Kunitz-type inhibitors as antihemostatic and Clinical Medicine, 115 (1), 89–97. factors. Insect Biochemistry and Molecular Biology, 39 (9), 579–595. Braun, N. J., Bodmer, J. L., Virca, G. D. et al. (1987) Kinetic studies on the Delpieto, H. A. and Russo, R. G. (2009) Acquired resistance to saliva interaction of eglin c with human leukocyte elastase and cathepsin anticoagulants by prey previously fed upon by vampire bats G. Biological Chemistry Hoppe-Seyler, 368 (4), 299–308. (Desmodus rotundus): evidence for immune response. Journal of Brossard, M. and Wikel, S. K. (2004) Tick immunobiology. Parasitology, Mammalogy, 90 (5), 1132–1138. 129 (Suppl), S161–S176. Dickinson, R. G., O’Hagan, J. E., Schotz, M. et al. (1976) Prostaglandin in Buchner, H. (1891) Zur Nomenklatur der schützenden Eiweisskörper. the saliva of the cattle tick Boophilus microplus. The Australian Centr Bakteriol Parasitenk, 10, 699–701. Journal of Experimental Biology and Medical Science, 54 (5), 475–486. Campbell, C. L., Vandyke, K. A., Letchworth, G. J. et al. (2005) Midgut and sal- Due, C., Fox, W., Medlock, J. M. et al. (2013) Tick bite prevention and ivary gland transcriptomes of the arbovirus vector Culicoides sonorensis tick removal. British Medical Journal (Clinical Research ed.), 347, (Diptera: Ceratopogonidae). Insect Molecular Biology, 14 (2), 121–136. f7123. Caljon, G., De Ridder, K., De Baetselier, P. et al. (2010) Identification of a Erwin, D. H. E. and Davidson, E. H. (2002) The last common bilaterian tsetse fly salivary protein with dual inhibitory action on human ancestor. Development, 129, 3021–3032. platelet aggregation. PLoS One, 5 (3), e9671. Electricwala, A., Sawyer, R. T., Jones, C. P. et al. (1991) Isolation of throm- Cartwright, T. (1974) The plasminogen activator of vampire bat saliva. bin inhibitor from the leech Hirudinaria manillensis. Blood Blood, 43 (3), 317–326. Coagulation and Finbrinolysis, 2 (1), 83–89. Cavalcante, R. R., Pereira, M. H. and Gontijo, N. F. (2003) Anti- Faria, F., Kelen, E. M., Sampaio, C. A. et al. (1999) A new factor Xa inhibi- complement activity in the saliva of phlebotomine sand flies and tor (lefaxin) from the Haementeria depressa leech. Thrombosis and other haematophagous insects. Parasitology, 127, 87–93. Haemostasis, 82 (5), 1469–1473. Champagne, D. E. (2005) Antihemostatic molecules from saliva of Fernandez, A. Z., Tablante, A., Bartoli, F. et al. (1998) Expression of bio- blood-feeding arthropods. Pathophysiology of Haemostasis and logical activity of draculin, the anticoagulant factor from vampire Thrombosis,34 (4–5), 221–227. bat saliva, is strictly dependent on the appropriate glycosylation of the native molecule. Biochimica et Biophysica acta, 1425 (2), Champagne, D. E., Nussenzveig, R. H. and Ribeiro, J. M. C. (1995) 291–299. Purification, partial characterization, and cloning of nitric oxide- carrying heme proteins (nitrophorins) from salivary glands of the Fernandez, A. Z., Tablante, A., Beguín, S. et al. (1999) Draculin, the anti- blood-sucking insect Rhodnius prolixus. The Journal of Biological coagulant factor in vampire bat saliva, is a tight-binding, non- Chemistry, 270 (15), 8691–8695. competitive inhibitor of activated factor X. Biochimica et Biophysica acta, 1434 (1), 135–142. Chapin, J. C. and Hajjar, K. A. (2015) Fibrinolysis and the control of blood coagulation. Blood Reviews, 29 (1), 17–24. Fogaça, A. C., Almeida, I. C., Eberlin, M. N. et al. (2006) Ixodidin, a novel antimicrobial peptide from the hemocytes of the cattle tick Cherniack, E. P. (2011) Bugs as drugs, part two: worms, leeches, scor- Boophilus microplus with inhibitory activity against serine protei- pions, snails, ticks, centipedes, and spiders. Alternative Medicine nases. Peptides, 27 (4), 667–674. Review: A Journal of Clinical Therapeutic, 16 (1), 50–58. Chopin, V., Salzet, M., Baert, Jl et al. (2000) Therostasin, a novel clotting Francischetti, I. M., Andersen, J. F. and Ribeiro, J. M. (2002a) Biochemical factor Xa inhibitor from the rhynchobdellid leech, Theromyzon tes- and functional characterization of recombinant Rhodnius prolixus sulatum. Journal of Biological Chemistry, 275 (42), 32701–32707. platelet aggregation inhibitor 1 as a novel lipocalin with high ............................................................................................... .................................................................. 15 Review article Bioscience Horizons � Volume 10 2017 ............................................................................................... .................................................................. affinity for adenosine diphosphate and other adenine nucleotides. Hawkey, C. (1966) Plasminogen activator in saliva of the vampire bat Biochemistry, 41, 3810–3818. Desmodus rotundus. Nature, 211 (5047), 434–435. Francischetti, I. M. B., Assumpção, T. C. F., Ma, D. et al. (2013) The Hawkey, C. (1967) Inhibitor of platelet aggregation present in saliva of ‘Vampirome’: Transcriptome and proteome analysis of the principal the vampire bat Desmodus rotundus. British Journal of Haematology, and accessory submaxillary glands of the vampire bat Desmodus 13 (6), 1014–1020. rotundus,avector of human rabies. Journal of Proteomics,82, 288–319. Hawkins, R. I. (1966) Factors affecting blood clotting from salivary Francischetti, I. M. B., Ribeiro, J. M., Champagne, D. et al. (2000) glands and crop of Glossina austeni. Nature (London), 212, 738. Purification, cloning, expression, and mechanism of action of a Hawkins, R. I. and Hellman, K. (1966) Investigations on a plasminogen novel platelet aggregation inhibitor from the salivary gland of the activator in Two-blood-Suckers, Rhodnius prolixus Stål and Hirudo bloodsucking bug, Rhodnius prolixus. Journal of Biological medicinalis;. British Journal of Haematology, 12, 86. Chemistry, 275, 12639–12650. Haycraft, J. B. (1884) On the action of a secretion obtained from the Francischetti, I. M. B., Sa-Nunes, A., Mans, B. J. et al. (2009) The role of medicinal leech on the coagulation of the blood. Proceedings of the saliva in tick feeding. Frontiers in Bioscience (Landmark Edition), 14, Royal Society of London, 36, 478–487. 2051–2088. Hellenius, W. J. and Ruben, J. A. (2004) The Evolution of endothermy in Francischetti, I. M. B., Valenzuela, J. G., Andersen, J. F. et al. (2002b) terrestrial vertebrates: Who? When? Why? in Physiological and Ixolaris, a novel recombinant tissue factor pathway inhibitor (TFPI) Biochemical Zoology, 77, pp. 1019–1042. from the salivary gland of the tick, Ixodes scapularis: identification of factor X and factor Xa as scaffolds for the inhibition of factor VIIa/ Hellmann, K. and Hawkins, R. I. (1964) Anticoagulant and fibrinolytic tissue factor complex. Blood, 99 (10), 3602–3612. activities from Rhodnius Prolixus Stal. Nature, 201, 1008–1009. Kraut, H., Frey, F. K. and Werle, E. (1930) Der Nachweis eines Hellmann, K. and Hawkins, R. I. (1965) Prolixins-S and prolixin-G; two Kreislaufhormons in der Pankreasdrüse. (IV. Mitteilung über dieses anticoagulants from Rhodnius prolixus Stål. Nature, 207 (994), Kreislaufhormon.). Hoppe-Seyler´s Zeitschrift für Physiologische 265–267. Chemie, 189 (3–4), 97–106. Hellmann, K. and Hawkins, R. I. (1966) An antithrombin (maculatin) and Friedrich, T., Kroger, B., Bialojan, S. et al. (1993) A Kazal-type inhibitor a plasminogen activator extractable from the blood-sucking hem- with thrombin specificity from Rhodnius prolixus. Journal of ipteran, Eutriatoma maculatus. British Journal of Haematology,12 Biological Chemistry, 268 (1993), 16216–16222. (4), 376–384. Fu, Z., Zhang, L., Liu, X. et al. (2013) Comparative proteomic analysis of Higgs, G. A., Vane, J. R., Hart, R. J. et al. (1976) Prostaglandins in the sal- the sun- and freeze-dried earthworm Eisenia fetida with differen- iva of cattle tick, Boophilus microplus (Canestrini) (Acarina, Ixodidae). tially thrombolytic activities. Journal of Proteomics, 83, 1–14. Bulletin of Entomological Research, 66, 665–670. Gardell, S. J., Duong, L. T., Diehl, R. E. et al. (1989) Isolation, characteriza- Hildebrandt, J. P. and Lemke, S. (2011) Small bite, large impact–saliva tion, and cDNA cloning of a vampire bat salivary plasminogen acti- and salivary molecules in the medicinal leech, Hirudo medicinalis. vator. The Journal of Biological Chemistry, 264 (30), 17947–17952. Naturwissenschaften, 98, 995–1008. Geng, P., Lin, L., Li, Y. et al. (2014) A novel fibrin(ogen)olytic trypsin-like Hovingh, P. and Linker, A. (1999) Hyaluronidase activity in leeches protease from Chinese oak silkworm (Antheraea pernyi): purification (Hirudinea). Comparative Biochemistry and Physiology. Part B, and characterization. Biochemical and Biophysical Research Biochemistry and Molecular Biology, 124 (3), 319–326. Communications, 445 (1), 64–70. Ibrahim, M. A., Ghazy, A. H., Maharem, T. et al. (2001) Isolation and Greenhall, A. M. and Schutt, W. A. (1996) Diaemus youngi. Mammalian properties of two forms of thrombin inhibitor from the nymphs of Species, 533, 1–7. the camel tick Hyalomma dromedarii (Acari: Ixodidae). Experimental and Applied Acarology, 25 (8), 675–698. Gregson, J. D. (1967) Observations on the movement of fluids in the vicinity of the mouthparts of naturally feeding Dermacentor ander- Isawa, H., Orito, Y., Jingushi, N. et al. (2007) Identification and character- soni Stiles. Parasitology, 57, 1–8. ization of plasma kallikrein-kinin system inhibitors from salivary glands of the blood-sucking insect Triatoma infestans. The FEBS Gulba, D. C., Praus, M. and Witt, W. (1995) DSPA alpha—Properties of Journal, 274 (16), 4271–4286. the plasminogen activators of the vampire bat Desmodus rotundus. Fibrinolysis, 9 (Suppl 1), 91–96. Ishimaru, Y., Gomez, E. A., Zhang, F. et al. (2012) Dimiconin, a novel coagulation inhibitor from the kissing bug, Triatoma dimidiata,a Hajnická, V., Kocáková, P., Sláviková, M. et al. (2001) Anti-interleukin-8 vector of Chagas disease. Journal of Experimental Biology, 215 (Pt activity of tick salivary gland extracts. Parasite Immunology, 23 (9), 20), 3597–3602. 483–489. Jacobi, Y. (1904) Über Hirudin. Deutsche Medizinische Wochenschrift, 30, Hall, J. E. (2011) Guyton and Hall’s Textbook of Medical Physiology, 12th 1786–1787. edn, Sauders Elsevier, Philadelphia, PA, USA, pp. 451–460. ............................................................................................... .................................................................. 16 Bioscience Horizons � Volume 10 2017 Review article ............................................................................................... .................................................................. Jacobs, J. W., Cupp, E. W., Sardana, M. et al. (1990) Isolation and charac- Li, G., Wang, K. Y., Li, D. et al. (2012) Cloning, expression and character- terization of a coagulation factor Xa inhibitor from black fly salivary ization of a gene from earthworm Eisenia fetida encoding a blood- glands. Thrombosis and Haemostasis, 64 (2), 235–238. clot dissolving protein. PLoS One, 7 (12), e53110. Jiang, Sy., Jiao, J., Zhang, Tt. et al. (2013) Pharmacokinetics study of Liao, M., Zhou, J., Gong, H. et al. (2009) Hemalin, a thrombin inhibitor iso- recombinant hirudin in the plasma of rats using chromogenic lated from a midgut cDNA library from the hard tick Haemaphysalis substrate, ELISA, and radioisotope assays. PLoS One,8(6), longicornis. Journal of Insect Physiology, 55 (2), 164–173. e64336. Liberatore, G. T., Samson, A., Bladin, C. et al. (2003) Vampire bat salivary Jones, G., Teeling, E. C. and Rossiter, S. J. (2013) From the ultrasonic to plasminogen activator (desmoteplase): a unique fibrinolytic the infrared: molecular evolution and the sensory biology of bats. enzyme that does not promote neurodegeneration. Stroke, 34 (2), Frontiers in Physiology, 4, 117. 537–543. Jongejan, F. and Uilenberg, G. (2004) The global importance of ticks. Linker, A., Hoffman, P. and Meyer, K. (1957) The hyaluronidase of the Parasitology, 129, S3–14. leech: an endoglucuronidase. Nature, 180 (4590), 810–811. Juckett, G. (2013) Arthropod bites. American Family Physician, 88 (12), Lloyd, C. M. and Walker, A. R. (1995) Salivary glands and saliva of 841–847. Amblyomma variegatum ticks: comparison of immatures and adults in relation to the pathogenesis of dermatophilosis. Veterinary Kazimírová, M. and Štibrániová, I. (2013) Tick salivary compounds: their Parasitology, 59, 59–67. role in modulation of host defences and pathogen transmission. Frontiers in Cellular and Infection Microbiology, 3, 43. Low, D. H. W., Sunagar, K., Undheim, E. A. B. et al. (2013) Dracula’s chil- dren: Molecular evolution of vampire bat venom. Journal of Koh, C. Y., Kazimirova, M., Trimnell, A. et al. (2007) Variegin, a novel fast Proteomics, 89, 95–111. and tight binding thrombin inhibitor from the tropical bont tick. Journal of Biological Chemistry, 282 (40), 29101–29113. Ma, D., Mizurini, D. M., Assumpção, T. C. F. et al. (2013) Desmolaris, a novel factor XIa anticoagulant from the salivary gland of the vam- Koh, C. Y. and Kini, R. M. (2008) Anticoagulants from hematophagous pire bat (Desmodus rotundus) inhibits inflammation and thrombosis animals. Expert Review of Hematology, 1 (2), 135–139. in vivo. Blood, 122 (25), 4094–4106. Kong, Y., Shao, Y., Chen, H. et al. (2013) A novel factor Xa-inhibiting Macedo-Ribeiro, S., Almeida, C., Calisto, B. M. et al. (2008) Isolation, peptide from Centipedes Venom. International Journal of Peptides cloning and structural characterisation of boophilin, a multifunc- Research and Therapeutics, 19, 303–311. tional Kunitz-type proteinase inhibitor from the cattle tick. PLoS One, 3 (2), e1624. Krätzschmar, J., Haendler, B., Langer, G. et al. (1991) The plasminogen activator family from the salivary gland of the vampire bat Maina, A. N., Jiang, J., Omulo, S. A. et al. (2014) High prevalence of Desmodus rotundus: cloning and expression. Gene, 105 (2), Rickettsia Africae variants in Amblyomma variegatum.Ticks from 229–237. domestic mammals in rural western Kenya: implications for human health. Vector-Bourne and Zoonotic Diseases (Larchmont, N.Y.),14(10), Lai, R., Takeuchi, H., Jonczy, J. et al. (2004) A thrombin inhibitor from 693–702. the ixodid tick, Amblyomma hebraeum. Gene, 342 (2), 243–249. Mans, B. J., Gasper, A. R. M. D., Louw, A. I. et al. (1998a) Apyrase activity Laurin M. and Reisz R. R. (2011) Synapsida. Mammals and their extinct and platelet aggregation inhibitors in the tick Ornithodoros savignyi relatives, accessed at: http://tolweb.org/Synapsida/14845 in Tree of (Acari: Argasidae). Experimental and Applied Acarology, 22, 353–366. Life Web Project [Last updated 14/8/11]. (9 March 2015). Mans, B. J., Gasper, A. R. M. D., Louw, A. I. et al. (1998b) Purification and Law, J. H., Ribeiro, J. M. C. and Wells, M. A. (1992) Biochemical insights characterisation of apyrase from the tick, Ornithodoros savignyi. derived from diversity in insects. Annual Review of Biochemistry, 61, Comparative Biochemistry and Physiology. Part B: Biochemistry & 87–111. Molecular Biology, 120, 617–624. Lent, C. M., Fliegner, K. H., Freedman, E. et al. (1988) Ingestive behaviour Mant, M. J. and Parker, K. R. (1981) Two platelet aggregation inhibitors and physiology of the medicinal leech. Journal of Experimental in tsetse (Glossina) saliva with studies of roles of thrombin and cit- Biology, 137, 513–527. rate in in vitro platelet aggregation. British Journal of Haematology, Lerner, E. A., Ribeiro, J. M., Nelson, R. J. et al. (1991) Isolation of maxadi- 48 (4), 601–608. lan, a potent vasodilatory peptide from the salivary glands of the Maraganore, J. M., Chao, B., Joseph, M. L. et al. (1989) Anticoagulant sand fly Lutzomyia longipalpis. Journal of Biological Chemistry, 266 activity of synthetic hirudin peptides. Journal of Biological (17), 11234–11236. Chemistry, 264, 8692–8698. Li, D., Scherfer, C., Korayem, A. M. et al. (2002) Insect hemolymph clot- Markwardt, F. (1957) Die Isolierung und chemische Charakterisierung ting: evidence for interaction between the coagulation system and des Hirudins. Hoppe-Seyler’s Zeitschrift für Physiologische Chemie, the prophenoloxidase activating cascade. Insect Biochemistry and 308, 147–156. Molecular Biology, 32 (8), 919–928. ............................................................................................... .................................................................. 17 Review article Bioscience Horizons � Volume 10 2017 ............................................................................................... .................................................................. Marshall, C. G. and Lent, C. M. (1988) Excitability and secretory activity Biochemica et Biophysica Acta (BBA) – Protein Structure and in the salivary gland cells of jawed leeches (Hirudinea: Molecular Enzymology, 1482 (1–2), 92–101. Gnathobdellida). Journal of Experimental Biology, 137, 89–105. Palta, S., Saroa, R. and Palta, A. (2014) Overview of the coagulation sys- Mihara, H., Sumi, H. and Yoneta, T. et al. (1991) A novel fibrinolytic tem. Indian Journal of anaesthesia, 58 (5), 515–523. enzyme extracted from the earthworm, Lumbricus rubellus. The Park, S. Y., Kye, K. C., Lee, M. H. et al. (1989) Fibrinolytic activity of the Japanese Journal of Physiology, 41 (3), 461–472. earthworm extract. Thrombosis and Haemostasis, 62, 545–550. Mϋhlhahn, P., Czisch, M., Morenweiser, R. et al. (1994) Structure of leech Parker, K. R. and Mant, M. J. (1979) Effects of tsetse (Glossina morsitans derived tryptase inhibitor (LDTI-C) in solution. FEBS Letters, 355 (3), morsitans Westw.) (Diptera: Glossinidae) salivary gland homogenate 290–296. on coagulation and fibrinolysis. Thrombosis and Haemostasis, 42 (2), Munro, R., Jones, C. P. and Sawyer, R. T. (1991) Calin–a platelet adhesion 743–751. inhibitor from the saliva of the medicinal leech. Blood Coagulation Patel, R., Ispoglou, S. and Apostolakis, S. (2014) Desmoteplase as a and Fibrinolysis: An International Journal in Haemostasis and potential treatment for cerebral ischaemia. Expert Opinion on Thrombosis, 2 (1), 179–184. Investigational Drugs, 23 (6), 865–873. Munshi, Y., Ara, I., Rafique, H. et al. (2008) Leeching in the history–a Peterson, K. J., Cotton, J. A., Gehling, J. G. et al. (2008) The Ediacaran emer- review. Pakistan Journal of Biological Sciences, 11 (13), 1650–1653. gence of bilaterians: congruence between the genetic and the geo- Murphy, K. (2012) Janeway’s Immunobiology, 8th edn, Garland Science, logical fossil records. Philosophical Transactions of the Royal Society of New York, USA, p.7. London. Series B, Biological Sciences, 363 (No. 1496), 1435–1443. Naski, M. C., Fenton, J. W., Maraganore, J. M. et al. (1990) The COOH- Piechowski-Jozwiak, B. and Bogousslavsky, J. (2013) The use of desmo- terminal domain of hirudin. An exosite-directed competitive inhibi- teplase (bat saliva) in the treatment of ischaemia. Expert Opinion on tor of the action of alpha-thrombin on fibrinogen. Journal of Biological Therapy, 13 (3), 447–453. Biological Chemistry, 265, 13484–13489. Ponczek, M. B., Bijak, M. Z. and Nowak, P. Z. (2012) Evolution of throm- Nawarskas, J. J. and Anderson, J. R. (2001) Bivalirudin: a new approach bin and other hemostatic proteases by survey of protochordate, to anticoagulation. Heart Disease (Hagerstown, Md), 3 (2), 131–137. hemichordate, and echinoderm genomes. Journal of Molecular Evolution,74 (5–6), 319–331. Nienaber, J., Gaspar, A. R. and Neitz, A. W. H. (1999) Savignin, a potent thrombin inhibitor isolated from the salivary glands of the tick Qiu, Y., Lee, K. S., Choo, Y. M. et al. (2013) Molecular cloning and antifi- Ornithodoros savignyi (Acari: Argasidae). Experimental Parasitology, brinolytic activity of a serine protease inhibitor from bumblebee 93 (2), 82–91. (Bombus terrestris) venom. Toxicon, 63, 1–6. Noeske-Jungblut, C., Haendler, B., Donner, P. et al. (1995) Triabin, a Ramachandra, R. N. and Wikel, S. K. (1995) Effects of Dermacentor highly potent exosite inhibitor of thrombin. Journal of Biochemical andersoni (Acari: Ixodidae) salivary gland extracts on Bos indicus and Chemistry, 270 (48), 28629–28634. B. taurus lymphocytes and macrophages: in vitro cytokine elabor- ation and lymphocyte blastogenesis. Journal of Medical Noeske-Jungblut, C., Krätzschmar, J., Haendler, B. et al. (1994) An inhibi- Entomology, 32, 338–345. tor of collagen-induced platelet aggregation from the saliva of Triatoma pallidipennis. The Journal of Biological Chemistry, 269 (7), Reverter, D., Vendrell, J., Canals, F. et al. (1998) A carboxypeptidase inhibi- 5050–5053. tor from the medical leech Hirudo medicinalis. Isolation, sequence analysis, cDNA cloning, recombinant expression, and characteriza- Oliveira, D. G., Alvarez-Flores, M. P., Lopes, A. R. et al. (2012) Functional tion. The Journal of Biological Chemistry, 273 (49), 32927–32933. characterisation of vizottin, the first factor Xa inhibitor purified from the leech Haementeria vizottoi. Thrombosis and Haemostasis, Ribeiro, J. M. C. (1995) Blood-feeding arthropods: live syringes or inver- 108 (3), 570–578. tebrate pharmacologists? Infectious Agents and Disease, 4, 143–152. Oren, M., Escande, Ml., Paz, G. et al. (2008) Urochordate histoincompati- Ribeiro, J. M., Evans, P. M., MacSwain, J. L. et al. (1992) Amblyomma ble interactions activate vertebrate-like coagulation system compo- americanum: characterization of salivary prostaglandins E2 and F2 nents. PLoS One, 3 (9), e3123. alpha by RP-HPLC/bioassay and gas chromatography-mass spec- trometry. Experimental Parasitology, 74 (1), 112–116. Paciaroni, M., Medeiros, E. and Bogousslavsky, J. (2009) Desmoteplase. Expert Opinion on Biological Therapy, 9 (6), 773–778. Ribeiro, J. M. C., Modi, G. B., Tesh, R. B. et al. (1989a) Salivary apyrase activity of some old world phlebotomine sand flies. Insect Paesen, G. C., Adams, P. L., Harlos, K. et al. (1999) Tick histamine- Biochemistry, 19 (4), 409–412. binding proteins: isolation cloning and three-dimensional structure. Molecular Cell, 3 (5), 661–671. Ribeiro, J. M., Makoul, G. T., Levine, J. et al. (1985) Antihemostatic, anti- inflammatory, and immunosuppressive properties of the saliva of a Paesen, G. C., Adams, P. L., Nuttall, P. A. et al. (2000) Tick histamine- tick, Ixodes dammini. Journal of Experimental Medicine, 161, binding proteins: lipocalins with a second binding cavity. 332–344. ............................................................................................... .................................................................. 18 Bioscience Horizons � Volume 10 2017 Review article ............................................................................................... .................................................................. Ribeiro, J. M., Makoul, G. T. and Robinson, D. R. (1988) Ixodes dammini: Schleuning, W. D., Alagon, A., Biodol, W. et al. (1992) Plasminogen acti- evidence for salivary prostacyclin secretion. The Journal of vators from the saliva of Desmodus rotundus (common vampire Parasitology, 74 (6), 1068–1069. bat): unique fibrin specificity. Annals of the New York Academy of Sciences, 667, 395–403. Ribeiro, J. M., Marinotti, O. and Gonzales, R. (1990a) A salivary vasodila- tor in the blood-sucking bug, Rhodnius prolixus. British Journal of Schutt, B. (2008) Dark Banquet; Blood and the Curious Lives of Blood- Pharmacology, 101 (4), 932–936. Feeding Creatures, Three Rivers Press, New York, USA, pp. 49–58. Ribeiro, J. M. and Mather, T. N. (1998) Ixodes scapularis: salivary kininase Seemüller, U., Meier, M., Ohlsson, K. et al. (1977) Isolation and charac- activity is a metallo dipeptidyl carboxypeptidase. Experimental terisation of a low molecular weight inhibitor (of chymotrypsin and Parasitology, 89 (2), 213–221. human granulocytic elastase and cathepsin G) from leeches. Hoppe-Seyler’s Zeitschrift für Physiologische Chemie, 358 (9), Ribeiro, J. M., Rossignol, P. A. and Spielman, A. (1986) Blood-finding 1105–1107. strategy of a capillary-feeding sandfly, Lutzomyia longipalpis. Comparative Biochemistry and Physiology. A, Comparative Sharma, A., Sonah, H., Deshmukh, R. K. et al. (2011) Cloning of fibrino- Physiology, 83 (4), 683–686. lytic protease-0 (Efp-0) gene from diverse earthworm individuals. Indian Journal of Biotechnology, 10 (3), 270–273. Ribeiro, J. M., Sarkis, J. J., Rossignol, P. A. et al. (1984) Salivary apyrase of Aedes aegypti: characterization and secretory fate. Comparative Snider, G. L., Stone, P. J., Lucey, E. C. et al. (1985) Eglin-c, a polypeptide Biochemistry and Physiology. B, Biochemistry and Molecular Biology, derived from the medicinal leech, prevents human neutrophil 79 (1), 81–86. elastase-induced emphysema and bronchial secretory cell metapla- sia in the hamster. The American Review of Respiratory Disease, 132 Ribeiro, J. M., Schneider, M. and Guimarães, J. A. (1995) Purification and (6), 1155–1161. characterization of prolixin S (nitrophorin 2), the salivary anticoagu- lant of the blood-sucking bug Rhodnius prolixus. The Biochemical Söllner, C., Mentele, R., Eckerskorn, C. et al. (1994) Isolation and charac- Journal, 308 (Pt 1), 243–249. terization of hirustasin, an antistasin-type serine-proteinase inhibi- tor from the medical leech Hirudo medicinalis. European Journal of Ribeiro, J. M., Vachereau, A., Modi, G. B. et al. (1989b) A novel vasodila- Biochemistry, 219 (3), 937–943. tory peptide from the salivary glands of the sand fly Lutzomyia longipalpis. Science (New York, N.Y.), 243 (4888), 212–214. Sommerhoff, C. P., Söllner, C., Mentele, R. et al. (1994) A Kazal-type inhibitor of human mast cell tryptase: isolation from the medical Ribeiro, J. M., Vaughan, J. A. and Azad, A. F. (1990b) Characterization of leech Hirudo medicinalis, characterization, and sequence analysis. the salivary apyrase activity of three rodent flea species. Biological Chemistry Hoppe Seyler, 375, 685–694. Comparative Biochemistry and Physiology. B, Comparative Biochemistry, 95 (2), 215–219. Stark, K. R. and James, A. A. (1995) A factor Xa-directed anticoagulant from the salivary glands of the yellow fever mosquito Aedes aegypti. Rigbi, M., Levy, H., Iraqi, F. et al. (1987) The saliva of the medicinal Experimental Parasitology, 81 (3), 321–331. leech Hirudo medicinalis–I. Biochemical characterization of the high molecular weight fraction. Comparative Biochemistry and Steranka, L. R., Manning, D. C., DeHaas, C. J. et al. (1988) Bradykinin as a Physiology. B, Comparative Biochemistry,87 (3),567–573. pain mediator: receptors are localized to sensory neurons, and antagonists have analgesic actions. Proceedings of the National Rigbi, M., Orevi, M. and Eldor, A. (1996) Platelet aggregation and coagu- Academy of Sciences of USA, 85, 3245–3249. lation inhibitors in leech saliva and their roles in leech therapy. Seminars in Thrombosis and Hemostasis, 22, 273–278. Stubbs, M. T., Morenweiser, R., Stürzebecher, J. et al. (1997) The three- dimensional structure of recombinant leech-derived tryptase Salzet, M., Chopin, V., Baert, Jl. et al. (2000) Theromin, a novel leech inhibitor in complex with trypsin. Implications for the structure of thrombin inhibitor. Journal of Biological Chemistry, 275 (40), human mast cell tryptase and its inhibition. Journal of Biological 30774–30780. Chemistry, 272, 19931–19937. Sanetra, M., Begemann, G., Becker, Mb. et al. (2005) Conservation and Su, J. B. (2014) Different cross-talk sites between the renin−angiotensin co-option in developmental programmes: the importance of hom- and the kallikrein−kinin systems. Journal of the Renin-Angiotensin- ology relationships. Frontiers in Zoology, 2, 15. Aldosterone System, 15 (4), 319–328. Santos, A., Ribeiro, J. M., Lehane, M. J. et al. (2007) The sialotranscriptome of Swadesh, J. K., Huang, I. Y. and Budzynski, A. Z. (1990) Purification and the blood-sucking bug Triatoma brasiliensis (Hemiptera, Triatominae). characterization of hementin, a fibrinogenolytic protease from the Insect Biochemistry and Molecular Biology, 37 (7), 702–712. leech Haementeria ghilianii. Journal of Chromotography, 502 (2), 359–369. Sauer, J. R. (1977) Acarine salivary glands—Physiological relationships. Journal of Medical Entomology, 14, 1–9. Tasiemski, A., Vandenbulcke, F., Mitta, G. et al. (2004) Molecular charac- terization of two novel antibacterial peptides inducible upon bac- Schleuning, W. D. (2001) Vampire bat plasminogen activator DSPA- terial challenge in an annelid, the leech Theromyzon tessulatum. The alpha-1 (desmoteplase): a thrombolytic drug optimized by natural Journal of Biological Chemistry, 279 (30), 30973–30982. selection. Haemostasis,31 (3–6), 118–122. ............................................................................................... .................................................................. 19 Review article Bioscience Horizons � Volume 10 2017 ............................................................................................... .................................................................. Tatchell R. J. and Binnington K. C. (1973) An active constituent of the Wan, H., Lee, K. S., Kim, B. Y. et al. (2013) A spider-derived Kunitz-type saliva of the cattle tick, Boophilus microplus; Proceedings of the 3rd serine protease inhibitor that acts as a plasmin inhibitor and an International Conference of Acarology; 1971, 745. elastase inhibitor. PLoS One, 8 (1), e53343. Tellgren-Roth, A., Dittmar, K., Massey, S. E. et al. (2009) Keeping the Wang, X., Coons, L. B., Taylor, D. B et al. (1996) Variabilin, a novel RGD- blood flowing-plasminogen activator genes and feeding behavior containing antagonist of glycoprotein IIb-IIIa and platelet aggrega- in vampire bats. Naturwissenschaften, 96 (1), 39–47. tion inhibitor from the hard tick Dermacentor variabilis. The Journal of Biological Chemistry, 271 (30), 17785–17790. Theopold, U., Schmidt, O., Söderhall, K. et al. (2004) Coagulation in arthropods: defence, wound closure and healing. Trends in Waxman, L. and Connolly, T. M. (1993) Isolation of an inhibitor selective Immunology, 25 (6), 289–294. for collagen-stimulated platelet aggregation from the soft tick Ornithodoros moubata. The Journal of Biological Chemistry, 268 (8), Tuszynski, G. P., Gasic, T. B. and Gasic, G. J. (1987) Isolation and charac- 5445–5449. terization of antistasin. An inhibitor of metastasis and coagulation. Journal of Biological Chemistry, 262 (20), 9718–9723. Waxman, L., Smith, D. E., Arcuri, K. E. et al. (1990) Tick anticoagulant peptide (TAP) is a novel inhibitor of blood coagulation factor Xa. Vachereau, A. and Ribeiro, J. M. C. (1989) Immunoreactivity of salivary Science, 248 (4955), 593–596. gland apyrase of Aedes aegypti with antibodies against a similar hydrolase present in the pancreas of mammals. Insect Biochemistry, Wikel, S. K. (1982) Histamine content of tick attachment sites and the 19 (6), 527–534. effect of H1 and H2 histamine antagonists on the expression of resistance. Annals of Tropical Medicine and Parasitology, 76, Valenzuela, J. G. (2004) Exploring tick saliva: from biochemistry to ‘sia- 179–185. lomes’ and functional genomics. Parasitology, 129 (Suppl), S83–S94. Wimsatt, W. A. (1959) Portrait of a vampire. Ward’s Natural Science Valenzuela, J. G., Francischetti, I. M. B., Pham, V. M. et al. (2002) Bulletin, 32, 35. Exploring the sialome of the tick Ixodes scapularis. Journal of Experimental Biology, 205, 2843–2864. You, W. K., Sohn, Y. D., Kim, K. Y. et al. (2004) Purification and molecular cloning of a novel serine protease from the centipede, Scolopendra van de Locht, A., Stubbs, M. T. and Bode, W. (1996) The ornithodorin- subspinipes mutilans. Insect Biochemistry and Molecular Biology,34 thrombin crystal structure, a key to the TAP enigma? The EMBO (3), 239–250. Journal, 15 (22), 6011–6017. Zaidi, S. M., Jameel, S. S., Zaman, F. et al. (2011) A systematic overview Viljoen, G. J., Bezuidenhout, J. D., Oberem, P. T. et al. (1986) Isolation of of the medicinal importance of Sanguivorous leeches. Alternative a neurotoxin from the salivary glands of female Rhipicephalus ever- Medicine Review, 16 (1), 59–65. tsi evertsi. Journal of Parasitology, 72, 865–874. ............................................................................................... .................................................................. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Bioscience Horizons Oxford University Press

Evolution of salivary secretions in haematophagous animals

Loading next page...
 
/lp/oxford-university-press/evolution-of-salivary-secretions-in-haematophagous-animals-hztCSfBzfq

References (195)

Publisher
Oxford University Press
Copyright
© The Author 2017. Published by Oxford University Press.
eISSN
1754-7431
DOI
10.1093/biohorizons/hzw015
Publisher site
See Article on Publisher Site

Abstract

BioscienceHorizons Volume 10 2017 10.1093/biohorizons/hzw015 .............................................. .................................................. .................................................. ............... Review article Evolution of salivary secretions in haematophagous animals Francesca L. Ware and Martin R. Luck School of Biosciences, University of Nottingham, Sutton Bonington Campus, Loughborough, Leicester LE12 5RD, UK *Corresponding author: 27 Braishfield Gardens, Bournemouth, Dorset BH8 0QA, UK. Email: francesca.ware@yahoo.co.uk Supervisor: Prof. Martin Luck, School of Biosciences, University of Nottingham, Sutton Bonington Campus, Loughborough, Leicester LE12 5RD, UK. .............................................. .................................................. .................................................. ............... Haemostasis is the prevention of blood fluidity in vertebrates and is the first stage of wound healing. Haematophagous animals use the blood of vertebrates as their sole source of nutrition and have evolved many salivary constituents to counteract the haemostatic response of their prey. These animals and their saliva have been studied for many years, with some applications in medicine. The purpose of this study is to compare the salivary constituents of leeches (Hirudinae), ticks (Argasidae and Ixodidae) and vampire bats (Desmodontinae) and to consider their evolutionary origin. Salivary constituents include plasminogen activa- tors (PAs), anticoagulants (activated factor X, FXa; inhibitors), vasodilators, platelet aggregation inhibitors (PAgI) and thrombin inhibitors. The animals studied all tend to possess an anticoagulant and a form of apyrase (PAgI) to assist with blood feeding. Ticks and vampire bats have a form of PA but the leech does not. The vampire bat has a PAgI but no vasodilator. The animals studied are from taxonomically unrelated groups but exploit similar mechanisms of action to facilitate their haematophagy. Given that the haematophagous lifestyle of these animals developed much later than their common ancestors, we conclude that their mechanisms for haematophagy have arisen by convergent evolution. Some molecules, e.g. serine proteases found in invertebrate saliva, are probably derived from a common ancestral gene. The possible paths that have led to evolution of vam- pire bat salivary components are considered. Further research into the homology of these salivary constituents is required to give insight into how these animals adapted to haematophagy and their further therapeutic potential. Key words: haematophagous, saliva, evolution, leech, tick, vampire bat Submitted on 21 September 2015; editorial decision on 28 November 2016 .............................................. .................................................. .................................................. ............... number of species, we have chosen to examine examples Introduction representing distinctly different phyla. We describe and com- pare the range of active components in the saliva of haema- Haematophagous animals are those which rely on blood from tophagous animals in the following three groups: leeches other animals as their only source of nutrition. These parasitic (Hirudinae), ticks (Ixodidae and Argasidae) and vampire bats creatures have evolved highly specific salivary molecules that (Desmodontinae). We then consider whether these components counteract the haemostatic response of the host and also exert may have arisen through convergent or divergent evolution. limited behavioural control. Whilst considerable research has been carried out on the salivary components and their poten- Haemostasis (Fig. 1) is the process by which blood flow stops tial for the treatment of human disease, few papers have at the site of an injury. It is one of the first responses to vascular explored their evolution. Because this unusual feeding method damage and initiates further processes including wound repair. is displayed across the animal kingdom but in a limited Briefly, coagulation (reviewed by Palta, Saroa and Palta, 2014) ............................................................................................... .................................................................. © The Author 2017. Published by Oxford University Press. This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/ licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com Review article Bioscience Horizons � Volume 10 2017 ............................................................................................... .................................................................. serine proteases). Plasmin is the active component of the plas- minogen (fibrinolytic) system (reviewed by Chapin and Hajjar, 2015), responsible for the eventual breakdown of fibrin fibres, fibrinogen, FV, FVIII, FXII and prothrombin at the start of the wound repair process (Tellgren-Roth et al., 2009; Hall, 2011; Barrett et al., 2012). Leeches Leeches are invertebrate, segmented worms from the phylum Annelida, class Clitellata, subclass Hirudinae (reviewed by Abdualkader et al., 2013). Leeches live in slow flowing streams and fresh water ponds (reviewed by Hildebrandt and Lemke, 2011). Young leeches, having left the safety of the cocoon, feed on amphibians. When their mouth parts have matured, they move on to feed on animals with thicker skin and more nutritious blood, including birds and mammals (Hildebrandt and Lemke, 2011). Leeches, specifically the medicinal leech Hirudo medicina- lis, have been used by physicians as a medicinal therapy for various diseases since early civilization (reviewed by Munshi et al., 2008). Haycraft (1884) reported that the H. medicinalis produced a substance with anticoagulant properties. In fact, Figure 1. Haemostasis in vertebrates. A simplified account of the leech produces several anticoagulants and thrombolytics, haemostasis in vertebrates, including mammals, identifying possible stored in the salivary glands (Chopin et al., 2000). targets for the salivary molecules of haematophagous animals. Adapted from Law, Ribeiro and Wells (1992, review article) and Barrett The leech attaches to its prey using the anterior portion of et al. (2012). Dashed arrows indicate inhibition. 5-HT, serotonin; TXA , its sucker. It begins periodic tilting movements of its three jaws, thromboxane A2. in order to slice open the skin. The pumping action of the pha- ryngeal muscles sucks the blood from the destroyed blood ves- sels and lymph of the host into the leech’scrop (Lent et al., involves the formation of a loose haemostatic plug, which is 1988). These jaw movements also initiate the secretion of saliva then converted into a definitive clot (thrombus) containing from its unicellular salivary gland cells, located anteriorly in fibrin. The formation of fibrin is initiated by a cascade of segments three and nine (Hildebrandt and Lemke, 2011). The enzymic reactions (clotting factors of the intrinsic and extrinsic mechanism of protein release from gland cells and the biochem- pathways), the last element of which is a complex of activated ical events needed to synthesize salivary proteins are unknown substances, collectively termed the prothrombin activator. This (Hildebrandt and Lemke, 2011). Serotonin may stimulate sal- complex catalyses the conversion of prothrombin to thrombin iva excretion (Marshall and Lent, 1988; Hildebrandt and and is considered to be the rate limiting step in blood coagula- Lemke, 2011) and pharyngeal peristalsis (Lent et al.,1988). tion (Hall, 2011; Barrett et al., 2012). The ingestion of blood lasts roughly 25 min (Lent et al., Thrombin is a serine protease, which activates and poly- 1988). Only the red blood cells and plasma proteins are of merizes fibrinogen to form fibrin. Fibrin stabilizing factor nutritious value to the leech. Plasma and haem derivatives are (Factor XIII or FXIII; we use this system of abbreviation excreted over the following 4–6days, ensuring efficient diges- throughout), released from platelets within the developing tion. Table 1 shows the haematophagy-relevant components of thrombus, is activated by thrombin and produces a stable saliva in various leech species and their function in the host. cross-linked fibrin meshwork containing trapped blood cells, platelets and plasma. After the clot has formed (3–6 min), it begins to contract due to the release of pro-coagulant (clot- Anticoagulants encouraging) substances, which cross-link more adjacent The majority of the anticoagulants found in leeches are inhibi- fibrin fibres. The retracting fibrin meshwork pulls the edges of tors of FXa preventing the conversion of prothrombin to the damaged blood vessel together, assisting haemostasis thrombin of the vertebrate coagulation cascade (common path- (Hall, 2011; Barrett et al., 2012). way). Examples are Therostatin (isolated from Theromyzon A euglobulin called plasminogen (profibrinolysin) is found tessulatum; Chopin et al., 2000) and Antistatin. Antistatin within the plasma proteins trapped in the clot. Plasminogen is has also been proposed to have anti-metastatic properties converted into plasmin (fibrinolysin) by tissue-type (t-PA) or (Tuszynski, Gasic and Gasic, 1987). Ghilanten is an anticoagu- urokinase-type (u-PA) plasminogen activators (PAs; both are lant (FXa inhibitor) obtained from the leech Haementeria ............................................................................................... .................................................................. 2 Bioscience Horizons � Volume 10 2017 Review article ............................................................................................... .................................................................. ............................................................................................... .................................................................. Table 1. Haematophagy-related salivary constituents of different leech species and their functions in the host Leech species Molecule Function in host Reference Hirudo medicinalis Hirudin Thrombin inhibitor Jacobi (1904), Markwardt (1957), Nawarskas and (European medicinal Anderson (2001), Coppens et al. (2012), Jiang et al. leech) (2013) Bufrudin Thrombin inhibitor Electricwala et al. (1991), Abdualkader et al. (2013) Apyrase Agonist of platelets Rigbi, Orevi and Eldor (1996), Hildebrandt and Lemke (2011) Theromin Thrombin inhibitor Salzet et al. (2000) Mammalian-type Reduce platelet adherence Rigbi et al. (1987) collagenase Calin Platelet adhesion and activation inhibition Munro, Jones and Sawyer (1991), Abdualkader et al. (2013) Haementeria vizottoi Vizottin Anticoagulant (FXa inhibitor) Oliveira et al. (2012) Hyaluronidase Digests hyaluronic acid present in the ECM Linker, Hoffman and Meyer (1957), Hovingh and Linker (1999), Hildebrandt and Lemke (2011) Inhibitor of C1-complement Anti-inflammatory Baskova et al. (1988), Boskova and Zavalova (2001) system component LCI Regulator of fibrinolysis rate or inhibits carboxypeptidase Reverter et al. (1998), Hildebrandt and Lemke (2011) Eglin-c Impairment of neutrophils (alpha-chymotrypsin, subtilisin, Seemüller et al. (1977), Snider et al. (1985), Braun et al. chymosin, granulocyte proteinases, elastase and (1987), Zaidi et al. (2011) cathepsin G inhibitor) LDTI Possible role in supressing cell-mediated inflammatory Mϋhlhahn et al. (1994), Sommerhoff et al. (1994), reactions Stubbs et al. (1997); Hildebrandt and Lemke (2011) Hirustasin Antistatin-type serine protease inhibitor. Also trypsin, Söllner et al. (1994), Boskova and Zavalova (2001) alpha-chymotrypsin and neutrophil cathepsin G inhibitor Haementeria officinalis Antistatin Anticoagulant (FXa inhibitor) and anti-metastasis Tuszynski, Gasic and Gasic (1987) (Mexican Leech) Haementeria ghilianii Ghilanten Anticoagulant (FXa inhibitor) and anti-metastasis Brankamp et al. (1990), Blankenship et al. (1990) (Great/Giant Amazon Saratin PAgI Barnes et al. (2001), Abdualkader et al. (2013) Leech) Hementin Fibrinogenolytic metalloproteinase Swadesh, Huang and Budzynski (1990) Continued Review article Bioscience Horizons � Volume 10 2017 ............................................................................................... .................................................................. ghilianii (Blankenship et al.,1990; Brankamp et al.,1990), and is also believed to have anti-metastasic properties. Lefaxin is another anticoagulant (FXa inhibitor) from Haementeria depressa (Faria et al.,1999). The N-terminal chain of lefaxin shows no homology to antistatin or ghilianten. Another FXa inhibitor isolated from Haementeria vizottoi, called Vizottin, displays a different effect compared with other leech antistatin-like inhibitors (Oliveira et al., 2012). Vizottin prevents the formation of FXa by the extrinsic tenase complex (FVIIa: tissue factor) and also inhibits free and bound FXa, probably by interacting with the active site of FXa (Oliveira et al., 2012). Thrombin inhibitors A direct thrombin inhibitor is an anticoagulant, which binds to thrombin directly and blocks its activity (reviewed by Coppens et al.,2012). Hirudin (Jacobi, 1904) was isolated from H. med- icinalis by Markwardt (1957; reviewed by Zaidi et al.,2011). Hirudin is the most potent naturally occurring thrombin inhibi- tor (Zaidi et al.,2011; Jiang et al., 2013) and many of its syn- thetic derivatives are used in clinics on a daily basis (reviewed by Nawarskas and Anderson, 2001; Zaidi et al.,2011). Subsequently, a similar anti-thrombin called Bufrudin has also been isolated (Electricwala et al.,1991; Abdualkader et al., 2013). Hirudin and bufrudin have slightly different structural (N-terminal amino acid sequence) and immunological proper- ties (Electricwala et al.,1991; Zaidi et al.,2011). Hirustasin is an antistatin-type serine protease inhibitor and the first tissue kallikrein inhibitor identified in the leech (Söllner et al.,1994; Kallikrein-kinin system discovered by Abelous and Bardier, 1909, as cited by review Su, 2014 and Kraut, Frey and Werle, 1930). Despite similarities to antistatin (FXa inhibitor; differing reactive site sequence and proteinase activity), hirusta- sin does not inhibit blood coagulation in vitro, nor is it amido- lytic of isolated FXa. This suggests that the specificity of antistatin-type proteinases influences coagulation (reviewed by Söllner et al.,1994; Boskova and Zavalova, 2001). A further thrombin inhibitor, Theromin, has been isolated from the Duck Leech Theromyzon tessulatum. It has no sequence homology with any other thrombin inhibitor (Salzet et al., 2000). Analgesia Some kinins are potent activators of nociceptive nerve cells, which induce or enhance pain sensations (Steranka et al., 1988; Hildebrandt and Lemke, 2011). Tissue kallikreins are proteases that cleave inactive kininogens to become active kinins (Hildebrandt and Lemke, 2011). The presence of antistatin-type substances (e.g. Hirustasin) in leech saliva may be an indication that it could reduce local tissue kallikrein activity in host tissue around the feeding site, therefore pre- venting the production of pain-inducing kinins. It may have an analgesic effect on the host (Hildebrandt and Lemke, 2011). An alternative explanation for kinin suppression is the ............................................................................................... .................................................................. Table 1. Continued Leech species Molecule Function in host Reference Haementeria depressa Lefaxin Anticoagulant (FXa inhibitor) Faria et al. (1999), Abdualkader et al. (2013) Hementerin Fibrinogenolytic metalloproteinase Chudzinski-Tavassi et al. (1998) Theromyson tessulatum Therostatin Anticoagulant (FXa inhibitor) Chopin et al. (2000) (Duck leech) Theromacin Antimicrobial Tasiemski et al. (2004), Abdualkader et al. (2013) Theromyzin Antimicrobial Tasiemski et al. (2004), Abdualkader et al. (2013) ECM, extracellular matrix; LCI, leech carboxypeptidase inhibitor; LDTI, leech derived tryptase inhibitor; PAgI, platelet aggregation inhibitor. Bioscience Horizons � Volume 10 2017 Review article ............................................................................................... .................................................................. release of kininases into the wound to inactivate kinins by an adenosine 5’-diphosphate (ADP) diphosphohydrolase. The proteolytic cleavage. This removal of pain-inducing or sensi- latter removes the terminal phosphate group from ADP, gen- tizing agents from the feeding site would explain the analgesic erating AMP that does not bind to purinergic receptors, effect of leeches. thereby suppressing platelet adhesion (Rigbi, Orevi and Eldor, 1996; Hildebrandt and Lemke, 2011). Fibrinolysis/increasing blood flow Leech carboxypeptidase inhibitor (LCI; Reverter et al.,1998), Ticks inhibits a metalloproteinase (Carboxypeptidase B) responsible for the cleavage of kinin and therefore its inactivation in blood Ticks are ectoparasites of the order Parasitiformes, sub-order plasma. By inhibiting carboxypeptidase B, this increases kinin Ixodida (Black and Piesman, 1994), which have an obligate presence, which may result in increased pain and blood flow haematophagous lifestyle (reviewed by Kazimírová and to the feeding site. Whilst the increased blood flow would be Štibrániová, 2013). There are two major families, the beneficial to the leech, the increase in pain (kinin) would not. Ixodidae (hard; Black and Piesman, 1994; reviewed by Therefore, the LCI effect may be more relevant to inhibiting Francischetti et al., 2009) and Argasidae (soft). Ixodidae ticks carboxypeptidase B’ effect on fibrinolysis (Hildebrandt and are further partitioned into the metastriate (Dermacentor or Lemke, 2011). Rhipicephalusgenera spp.), which have short mouth parts and secrete a cement for attachment to the host (Francischetti Inhibitors of host immune system et al., 2009; Kazimírová and Štibrániová, 2013; reviewed by Sauer, 1977), and prostriate ticks, which use long, barbed The complement system is part of the innate immune system of mouth parts to stay attached to their host (Francischetti et al., vertebrates and is the first defence against invaders (e.g. bac- 2009; Kazimírová and Štibrániová, 2013). teria; Buchner, 1891). An inhibitor of complement component C1 (60–70 kDa), from leech saliva, blocks both the classic and Adult Ixodidae ticks prefer large mammals like humans alternative pathways of the complement system (Baskova et al., and especially, ruminant livestock (Maina et al., 2014). 1988; as cited and by Boskova and Zavalova, 2001)and is Immature Ixodidae ticks also feed on smaller mammals, birds anti-inflammatory (Hildebrandt and Lemke, 2011). and reptiles (Maina et al., 2014). Soft ticks (Argasidae) feed on a wide range of vertebrates including amphibians, reptiles Leech saliva contains a specific inhibitor of mast cells, leech- and birds (Kazimírová and Štibrániová, 2013). derived tryptase inhibitor (LDTI; Mϋhlhahn et al., 1994; Sommerhoff et al., 1994; Stubbs et al.,1997; Hildebrandt and The constituents of tick saliva diversify as they mature, Lemke, 2011). It exists in three isoforms with differing C- and quantities of various substances vary between each moult termini. Its biological function is not yet characterized, but it (instar) until sexual maturity is reached (Lloyd and Walker, may supress cell-mediated inflammation in host tissues at the 1995; Due et al., 2013; Juckett, 2013). feeding site (Hildebrandt and Lemke, 2011). The active constituents of tick saliva are shown in Table 2. A secretory protein found in leeches named Eglin-c Ticks form a haemorrhagic pool within the tissues of the host (Seemüller et al., 1977) binds to human neutrophils (Snider from which they feed (reviewed by Ribeiro, 1995). When a et al.,1985; Braun et al., 1987). Neutrophils are the most abun- tick is not attached to a host, salivary gland lobes produce dant white blood cell in mammals and part of the innate hygroscopic saliva. This helps the tick remain hydrated whilst immune system (Murphy, 2012). Eglin-c stops neutrophils at it waits for a host, sometimes for years (reviewed by Bowman the feeding site from entering the surrounding tissue, preventing and Sauer, 2004; Francischetti et al., 2009). Salivation is inflammation. It is therefore proposed as an anti-inflammatory believed to be under nervous control, involving cAMP and agent, which protects host tissues from destruction by endogen- calcium (Sauer, 1977). Ticks alternate in cycles of feeding and ous neutrophils (Hildebrandt and Lemke, 2011). salivation, each lasting 5–20 min (Gregson, 1967). After feed- ing, they fall off their hosts and become inactive and unwilling Hyaluronidase (Linker, Hoffman and Meyer, 1957; to reattach (Bowman and Sauer, 2004). It was long believed, Hovingh and Linker, 1999) digests hyaluronic acid present in as early as 27–79 AD, that ticks would die after gorging on the extracellular matrix of host tissues. Its secretion in leech blood due to lack of an anus. However, ticks do possess an saliva facilitates the distribution of other salivary molecules. anus and excrete mainly guanine and other nitrogenous waste These interfere with immune cell function in the host by in small amounts (Bowman and Sauer, 2004). mobilizing water molecules from proteoglycans and destabil- izing the matrix (Hildebrandt and Lemke, 2011). Vasodilators and inhibitors of immune Inhibitors of platelet activation and system function adherence A vasodilator assists the tick in its blood feeding by increasing Leech saliva contains two substances, which reduce host blood flow to feeding site (Ribeiro, Makoul and Robinson, platelet adherence to vessel walls at the feeding site: a 1988). The salivary vasodilators from Ixodes scapularis/dam- mammalian-type collagenase (Rigbi et al., 1987) and apyrase, mini are the arachidonic acid derivatives prostacyclin (PGI ; ............................................................................................... .................................................................. 5 Review article Bioscience Horizons � Volume 10 2017 ............................................................................................... .................................................................. Table 2. Haematophagy-related substances found in saliva of hard (Ixodidae) and soft (Argasidae) ticks and their functions in the host Tick species Molecule Function in host Reference Hard ticks (Ixodidae) Ixodes scapularis or Ixodes dammini (Deer/ PGI Platelet inhibitor Ribeiro, Makoul and Robinson (1988), Blacklegged tick) Kazimírová and Štibrániová (2013) PGE Vasodilator Ribeiro et al. (1985), Law, Ribeiro and Wells (1992) Metalloprotease Inhibits angiogenesis Valenzuela et al. (2002), Kazimírová and Štibrániová (2013) Ixolaris TFPI Lai et al. (2004), Francischetti et al. (2002b) - Kininase enzyme Ribeiro et al. (1985), Ribeiro and Mather (analgesic effect) (1998), Francischetti et al. (2009) Amblyomma americanum (Lone Star tick) PGE Vasodilator Ribeiro et al. (1992), Law, Ribeiro and Wells (1992), Ribeiro (1995) PGF Vasodilator Ribeiro et al. (1992) 2α Amblyomma hebraeum (South African Bont Tick) Amblin Thrombin inhibitor Lai et al. (2004) Amblyomma variegatum (Tropical Bont tick) Variegin Thrombin inhibitor Koh et al. (2007), Koh and Kini (2008) Peptide AP18 Enhances thrombin Koh et al. (2007) amidolytic activity - Anti-IL-8 Hajnická et al. (2001) Rhipicephalus/Boophilus microplus (Cattle/ PGE Vasodilator Dickinson et al. (1976), Higgs et al. Southern cattle tick) (1976), Law, Ribeiro and Wells (1992), Tatchell and Binnington (1973) Boophilin Thrombin inhibitor; Macedo-Ribeiro et al. (2008); Liao et al. serine protease (2009) inhibitor; potential anticoagulant Ixodidin Antimicrobial single- Fogaça et al. (2006) domain inhibitor/ protease inhibitor Rhipicephalus evertsi evertsi (Red-legged tick) Neurotoxin Paralysis of host Viljoen et al. (1986), Lloyd and Walker (1995) Hyalomma dromedarii (Camel tick) NTI-1 Thrombin inhibitor Ibrahim et al. (2001) NTI-2 Haemaphysalis Longicornis (Cattle/Bush/Shrub Longistatin PA and anticoagulant Anisuzzaman et al. (2010, 2011) tick) properties. (isolated from midgut) Hemalin Thrombin inhibitor Liao et al. (2009) Dermacentor variabilis (American dog/Wood Variabilin Platelet inhibitor Wang et al. (1996), Francischetti et al. tick) (Disintegrin) (2009) Dermacentor andersoni (Rocky Mountain - Anti-TNFα Ramachandra and Wikel (1995) wood tick) Continued ............................................................................................... .................................................................. 6 Bioscience Horizons � Volume 10 2017 Review article ............................................................................................... .................................................................. Table 2. Continued Tick species Molecule Function in host Reference Dermacentor reticulatus (Marsh/Ornate Cow/ - Anti-IL-8 Hajnická et al. (2001) American Levi tick), Rhipicephalus appendiculatus (Brown/Brown Ear tick), Haemaphysalis inermis (Winter tick), Ixodes ricinus (Sheep/Castor Bean/Deer tick) Soft ticks (Argasidae) Ornithodoros moubata (Soft/Argasid/African Ornithodorin Thrombin inhibitor van de Locht, Stubbs and Bode (1996), relapsing fever/Tampan tick/Eyeless Kazimírová and Štibrániová (2013) tampan) Moubatin PAgI Waxman and Connolly (1993), Basanova, Baskova and Zavalova (2002) TAP Anticoagulant (FXa Waxman et al. (1990) inhibitor) Ornithodoros savignyi (Sand tampan) Savignin Inhibits thrombin- Nienaber, Gaspar and Neitz (1999) induced platelet aggregation Apyrase Platelet inhibitor Mans et al. (1998a, 1998b), Kazimírová and Štibrániová (2013) Table includes all known putative secreted proteins, excluding housekeeping proteins or transcripts. PGI , prostaglandin I or prostacyclin; PGE , prostaglandin E ; ‘-’, name not specified; PGF , prostaglandin F ; NTI, non-competitive thrombin inhibitor; TFPI, 2 2 2 2 2α 2alpha tissue factor pathway inhibitor; TAP, tick anticoagulant peptide; PAgI, platelet aggregation inhibitor; PA, plasminogen activator. Ribeiro, Makoul and Robinson, 1988) and PGE (Ribeiro Anticoagulant and PA et al.,1985, reviewed by Valenzuela, 2004). PGE is also Haemaphysalis longicornis secretes a PA, characterized and foundinthe saliva of Amblyomma americanum (Ribeiro named Longistatin (Anisuzzaman et al.,2010, 2011). This has et al.,1992)and Boophilus microplus (Dickinson et al., high specificity to fibrin clot-bound plasminogen and has been 1976; Higgs et al., 1976; reviewed by Tatchell and proposed to have anticoagulant function also (Anisuzzaman Binnington, 1973; Law, Ribeiro and Wells, 1992). These et al.,2011). This combination of functions suggests that it is a may be involved in the generalized lymphocyte suppression critical component in the maintenance of blood pools in the of tick-infested species (Ribeiro et al., 1985; Law, Ribeiro feeding process of Ixodid ticks (Anisuzzaman et al., 2011). and Wells, 1992). Tick histamine-binding proteins (HBPs) are lipocalins, Platelet aggregation inhibitors which trap cationic, hydrophilic molecules. This is in contrast Tick saliva contains apyrase, a platelet agonist (Kazimírová and to most lipocalins that bind lipophilic compounds. This is pre- Štibrániová, 2013) and platelet inhibitors such as disintegrins sumably an adaptation to haematophagy; the salivary glands and prostacyclin. These inhibitors, for example Variabilin, con- of most if not all species of tick contain highly histamine- tain the arginine-glycine-aspartic acid (RGD) motif, which pre- specific binding proteins (Paesen et al., 1999, 2000). Evidence vents the binding of fibrinogen to platelets (Wang et al., 1996; suggests that expression of histamine is linked to an acquired Francischetti et al., 2009). A further platelet inhibitor called tick resistance (D. andersoni; reviewed by Wikel, 1982; Moubatin is capable of weakly inhibiting collagen-induced pla- Brossard and Wikel, 2004). telet aggregation (reviewed by Waxman and Connolly, 1993; The metalloprotease found in I. scapularis inhibits angio- Basanova, Baskova and Zavalova, 2002). genesis, the process of forming new blood capillaries (Valenzuela et al., 2002; Kazimírová and Štibrániová, Thrombin inhibitors 2013). Cytokines, including chemokines, interferons and tumour necrosis factor (TNF), are important in vertebrate Two non-competitive thrombin inhibitors, NTI-1 (3.2 kDa) and host immune responses, its development and expression. NTI-2 (14.9 kDa), have been isolated from Hyalomma drome- Tick saliva also contains anti-TNF (Ramachandra and darii (Ibrahim et al., 2001). A potent thrombin inhibitor found Wikel, 1995), anti-IL-8 (interleukin 8; Hajnická et al., 2001) in the tropical bont tick (Amblyomma variegatum), called and blocks other cytokine binding activities (Brossard and Variegin is a 32 residue polypeptide and one of the smallest Wikel, 2004). thrombin inhibitors discovered in nature (Koh et al., 2007, ............................................................................................... .................................................................. 7 Review article Bioscience Horizons � Volume 10 2017 ............................................................................................... .................................................................. reviewed by Koh and Kini, 2008). It has no structural similar- individuals (Greenhall and Schutt, 1996; Altringham, 2011; ities with thrombin inhibitors found in other haematophagous reviewed by Wimsatt, 1959). animals, including hirudin, Rhodniin and Theromin, even Vampire bats often roost in a group of 8–12 females; these though they have the same binding site on thrombin (Koh et al., are related and unrelated individuals with whom they per- 2007). Peptide AP18 (a synthetic peptide of variegin) has been form reciprocal altruism, the reduction of one animal’s fitness reportedto bindtoexosite-I, slightly enhancing thrombin ami- to increase that of another with the expectation that this dolytic activity (Koh et al., 2007). behaviour will be returned (Altringham, 2011). The fact that Boophilius are one-host ticks, which feed on cattle vampire bats perform this with those to whom they are not (Jongejan and Uilenberg, 2004). Boophilin, a proteinase closely related suggests a highly stable, social composition inhibitor, inhibits thrombin and interferes with serine pro- within the colony (Altringham, 2011). teases: trypsin and plasmin (Macedo-Ribeiro et al., 2008). Once the bat has located its prey (using long-range vision, Hemalin (Liao et al., 2009), a thrombin inhibitor of the olfaction, acute hearing and echolocation), it uses close-range Kunitz-type family, has high homology with boophilin thermal and mechanical sensitivity strategies to locate blood (Macedo-Ribeiro et al., 2008). The thrombin inhibitor capillaries below the skin surface (Jones, Teeling and Amblin (Lai et al., 2004) has been isolated from the haemo- Rossiter, 2013; Low et al., 2013). They attack only resting or lymph of ixodid tick Amblyomma hebraeum and displays sleeping prey and generally feed without disturbance sequence similarities to boophilin (Macedo-Ribeiro et al., (Wimsatt, 1959). The process must therefore be painless 2008) and Ixolaris (a tissue factor pathway inhibitor, TFPI; (Wimsatt, 1959), suggesting an analgesic in the saliva. Each Francischetti et al., 2002b). Inhibitors of the Kunitz family species possesses a characteristic lower lip median groove, with bovine-pancreatic-trypsin inhibitor domains are also which facilitates feeding (Wimsatt, 1959) but with small dif- common in ticks (reviewed by Corral-Rodríguez et al., 2009). ferences in anatomy (Greenhall and Schutt, 1996). Savignin is a thrombin inhibitor found in Ornithodoros savignyi, which inhibits thrombin-induced platelet aggrega- Desmodus rotundus possess upper and lower incisors. The tion (Nienaber, Gaspar and Neitz, 1999). It is a competitive, upper incisors diverge to create an upside-down ‘V’ shape. slow, tight-binding inhibitor that interacts with thrombin This creates a crater-like ‘divot’ 1–2 mm deep in the host’s tis- exosite-I and blocks its catalytic site (as does leech hirudin; sue. The bat laps up the blood via two straw-like ducts on the Nienaber, Gaspar and Neitz, 1999). ventral side of the tongue (Wimsatt, 1959), whilst saliva is released from the dorsal side of the tongue from the sub- maxillary gland (Low et al., 2013). It takes 20–30 min for a Other bat to take in ~25 ml, which is approximately 60% of their Neurotoxins found in the saliva of Rhipicephalus evertsi ever- bodyweight (Altringham, 2011). To prevent themselves from tsi cause host paralysis (Viljoen et al., 1986; Lloyd and being grounded by the weight and open to potential preda- Walker, 1995) and have presumably evolved to prevent the tors, their kidneys excrete excess plasma from the already host interrupting feeding sessions. Tick anticoagulant peptide ingested blood before they finish a feed (Altringham, 2011). (TAP), isolated from Ornithodoros moubata, functions as an The concoction of anticoagulants and other biochemicals inhibitor of FXa (Waxman et al., 1990). Ixodes scapularis in the bat’s saliva causes the normal response to this type of contains a kininase enzyme, which may contribute to the injury in the host to be delayed from minutes to hours (Low analgesic effect of bites (Ribeiro et al., 1985; Ribeiro and et al., 2013). The overall anticoagulant activity of the saliva Mather, 1998; Francischetti et al., 2009). decreases progressively after daily salivation and is restored after 4 days (Fernandez et al., 1998). Multiple feedings from an individual prey animal may increase its resistance to the Vampire bats anticoagulants, indicating that an immune response can be Three species of bat have evolved a haematophagous lifestyle: acquired (Delpieto and Russo, 2009). Diphylla ecaudata, Desmodus rotundus and Diaemus youngi. Anticoagulants These ‘vampire’ bats are mammals of the order Chiroptera, sub-order Microchiroptera, sub-family Desmodontinae Draculin is an anticoagulant found in vampire bat saliva (other members of the sub-order are insectivores). Avian (Apitz-Castro et al.,1995; Fernandez et al.,1998, 1999). It hosts are the only prey of the hairy-legged vampire bat (D. requires glycosylation of the native molecule to become a bio- ecaudata; Tellgren-Roth et al., 2009; Low et al., 2013). The logically active, non-competitive, tight-binding inhibitor of common vampire bat (D. rotundus) feeds substantially only FXa (Fernandez et al.,1998, 1999). It is also an inhibitor of on mammalian hosts, including livestock and humans (Low FIXa of the intrinsic pathway of blood coagulation (Fernandez et al., 2013). The white-winged vampire bat (D. youngi) et al., 1998; Basanova, Baskova and Zavalova, 2002; Oliveira feeds on both mammalian and avian blood (Low et al., et al.,2012). 2013). They are distributed widely over central and South America, typically living in caves, tree hollows and aban- Recent transcriptome and proteome studies of D. rotundus doned mines (Low et al., 2013)incoloniesof30–100 glands (Ma et al., 2013) revealed a 2-Kunitz-domain type ............................................................................................... .................................................................. 8 Bioscience Horizons � Volume 10 2017 Review article ............................................................................................... .................................................................. inhibitor, named Desmolaris (Ma et al., 2013). Desmolaris is This raises the question of whether divergent or convergent evo- a naturally deleted form of Kunitz-I-domainless TFPI and has lutionary mechanisms may have been in play. similar actions: it tightly binds FXIa of the extrinsic pathway We take divergent evolution to be the emergence in a spe- of the coagulation cascade (Ma et al., 2013). cies of an apomorphy not present in an ancestor (or the reten- tion of a characteristic in one descendant and its loss in Platelet aggregation inhibition and another descendant). This mechanism can be reliably invoked fibrinolytic activity when unique characteristics and mechanisms occur in a spe- cies or higher taxonomic group. We take convergent evolu- A platelet aggregation inhibitor (PAgI) (ADP-induced and tion to be evidenced by the presence of an apomorphy in thrombin-induced) in vampire bat saliva exerts its effects by more than one species when it is unlikely to have been present blocking ADP-binding sites or by specifically acting on the in their closest common ancestor. plasma cofactor involved in the platelet aggregation reaction (Hawkey, 1967). This molecule could be the apyrase or phos- The three animal groups represented here (mammals, phatase recently found in the saliva of D. rotundus arthropods and annelids) apparently diverged early in animal (Francischetti et al., 2013). evolution (reviewed by Sanetra et al., 2005). Mammals are deuterostomes while arthropods and annelids are proto- PA/fibrinolytic activity stomes (diverged from bilateria); arthropods are ecdysozoa and annelids are lophotrochozoa (diverged from protosto- The fibrinolytic activity of vampire bat saliva has been known mia). These major divergences probably occurred during the since 1932 (Bier, 1932). A PA (a serine protease) has been pre-Cambrian, Ediacaran era around 600 million years ago found and named D. rotundus salivary plasminogen activator (m.y.a.; Erwin and Davidson 2002; Peterson et al., 2008). In (DSPA), Desmokinase (Cartwright, 1974; Low et al.,2013)or addition, all three animals prey on amniote homeotherms— Desmoteplase (reviewed by Paciaroni, Medeiros and mammals and birds—which emerged considerably later than Bogousslavsky, 2009). It has been characterized (Hawkey, the parasite clades: 300–250 m.y.a. (Hellenius and Ruben 1966; Gardell et al.,1989) and cloned (Krätzschmar et al., 2004; Laurin and Reisz, 2011). 1991; Gulba, Praus and Witt, 1995; Francischetti et al.,2013). Four DSPAs (α1, α2, β and γ) have been isolated and character- Given that haematophagy is a rare phenomenon in each of ized (reviewed by Piechowski-Jozwiak and Bogousslavsky, the three groups, the likelihood that it is a characteristic which 2013); they are encoded by four different highly conserved has been retained from nearest common ancestors is extremely genes, each with distinct structure and properties (Krätzschmar small. Furthermore, assuming that the vertebrate hosts for the et al., 1991). All are homologous to human t-PA, except that haematophagous leeches and ticks emerged much later than the DSPAs are single-chain molecules and are dependent on the common ancestor of these parasites and that the mamma- fibrin as a cofactor (Liberatore et al.,2003). lian parasite–host relationships of the vampire bats must also have emerged later, we take the haematophagy of our three DSPA-α1 has been investigated for its pharmaceutical animals groups to be a convergently evolved lifestyle. potential and was in phase III clinical trials (Piechowski- Jozwiak and Bogousslavsky, 2013). Several substances found Given this conclusion, it would appear that any similarities in D. rotundus saliva (Table 3) could be of potential use in between the species in the mechanisms they employ to achieve thrombolytic therapy. haematophagy (preventing blood coagulation, nociception, would healing, etc.) must also have arisen convergently. Note, however, that the fundamental processes of coagulation, noci- Discussion ception and wound healing, against which haematophagous animals work, are largely common to mammals and birds. Haematophagous animals have evolved mechanisms, which They therefore define and limit the biochemical mechanisms, counteract the haemostatic response of their hosts. The which must have evolved for the haematophagous lifestyle to Triatominae sub-family of haematophagous bugs possesses be successful. A further possibility is that the mechanisms many beneficial compounds in their saliva to counteract haematophagous animals use are co-opted from, and adapta- haemostasis. Lipocalins, for example, also found in many other tions of, commonly existing biochemical processes. To this species of blood-feeding arthropods (Santos et al.,2007). The extent, they represent divergent evolution from their non- variety of mechanisms is extensive and several have been haematophagous cousins as well as convergent evolution exploited in medical applications (Abdualkader et al.,2013; towards unrelated similarity. reviewed by Cherniack, 2011; Piechowski-Jozwiak and Bogousslavsky, 2013). The three types of animal discussed in Most haematophagous species possesses at least one anti- this review represent taxonomically distinct groups and it is coagulant, which inhibits one or multiple factors involved clear that they exhibit similarities and differences in the in the mammalian coagulation cascade (Tables 1–4). mechanisms they employ. Other haematophagous parasites not Anticoagulants, most of which are FXa inhibitors, are the discussed (Table 4)such as sandflies (Lutzomyia longipalpis) dominant constituents of leech saliva. Although the N- also possess anti-coagulation mechanisms (Collin et al.,2012). terminal sequence of lefaxin showed no homology with other ............................................................................................... .................................................................. 9 Review article Bioscience Horizons � Volume 10 2017 ............................................................................................... .................................................................. Table 3. Haematophagy-related molecules found in the sub-maxillary and accessory glands of the vampire bat (D. rotundus) Molecule Function in host Reference Draculin Anticoagulant (FXa and FIXa Apitz-Castro et al. (1995), Fernandez et al. (1998, 1999), Basanova, inhibitor) Baskova and Zavalova (2002) Desmolaris Anticoagulant (FXIa Ma et al. (2013) inhibitor) Desmoteplase (DSPA-alpha-1) PA (fibrinolytic/ Bier (1932), Gardell et al. (1989), Krätzschmar et al. (1991), thrombolytic) Schleuning et al. (1992), Gulba, Praus and Witt (1995), Paciaroni, Merdeiros and Bogoussalvsky (2009); Tellgren-Roth et al. (2009); Francischetti et al. (2013), Piechowski-Jozwiak and Bogousslavsky (2013), reviewed by Patel, Ispoglou and Apostolakis (2014) Phosphatase Anti-platelet Hawkey (1967) Apyrase Anti-platelet Hawkey (1967) Pituitary adenylate cyclase Vasodilator activating peptide (PACAP) C-type natriuretic peptide (CNP) Vasodilator TNFα-stimulated gene 6 (TSG-6) Anti-inflammatory Lipophilin/secretoglobin precursors Anti-inflammatory (several) Lipocalin and other lipid carriers Anti-inflammatory Cystatin Anti-inflammatory (cycteine- type inhibitor) Chemokine CCL28 Antimicrobial (broad spectrum) β-Defensin, BPI/LBP/CETP family Antimicrobial lymphotoxin Kunitz domain (protease inhibitor Kunitz inhibitor domain) Neuroserpins Modulates activation of fibrinolysis triggered by DSPA Chymase Serine protease Clotting pathways serine proteases Protease DNAse Effects neutrophil function Source: Francischetti et al. (2013) unless otherwise stated specified. Further research is needed to confirm the function of some of these molecules in the host. Serine proteases are included because PAs belong to this family of enzymes. FIXa, activated coagulation factor IX; FXIa, activated coagulation factor XI; CCL28, (C-C motif) ligand 28; BPI/LBP/CETP family, bactericidal permeability-increasing protein/lipopolysaccharide-binding protein/cholesteryl ester transfer protein family; DNAse, deoxyribonuclease; PA, plasminogen activator; DSPA, D rotundus salivary plasminogen activator. leech anticoagulants it does show homology with Prolixin S Whilst evidence of analgesia is currently not available for derived from Rhodnius prolixus (Kissing bug; Hellmann and leeches, it is for Ixodes ticks (Ribeiro et al., 1985; Hawkins, 1965). The anticoagulant and anti-metastatic Hildebrandt and Lemke, 2011). Similar feeding physiology of agents antistatin and ghilanten are found in different species leeches has been observed in R. prolixus, suggesting that simi- of leech so it is most likely that these properties have devel- lar processes may be required for efficient blood feeding, also oped separately to facilitate blood feeding in these species. acquired convergently (Lent et al., 1988). ............................................................................................... .................................................................. 10 Bioscience Horizons � Volume 10 2017 Review article ............................................................................................... .................................................................. Table 4. Molecules in the saliva of haematophagous and non-haematophagous animals other than the three groups considered in detail in this review Species Molecule Function in host Reference Haematophagous animals Lutzomyia longipalpis (Sand fly) Maxadilan Vasodilator Ribeiro et al. (1989b), Law, Ribeiro and Wells (1992), Lerner et al. (1991). Lufaxin Anticoagulant (FXa Collin et al. (2012). inhibitor) - Anti-complement Cavalcante, Pereira and Gontijo (2003). Apyrase PAgI Ribeiro, Rossignol and Spielman (1986), Ribeiro (1995). Phlebotomus papatasi (Sand fly) Apyrase PAgI Ribeiro et al. (1989a), Ribeiro (1995). Phlebotomus argentipes Apyrase PAgI Ribeiro et al. (1989a), Ribeiro (1995). Phlebotomus perniciosus Apyrase PAgI Ribeiro et al. (1989a), Ribeiro (1995). Glossina morsitans (Savannah Tsetse fly) 5′nucleotidase-related PAgI Mant and Parker (1981), Caljon et al. Apyrase (2010). - Thrombin inhibitor Parker and Mant (1979), Ribeiro (1995). Glossina austeni (Savannah Tsetse fly) - PA Hawkins (1966), Hawkey (1967). Simulium vittatum (Black fly/Buffalo gnat) - Thrombin inhibitor Jacobs et al. (1990), Ribeiro (1995). - Anticoagulant (FVII Makonnen et al., unpublished inhibitor) observations as cited by Ribeiro (1995) - Anticoagulant (FXa Jacobs et al. (1990) inhibitor) - Anticoagulant (FVa Abebe et al. (1996), Basanova, Baskova inhibitor) and Zavalova (2002) Aedes aegypti (Yellow-fever mosquito) - Anticoagulant Stark and James (1995) (serine protease inhibitor of FXa) Apyrase PAgI Ribeiro et al. (1984), Vachereau and Ribeiro (1989) Oropsylla bacchii (Flea) Apyrase PAgI Ribeiro, Vaughan and Azad (1990b), Ribeiro (1995) Orchopea howardii (Squirrel Flea) Apyrase PAgI Ribeiro, Vaughan and Azad (1990b), Ribeiro (1995) Xenopsylla cheopis (Oriental Rat Flea) Apyrase PAgI Ribeiro, Vaughan and Azad (1990b), Ribeiro (1995) Rhodniini/Triatomini (Kissing/ Apyrase PAgI Ribeiro, Marinotti and Gonzales (1990a), Traitomines/Assassin/Conenose bug) Law, Ribeiro and Wells (1992), Santos et al. (2007) Dimiconin Anticoagulant (FXII Ishimaru et al. (2012) inhibitor) Continued ............................................................................................... .................................................................. 11 Review article Bioscience Horizons � Volume 10 2017 ............................................................................................... .................................................................. Table 4. Continued Species Molecule Function in host Reference Prolixin S (NP2) Anticoagulant Hellmann and Hawkins (1965), Ribeiro, Schneider and Guimarães (1995) Triabin Thrombin inhibitor Noeske-Jungblut et al. (1995), Ishimaru et al. (2012) Rhodniin Thrombin Inhibitor Friedrich et al. (1993) Pallidipin 1 and 2 Collagen-induced Noeske-Jungblut et al. (1994), Basanova, PAgI Baskova and Zavalova (2002), Ishimaru et al. (2012) R. prolixus aggregation PAgI Francischetti et al. (2000), Francischetti, inhibitor-1 (RPAI-1) Andersen and Ribeiro (2002a), Francischetti et al. (2009), Ishimaru et al. (2012) Triafestin-1 and -2 Plasma kallikrein- Isawa et al. (2007), Ishimaru et al. (2012). kinin system inhibitors (FXII and kininogen inhibitor) NP1-4 Vasodilator Ribeiro, Marinotti and Gonzales (1990a), Champagne, Nussenzveig and Ribeiro (1995), Basanova, Baskova and Zavalova (2002), reviewed by Champagne (2005) Lipocalins Transports small Santos et al. (2007) hydrophobic molecules Serine protease Protease Santos et al. (2007) -PA Hellmann and Hawkins (1964), Hawkins and Hellman (1966), Hawkey (1967) - Anticoagulant (FVIII Hellmann and Hawkins (1965, 1966) inhibitor) Eutriatoma maculatus (Assasin bug) Maculatin Thrombin Inhibitor Hellmann and Hawkins (1966) -PA Hellmann and Hawkins (1966) Culicoides sonorensis (Biting midge) TFPI1 and TFPI2 Protease inhibitors Campbell et al. (2005) - Hyaluronidase Campbell et al. (2005) Non-haematophagous animals Bombus terrestris (Buff-tailed/Large earth Bt-KTI Serine protease Qiu et al. (2013) Bumblebee) inhibitor (acts as a plasmin inhibitor) Scolopendra subspinipes mutilans - Anticoagulant (FXa Kong et al. (2013) (Chinese red-head/Chinese inhibitor) red-headed centipede) Scolonase Serine protease You et al. (2004) Continued ............................................................................................... .................................................................. 12 Bioscience Horizons � Volume 10 2017 Review article ............................................................................................... .................................................................. Table 4. Continued Species Molecule Function in host Reference Eisenia fetida (Redworm/Brandling/ Earthworm Fibrinolytic Thrombolytic/ Park et al. (1989) as cited by Sharma Panfish/Trout/Tiger/Red Wiggler/Red enzyme (EFE)/ Fibrinolytic et al. (2011), Mihara et al. (1991), Californian Earthworm), Lumbricus Lumbrokinase Sharma et al. (2011), Fu et al. (2013), rubellus, Eudrilus eugeniae (various Li et al. (2012) Earthworms) Antheraea pernyi (Chinese oak silkworm) Cocoonase Thrombolytic Geng et al. (2014) The table excludes putatively secreted proteins, housekeeping proteins/transcripts, substances with unknown function and substances not related to wound repair, thrombolysis or blood feeding. Most potent known to man (greater than calcitonin gene-related peptide). Serine proteases are included because PAs belong to this family of enzymes. ‘-’, name not specified; NP, nitroporins; TFPI1 and TFPI2, tissue factor pathway inhibitor 1 and 2; FVa, FVIIa, FXa, FXIIa, FXIIIa, activated coagulation factors V, VII, X, XII, XIII; PAgI, platelet aggregation inhibitor; PA, plasminogen activator. The PAs in the vampire bats are highly conserved, espe- Both haematophagous animals and non-haematophagous cially between the two species which feed upon mammals animals possess some sort of serine protease and serine prote- (D. rotundus and D. youngi; Tellgren-Roth et al., 2009). ase inhibitor (Qiu et al., 2013). The presence of serine prote- Evolution of the plasminogen gene may have led to the wide ase inhibitors in diverse groups of insects (honey bee, utilization of mammalian livestock by these two species and mosquito; Table 4; Qiu et al., 2013) suggests that endogenous consequently led to the rise of D. rotundus as the common suppressors of serine proteases, of which PA is a family mem- vampire bat (Tellgren-Roth et al., 2009). Absence of a PA in ber, are required at some stage of insect life and have been leeches suggests that the PA activity in the saliva of H. longi- conserved for reasons unconnected with blood feeding. cornis, Eutriatoma maculatus, R. prolixus (Hellmann and Insects heal wounds using cross-linking enzymes including Hawkins, 1964; Hawkins and Hellman, 1966) and D. rotun- transglutaminase, phenoloxidase (reviewed by Theopold dus is the result of convergent evolution. If it originated from et al., 2004) and lipophorin (Li et al., 2002) to form haemo- a common ancestral gene, it is unclear why the leech would lymph clots. No true orthologues of vertebrate blood clotting lose a potentially beneficial feature during its evolution. factors have been found in insects but several proteins with similar functional domains have been detected (Theopold The dual function of tick Longistatin, as a PA and anticoagu- et al., 2004). lant, indicates the importance of these components to species survival. It also suggests that other PAs are the result of replica- In line with the similarities between serine proteases and tion and division over the course of evolution. The evolutionary domains of coagulation factors, some serine proteases in the split of the PAs and anticoagulants might have led to the now protochordate Botryllus schosseri (star ascidian/golden star separate entities which can be observed in other invertebrates. tunicate) are homologous to vertebrate blood-coagulation Further investigation into the homology between Longistatin proteases (Ponczek, Bijak and Nowak, 2012). They par- and other PAs and anticoagulants may provide further insight ticipate in reactions involving the provoked aggregation (Bowman and Sauer 2004; Anisuzzaman et al., 2011). The of different cell-type colonies leading to cell clumping at thrombin inhibitor variegin, from the tick A. variegatum,has the site of contact. This suggests that a cascade of acti- no homology with other haematophagous animal thrombin vated serine proteases was initially a defence mechanism inhibitors and presumably evolved independently. Peptide (not associated with vascular injury; Wan et al.,2013), AP18 binding to exosite-I also in A. variegatum ticks is compar- which evolved into the vertebrate complement system and able to hirudin C terminus behaviour of leeches (Maraganore haemostatic response (Oren et al., 2008; Ponczek, Bijak et al., 1989; Naski et al.,1990), suggesting that these two and Nowak, 2012). sequences have a similar mechanism of action even though the Although the evidence suggests that PAs in invertebrates two species are phylogenetically distant (Koh et al., 2007). were derived from a common ancestor, this does not seem The vampire bat uses a distinct PAgI to block ADP- likely for the vampire bats. One hypothesis for the transition induced platelet aggregation. The PAgI of other haema- of vampire bats to haematophagy is arboreal feeding (Schutt, tophagous animals uses different biochemical mechanisms. 2008): they may have evolved from carnivorous bats whose Apyrase, which hydrolyses ATP and ADP, is commonly prey had grown too large for them to hunt by their usual found in haematophagous insects including the tsetse fly methods. A behavioural shift allowed the proto-vampires to Glossina morsitans (Mant and Parker, 1981; Caljon et al., utilize these larger animals as a food resource. 2010). Amounts of apyrase activity are elevated in blood- We hypothesize two alternative mechanisms for the devel- feeding animals compared with non-blood feeding insects, opment of the necessary salivary biochemistry in vampire suggesting exploitation of a pre-existing mechanism bats to support this transition: (Francischetti et al., 2009). ............................................................................................... .................................................................. 13 Review article Bioscience Horizons � Volume 10 2017 ............................................................................................... .................................................................. (1) Adaptation of blood haemostatic molecules into anti- completing a PhD, to ultimately become a lecturer in immun- haemostatic salivary molecules. Because PAs have been ology. F.L.W. designed the study, carried out the research and found in all vertebrates investigated so far (reviewed by wrote up the paper. She has primary responsibility for the Schleuning, 2001), it is possible that molecules present in paper. M.R.L. assisted in writing up the paper. a common ancestor could have evolved separate but par- allel functions. The mechanism for this would be gene duplication followed by mutation and selection. Since References other haematophagous animals possess different salivary components, the convergence to haematophagy, facili- Abdualkader, A. M., Ghawi, A. M., Alaama, M. et al. (2013) Leech thera- tated by duplication, must have resulted from selection peutic applications. Indian Journal of Pharmaceutical Sciences,75 according to different mammalian lifestyles or habitat. (2), 127–137. Whether this would have been possible in the relatively short evolutionary time available (between 6000 and Abebe, M., Ribeiro, J. M., Cupp, M. S. et al. (1996) Novel anticoagulant 2 000 000 years, with the Phyllostomidae family diver- from salivary glands of Simulium vittatum (Diptera: Simuliidae) inhi- ging at 35 m.y.a.; Schutt, 2008; Jones, Teeling and bits activity of coagulation factor V. Journal of Medical Entomology, Rossiter, 2013) is not clear. 33 (1), 173–176. (2) Exploitation of host blood proteins. Anti-haemostatic Abelous, J. E. and Bardier, E. (1909) ‘Les substances hypotensives de proteins could be obtained directly from the blood of the l’urine humaine normale’. CR Société de Biologie (in French), 66, hosts and exploited adaptively. For this to occur, bats 511–520. would need to absorb the materials intact and without destruction by their immune system, and be able to Altringham, J. D. (2011) Bats: From Evolution to Conservation, 2nd edn, exploit them in subsequent feeding. The blood coagula- Oxford University Press Inc, New York, USA, pp. 29–30 , 160–161. tion factors FVII, FX and FIX have similar sequence and Anisuzzaman, Islam, M. K., Alim, M. A. et al. (2011) Longistatin, a plas- domain arrangements in all vertebrates (Ponczek, Bijak minogen activator, is key to the availability of blood-meals for ixo- and Nowak, 2012) so it is possible that those derived did ticks. PLoS Pathogens, 7 (3), e1001312. from the host were well tolerated. Anisuzzaman, Islam, M. K., Miyoshi, T. et al. (2010) Longistatin, a novel EF-hand protein from the ixodid tick Haemaphysalis longicornis,is Conclusion required for acquisition of host blood-meals. International Journal for Parasitology, 40 (6), 721–729. Haematophagous animals possess many salivary constituents, which are capable of preventing haemostasis in their host, Apitz-Castro, R., Béguin, S., Tablante, A. et al. (1995) Purification and exploiting a range of different mechanisms. Given that hae- partial characterization of draculin, the anticoagulant factor present matophagy is considered to have evolved independently sev- in the saliva of vampire bats (Desmodus rotundus). Thrombosis and eral times, the active salivary components presumably Haemostasis, 73 (1), 94–100. emerged by convergent evolution. The derivation of these Barnes, C. S., Krafft, B., Frech, M. et al. (2001) Production and character- molecules has yet to be defined, but the salivary components ization of saratin, an inhibitor of von Willebrand factor-dependent of leeches and ticks at least could be adaptations of common platelet adhesion to collagen. Seminars in Thrombosis and ancestral genes. Hemostasis, 27 (4), 337–348. Recent advances in large-scale transcriptome analysis and Barrett, K. E., Barman, S. M., Boitano, S. et al. (2012) Ganong’s Review of structural proteomic analysis have made the identification of Medical Physiology, 24th edn, McGraw-Hill Companies Inc, China, such molecules easier. Identification of new molecules and pp. 564–569. comparative homology of current proteins, especially those with anti-metastasis and analgesic properties, may help Basanova, A. V., Baskova, I. P. and Zavalova, L. L. (2002) Vascular- explain their origin as well as the evolution to haematophagy. platelet and plasma hemostasis regulators from bloodsucking ani- mals. Biochemistry (Moscow), 67 (1), 143–150. Baskova, I. P., Nikonov, G. I., Mirkamali, E. G. et al. (1988) Influence of a Author biography preparation from the medicinal leech on phagocytosis and comple- ment system. Kazansky Medicinsky Zhurnal, 5, 334–336. Francesca graduated with a 2:1 BSc (Hons) Animal Science degree from the University of Nottingham in 2014. It was Bier, O. G. (1932) Action anticoagulante et fibrinolytique de l’extract during this time that she developed a passionate interest in des glands salivaires d’une chauve-souris hèmatophage (Desmodus immunology, infection and wound healing. Francesca was rufus). Comptes Rendus Hebdomadaires des Séances – Société de awarded one of fifteen School of Life Sciences Taught MSc Biologie, 110, 129–131. Scholarship and has now completed her MSc in Immunology and Allergy at the University of Nottingham. She hopes to Black, W. C. 4th and Piesman, J. (1994) Phylogeny of hard- and soft-tick continue in the field of immunology research, by one day taxa (Acari: Ixodida) based on mitochondrial 16S rDNA sequences. ............................................................................................... .................................................................. 14 Bioscience Horizons � Volume 10 2017 Review article ............................................................................................... .................................................................. Proceedings of the National Academy of Sciences of the United States Chudzinski-Tavassi, A. M., Kelen, E. M., de Paula Rosa, A. P. et al. (1998) of America, 91 (21), 10034–10038. Fibrino(geno)lytic properties of purified hementerin, a metallopro- teinase from the leech Haementeria depressa. Thrombosis and Blankenship, D. T., Brankamp, R. G., Manley, G. D. et al. (1990) Amino Hemostasis, 80 (1), 155–160. acid sequence of ghilanten: anticoagulant-antimetastatic principle of the South American leech, Haementeria ghilianii. Biochemical and Collin, N., Assumpção, T. C. F., Mizurini, D. M. et al. (2012) Lufaxin, a Biophysical Research Communications, 166 (3), 1384–1389. novel factor Xa inhibitor from the salivary gland of the sand fly Lutzomyia longipalpis blocks protease-activated receptor 2 activa- Boskova, I. P. and Zavalova, L. L. (2001) Proteinase inhibitors from the medi- tion and inhibits inflammation and thrombosis in vivo. cinal leech Hirudo medicinalis. Biochemistry (Moscow), 66 (7), 703–714. Arteriosclerosis, Thrombosis and Vascular Biology, 32 (9), 2185–2198. Bowman, A. S. and Sauer, J. R. (2004) Tick salivary glands: function, Coppens, M., Eikelboom, J. W., Gustafsson, D. et al. (2012) Translational physiology and future. Parasitology, 129 (Suppl), S67–S81. success stories: development of direct thrombin inhibitors. Brankamp, R. G., Blankenship, D. T., Sunkara, P. S. et al. (1990) Circulation Research, 111, 920–929. Ghilantens: anticoagulant-antimetastatic proteins from the South Corral-Rodríguez, M. A., Macedo-Ribeiro, S., Barbosa Pereira, P. J. B. American leech, Haementeria ghilianii. The Journal of Laboratory et al. (2009) Tick-derived Kunitz-type inhibitors as antihemostatic and Clinical Medicine, 115 (1), 89–97. factors. Insect Biochemistry and Molecular Biology, 39 (9), 579–595. Braun, N. J., Bodmer, J. L., Virca, G. D. et al. (1987) Kinetic studies on the Delpieto, H. A. and Russo, R. G. (2009) Acquired resistance to saliva interaction of eglin c with human leukocyte elastase and cathepsin anticoagulants by prey previously fed upon by vampire bats G. Biological Chemistry Hoppe-Seyler, 368 (4), 299–308. (Desmodus rotundus): evidence for immune response. Journal of Brossard, M. and Wikel, S. K. (2004) Tick immunobiology. Parasitology, Mammalogy, 90 (5), 1132–1138. 129 (Suppl), S161–S176. Dickinson, R. G., O’Hagan, J. E., Schotz, M. et al. (1976) Prostaglandin in Buchner, H. (1891) Zur Nomenklatur der schützenden Eiweisskörper. the saliva of the cattle tick Boophilus microplus. The Australian Centr Bakteriol Parasitenk, 10, 699–701. Journal of Experimental Biology and Medical Science, 54 (5), 475–486. Campbell, C. L., Vandyke, K. A., Letchworth, G. J. et al. (2005) Midgut and sal- Due, C., Fox, W., Medlock, J. M. et al. (2013) Tick bite prevention and ivary gland transcriptomes of the arbovirus vector Culicoides sonorensis tick removal. British Medical Journal (Clinical Research ed.), 347, (Diptera: Ceratopogonidae). Insect Molecular Biology, 14 (2), 121–136. f7123. Caljon, G., De Ridder, K., De Baetselier, P. et al. (2010) Identification of a Erwin, D. H. E. and Davidson, E. H. (2002) The last common bilaterian tsetse fly salivary protein with dual inhibitory action on human ancestor. Development, 129, 3021–3032. platelet aggregation. PLoS One, 5 (3), e9671. Electricwala, A., Sawyer, R. T., Jones, C. P. et al. (1991) Isolation of throm- Cartwright, T. (1974) The plasminogen activator of vampire bat saliva. bin inhibitor from the leech Hirudinaria manillensis. Blood Blood, 43 (3), 317–326. Coagulation and Finbrinolysis, 2 (1), 83–89. Cavalcante, R. R., Pereira, M. H. and Gontijo, N. F. (2003) Anti- Faria, F., Kelen, E. M., Sampaio, C. A. et al. (1999) A new factor Xa inhibi- complement activity in the saliva of phlebotomine sand flies and tor (lefaxin) from the Haementeria depressa leech. Thrombosis and other haematophagous insects. Parasitology, 127, 87–93. Haemostasis, 82 (5), 1469–1473. Champagne, D. E. (2005) Antihemostatic molecules from saliva of Fernandez, A. Z., Tablante, A., Bartoli, F. et al. (1998) Expression of bio- blood-feeding arthropods. Pathophysiology of Haemostasis and logical activity of draculin, the anticoagulant factor from vampire Thrombosis,34 (4–5), 221–227. bat saliva, is strictly dependent on the appropriate glycosylation of the native molecule. Biochimica et Biophysica acta, 1425 (2), Champagne, D. E., Nussenzveig, R. H. and Ribeiro, J. M. C. (1995) 291–299. Purification, partial characterization, and cloning of nitric oxide- carrying heme proteins (nitrophorins) from salivary glands of the Fernandez, A. Z., Tablante, A., Beguín, S. et al. (1999) Draculin, the anti- blood-sucking insect Rhodnius prolixus. The Journal of Biological coagulant factor in vampire bat saliva, is a tight-binding, non- Chemistry, 270 (15), 8691–8695. competitive inhibitor of activated factor X. Biochimica et Biophysica acta, 1434 (1), 135–142. Chapin, J. C. and Hajjar, K. A. (2015) Fibrinolysis and the control of blood coagulation. Blood Reviews, 29 (1), 17–24. Fogaça, A. C., Almeida, I. C., Eberlin, M. N. et al. (2006) Ixodidin, a novel antimicrobial peptide from the hemocytes of the cattle tick Cherniack, E. P. (2011) Bugs as drugs, part two: worms, leeches, scor- Boophilus microplus with inhibitory activity against serine protei- pions, snails, ticks, centipedes, and spiders. Alternative Medicine nases. Peptides, 27 (4), 667–674. Review: A Journal of Clinical Therapeutic, 16 (1), 50–58. Chopin, V., Salzet, M., Baert, Jl et al. (2000) Therostasin, a novel clotting Francischetti, I. M., Andersen, J. F. and Ribeiro, J. M. (2002a) Biochemical factor Xa inhibitor from the rhynchobdellid leech, Theromyzon tes- and functional characterization of recombinant Rhodnius prolixus sulatum. Journal of Biological Chemistry, 275 (42), 32701–32707. platelet aggregation inhibitor 1 as a novel lipocalin with high ............................................................................................... .................................................................. 15 Review article Bioscience Horizons � Volume 10 2017 ............................................................................................... .................................................................. affinity for adenosine diphosphate and other adenine nucleotides. Hawkey, C. (1966) Plasminogen activator in saliva of the vampire bat Biochemistry, 41, 3810–3818. Desmodus rotundus. Nature, 211 (5047), 434–435. Francischetti, I. M. B., Assumpção, T. C. F., Ma, D. et al. (2013) The Hawkey, C. (1967) Inhibitor of platelet aggregation present in saliva of ‘Vampirome’: Transcriptome and proteome analysis of the principal the vampire bat Desmodus rotundus. British Journal of Haematology, and accessory submaxillary glands of the vampire bat Desmodus 13 (6), 1014–1020. rotundus,avector of human rabies. Journal of Proteomics,82, 288–319. Hawkins, R. I. (1966) Factors affecting blood clotting from salivary Francischetti, I. M. B., Ribeiro, J. M., Champagne, D. et al. (2000) glands and crop of Glossina austeni. Nature (London), 212, 738. Purification, cloning, expression, and mechanism of action of a Hawkins, R. I. and Hellman, K. (1966) Investigations on a plasminogen novel platelet aggregation inhibitor from the salivary gland of the activator in Two-blood-Suckers, Rhodnius prolixus Stål and Hirudo bloodsucking bug, Rhodnius prolixus. Journal of Biological medicinalis;. British Journal of Haematology, 12, 86. Chemistry, 275, 12639–12650. Haycraft, J. B. (1884) On the action of a secretion obtained from the Francischetti, I. M. B., Sa-Nunes, A., Mans, B. J. et al. (2009) The role of medicinal leech on the coagulation of the blood. Proceedings of the saliva in tick feeding. Frontiers in Bioscience (Landmark Edition), 14, Royal Society of London, 36, 478–487. 2051–2088. Hellenius, W. J. and Ruben, J. A. (2004) The Evolution of endothermy in Francischetti, I. M. B., Valenzuela, J. G., Andersen, J. F. et al. (2002b) terrestrial vertebrates: Who? When? Why? in Physiological and Ixolaris, a novel recombinant tissue factor pathway inhibitor (TFPI) Biochemical Zoology, 77, pp. 1019–1042. from the salivary gland of the tick, Ixodes scapularis: identification of factor X and factor Xa as scaffolds for the inhibition of factor VIIa/ Hellmann, K. and Hawkins, R. I. (1964) Anticoagulant and fibrinolytic tissue factor complex. Blood, 99 (10), 3602–3612. activities from Rhodnius Prolixus Stal. Nature, 201, 1008–1009. Kraut, H., Frey, F. K. and Werle, E. (1930) Der Nachweis eines Hellmann, K. and Hawkins, R. I. (1965) Prolixins-S and prolixin-G; two Kreislaufhormons in der Pankreasdrüse. (IV. Mitteilung über dieses anticoagulants from Rhodnius prolixus Stål. Nature, 207 (994), Kreislaufhormon.). Hoppe-Seyler´s Zeitschrift für Physiologische 265–267. Chemie, 189 (3–4), 97–106. Hellmann, K. and Hawkins, R. I. (1966) An antithrombin (maculatin) and Friedrich, T., Kroger, B., Bialojan, S. et al. (1993) A Kazal-type inhibitor a plasminogen activator extractable from the blood-sucking hem- with thrombin specificity from Rhodnius prolixus. Journal of ipteran, Eutriatoma maculatus. British Journal of Haematology,12 Biological Chemistry, 268 (1993), 16216–16222. (4), 376–384. Fu, Z., Zhang, L., Liu, X. et al. (2013) Comparative proteomic analysis of Higgs, G. A., Vane, J. R., Hart, R. J. et al. (1976) Prostaglandins in the sal- the sun- and freeze-dried earthworm Eisenia fetida with differen- iva of cattle tick, Boophilus microplus (Canestrini) (Acarina, Ixodidae). tially thrombolytic activities. Journal of Proteomics, 83, 1–14. Bulletin of Entomological Research, 66, 665–670. Gardell, S. J., Duong, L. T., Diehl, R. E. et al. (1989) Isolation, characteriza- Hildebrandt, J. P. and Lemke, S. (2011) Small bite, large impact–saliva tion, and cDNA cloning of a vampire bat salivary plasminogen acti- and salivary molecules in the medicinal leech, Hirudo medicinalis. vator. The Journal of Biological Chemistry, 264 (30), 17947–17952. Naturwissenschaften, 98, 995–1008. Geng, P., Lin, L., Li, Y. et al. (2014) A novel fibrin(ogen)olytic trypsin-like Hovingh, P. and Linker, A. (1999) Hyaluronidase activity in leeches protease from Chinese oak silkworm (Antheraea pernyi): purification (Hirudinea). Comparative Biochemistry and Physiology. Part B, and characterization. Biochemical and Biophysical Research Biochemistry and Molecular Biology, 124 (3), 319–326. Communications, 445 (1), 64–70. Ibrahim, M. A., Ghazy, A. H., Maharem, T. et al. (2001) Isolation and Greenhall, A. M. and Schutt, W. A. (1996) Diaemus youngi. Mammalian properties of two forms of thrombin inhibitor from the nymphs of Species, 533, 1–7. the camel tick Hyalomma dromedarii (Acari: Ixodidae). Experimental and Applied Acarology, 25 (8), 675–698. Gregson, J. D. (1967) Observations on the movement of fluids in the vicinity of the mouthparts of naturally feeding Dermacentor ander- Isawa, H., Orito, Y., Jingushi, N. et al. (2007) Identification and character- soni Stiles. Parasitology, 57, 1–8. ization of plasma kallikrein-kinin system inhibitors from salivary glands of the blood-sucking insect Triatoma infestans. The FEBS Gulba, D. C., Praus, M. and Witt, W. (1995) DSPA alpha—Properties of Journal, 274 (16), 4271–4286. the plasminogen activators of the vampire bat Desmodus rotundus. Fibrinolysis, 9 (Suppl 1), 91–96. Ishimaru, Y., Gomez, E. A., Zhang, F. et al. (2012) Dimiconin, a novel coagulation inhibitor from the kissing bug, Triatoma dimidiata,a Hajnická, V., Kocáková, P., Sláviková, M. et al. (2001) Anti-interleukin-8 vector of Chagas disease. Journal of Experimental Biology, 215 (Pt activity of tick salivary gland extracts. Parasite Immunology, 23 (9), 20), 3597–3602. 483–489. Jacobi, Y. (1904) Über Hirudin. Deutsche Medizinische Wochenschrift, 30, Hall, J. E. (2011) Guyton and Hall’s Textbook of Medical Physiology, 12th 1786–1787. edn, Sauders Elsevier, Philadelphia, PA, USA, pp. 451–460. ............................................................................................... .................................................................. 16 Bioscience Horizons � Volume 10 2017 Review article ............................................................................................... .................................................................. Jacobs, J. W., Cupp, E. W., Sardana, M. et al. (1990) Isolation and charac- Li, G., Wang, K. Y., Li, D. et al. (2012) Cloning, expression and character- terization of a coagulation factor Xa inhibitor from black fly salivary ization of a gene from earthworm Eisenia fetida encoding a blood- glands. Thrombosis and Haemostasis, 64 (2), 235–238. clot dissolving protein. PLoS One, 7 (12), e53110. Jiang, Sy., Jiao, J., Zhang, Tt. et al. (2013) Pharmacokinetics study of Liao, M., Zhou, J., Gong, H. et al. (2009) Hemalin, a thrombin inhibitor iso- recombinant hirudin in the plasma of rats using chromogenic lated from a midgut cDNA library from the hard tick Haemaphysalis substrate, ELISA, and radioisotope assays. PLoS One,8(6), longicornis. Journal of Insect Physiology, 55 (2), 164–173. e64336. Liberatore, G. T., Samson, A., Bladin, C. et al. (2003) Vampire bat salivary Jones, G., Teeling, E. C. and Rossiter, S. J. (2013) From the ultrasonic to plasminogen activator (desmoteplase): a unique fibrinolytic the infrared: molecular evolution and the sensory biology of bats. enzyme that does not promote neurodegeneration. Stroke, 34 (2), Frontiers in Physiology, 4, 117. 537–543. Jongejan, F. and Uilenberg, G. (2004) The global importance of ticks. Linker, A., Hoffman, P. and Meyer, K. (1957) The hyaluronidase of the Parasitology, 129, S3–14. leech: an endoglucuronidase. Nature, 180 (4590), 810–811. Juckett, G. (2013) Arthropod bites. American Family Physician, 88 (12), Lloyd, C. M. and Walker, A. R. (1995) Salivary glands and saliva of 841–847. Amblyomma variegatum ticks: comparison of immatures and adults in relation to the pathogenesis of dermatophilosis. Veterinary Kazimírová, M. and Štibrániová, I. (2013) Tick salivary compounds: their Parasitology, 59, 59–67. role in modulation of host defences and pathogen transmission. Frontiers in Cellular and Infection Microbiology, 3, 43. Low, D. H. W., Sunagar, K., Undheim, E. A. B. et al. (2013) Dracula’s chil- dren: Molecular evolution of vampire bat venom. Journal of Koh, C. Y., Kazimirova, M., Trimnell, A. et al. (2007) Variegin, a novel fast Proteomics, 89, 95–111. and tight binding thrombin inhibitor from the tropical bont tick. Journal of Biological Chemistry, 282 (40), 29101–29113. Ma, D., Mizurini, D. M., Assumpção, T. C. F. et al. (2013) Desmolaris, a novel factor XIa anticoagulant from the salivary gland of the vam- Koh, C. Y. and Kini, R. M. (2008) Anticoagulants from hematophagous pire bat (Desmodus rotundus) inhibits inflammation and thrombosis animals. Expert Review of Hematology, 1 (2), 135–139. in vivo. Blood, 122 (25), 4094–4106. Kong, Y., Shao, Y., Chen, H. et al. (2013) A novel factor Xa-inhibiting Macedo-Ribeiro, S., Almeida, C., Calisto, B. M. et al. (2008) Isolation, peptide from Centipedes Venom. International Journal of Peptides cloning and structural characterisation of boophilin, a multifunc- Research and Therapeutics, 19, 303–311. tional Kunitz-type proteinase inhibitor from the cattle tick. PLoS One, 3 (2), e1624. Krätzschmar, J., Haendler, B., Langer, G. et al. (1991) The plasminogen activator family from the salivary gland of the vampire bat Maina, A. N., Jiang, J., Omulo, S. A. et al. (2014) High prevalence of Desmodus rotundus: cloning and expression. Gene, 105 (2), Rickettsia Africae variants in Amblyomma variegatum.Ticks from 229–237. domestic mammals in rural western Kenya: implications for human health. Vector-Bourne and Zoonotic Diseases (Larchmont, N.Y.),14(10), Lai, R., Takeuchi, H., Jonczy, J. et al. (2004) A thrombin inhibitor from 693–702. the ixodid tick, Amblyomma hebraeum. Gene, 342 (2), 243–249. Mans, B. J., Gasper, A. R. M. D., Louw, A. I. et al. (1998a) Apyrase activity Laurin M. and Reisz R. R. (2011) Synapsida. Mammals and their extinct and platelet aggregation inhibitors in the tick Ornithodoros savignyi relatives, accessed at: http://tolweb.org/Synapsida/14845 in Tree of (Acari: Argasidae). Experimental and Applied Acarology, 22, 353–366. Life Web Project [Last updated 14/8/11]. (9 March 2015). Mans, B. J., Gasper, A. R. M. D., Louw, A. I. et al. (1998b) Purification and Law, J. H., Ribeiro, J. M. C. and Wells, M. A. (1992) Biochemical insights characterisation of apyrase from the tick, Ornithodoros savignyi. derived from diversity in insects. Annual Review of Biochemistry, 61, Comparative Biochemistry and Physiology. Part B: Biochemistry & 87–111. Molecular Biology, 120, 617–624. Lent, C. M., Fliegner, K. H., Freedman, E. et al. (1988) Ingestive behaviour Mant, M. J. and Parker, K. R. (1981) Two platelet aggregation inhibitors and physiology of the medicinal leech. Journal of Experimental in tsetse (Glossina) saliva with studies of roles of thrombin and cit- Biology, 137, 513–527. rate in in vitro platelet aggregation. British Journal of Haematology, Lerner, E. A., Ribeiro, J. M., Nelson, R. J. et al. (1991) Isolation of maxadi- 48 (4), 601–608. lan, a potent vasodilatory peptide from the salivary glands of the Maraganore, J. M., Chao, B., Joseph, M. L. et al. (1989) Anticoagulant sand fly Lutzomyia longipalpis. Journal of Biological Chemistry, 266 activity of synthetic hirudin peptides. Journal of Biological (17), 11234–11236. Chemistry, 264, 8692–8698. Li, D., Scherfer, C., Korayem, A. M. et al. (2002) Insect hemolymph clot- Markwardt, F. (1957) Die Isolierung und chemische Charakterisierung ting: evidence for interaction between the coagulation system and des Hirudins. Hoppe-Seyler’s Zeitschrift für Physiologische Chemie, the prophenoloxidase activating cascade. Insect Biochemistry and 308, 147–156. Molecular Biology, 32 (8), 919–928. ............................................................................................... .................................................................. 17 Review article Bioscience Horizons � Volume 10 2017 ............................................................................................... .................................................................. Marshall, C. G. and Lent, C. M. (1988) Excitability and secretory activity Biochemica et Biophysica Acta (BBA) – Protein Structure and in the salivary gland cells of jawed leeches (Hirudinea: Molecular Enzymology, 1482 (1–2), 92–101. Gnathobdellida). Journal of Experimental Biology, 137, 89–105. Palta, S., Saroa, R. and Palta, A. (2014) Overview of the coagulation sys- Mihara, H., Sumi, H. and Yoneta, T. et al. (1991) A novel fibrinolytic tem. Indian Journal of anaesthesia, 58 (5), 515–523. enzyme extracted from the earthworm, Lumbricus rubellus. The Park, S. Y., Kye, K. C., Lee, M. H. et al. (1989) Fibrinolytic activity of the Japanese Journal of Physiology, 41 (3), 461–472. earthworm extract. Thrombosis and Haemostasis, 62, 545–550. Mϋhlhahn, P., Czisch, M., Morenweiser, R. et al. (1994) Structure of leech Parker, K. R. and Mant, M. J. (1979) Effects of tsetse (Glossina morsitans derived tryptase inhibitor (LDTI-C) in solution. FEBS Letters, 355 (3), morsitans Westw.) (Diptera: Glossinidae) salivary gland homogenate 290–296. on coagulation and fibrinolysis. Thrombosis and Haemostasis, 42 (2), Munro, R., Jones, C. P. and Sawyer, R. T. (1991) Calin–a platelet adhesion 743–751. inhibitor from the saliva of the medicinal leech. Blood Coagulation Patel, R., Ispoglou, S. and Apostolakis, S. (2014) Desmoteplase as a and Fibrinolysis: An International Journal in Haemostasis and potential treatment for cerebral ischaemia. Expert Opinion on Thrombosis, 2 (1), 179–184. Investigational Drugs, 23 (6), 865–873. Munshi, Y., Ara, I., Rafique, H. et al. (2008) Leeching in the history–a Peterson, K. J., Cotton, J. A., Gehling, J. G. et al. (2008) The Ediacaran emer- review. Pakistan Journal of Biological Sciences, 11 (13), 1650–1653. gence of bilaterians: congruence between the genetic and the geo- Murphy, K. (2012) Janeway’s Immunobiology, 8th edn, Garland Science, logical fossil records. Philosophical Transactions of the Royal Society of New York, USA, p.7. London. Series B, Biological Sciences, 363 (No. 1496), 1435–1443. Naski, M. C., Fenton, J. W., Maraganore, J. M. et al. (1990) The COOH- Piechowski-Jozwiak, B. and Bogousslavsky, J. (2013) The use of desmo- terminal domain of hirudin. An exosite-directed competitive inhibi- teplase (bat saliva) in the treatment of ischaemia. Expert Opinion on tor of the action of alpha-thrombin on fibrinogen. Journal of Biological Therapy, 13 (3), 447–453. Biological Chemistry, 265, 13484–13489. Ponczek, M. B., Bijak, M. Z. and Nowak, P. Z. (2012) Evolution of throm- Nawarskas, J. J. and Anderson, J. R. (2001) Bivalirudin: a new approach bin and other hemostatic proteases by survey of protochordate, to anticoagulation. Heart Disease (Hagerstown, Md), 3 (2), 131–137. hemichordate, and echinoderm genomes. Journal of Molecular Evolution,74 (5–6), 319–331. Nienaber, J., Gaspar, A. R. and Neitz, A. W. H. (1999) Savignin, a potent thrombin inhibitor isolated from the salivary glands of the tick Qiu, Y., Lee, K. S., Choo, Y. M. et al. (2013) Molecular cloning and antifi- Ornithodoros savignyi (Acari: Argasidae). Experimental Parasitology, brinolytic activity of a serine protease inhibitor from bumblebee 93 (2), 82–91. (Bombus terrestris) venom. Toxicon, 63, 1–6. Noeske-Jungblut, C., Haendler, B., Donner, P. et al. (1995) Triabin, a Ramachandra, R. N. and Wikel, S. K. (1995) Effects of Dermacentor highly potent exosite inhibitor of thrombin. Journal of Biochemical andersoni (Acari: Ixodidae) salivary gland extracts on Bos indicus and Chemistry, 270 (48), 28629–28634. B. taurus lymphocytes and macrophages: in vitro cytokine elabor- ation and lymphocyte blastogenesis. Journal of Medical Noeske-Jungblut, C., Krätzschmar, J., Haendler, B. et al. (1994) An inhibi- Entomology, 32, 338–345. tor of collagen-induced platelet aggregation from the saliva of Triatoma pallidipennis. The Journal of Biological Chemistry, 269 (7), Reverter, D., Vendrell, J., Canals, F. et al. (1998) A carboxypeptidase inhibi- 5050–5053. tor from the medical leech Hirudo medicinalis. Isolation, sequence analysis, cDNA cloning, recombinant expression, and characteriza- Oliveira, D. G., Alvarez-Flores, M. P., Lopes, A. R. et al. (2012) Functional tion. The Journal of Biological Chemistry, 273 (49), 32927–32933. characterisation of vizottin, the first factor Xa inhibitor purified from the leech Haementeria vizottoi. Thrombosis and Haemostasis, Ribeiro, J. M. C. (1995) Blood-feeding arthropods: live syringes or inver- 108 (3), 570–578. tebrate pharmacologists? Infectious Agents and Disease, 4, 143–152. Oren, M., Escande, Ml., Paz, G. et al. (2008) Urochordate histoincompati- Ribeiro, J. M., Evans, P. M., MacSwain, J. L. et al. (1992) Amblyomma ble interactions activate vertebrate-like coagulation system compo- americanum: characterization of salivary prostaglandins E2 and F2 nents. PLoS One, 3 (9), e3123. alpha by RP-HPLC/bioassay and gas chromatography-mass spec- trometry. Experimental Parasitology, 74 (1), 112–116. Paciaroni, M., Medeiros, E. and Bogousslavsky, J. (2009) Desmoteplase. Expert Opinion on Biological Therapy, 9 (6), 773–778. Ribeiro, J. M. C., Modi, G. B., Tesh, R. B. et al. (1989a) Salivary apyrase activity of some old world phlebotomine sand flies. Insect Paesen, G. C., Adams, P. L., Harlos, K. et al. (1999) Tick histamine- Biochemistry, 19 (4), 409–412. binding proteins: isolation cloning and three-dimensional structure. Molecular Cell, 3 (5), 661–671. Ribeiro, J. M., Makoul, G. T., Levine, J. et al. (1985) Antihemostatic, anti- inflammatory, and immunosuppressive properties of the saliva of a Paesen, G. C., Adams, P. L., Nuttall, P. A. et al. (2000) Tick histamine- tick, Ixodes dammini. Journal of Experimental Medicine, 161, binding proteins: lipocalins with a second binding cavity. 332–344. ............................................................................................... .................................................................. 18 Bioscience Horizons � Volume 10 2017 Review article ............................................................................................... .................................................................. Ribeiro, J. M., Makoul, G. T. and Robinson, D. R. (1988) Ixodes dammini: Schleuning, W. D., Alagon, A., Biodol, W. et al. (1992) Plasminogen acti- evidence for salivary prostacyclin secretion. The Journal of vators from the saliva of Desmodus rotundus (common vampire Parasitology, 74 (6), 1068–1069. bat): unique fibrin specificity. Annals of the New York Academy of Sciences, 667, 395–403. Ribeiro, J. M., Marinotti, O. and Gonzales, R. (1990a) A salivary vasodila- tor in the blood-sucking bug, Rhodnius prolixus. British Journal of Schutt, B. (2008) Dark Banquet; Blood and the Curious Lives of Blood- Pharmacology, 101 (4), 932–936. Feeding Creatures, Three Rivers Press, New York, USA, pp. 49–58. Ribeiro, J. M. and Mather, T. N. (1998) Ixodes scapularis: salivary kininase Seemüller, U., Meier, M., Ohlsson, K. et al. (1977) Isolation and charac- activity is a metallo dipeptidyl carboxypeptidase. Experimental terisation of a low molecular weight inhibitor (of chymotrypsin and Parasitology, 89 (2), 213–221. human granulocytic elastase and cathepsin G) from leeches. Hoppe-Seyler’s Zeitschrift für Physiologische Chemie, 358 (9), Ribeiro, J. M., Rossignol, P. A. and Spielman, A. (1986) Blood-finding 1105–1107. strategy of a capillary-feeding sandfly, Lutzomyia longipalpis. Comparative Biochemistry and Physiology. A, Comparative Sharma, A., Sonah, H., Deshmukh, R. K. et al. (2011) Cloning of fibrino- Physiology, 83 (4), 683–686. lytic protease-0 (Efp-0) gene from diverse earthworm individuals. Indian Journal of Biotechnology, 10 (3), 270–273. Ribeiro, J. M., Sarkis, J. J., Rossignol, P. A. et al. (1984) Salivary apyrase of Aedes aegypti: characterization and secretory fate. Comparative Snider, G. L., Stone, P. J., Lucey, E. C. et al. (1985) Eglin-c, a polypeptide Biochemistry and Physiology. B, Biochemistry and Molecular Biology, derived from the medicinal leech, prevents human neutrophil 79 (1), 81–86. elastase-induced emphysema and bronchial secretory cell metapla- sia in the hamster. The American Review of Respiratory Disease, 132 Ribeiro, J. M., Schneider, M. and Guimarães, J. A. (1995) Purification and (6), 1155–1161. characterization of prolixin S (nitrophorin 2), the salivary anticoagu- lant of the blood-sucking bug Rhodnius prolixus. The Biochemical Söllner, C., Mentele, R., Eckerskorn, C. et al. (1994) Isolation and charac- Journal, 308 (Pt 1), 243–249. terization of hirustasin, an antistasin-type serine-proteinase inhibi- tor from the medical leech Hirudo medicinalis. European Journal of Ribeiro, J. M., Vachereau, A., Modi, G. B. et al. (1989b) A novel vasodila- Biochemistry, 219 (3), 937–943. tory peptide from the salivary glands of the sand fly Lutzomyia longipalpis. Science (New York, N.Y.), 243 (4888), 212–214. Sommerhoff, C. P., Söllner, C., Mentele, R. et al. (1994) A Kazal-type inhibitor of human mast cell tryptase: isolation from the medical Ribeiro, J. M., Vaughan, J. A. and Azad, A. F. (1990b) Characterization of leech Hirudo medicinalis, characterization, and sequence analysis. the salivary apyrase activity of three rodent flea species. Biological Chemistry Hoppe Seyler, 375, 685–694. Comparative Biochemistry and Physiology. B, Comparative Biochemistry, 95 (2), 215–219. Stark, K. R. and James, A. A. (1995) A factor Xa-directed anticoagulant from the salivary glands of the yellow fever mosquito Aedes aegypti. Rigbi, M., Levy, H., Iraqi, F. et al. (1987) The saliva of the medicinal Experimental Parasitology, 81 (3), 321–331. leech Hirudo medicinalis–I. Biochemical characterization of the high molecular weight fraction. Comparative Biochemistry and Steranka, L. R., Manning, D. C., DeHaas, C. J. et al. (1988) Bradykinin as a Physiology. B, Comparative Biochemistry,87 (3),567–573. pain mediator: receptors are localized to sensory neurons, and antagonists have analgesic actions. Proceedings of the National Rigbi, M., Orevi, M. and Eldor, A. (1996) Platelet aggregation and coagu- Academy of Sciences of USA, 85, 3245–3249. lation inhibitors in leech saliva and their roles in leech therapy. Seminars in Thrombosis and Hemostasis, 22, 273–278. Stubbs, M. T., Morenweiser, R., Stürzebecher, J. et al. (1997) The three- dimensional structure of recombinant leech-derived tryptase Salzet, M., Chopin, V., Baert, Jl. et al. (2000) Theromin, a novel leech inhibitor in complex with trypsin. Implications for the structure of thrombin inhibitor. Journal of Biological Chemistry, 275 (40), human mast cell tryptase and its inhibition. Journal of Biological 30774–30780. Chemistry, 272, 19931–19937. Sanetra, M., Begemann, G., Becker, Mb. et al. (2005) Conservation and Su, J. B. (2014) Different cross-talk sites between the renin−angiotensin co-option in developmental programmes: the importance of hom- and the kallikrein−kinin systems. Journal of the Renin-Angiotensin- ology relationships. Frontiers in Zoology, 2, 15. Aldosterone System, 15 (4), 319–328. Santos, A., Ribeiro, J. M., Lehane, M. J. et al. (2007) The sialotranscriptome of Swadesh, J. K., Huang, I. Y. and Budzynski, A. Z. (1990) Purification and the blood-sucking bug Triatoma brasiliensis (Hemiptera, Triatominae). characterization of hementin, a fibrinogenolytic protease from the Insect Biochemistry and Molecular Biology, 37 (7), 702–712. leech Haementeria ghilianii. Journal of Chromotography, 502 (2), 359–369. Sauer, J. R. (1977) Acarine salivary glands—Physiological relationships. Journal of Medical Entomology, 14, 1–9. Tasiemski, A., Vandenbulcke, F., Mitta, G. et al. (2004) Molecular charac- terization of two novel antibacterial peptides inducible upon bac- Schleuning, W. D. (2001) Vampire bat plasminogen activator DSPA- terial challenge in an annelid, the leech Theromyzon tessulatum. The alpha-1 (desmoteplase): a thrombolytic drug optimized by natural Journal of Biological Chemistry, 279 (30), 30973–30982. selection. Haemostasis,31 (3–6), 118–122. ............................................................................................... .................................................................. 19 Review article Bioscience Horizons � Volume 10 2017 ............................................................................................... .................................................................. Tatchell R. J. and Binnington K. C. (1973) An active constituent of the Wan, H., Lee, K. S., Kim, B. Y. et al. (2013) A spider-derived Kunitz-type saliva of the cattle tick, Boophilus microplus; Proceedings of the 3rd serine protease inhibitor that acts as a plasmin inhibitor and an International Conference of Acarology; 1971, 745. elastase inhibitor. PLoS One, 8 (1), e53343. Tellgren-Roth, A., Dittmar, K., Massey, S. E. et al. (2009) Keeping the Wang, X., Coons, L. B., Taylor, D. B et al. (1996) Variabilin, a novel RGD- blood flowing-plasminogen activator genes and feeding behavior containing antagonist of glycoprotein IIb-IIIa and platelet aggrega- in vampire bats. Naturwissenschaften, 96 (1), 39–47. tion inhibitor from the hard tick Dermacentor variabilis. The Journal of Biological Chemistry, 271 (30), 17785–17790. Theopold, U., Schmidt, O., Söderhall, K. et al. (2004) Coagulation in arthropods: defence, wound closure and healing. Trends in Waxman, L. and Connolly, T. M. (1993) Isolation of an inhibitor selective Immunology, 25 (6), 289–294. for collagen-stimulated platelet aggregation from the soft tick Ornithodoros moubata. The Journal of Biological Chemistry, 268 (8), Tuszynski, G. P., Gasic, T. B. and Gasic, G. J. (1987) Isolation and charac- 5445–5449. terization of antistasin. An inhibitor of metastasis and coagulation. Journal of Biological Chemistry, 262 (20), 9718–9723. Waxman, L., Smith, D. E., Arcuri, K. E. et al. (1990) Tick anticoagulant peptide (TAP) is a novel inhibitor of blood coagulation factor Xa. Vachereau, A. and Ribeiro, J. M. C. (1989) Immunoreactivity of salivary Science, 248 (4955), 593–596. gland apyrase of Aedes aegypti with antibodies against a similar hydrolase present in the pancreas of mammals. Insect Biochemistry, Wikel, S. K. (1982) Histamine content of tick attachment sites and the 19 (6), 527–534. effect of H1 and H2 histamine antagonists on the expression of resistance. Annals of Tropical Medicine and Parasitology, 76, Valenzuela, J. G. (2004) Exploring tick saliva: from biochemistry to ‘sia- 179–185. lomes’ and functional genomics. Parasitology, 129 (Suppl), S83–S94. Wimsatt, W. A. (1959) Portrait of a vampire. Ward’s Natural Science Valenzuela, J. G., Francischetti, I. M. B., Pham, V. M. et al. (2002) Bulletin, 32, 35. Exploring the sialome of the tick Ixodes scapularis. Journal of Experimental Biology, 205, 2843–2864. You, W. K., Sohn, Y. D., Kim, K. Y. et al. (2004) Purification and molecular cloning of a novel serine protease from the centipede, Scolopendra van de Locht, A., Stubbs, M. T. and Bode, W. (1996) The ornithodorin- subspinipes mutilans. Insect Biochemistry and Molecular Biology,34 thrombin crystal structure, a key to the TAP enigma? The EMBO (3), 239–250. Journal, 15 (22), 6011–6017. Zaidi, S. M., Jameel, S. S., Zaman, F. et al. (2011) A systematic overview Viljoen, G. J., Bezuidenhout, J. D., Oberem, P. T. et al. (1986) Isolation of of the medicinal importance of Sanguivorous leeches. Alternative a neurotoxin from the salivary glands of female Rhipicephalus ever- Medicine Review, 16 (1), 59–65. tsi evertsi. Journal of Parasitology, 72, 865–874. ............................................................................................... ..................................................................

Journal

Bioscience HorizonsOxford University Press

Published: Feb 14, 2017

There are no references for this article.