Fish & Shellfish Immunology 34 (2013) 968e972
Contents lists available at SciVerse ScienceDirect
Fish & Shellfish Immunology
journal homepage: www.elsevier.com/locate/fsi
Molecular mechanisms of the shrimp clotting system
Mary Beth B. Maningas a, b, Hidehiro Kondo a, Ikuo Hirono a, *
a
Graduate School of Marine Science and Technology, Tokyo University of Marine Science and Technology, Konan 4-5-7, Minato-ku, Tokyo 108-8477, Japan
Department of Biological Sciences, College of Science, Molecular Biology and Biotechnology Laboratory, Research Center for the Natural and Applied Sciences,
University of Santo Tomas, Espana, 1015 Manila, Philippines
b
a r t i c l e i n f o
a b s t r a c t
Article history:
Received 10 May 2012
Received in revised form
3 September 2012
Accepted 18 September 2012
Available online 5 October 2012
Shrimp, like other invertebrates, relies solely on its innate immune system, to combat invading pathogens. The invertebrate immune system has ancient origins that involve cellular and humoral responses.
The clotting system of the humoral immune response is the first line of defense against pathogens and
also serves to prevent blood loss during injury and wound healing. Tranglutaminase and clotting protein
are molecules involved in the blood clotting system of crayfish and shrimp. Studies have shown that the
shrimp clotting system is linked with the activation of antimicrobial peptides, similar to that of the
horseshoe crab. Unlike the horseshoe crab and crayfish blood coagulation which are well studied
systems, blood clotting in shrimp remains poorly understood. Here we review the shrimp clotting system
and its involvement in innate immunity.
Ó 2012 Elsevier Ltd. All rights reserved.
Keywords:
Shrimp
Clotting system
Clotting protein
Transglutaminase
1. Introduction
Shrimp is one of the most important food products in the
international market, accounting for almost 20% of the overall
aquaculture commodities in trade [1]. Shrimp aquaculture has been
an indispensable source of revenue in the inter-tropical countries of
Southeast Asia and other developing countries. In the last two
decades, the shrimp aquaculture industry has grown rapidly and is
now generating over billion dollars a year in trade and employs
over 1 million people globally. Unfortunately, the industry has been
plagued by outbreaks of bacterial or viral diseases. Completion of
sequencing of the shrimp genome has been hindered by its
redundancy and high AT content and the lack of stable cell lines
renders it difficult to perform gene functional studies [2e6]. Hence,
the shrimp immune system remains poorly understood.
The shrimp immune system like other invertebrates lacks an
adaptive immune system and relies solely on its innate immunity
against invading pathogens. Innate immunity is an ancient
protective mechanism that appeared early in the evolution of
metazoans and is divided into humoral and cellular responses [7,8].
The humoral response includes clotting, melanization of damaged
tissues and pathogens by the prophenoloxidase oxidase system
(PPO), activation of antimicrobial peptides (AMPs) and the release
of stress responsive proteins and molecules believed to function in
* Corresponding author. Tel./fax: þ81 35463 0689.
E-mail address: [email protected] (I. Hirono).
1050-4648/$ e see front matter Ó 2012 Elsevier Ltd. All rights reserved.
http://dx.doi.org/10.1016/j.fsi.2012.09.018
opsonization and iron sequestration. Perhaps among the four
components, the clotting system is the most complex as it has been
implicated as an immune effector in both the vertebrate and the
invertebrate system [8]. The clotting system in vertebrates and
other invertebrates including insects [9e13], and chelicerates [14e
17], and other crustacean like crayfish [18e20] has been reviewed.
To date, there is no specific review yet in shrimp blood coagulation.
This paper aims to present the mechanism of shrimp clotting
system and its involvement in immunity.
2. Molecular mechanism of clotting systems
Clotting systems are essential components of the immune
system of almost all metazoans. The clotting system of the humoral
immune response is the first line of defense and an integral part of
the overall invertebrate immune system. It is also important in the
prevention of blood loss during injury and wound healing.
Understanding the molecular mechanism of the shrimp clotting
system, might help to clarify the origin of vertebrate immunity and
might pave the way for a more unified concept on innate immunity
among the vertebrate and invertebrate systems. The clotting
system in invertebrates was first described in horseshoe crab,
a chelicerate, and later in crayfish, a crustacean, and insects as
model organisms. Different mechanisms have evolved in each of
these groups as well as vertebrates [10,11,15]. The blood coagulation
processes in chelicerates and vertebrates differ but both involve
a proteolytic cascade in order to activate clotting enzymes [14]. A
calcium-dependent TGase known as factor XIIIa is involved in the
M.B.B. Maningas et al. / Fish & Shellfish Immunology 34 (2013) 968e972
final steps of the vertebrate blood clotting system, in which the
TGase stabilizes the blood clot by cross-linking aggregates of fibrin
[14]. In the horseshoe crab, TGase from hemocytes promotes the
cross-linking of coagulin, with hemocyte cell surface proteins [14].
Crustaceans appear to utilize the simplest coagulation process,
involving cross-linking aggregates of clotting proteins (CPs) that is
catalyzed by a Calcium-dependent transglutaminase (TGase),
which is released from the hemocytes under foreign particle
stimulus or tissue damage [21]. The TGase, under this process,
forms g-glutamyl- ε-lysine crosslinks between glutamine and
lysine residues of the CP [21,22]. TGase was cloned and localized in
crayfish [23], and was found to be involved in the blood coagulation
of tiger shrimp [24e26].
Clearly, the shrimp clotting system involves TGase and clotting
proteins and seems to follow the cascade reported in crayfish
[19,20]. Recently, expression of antimicrobial peptides (AMPs);
lysozyme and crustine were significantly down-regulated in TGase
depleted shrimp [27] which is similar to the earlier report in
horseshoe crab where the clotting system is linked with the activation of AMPs [16]. Inhibition of lysozyme and crustine expression
indicates that the shrimp clotting system is linked to the expression
of anti-microbial peptide genes. This finding provides evidence that
genes involved in the clotting system affect the expression of
antimicrobial peptides (AMPs) in shrimp. The shrimp blood coagulation system may be linked with the production of AMPs similar
with the horseshoe crab coagulation model rather than the crayfish. The horseshoe crab model speculates that the cascade
provides a dual action: clotting and production of antimicrobial
substances against invading agents [14e16]. Furthermore, the NH2
terminal portions of factor B and of proclotting enzyme contain clip
domains identical to that of big defensin, an AMP in horseshoe crab
[16]. The exact mechanism on the activation of AMPs in chelicerates
and crustaceans remains to be elucidated.
The blood coagulation processes in the chelicerate (horseshoe
crab) and the crustacean (crayfish) models are different. The
horseshoe crab utilizes a proteolytic cascade similar to that of
human. The proteolytic cleavage in horseshoe crab not only activates the downstream proteins but also leads to the release of small
cleavage products which act as anti-microbial peptides [14,15].
Clots from horseshoe crabs bind bacteria and kill them in collaboration with a plasma factor [16]. In crayfish on the other hand,
hemolymph is clotted primarily by one system. This system
consists of a clotting protein and a TGase. The TGase appears to be
released from the hemocytes or other tissues [23e26] and is
apparently the concluding phase of the cascade and this polymerization step has been conserved through evolution [19,20].
969
In insects, the prophenoloxidase cascade appears to be involved
in blood coagulation system [9e13]. Studies showed that expression of AMPs was inhibited in TGase and CP depleted shrimp
[27,28], which suggests that there is a link between the clotting
system activation of AMPS. In a review, it was suggested that the
clotting proteins of crayfish and shrimp do not share any structural
similarities with mammalian fibrinogen or horseshoe crab coagulogen [20], this suggest that the crustacean clotting proteins
constitute a separate group of clotting factors. Recently, hemolymph clotting in insects localized the immune effectors in the
vicinity of a breach of the cuticle and restricted the spread of
invasive particles across the hemocoel [11]. A quite old paper in
crayfish revealed that the clotting process and the prophenoloxydase system seemed to be triggered by endogenous proteases [29].
In another effort to elucidate the clotting process in crayfish, alpha2-macroglobulin was tested as substrate for TGase [30]. Alpha-2macroglobulins belongs to a group of high-molecular mass, broad
spectrum proteinase inhibitor and was shown to be capable of
crosslinking with CP by TGase to form clots [30]. This process might
prevent microorganism to enter a wound since these Alpha-2macroglobulins can inhibit their proteinases [30]. This review
paper proposed a blood clotting cascade in shrimp (Fig. 1) in which
the clotting system is linked with the activation of AMPS [34,35,39].
Whether these findings, present a unifying concept on blood clotting and its association with AMPs among invertebrates needs
further investigation.
3. TGase: multifunctional properties
TGase has been documented to be involved in blood coagulation, which is a conserved defense mechanism among invertebrates. TGase was localized in the hemocytes of decapod
crustaceans in the early 1990s [31]. Later on, it was cloned and
localized in the hemocytes of crayfish [23], and found to be
involved in cell proliferation [32] and immune response against
bacterial and viral infection [28]. Another type of TGase was cloned
but was also found to be involved in the tiger shrimp [25] and
chinese shrimp [26] blood coagulation system. Recently,
biochemical assays were utilized to establish the involvement of
TGase in coagulating plasma clotting protein [33] and its participation in bacterial (Vibrio alginolyticus) infection [34]. Depletion of
TGase in shrimp by RNAi efficiently blocked blood coagulation, at
the phenotypic level, highlighting its essential function in the blood
clotting system of shrimp [28].
However, several studies have implicated TGase in different
immune mechanisms in invertebrates. TGase is involved in
Fig. 1. Proposed clotting system in shrimp and its association with the activation of AMPs.
970
M.B.B. Maningas et al. / Fish & Shellfish Immunology 34 (2013) 968e972
biochemical process particularly in post-translational protein
remodelling, cell proliferation and is widely distributed in tissues
and body fluid of animals [35,36]. Recently, TGase was not only
implicated in blood coagulation but also in pathogen entrapment
both in human and in drosophila [37]. In vivo gene silencing of
TGase in shrimp significantly increased mortality following bacterial (Vibrio peanecida) and viral infection (white spot virus) [28]. In
the Pacific white shrimp (Litopenaeus vannamei), TGase activity
declined after infection with Taura Syndrome Virus (TSV), which
also led to poor hemolymph coagulation [38], while a bacterial
challenge using Vibrio harveyi caused the up-regulation of TGase
gene expression [39]. In drosophila, reduced TGase activity after
sepsis injury significantly increased larvae mortality [37]. In a study
using RNAi and microarray technologies, the absence of TGase in
shrimp, significantly down-regulated the expression of lysozyme
and crustins, which are both antimicrobial peptides believed to be
involved in combating microbial pathogens [27]. The down regulation of crustine and lysozyme were corroborated by a significant
decrease in the total bacterial count. Moreover, TGase- depleted
shrimp were found to have lower total hemocyte counts, indicating
the possible role of TGase in hemocyte homeostasis. In crayfish,
blocking of TGase using hematopoietic cell lines (HPT) promoted
cell migration and prevented cell differentiation, underscoring its
functional involvement in hematopoiesis [32,40]. Cell surface
activity was recently found to be high in the center of the HPT and
to be low or absent in migrating cells, suggesting that TGase activity
is involved in the crosslinking of extracellular matrix (ECM) [40].
This suggestion is supported by a recent report that Drosophila has
a TGase-dependent protein crosslinking which plays a key role in
cuticle morphogenesis and sclerotization [41]. All together, these
findings indicate that, crustacean TGases, besides being involved in
clotting, also play a crucial role in the regulation of immune genes
and hematopoiesis. TGase is the only clotting factor that has
homologues across different species and is therefore, the best
conserved protein in the coagulation system [8,42,43]. Although
the functions of TGase are becoming better understood, it is still
unclear how it is released from the hemocytes in the presence of
plasma clotting proteins.
4. Functions of clotting proteins
The first crustacean clotting protein (CP) identified to be
responsible for clot formation was reported in crayfish [21]. Crustacean CPs have been cloned and characterized from several
species, including the fresh water crayfish, Pacificus leniusculus
[22,44], tiger shrimp, Penaeus monodon [45], white shrimp,
L. vannamei [46,47] and pink shrimp, Farnfantepenaeus paulensis
[48]. This CP was found to be a very high-density lipoprotein
(VHDL) purified from the hemolymph of crayfish [49] and in white
shrimp it was proposed to have a role in lipid transport but this has
not been tested in vivo [50]. Analysis of different CP N-terminal
amino acid sequence exhibits a conserved region with 60e80%
similarity across different crustaceans [44]. However, other CP
regions are highly variable and this might explain why other
crustacean CPs are unrelated to the CPs found in horseshoe crab,
vertebrates and insects, which are also unrelated to one another [9].
Moreover, studies on hemolymph clotting revealed a high level of
variation of clot components between different arthropod classes
and even within them [9,10].
Gene silencing of CP in vivo rendered the kuruma shrimps very
susceptible to bacterial (Vibrio penaecida) and viral (WSSV) infection
[28]. Moreover, depletion of CP by gene silencing even without
a challenge triggered a dramatic increase in mortality (our unpublished data), highlighting its vital role in shrimp survival. Exposure of
shrimp to trauma such as eye stalk- and europod-ablation increased
CP concentration to compensate for lost body fluids, underscoring its
role in shrimp hemostasis [50]. These results are congruent with an
earlier study that a VHDL/CP in crayfish has multiple physiological
functions and is crucial to the animals’ defense mechanism [49e51].
All these findings pointed the unequivocal role of CP in blood coagulation as well as in shrimp hemostasis.
5. Association of antimicrobial peptides and clotting systems
Antimicrobial peptides (AMPS) and clotting system are key
components of the crustacean innate immunity in particular the
humoral system. AMPs are ubiquitously found in all kingdoms from
bacteria to mammals and are naturally known as natural antibiotics
because of their rapid and efficient antimicrobial effects against
invading pathogens [52]. Evidence showed that AMPs have multifunctional diverse biological roles [53e56]. Several reviews have
been published in crustacean AMPs [57e60], in particular in
penaeid shrimp [61,62]. Evidence showed that AMPs in shrimp are
involved in host defense and survival [63,64] and that they are upregulated after both WSSV and V. harveyi challenge [65].
In the well studied horseshoe crab, evidence pointed out that
blood coagulation has been linked to the release of AMPs [14e17].
In TGase depleted shrimp, expression of two AMPS; crustine and
lysozyme was significantly down-regulated, suggesting an interaction between the blood coagulation system and AMPs expression [27]. In horseshoe crab, the amino- terminal L chains of factor B
and pro-clotting enzyme contain a small compact domain with
three disulfide bonds called the “clip domain theory” [17]. A similar
“clip domain” has also been reported in the NH2-terminal proenzyme regions of Drosophila-derived serine proteases [66]. The
folding pattern of the three disulfide bridges located in the “clip
domain” is similar to that of “big defensin”, which was identified as
an antimicrobial peptide in Tachypleus tridentatus hemocytes [16].
Moreover, the COOH-terminal end of the “clip domain” in proclotting enzyme constitutes a hinge region susceptible to proteolysis, in the same way as defensin, and might be released during the
activations of serine protease zymogens, to act as an antimicrobial
substance. On the other hand, the “clip domain” from the prophenoloxidase activated serine protease of fresh water crayfish has an
antimicrobial activity similar to that of human b-defensin [67].
These provides evidence that the clotting cascade could also
produce antimicrobial agents, and thus provide a dual action clotting and killing system against invaders similar to insects [10,68].
Hemolymph coagulation in decapods is a powerful reaction that
contributes not just in preventing blood loss but also in immobilizing and inactivating microorganisms by entrapping them
together with immune effectors [57]. In humans, the coagulation
system plays an important role in host/pathogen interactions and
host responses to infections [69] and was affected during inflammation [70]. Taken all together, studies revealed that molecules
involved in clotting system might also be involved in some immune
defense and processes.
6. Conclusions
The shrimp clotting system like other invertebrates is an
essential part of the shrimp biodefense system. There is evidence
that the absence of genes involved in the shrimp blood clotting
system impairs the expression of AMPs. This is also the case in the
downstream of the blood coagulation cascade in horseshoe crab,
where blood clotting was linked with the activation of AMPs.
Whether, the blood coagulation cascade of shrimp represents
another or an intermediate system, or supports a unified model in
crustaceans based on the horseshoe crab system remains to be
determined.
M.B.B. Maningas et al. / Fish & Shellfish Immunology 34 (2013) 968e972
References
[1] Fao: state of world aquaculture. Rome: FAO Fisheries Department; 2006.
[2] Huang SW, Lin YY, You EM, Liu TT, Shu HY, Wu KM, et al. Fosmid library end
sequencing reveals a rarely known genome structure of marine shrimp. BMC
Genomics 2011;12:242e61.
[3] Koyama T, Asakawa S, Katagiri T, Shimizu A, Fagutao FF, Mavichak R, et al.
Hyper-expansion of large DNA segments in the genome of kuruma shrimp,
Marsupenaeus japonicus. BMC Genomics 2010;1:141e52.
[4] Chow S, Dougherty WJ, Sandifer PA. Meiotic chromosome complements and
nuclear DNA contents of four species of shrimps of the genus Penaeus.
J Crustacean Biol 1990;10:29e36.
[5] Bachmann K, Rheinsmith EL. Nuclear DNA amounts in pacific Crustacea.
Chromosoma 1973;43:225e36.
[6] Maneeruttanarungroj C, Pongsomboon S, Wuthisuthimethavee S, Klinbunga S,
Wilson KJ, Swan J, et al. Development of polymorphic expressed sequence tagderived microsatellites for the extension of the genetic linkage map of the
black tiger shrimp (Penaeus monodon). Anim Genet 2006;37:363e8.
[7] Kimbrell DA, Beutler B. The evolution and genetics of innate immunity. Nat
Rev Genet 2001;2:256e67.
[8] Loof TG, Schmidt O, Herwald H, Theopold U. Coagulation systems of invertebrates and vertebrates and their roles in innate immunity: the same side of
two coins? J Innate Immun 2011;3:34e40.
[9] Theopold U, Li S, Fabbri M, Scherfer, Schmidt. The coagulation of insect
hemolymph. Cell Mol Life Sci 2002;59:263e372.
[10] Theopold U, Schmidt O, Söderhäll K, Dushay MS. Coagulation in arthropods:
defence, wound closure and healing. Trends Immunol 2004;25:289e94.
[11] Haine ER, Rolff J, Siva-Jothy MT. Functional consequences of blood clotting in
insects. Dev Comp Immunol 2007;31:456e64.
[12] Dushay M. Insect hemolymph clotting. Cell Mol Life Sci 2009;66:2643e50.
[13] Li D, Schefer C, Korayem AM, Zhao Z, Schmidt O, Theopold U. Insect hemolymph clotting: evidence for interaction between the coagulation system and
the prohenoloxidase activating cascade. Insect Biochem Mol Biol 2002;26:
177e84.
[14] Iwanaga S. The molecular basis of innate immunity in the horseshoe crab. Curr
Opin Immunol 2002;14:87e95.
[15] Osaki T, Kabawata S. Structure and function of coagulogen, a clottable protein
in horseshoe crab. Cell Mol Life Sci 2004;61:1257e65.
[16] Iwanaga S, Lee BL. Recent advances in the innate immunity of invertebrate
animals. J Biochem.Mol Biol 2005;38:128e50.
[17] Muta T, Iwanaga S. The role of hemolymph coagulation in innate immunity.
Curr Opin Immunol 1996;8:41e7.
[18] Sritunyalucksana K, Söderhäll K. The proPO and clotting system in crustaceans. Aquaculture 2000;101:53e69.
[19] Lee YS, Söderhäll K. Early events in crustacean innate immunity. Fish Shellfish
Immunol 2002;12:421e37.
[20] Jiravanichpaisal P, Lee BL, Söderhäll K. Cell mediated review in arthropods:
hematopoiesis, coagulation, melanization and opsonization. Immunobiology
2006;211:213e36.
[21] Kopacek P, Hall M, Söderhäll K. Characterization of a clotting protein, isolated
from plasma of the freshwater crayfish Pacifastacus leniusculus. Eur J Biochem
1993;213:519e97.
[22] Hall M, Ruigong W, Antwerpen R, Sottrup-Jensen V, Söderhäll K. The crayfish
plasma clotting protein: a vitellogenin-related protein responsible for clot
formation in crustacean blood. Proc Natl Acad Sci USA 1999;96:1965e70.
[23] Wang R, Liang Z, Hall M, Söderhäll K. A transglutaminase involved in the
coagulation system of the freshwater crayfish, Pacifastacus leniusculus. Tissue
localization and cDNA cloning. Fish Shellfish Immunol 2011;11:623e37.
[24] Huang CC, Sritunyalucksana K, Söderhäll K, Song YL. Molecular cloning and
characterization of tiger shrimp (Penaeus monodon) transglutaminase. Dev
Comp Immunol 2004;28:279e94.
[25] Chen MY, Hu KY, Huang CC, Song YL. A second type of transglutaminase is
involved in shrimp coagulation. Dev Comp Immunol 2005;29:1003e16.
[26] Liu YC, Li FH, Wang B, Dong B, Zhang QL, Luan W, et al. A transglutaminase
from Chinese shrimp (Fenneropenaeus chinensis), full-length cDNA cloning,
tissue localization and expression profile after challenge. Fish Shellfish
Immunol 2007;22:576e88.
[27] Fagutao F, Maningas MBB, Kondo H, Aoki T, Hirono I. Transglutaminase
regulates immune-related genes in shrimp. Fish Shellfish Immunol 2012;
32(5):711e5.
[28] Maningas MBB, Kondo H, Hirono I, Saito-Taki T, Aoki T. Essential function of
transglutaminase and clotting protein in shrimp immunity. Mol Immunol
2008;45:1269e75.
[29] Söderhäll K. Fungal cell wall â-1, 3-glucans induce clotting and phenoloxidase
attachment to foreign surfaces of crayfish hemocyte lysate. Dev Comp
Immunol 1981;5:565e73.
[30] Hall M, Söderhäll K. Crayfish alpha-2-macroglobulin as a substrate to for
TGase. Comp.Biochem.Physiol 1994;108B:65e72.
[31] Martin GG, Hose JE, Omori S, Chong C, Hoodbhoy T, McKrell N. Localization
and roles of transglutaminase in hemolymph coahulation in decapods crustaceans. Comp Biochem Physiol 1991;3:517e22.
[32] Lin X, Söderhäll K, Söderhäll. Transglutaminase activity in the hematopoietic
tissue of a crustacean, Pacifastacus leniusculus, importance in hemocyte
homeostasis. BMC Immunol 2008;9:58.
971
[33] Yeh MS, Kao LR, Huang CJ, Tsai IH. Biochemical characterization and cloning of
transglutaminases responsible for hemolymph clotting in Penaeus monodon
and Marsupenaeus japonicus. Biochim Biophys Acta 2006;1764:1167e78.
[34] Yeh MS, Liu CH, Hung CW, Cheng W. cDNA cloning, identification, tissue
localisation, and transcription profile of a transglutaminase from white
shrimp, Litopenaeus vannamei, after infection by Vibrio alginolyticus. Fish
Shellfish Immunol 2009;27(6):748e56.
[35] Griffin M, Casadio R, Bergamini CM. Transglutaminase: nature’s biological
glues. Biochem J 2002;368(Pt 2):377e96.
[36] Lorand L, Graham RM. Transglutaminases: crosslinking enzymes with pleiotropic functions. Nat Rev 2003;4(2):140e56.
[37] Wang Z, Wilhelmsson C, Hyrsl P, Loof TG, Dobes P, Klupp M, et al. Pathogen
entrapment by transglutaminasedA conserved early innate immune mechanism. PLoS Pathog 2010;6(2):e1000763. http://dx.doi.org/10.1371/journal.
ppat.1000763.
[38] Song YL, Yu CI, Lien TW, Huang CC, Lin MN. Haemolymph parameters of
Pacific white shrimp (Litopenaeus vannamei) infected with Taura syndrome
virus. Fish Shellfish Immunol 2003;14(4):317e31.
[39] Han-Ching Wang K, Tseng CE, Lin HY, Chen IT, Chen YH, Chen YM, et al. RNAi
knock-down of the Litopenaeus vannamei toll gene (LvToll) significantly
increases mortality and reduces bacterial clearance after challenge with Vibrio
harveyi. Dev Comp Immunol 2010;34(1):49e58.
[40] Lin X, Söderhäll I. Crustacean hematopoiesis and the astakine cytokines. Blood
2011;117(24):6417e24.
[41] Shibata T, Ariki S, Shinzawa N, Miyaji R, Suyama H, Sako M, et al. Protein
crosslinking by transglutaminase controls cuticle morphogenesis in drosophila.
PLoS One 2010;5(10):e13477. http://dx.doi.org/10.1371/journal.pone.0013477.
[42] Jiang Y, Doolittle RF. The evolution of vertebrate blood coagulation as viewed
from a comparison of puffer fish and sea squirt genomes. Proc Natl Acad Sci
USA 2003;100:7527e32.
[43] Cerenius L, Kawabata S, Lee BL, Nonaka M, Soderhall K. Proteolytic cascade
and their involvement in invertebrate immunity. Trends Biochem Sci 2010;
35(10):575e83.
[44] Yeh MS, Chen YL, Tsai IH. The hemolymph clottable proteins of tiger
shrimp, Penaeus monodon, and related species. Comp Biochem Physiol
1998;121:169e76.
[45] Yeh MS, Huang CJ, Leu JH, Lee YC, Tsai IH. Molecular cloning and characterization of a hemolymph clottable protein from tiger shrimp (Penaeus monodon). Eur J Biochem 1999;266:624e33.
[46] Montano-Perez K, Yepiz-Plascencia G, Higuera-Ciapara I, Vargas-Albores F.
Purification and characterization of clotting protein from the white shrimp
Penaeus vannamei. Comp Biochem Physiol B 1999;122:381e7.
[47] Reyes-Izquierdo T, Vargas-Albores F. Proteinase activity in the white shrimp
(Penaeus vannamei) clotting protein. Biochem Biophys Res Comm 2001;287:
332e6.
[48] Perazzolo LM, Lorenzini DM, Daffre S, Barracco MA. Purification and partial
characterization of the plasma clotting protein from pink shrimp Farfantepenaeus paulensis. Comp Biochem Physiol 2005;142(3):302e7.
[49] Hall M, van Heusden MC, Söderhäll K. Identification of the major lipoproteins
in crayfish hemolymph as proteins involved in immune recognition and
clotting. Biophys Res Commun 1995;216:939e46.
[50] Yepiz-Plascencia G, Jimenez-Vega F, Romo-Figueroa MG, Sotelo-Mundo RR,
Vargas-Albores F. Molecular characterization of the bi-functional VHDL-CP
from the hemolymph of white shrimp Penaeus vannamei. Comp Biochem
Physiol B 2002;132:585e92.
[51] Yeh MS, Huang CJ, Cheng JH, Tsai IH. Tissue-specific expression and regulation
of the hemolymph clottable protein of tiger shrimp (Penaeus monodon). Fish
Shellfish Immunol 2007;23:272e9.
[52] Bulet P, Stöcklin R, Menin L. Anti-microbial peptides: from invertebrates to
vertebrates. Immunol Rev 2004;198:169e84.
[53] Guaní-Guerra E, Santos-Mendoza T, Lugo-Reyes SO, Terán LM. Antimicrobial
peptides: general overview and clinical implications in human health and
disease. Clin Immunol 2010;135:1e11.
[54] Kamysz W, Okroj M, Lukasiak J. Novel properties of antimicrobial peptides.
Acta Biochim Pol 2003;50:461e9.
[55] Lai Y, Gallo RL. AMPed up immunity: how antimicrobial peptides have
multiple roles in immune defense. Trends Immunol 2009;30:131e41.
[56] Yount NY, Bayer AS, Xiong YQ, Yeaman MR. Advances in antimicrobial peptide
immunobiology. Biopolymers 2006;84:435e58.
[57] Bachere E, Gueguen Y, de Lorgeni J, Gamier J, Romestand B. Insights into the
anti-microbial defense of marine invertebrates: the penaeid shrimps and the
oyster Craccostrea gigas. Immunol Rev 2004;198:149e68.
[58] Destoumieux D, Bulet P, Loew D, VanDorsselaer A, Rodriguez J, Bachere E.
Penaeidins, a new family of antimicrobial peptides isolated from the shrimp
Penaeus vannamei (decapoda). J Biol Chem 1997;272:28398e406.
[59] Cuthbertson BJ, Deterding LJ, Williams JG, Tomer KB, Etienne K, Blackshear PJ,
et al. Diversity in penaeidin antimicrobial peptide form and function. Dev
Comp Immunol 2008;32:167e81.
[60] Rosa RD, Barracco. Antimicrobial peptides in crustaceans. Inv Surv J 2010;7:
262e84.
[61] Tassanakajon A, Amparyup P, Somboonwiwat K, Supungul P. Cationic antimicrobial peptides in penaeid shrimp. Mar Biotechnol 2010;12(5):487e505.
[62] Zhao XF, Wang JX. The antimicrobial peptides of the immune response of
shrimp. Inv Surv J 2008;5:162e79.
972
M.B.B. Maningas et al. / Fish & Shellfish Immunology 34 (2013) 968e972
[63] de Lorgeril J, Gueguen Y, Goarant C, Goyard E, Mugnier C, Fievet J, et al.
A relationship between antimicrobial peptide gene expression and capacity of
a selected shrimp line to survive a Vibrio infection. Mol Immunol 2008;
45(12):3438e45.
[64] Kaizu A, Fagutao FF, Kondo H, Aoki T, Hirono I. Functional analysis of C-type
lysozyme in Penaeid shrimp. J Biol Chem 2011;286(52):44344e9.
[65] Tassanakajon A, Somboonwiwat K. Antimicrobial peptides from the black tiger
shrimp Penaeus monodon - a review. In: Bondad-Reantaso MG, Jones JB, Corsin F, Aoki T, editors. Diseases in Asian aquaculture, vol. VII; 2011. p. 229e40.
[66] Smith CL, De Lotto R. A common domain within the pro-enzyme regions of
the Drosophila snake and easter proteins and Tachypleus pro-clotting
[67]
[68]
[69]
[70]
enzymes defines a new subfamily of serine proteases. Protein Sci 1992;1:
1225e6.
Wang R, Lee SY, Cerenius L, Söderhäll K. Properties of the prophenoloxidase
activating enzyme of the freshwater crayfish, Pacifastacus leniusculus. Eur J
Biochem 2001;268:895e902.
Krem MM, Cera ED. Evolution of enzyme cascades from embryonic development to blood coagulation. Trends Biochem Sci 2002;27:67e74.
Sun H. The interaction between pathogens and the host coagulation system.
Physiology 2005;21:281e8.
Esmon CT. The interactions between inflammation and coagulation. Br J
Haematol 2005;131:417e30.