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Chapter 6 ALF gene was found to be up-regulated on WSSV challenge even in the control group, especially in the heart. However, ALF was found to be down-regulated in the muscle and hepatopancreas of the control group of shrimps on WSSV challenge. In case of Bacillus treated group, a complete down-regulation of the ALF gene could be detected in all the target tissues. The only ALF expression detected in this group of shrimps was in the gills and intestine and hepatopancreas (Fig. 6. 19 B & D). 6.3.4.2. Expression profile of crustin-1 mRNAs of crustin-1 gene could be detected in all of the target tissues in the control group of animals prior to WSSV challenge. However, on Bacillus administration, a complete down-regulation of the crustin-1 gene could be detected in all the tissues. Gill, muscle, and intestine was found to support maximum expression of the crustin-1 gene in the control shrimps, whereas in case of Bacillus treated group of shrimps, expression could be detected only in the gills (Fig. 6. 20 A & C). Not much variation could be observed in the pattern of expression of crustin-1 for the control and Bacillus treated group of shrimps post-challenge WSSV. On challenge with WSSV, a complete down-regulation of the crustin- 1 gene could be observed in the various target tissues of both the control and Bacillus administered group of shrimps, with expressions limiting only to gills and intestine. When compared to the control group, Bacillus administered group supported up-regulation of the crustin-1 gene post- challenge WSSV in both the gills and intestine. Maximum level of expression was found to be in the intestine followed by gills. No transcripts of crustin-1 could be detected in muscle, hepatopancreas or heart of both the control group and Bacillus treated groups (Fig. 6. 20 B & D). 6.3.4.3. Expression profile of crustin-2 Crustin-2 was found to be expressed at very high levels in all the target tissues of the control group of shrimps pre-challenge WSSV. However, Molecular Characterization and Gene Expression Profiling of Antimicrobial Peptides in Penaeid Shrimps 349
Chapter 6 the gene was found to be down-regulated in the Bacillus treated group prior to WSSV challenge. In Bacillus administered group, maximum expression was found in the gill, followed by intestine and muscle (Fig. 6. 21 A & C). On challenge with WSSV, a general down-regulation of the crustin-2 gene could be noticed in all the target tissues of the control group. However, in case of Bacillus treated group of shrimps, the crustin-2 gene was found to be up-regulated considerably in all the tissues except in case of heart. Maximum expression was found in the gill and intestine (Fig. 6. 21 B & D). 6.3.4.4. Expression profile of crustin-3 Crustin-3 was found to be expressed in all the target tissues of the control group of shrimps pre-challenge WSSV. Maximum expression of the gene was found in gill and intestine. However, the gene was found to be completely down-regulated on administration of Bacillus, with expressions limiting to gill, intestine and muscle of the treated group (Fig. 6. 22 A & C). On challenge with WSSV, complete down-regulation of the crustin-3 gene could be noticed in various target tissues. In the control group; gill was the only tissue that was found to express the crustin-3 gene, that too at very low level. However, when compared to the control, the Bacillus treated group up-regulated the crustin-3 gene, with maximum expression in the gill followed by intestine (Fig. 6. 22 B & D). 6.3.4.5. Expression profile of penaeidin-3 Penaeidin-3 was found to be expressed at very high levels in all the target tissues, except gills in the control group of shrimps prior to WSSV challenge. On administration of Bacillus, the penaeidin-3 gene was found to be up-regulated considerably in the gills. However, the gene was found to express at the same level in the muscle and intestine of the experimental animals. In case of hepatopancreas and heart, no expression of penaeidin-3 gene could be found (Fig. 6. 23 A & C). Molecular Characterization and Gene Expression Profiling of Antimicrobial Peptides in Penaeid Shrimps 350
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Molecular Characterization and Gene
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Declaration I hereby do declare tha
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Acknowledgements This work was carr
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My Seniors, Dr. Annies Joseph, Dr.
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Contents Title Page Nos. 1 General
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in response to WSSV challenge 252-2
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α-2M Alpha-2-Macroglobulin ALF Ant
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ppA Prophenoloxidase-Activating Enz
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1.1 Introduction Chapter 1 Aquacult
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Chapter 1 the giant tiger shrimp (P
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Common names: Chapter 1 Giant tiger
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1.3.1. Diseases Chapter 1 Diseases
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Lymphoidal parvo-like virus (LPV) B
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Chapter 1 and protective occlusion
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Chapter 1 small brown masses in the
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1.4.6. Natural epidemics Chapter 1
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Chapter 1 can differentiate WSSV-in
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1.5.1.1. Probiotics Chapter 1 Alter
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Chapter 1 tried in P. monodon, resu
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Chapter 1 The first and essential i
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Chapter 1 1998). The innate immunit
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1.6.1 Haemocytes Chapter 1 The haem
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Chapter 1 In crustaceans, the sheet
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Chapter 1 Theopold et al., 2002). C
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Chapter 1 kill/remove the invading
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Chapter 1 forming complexes with a
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1.7 Antimicrobial Peptides (AMPs) 1
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Chapter 1 the response is very fast
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Chapter 1 Hirsch, 1947). While they
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Chapter 1 AMPs to various degrees u
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1.7.5 Biological activity Chapter 1
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Chapter 1 including the microbes as
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Tachyplesin I Chapter 1 Crustacea P
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Anionic and cationic peptides that
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Anionic and Cationic AMPs with cyst
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Chapter 1 Fig. 1.7. Structural clas
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Chapter 1 Group III: Linear peptide
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Chapter 1 through regular processes
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Chapter 1 ultimately signal to tran
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Chapter 1 divalent polyanionic cati
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Chapter 1 type of transmembrane por
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Chapter 1 other intracellular targe
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Chapter 1 Apidaecin, a short, proli
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Chapter 1 Hultmark, 1987; Gennaro a
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Chapter 1 quite opposite characteri
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Chapter 1 depletion of mitochondria
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Chapter 1 conformations adopted by
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Chapter 1 receptor may be a potenti
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Chapter 1 exposure to their targets
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Chapter 1 generating a rich spectru
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Chapter 1 by enkelytin (Goumon et a
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Lactoferricin Cow Mastitis control
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Chapter 1 trial, in which peptide T
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Chapter 1 a too limited spectrum to
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Chapter 1 number of research groups
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CHAPTER-2 Molecular Characterizatio
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Chapter 2 interaction with negative
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Chapter 2 inhibit the endotoxin or
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Chapter 2 structure be modified to
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Chapter 2 and the region might be l
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Chapter 2 been made of the spectra
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Chapter 2 their discovery and initi
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Chapter 2 terminal pyroglutamic aci
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Chapter 2 One can assume that the p
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Chapter 2 by degranulation into the
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2.2. Materials and Methods 2.2.1. E
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Chapter 2 washed by adding 1 ml 75
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Chapter 2 entry of recombinant DNA
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Chapter 2 and the deduced amino aci
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Chapter 2 Fenneropenaeus, Litopenae
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2.3.1.3. Crustin-1 (GQ334395) Chapt
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2.3.1.3.5. Phylogenetic analysis of
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Chapter 2 sequence also showed high
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Chapter 2 chinensis and F. indicus
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Chapter 2 monodon and Group III of
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Chapter 2 Tiger shrimp (P. monodon)
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Chapter 2 ALF-1 was a partial seque
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Chapter 2 As mentioned before, the
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Chapter 2 Munoz et al., 2004). The
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Table 2.1. Primers used for the stu
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Table 2.4. BLASTn analysis of ALF-2
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Table 2.8. BLASTn analysis of Crust
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Table 2.12. BLASTn analysis of Pena
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Fig.2.5. Agarose gel electrophoreto
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Chapter 2 Fig.2.9. Multiple alignme
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Chapter 2 Fig. 2.12 A bootstrapped
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Chapter 2 Fig. 2.15. Multiple align
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Chapter 2 Fig.2.18. A bootstrapped
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Chapter 2 Fig.2.20. Signal peptide
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Chapter 2 Fig.2.24 Multiple alignme
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Chapter 2 LITOPENAEUS SP. FARFANTEP
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Chapter 2 Fig. 2.28. Nucleotide and
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Chapter 2 LITOPENAEUS SP. FENNEROPE
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Chapter 2 Fig.2.43 A bootstrapped n
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Chapter 2 LITOPENAEUS SP. FENNEROPE
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Chapter 2 Fig.2.51 Signal peptide a
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Chapter 2 FENNEROPENAEUS SP. FARFAN
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CHAPTER-3 Molecular Characterizatio
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Chapter 3 certainly help us to unra
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Chapter 3 3.3.1.1. Anti-lipopolysac
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Chapter 3 was predicted out to be 1
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Chapter 3 random coiled structure t
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3.3.1.3.5. Phylogenetic analysis of
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Chapter 3 case has been reported in
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Chapter 3 Multiple alignments were
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Chapter 3 negative bacteria. In shr
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Chapter 3 indicated a random coiled
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Chapter 3 horseshoe crab big defens
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Table 3.4. BLASTn analysis of ALF-2
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Table.3.8. BLASTn analysis of Fi-pe
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Chapter 3 Fig.3.2. Nucleotide and a
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Chapter 3 Fig.3.5. Multiple alignme
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Chapter 3 Fig.3.8. Nucleotide and a
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Chapter 3 Fig.3.12. Multiple alignm
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Chapter 3 Fig.3.14. A bootstrapped
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SHRIMPS KIND CRAB LOBSTERS CRABS Ch
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Chapter 3 Fig. 3.28. Multiple align
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PENAEIDIN-2 Chapter 3 PENAEIDIN- 3
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Fig.3.31. Nucleotide and amino acid
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Chapter 3 Fig.3.37 Nucleotide and a
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4.1. Introduction Chapter 4 The aqu
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Chapter 4 appearance and have major
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Chapter 4 the tissues has recently
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Chapter 4 with the ability of shrim
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Chapter 4 with a low dose of WSSV b
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Chapter 4 molecules they are likely
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Chapter 4 for understanding gene ex
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Chapter 4 4.3.2. Expression profile
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Expression profile of endonuclease
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Chapter 4 bacterial and fungal infe
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Chapter 4 measuring TSV and YHV loa
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Chapter 4 Of particular interest he
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Chapter 4 large distribution and ab
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Chapter 4 WSSV genes viz. endonucle
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Table 4.2. Primers used for the stu
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(A) (B) B 28 1 2 3 4 5 6 7 8 9 10 C
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(A) (B) B 28 1 2 3 4 5 6 7 8 9 10 C
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(A) (B) B 28 1 2 3 4 5 6 7 8 9 10 C
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(A) (B) B 28 1 2 3 4 5 6 7 8 9 10 C
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(A) (B) B 28 1 2 3 4 5 6 7 8 9 10 F
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(A) (B) B 28 1 2 3 4 5 6 7 8 9 10 C
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(A) (B) B 28 1 2 3 4 5 6 7 8 9 10 C
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(A) (B) B 28 1 2 3 4 5 6 7 8 9 10 C
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CHAPTER-5 Expression Profile of Ant
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Chapter 5 1994). The emergence of b
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Chapter 5 (Bovill et al., 2001). In
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5.2.2. Test diets Chapter 5 Four ex
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5.2.2.4. Preparation of Glucan inco
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Chapter 5 genes (latency related ge
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Chapter 5 On WSSV challenge, crusti
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Chapter 5 5.3.4. Expression profile
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Page 323 and 324: Chapter 5 For the present study two
Page 325 and 326: Chapter 5 ameobocytes of two marine
Page 327 and 328: Chapter 5 variation in expression o
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Page 331 and 332: Chapter 5 manifested in terms of gr
Page 333 and 334: (A) (B) (C) C CHY CSY CHG CSG C CHY
Page 341 and 342: (A) (B) -VE CTRL +VE CTRL CHG CSG C
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Page 351 and 352: (A) PRE-CHALLENGE (B) (C) (D) CONTR
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Page 355 and 356: (A) (B) G M HP HR I Chapter 5 Fig.
Page 357 and 358: Chapter 5 Fig. 5.26. Post challenge
Page 359 and 360: 6.1. Introduction Chapter 6 Prophyl
Page 361 and 362: Chapter 6 production of inhibitory
Page 363 and 364: 6.2. Materials and methods 6.2.1. E
Page 365 and 366: Chapter 6 tissue cDNA was performed
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Page 379 and 380: Chapter 6 Detailed tissue-wise anal
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Page 383 and 384: (A) (B) Fig. 6.1. Experimental diet
Page 385 and 386: (A) (B) (C) Fig. 6.3. Expression pr
Page 387 and 388: (A) (B) (C) C B M BM C B M BM Fig.
Page 389 and 390: (A) (B) (C) Fig. 6.7. Expression pr
Page 391 and 392: (A) (B) (C) Chapter 6 Fig. 6.9. Exp
Page 393 and 394: (A) (B) -VE CTRL +VE CTRL B M BM Ch
Page 405 and 406: (A) (C) (D) PRE-CHALLENGE POST-CHAL
Page 407 and 408: (A) (B) G M HP HR I Chapter 6 Fig.
Page 409 and 410: Chapter 6 Fig. 6.27. Post challenge
Page 411 and 412: 7.1. Summary Chapter 7 Tropical shr
Page 413 and 414: Salient findings Chapter 7 � From
Page 415 and 416: Chapter 7 � Tissue-wise expressio
Page 417 and 418: Chapter 7 application in shrimp cul
Page 419 and 420: References Aboudy, Y., Mendelson, E
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References Anggraeni, M.S., Owens,
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References Bals, R., Wang, X., Zasl
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References derived from the N-termi
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References Breukink, E., de Kruijff
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References Burgents, J.E., Burnett,
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References 1,3-β-D-glucan-binding
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References Chen, J.Y., Chuang, H.,
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References Cole, A.M., Hong, T., Bo
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References Dathe, M., Wieprecht, T.
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References Diamond, G., Zasloff, M.
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References Faber, C., Stallmann, H.
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References Flint, S.J., Enquist, L.
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References Gennaro, R., Zanetti, M.
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References Gudmundsson, G.H., Lidho
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References Harwig, S.S., Swiderek,
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References Hirakura, Y., Kobayashi,
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References Huang, C.C., Song, Y.L.
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References Itami, T., Takahashi, Y.
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References Jiravanichpaisal, P. Whi
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References Kang, J.H., Shin, S.Y.,
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References Kono, T., Savan, R., Sak
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References rich peptide derived fro
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References Li, L., Wang, J.X., Zhao
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References Liu, C.H., Cheng, W., Ku
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References Lorenzon, S., de Guarrin
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References Medvinsky, A., Dzierzak,
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References Moriarty, D.J.W. 1996. M
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References Otta, S.K., Karunasagar,
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References Litopenaeus vannamei cha
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References experimental system. In:
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References the black tiger shrimp P
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References Tapay, L.M., Cesar, E.,
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References van Hulten, M.C., Reijns
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Penaeus monodon anti-lipopolysaccha
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Penaeus monodon crustin-like antimi
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Penaeus monodon crustin-like antimi
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Penaeus monodon penaeidin-like anti
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Penaeus monodon elongation factor m
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Fenneropenaeus indicus anti-lipopol
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Fenneropenaeus indicus penaeidin-li
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Fenneropenaeus indicus beta-actin m
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PUBLICATIONS
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Short sequence report Molecular cha
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Fig. 2. Multiple alignment of nucle
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220 The phylogenetic relationships
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Table 1 Rearing conditions and wate
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5 ′ cttgtggttgacaatggctccg3
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Expression of WSSV genes in the hae
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Fig. 4. Expression profile of crust
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S.P. Antony et al. / Immunobiology
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