Expression profiles of three crustin-like genes in Litopenaeus vannamei under Vibrio harveyi infection

  • Mao-Cang Yan
  • Yao-Hua Wang
  • De-Wei Ji
  • Fang Li
  • Li-Hua Hu
  • Min ZhangEmail author


Immune response of three crustin-like genes isolated from Litopenaeus vannamei under Vibrio harveyi infection was analyzed. Sequence analysis revealed that three crustin-like proteins contained whey acidic protein (WAP) domain, Crs 1 encoded 126 amino acids, Crs 2 encoded 156 amino acids, and Crs3 encoded 87 amino acids. Tissue expression pattern analysis showed that Crs 1 was not detected in the intestine, it was highest expressed in the muscle and lowest expressed in the hepatopancreas; Crs 2 was not detected in the hepatopancreas, intestine, and muscle but it was highest expressed in the hemocyte. Crs 3 was expressed in all tested tissues and was more abundant in the gill and muscle. Bacterial infection assay showed that mRNA level of Crs 1 in the gill was significantly upregulated at 24 h and 7 days; it was upregulated at 7 days in the muscle. The transcript level of Crs 2 in the hemocyte was upregulated at 24 h and significantly downregulated at 7 days, and it was downregulated in the heart at 24 h and 7 days. mRNA levels of Crs 3 in the hemocyte and hepatopancreas were significantly increased at 24 h; it was upregulated at 7 days in the heart and muscle. The results indicated that immune response of different crustins was tissue specific and the data provided us basic knowledge about the roles of three crustin-like genes in response to bacterial infection.


Litopenaeus vannamei Crustin-like Antimicrobial peptide Vibrio harveyi Gene expression 


Funding information

The present study was financially supported by the Science and Technology Program of Zhejiang Province (No. 2017F30041), Major Science and Technology Specific Program of Zhejiang Province (No. 2016C02055-5), and Science and Technology Program of Wenzhou City (No. N20160007).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

This article does not contain any studies with animals performed by any of the authors.


  1. Ananda Raja R, Sridhar R, Balachandran C, Palanisammi A, Ramesh S, Nagarajan K (2017) Pathogenicity profile of Vibrio parahaemolyticus in farmed Pacific white shrimp, Penaeus vannamei. Fish Shellfish Immunol 67:368–381CrossRefGoogle Scholar
  2. Austin B, Austin DA (2016) Bacterial fish pathogens: diseases of farmed and wild fish, 6th (revised). Springer International Publishing, LondonCrossRefGoogle Scholar
  3. Austin B, Zhang XH (2006) Vibrio harveyi: a significant pathogen of marine vertebrates and invertebrates. Lett Appl Microbiol 43:119–124CrossRefGoogle Scholar
  4. Bartlett TC, Cuthbertson BJ, Shepard EF, Chapman RW, Gross PS, Gregory WW (2002) Crustins, homologues of an 11.5-kDa antibacterial peptide, from two species of penaeid shrimp, Litopenaeus vannamei and Litopenaeus setiferus. Mar Biotechnol 4:278–293CrossRefGoogle Scholar
  5. Gross PS, Bartlett TC, Browdy CL, Chapman RW, Warr GW (2001) Immune gene discovery by expressed sequence tag analysis of hemocytes and hepatopancreas in the Pacific White Shrimp, Litopenaeus vannamei, and the Atlantic White Shrimp, L. setiferus. Dev Comp Immunol 25:565–577CrossRefGoogle Scholar
  6. Kim B, Kim M, Kim AR, Yi M, Choi JH, Park H, Park W, Kim HW (2013) Differences in gene organization between type I and type II crustins in the morotoge shrimp, Pandalopsis japonica. Fish Shellfish Immunol 35:1176–1184CrossRefGoogle Scholar
  7. Li F, Xiang JX (2013) Recent advances in researches on the innate immunity of shrimp in China. Dev Comp Immunol 39:11–26CrossRefGoogle Scholar
  8. Li M, Ma CX, Li HY, Peng JX, Zeng DG, Chen XH, Li CZ (2018) Molecular cloning, expression, promoter analysis and functional characterization of a new crustin from Litopenaeus vannamei. Fish Shellfish Immunol 73:42–49CrossRefGoogle Scholar
  9. Lightner DV (1983) Disease in cultured penaeid shrimp. In: McVey JP (ed) CRC handbook of mariculture. CRC Press, Boca Raton, pp 289–320Google Scholar
  10. Pisuttharachai D, Fagutao FF, Yasuike M, Aono H, Yano Y, Murakami K, Kondo H, Aoki T, Hirono I (2009) Characterization of crustin antimicrobial proteins from Japanese spiny lobster Panulirus japonicus. Dev Comp Immunol 33:1049–1054CrossRefGoogle Scholar
  11. Schnapp D, Kemp GD, Smith VJ (1996) Purification and characterization of a prolinerich antibacterial peptide, with sequence similarity to bactenecin-7, from the haemocytes of the shore crab, Carcinus maenas. Eur J Biochem 240:532–539CrossRefGoogle Scholar
  12. Schryver PD, Defoirdt T, Sorgeloos P (2014) Early mortality syndrome outbreaks: a microbial management issue in shrimp farming? PLoS Pathog 10:e1003919CrossRefGoogle Scholar
  13. Shockey JE, O’Leary NA, de la Vega E, Browdy CL, John EB, Gross PS (2009) The role of crustins in Litopenaeus vannamei in response to infection with shrimp pathogens: an in vivo approach. Dev Comp Immunol 33:668–673CrossRefGoogle Scholar
  14. Smith VJ, Fernandes JM, Kemp GD, Hauton C (2008) Crustins: enigmatic WAP domain-containing antibacterial proteins from crustaceans. Dev Comp Immunol 32:758–772CrossRefGoogle Scholar
  15. Supungul P, Tang S, Maneeruttanarungroj C, Rimphanitchayakit V, Hirono I, Aoki T, Tassanakajon A (2008) Cloning, expression and antimicrobial activity of crustinPm1, a major isoform of crustin, from the black tiger shrimp Penaeus monodon. Dev Comp Immunol 32:61–70CrossRefGoogle Scholar
  16. Tassanakajon A, Amparyup P, Somboonwiwat K, Supungul P (2011) Cationic antimicrobial peptides in penaeid shrimp. Mar Biotechnol 13:639–657CrossRefGoogle Scholar
  17. Tassanakajon A, Somboonwiwat K, Amparyup P (2015) Sequence diversity and evolution of antimicrobial peptides in invertebrates. Dev Comp Immunol 48:324–341CrossRefGoogle Scholar
  18. Vargas-Albores F, Yepiz-Plascencia G, Jimenez-Vega F, Avila-Villa A (2004) Structural and functional differences of Litopenaeus vannamei crustins. Comp Biochem Physiol B 138:415–422CrossRefGoogle Scholar
  19. Walker PJ, Winton JR (2010) Emerging viral diseases of fish and shrimp. Vet Res 41:51CrossRefGoogle Scholar
  20. Zhang J, Li F, Wang Z, Xiang J (2007) Cloning and recombinant expression of a crustin like gene from Chinese shrimp, Fenneropenaeus chinensis. J Biotechnol 127:605–614CrossRefGoogle Scholar
  21. Zhang M, Huang XK, Hu LH, Ji DW, Tang M, Li F, Luo K, Zhang JM, Chai XL, Yan MC (2018) Effects of salinity fluctuation on gene expression profiles of female Litopenaeus vannamei broodstocks. Isr J Aquacult–Bamid 70:1476–1484Google Scholar

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© Springer Nature Switzerland AG 2019

Authors and Affiliations

  1. 1.Zhejiang Mariculture Research InstituteZhejiangChina
  2. 2.Zhejiang Key Lab of Exploitation and Preservation of Coastal Bio-ResourceZhejiangChina

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