Molecular characterization of a crustin-like antimicrobial peptide in the giant tiger shrimp, Penaeus monodon, and its expression profile in response to various immunostimulants and challenge with WSSV

Abstract

a b s t r a c tA crustin-like antimicrobial peptide from the haemocytes of giant tiger shrimp, Penaeus monodon was partially characterized at the molecular level and phylogenetic analysis was performed. The partial coding sequence of 299 bp and 91 deduced amino acid residues possessed conserved cysteine residues characteristic of the shrimp crustins. Phylogenetic tree and sequence comparison clearly confirmed divergence of this crustin-like AMP from other shrimp crustins. The differential expression of the crustin-like AMP in P. monodon in response to the administration of various immunostimulants viz., two marine yeasts (Candida haemulonii S27 and Candida sake S165) and two ␤-glucan isolates (extracted from C. haemulonii S27 and C. sake S165) were noted during the study. Responses to the application of two gram-positive probiotic bacteria (Bacillus MCCB101 and Micrococcus MCCB104) were also observed. The immune profile was recorded pre-and post-challenge white spot syndrome virus (WSSV) by semi-quantitative RT-PCR. Expressions of seven WSSV genes were also observed for studying the intensity of viral infection in the experimental animals. The crustin-like AMP was found to be constitutively expressed in the animal and a significant down-regulation could be noted post-challenge WSSV. Remarkable down-regulation of the gene was observed in the immunostimulant fed animals pre-challenge followed by a significant up-regulation post-challenge WSSV. Tissue-wise expression of crustin-like AMP on administration of C. haemulonii and Bacillus showed maximum transcripts in gill and intestine. The marine yeast, C. haemulonii and the probiotic bacteria, Bacillus were found to enhance the production of crustin-like AMP and confer significant protection to P. monodon against WSSV infection.

Validation of Reference Genes for Real-Time PCR of Reproductive System in the Black Tiger Shrimp

“…In the Pacific blue shrimp Penaeus stylirostris , elongation factor 1 alpha ( EF-1α ) and glyceraldehydes-3 phosphate dehygrogenase ( GAPDH ) have been validated as reference genes for expression analysis of immune genes [13]. In the black tiger shrimp ( P. monodon), several housekeeping genes, such as β-actin [14], [15], [16], EF-1α [ 17 ], [18], [19], [ 20 ], GAPDH [19], 40S rRNA [21], 18S rRNA [16], [22], and elongation factor 2 [23] have been used as an internal control for qPCR. However, to date, no study has validated their suitability as an internal control for gene expression analysis using qPCR in P. monodon .…”

Host MicroRNA-217 Promotes White Spot Syndrome Virus Infection by Targeting Tube in the Chinese Mitten Crab (Eriocheir sinensis)

“…ALF from Fenneropeneaus chinensis played a key role in inhibiting WSSV replication (Li S. et al, 2015). In Penaeus monodon , the enhanced crustin-like AMP conferred significant protection from WSSV infection (Antony et al, 2011). The knockdown of Tube gene resulted in a reduction of ALF1, ALF2, ALF3, Crus1 , and Crus2 transcripts in crabs.…”

Respuesta inmune y expresion de genes en el camaron blanco (Litopenaeus vannamei) inducida por inmunoestimulantes microbianos

“…La crustina es un péptido antimicrobiano muy importante en los crustáceos decápodos. Antony et al (2010) caracterizaron la crustina en los hemocitos del camarón tigre gigante (Penaeus monodon) a nivel molecular. Los autores observaron una expresión diferencial del gen de la crustina en respuesta a la administración de diversos inmunoestimulantes (levaduras y bacterias).…”