Genomic Research for Increase Culture Efficiency of Shrimp: The Study of Genes and Proteins Associated with Disease Resistance

 

Functional genomic approach to identify candidate genes and proteins responding to pathogens in black tiger shrimp Penaeus monodon

 

The cultivation of black tiger shrimp encounters many problems. One major problem is the diseases. Among these diseases, viral and bacterial infections cause an extensively serious production loss in shrimp industry. In shrimp, their defensive response to diseases is believed to depend on an innate immune system that resists the invasion by pathogens. For immediate immune response to pathogens, several proteins and peptides are synthesized, stored in the hemocytes and released into the hemolymph upon infections. Subsequently, more biosynthesis of the proteins is required for continuing the defense or restoring the hemocyte defense proteins, leading to the changes in gene expression as the response progresses.

 

There may be several genes in shrimp involved in the interaction between host and viruses. The studies of changes in gene expression profile enable the identification of these genes. The microarray technique has been proved to be a powerful tool to investigate the expression of thousands of genes in a single hybridization. To identify the differentially expressed genes in P. monodon in response to virus and bacterial infections, microarray technique is used in the hunt for candidate genes by comparing the gene expression patterns in the hemocytes of healthy and infected shrimp. The identification of new viral and bacteria responsive genes will be identified. The studies are also extended to identify more genes regulated in response to viral and bacterial infections using a suppression subtractive hybridization approach. Responsive genes from the microarray and suppression subtractive hybridization results will be assembled in a stress chip, tailored for use in the future as biomarkers to measure the well-being of shrimp.

 

Researchers

Associate Professor Anchalee Tassanakajon, Ph.D.

Associate Professor Vichien Rimphanitchayakit, Ph.D.

Siriporn Pongsomboon, Ph.D.

Premruethai Supungul, Ph.D.

Piti Ampayup, Ph.D.

Sureerat Tang, M.Sc.

 

Sub-project I : Microarray analysis of white spot syndrome virus (WSSV), yellow head virus (YHV) and Vibrio harveyi infected P. monodon

by Siriporn Pongsomboon, Sureerat Tang, Anchalee Tassanakajon

 

Sub-project II : Construction of a viral- and bacterial-stress chip of P. monodon and the stress chip test for use as biomarkers to identify healthy shrimp

by Siriporn Pongsomboon, Sureerat Tang, Piti Ampayup, Anchalee Tassanakajon

 

Characterization and application of antimicrobial peptides from the back tiger shrimp Penaeus monodon

Concomitant with the growth of the shrimp culture industry has been the emerging of the diseases, especially those caused by the infectious agents, plaguing the world shrimp culture industry.  The important diseases have been the viral or bacterial etiologies.  Antibiotic uses are in many cases ineffective, result in the increases in virulence of pathogens, and, importantly, are the cause for concern in the possibility of transferring the antibiotic resistance to human pathogens.  Therefore, antimicrobial peptides (AMPs) are the promising resources for therapeutic and pharmaceutical applications in this field.

 

 

AMPs are a major component of the innate immune defense system.  AMPs provide an immediate and rapid response to invading microorganisms including Gram-positive and Gram-negative bacteria, yeast and fungi.  However, only a few AMPs have been isolated from the marine invertebrates.  In this project, we aim to isolate and characterize more AMPs from the shrimp Penaeus monodon by searching through the cDNA libraries in the Penaeus mondon EST database.  The putative AMP homologues will be expressed, characterized for their antimicrobial activities.  The results from this study provide not only more knowledge in shrimp immunity but also an alternative to antibiotic in shrimp culture industry.  The presence of AMPs can also be used as indication for the selection of high health broodstock.

 

Researchers

Associate Professor Anchalee Tassanakajon, Ph.D.

Associate Professor Vichien Rimphanitchayakit, Ph.D.

Premruethai Supungul, Ph.D.

Kunlaya Somboonwiwat, Ph.D.

 

Sub-project I: Identification and characterization of antimicrobial peptides from the black tiger shrimp Penaeus monodon

By Premruethai Supungul, Kunlaya Somboonwiwat, Tipachai Vatanavicharn, Vichien Rimphanitchayakit, Anchalee Tassanakajon

 

Sub-project II: Effect of recombinant anti-lipopolysaccharide factor type 3 on vibriosis in the tiger shrimp Penaeus monodon

By Sirikwan Pongrateep, Kunlaya Somboonwiwat

 

Serine Proteinase Inhibitors

In animal, proteinases are used in several important physiological processes, particularly the blood clotting reaction. Therefore, serine proteinase inhibitors (SPIs) are used to limit the level and extent of the proteinase activity. Several SPIs have been found in the black tiger shrimps during the course of expressed sequence tag analysis, mostly from the hemocytes. It is believed that some of these SPIs are involved in blood coagglulation, prophenoloxidase cascades and possibly the defense against certain phathogens. It is, therefore, our aim to uncover the characteristics and functions of these SPIs. There are, currently, two projects in this area of study.

 

Researchers

Associate Professor Anchalee Tassanakajon, Ph.D.

Associate Professor Vichien Rimphanitchayakit, Ph.D.

Kunlaya Somboonwiwat, Ph.D.

   

Sub-project I : Domain activities and gene organization of the 5 Kazal domain serine proteinase inhibitor from the black tiger shrimp Penaeus monodon

by Suchao Donpudsa, Anchalee Tassanakajon, Vichien Rimphanitchayakit

 

Sub-project II : Expression and characterizaton of SERPIN gene in the black tiger shrimp Penaeus monodon

by Kunlaya Somboonwiwat, Vichien Rimphanitchayakit, Anchalee Tassanakajon

 

Molecular cloning and characterization of genes controlling osmoregulation in black tiger shrimp (Penaeus monodon)

The black tiger shrimp (Penaeus monodon) belong to a group of euryhaline crustaceans that generally inhabit under a wide range of salinity levels, low salinity in mangrove swamp to high salinity in the off-shore sea. The matured shrimp naturally migrate from brackish water to the deep sea for mating and spawning, and the developing shrimp larva return to the nutrition-rich in-shore sea or mangrove swamp for growing. Therefore, shrimp farmers can expand their farms extensively in different areas/environments. However, extremely high or low salinities always give more problems than the suitable salinity, which range from 15 to 25 ppt. Cultivation of shrimp in extremely high salinity over 30 ppt may cause disease problems, particularly the white spot or yellow head viral and luminescent bacterial infections. After 70-90 days at low salinity, the shrimp can not molt properly, and stunning can be observed. An intensive effort has been made to produce a high quality broodstock under the pond-reared condition instead of catching them from the sea. The current outcome is determined as imperfect success because the shrimp can be reared successfully but the development of gonad and sperm is still dependent on the stimulation of high salinity level before mating. Genome information, particularly those obtained from the shrimp EST project, has accelerated the discovery of genes that play the important roles in various biological and physiological systems. The study of genes involved in osmoregulation will provide an understanding of mechanism of osmoregulation in shrimp, and will be useful for shrimp cultivation at low salinity condition and the production of pond-reared broodstock.

 

Researchers

Associate Professor Anchalee Tassanakajon, Ph.D.

Siriporn Pongsomboon, Ph.D.

Suwit Wuthisuthimethavee, Ph.D.

Piti Ampayup , Ph.D.

 

Sub-project I : Identification and characterization of Na +/K +-ATPase/carbonic anhydrase genes and proteins

by Siriporn Pongsomboon, Piti Ampayup, Suwit Wuthisuthimethavee, Anchalee Tassanakajon

 

Sub-project II : Identification of functionally relevant genes controlling osmoregulation in P. monodon by using differential display reverse transcription-PCR (DD RT-PCR)

by Suchonma Udomlertpreecha, Siriporn Pongsomboon, Suwit Wuthisuthimethavee, Piti Ampayup, Anchalee Tassanakajon