Skip to main content
Log in

Prevalence of microsporidian parasite, Enterocytozoon hepatopenaei in cultured Pacific White shrimp, Litopenaeus vannamei (Boone, 1931) in West Bengal, East Coast of India

  • Published:
Aquaculture International Aims and scope Submit manuscript

Abstract

Enterocytozoon hepatopenaei (EHP) is an emerging microsporidian pathogen in shrimp aquaculture. In the present study, prevalence rate of EHP in cultured Litopenaeus vennamei from West Bengal, east coast of India, was investigated. Histology of hepatopancreas of the infected animals showed eosinophilic to basophilic inclusions in the epithelial cells with moderate necrotic tubular detachment from the basal membrane. Extracted DNA from the hepatopancreatic tissues were subjected to PCR by using two different sets of primers targeting the ssu rRNA genes. An expected PCR product of 951 and 510 bp were yielded for the EHP-positive shrimp samples. Based on the BLASTP, the sequence of ssu rRNA gene (KU179095) of the collected samples showed 100% homology with EHP ssu rRNA gene sequences submitted in NCBI from different countries viz. Vietnam, Thailand, China and India. Further, all the field samples were found to be EHP+ by using a new second-generation primer targeting the spore wall protein gene (SWP) of EHP. For detection of EHP, 119 samples from East Midnapur district, 50 samples from North 24 Parganas district and 50 Samples from South 24 Parganas district were being screened. Overall prevalence rate of EHP in the cultured L. vennamei farm in West Bengal was estimated to be 84.9%, which was highest in comparison to earlier reports in India. Our finding showed that EHP could be detected from slow growing shrimps as well as in some cases from White Faeces Syndrome-affected animals. This is the first report on the occurrence of Enterocytozoon hepatopenaei in cultured L. vannamei in West Bengal, east coast of India.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

References

  • Amogan HP, Martinez JP, Ciuffetti LM (2007) Karyotype and genome size of Nadelspora canceri determined by pulsed field gel electrophoresis. Acta Protozool 45(3):249

    Google Scholar 

  • Canning EU, Curry A, Overstreet RM (2002) Ultrastructure of Tuzetia weidneri sp. n.(microsporidia: Tuzetiidae) in skeletal muscle of Litopenaeus setiferus and Farfantepenaeus aztecus (Crustacea: Decapoda) and new data on Perezia nelsoni (microsporidia: Pereziidae) in L. setiferus. Acta Protozool 41:63–77

    Google Scholar 

  • Chayaburakul K, Nash G, Pratanpipat P, Sriurairatana S, Withyachumnarnkul B (2004) Multiple pathogens found in growth-retarded black tiger shrimp Penaeus monodon cultivated in Thailand. Dis Aquat Organ 9 60(2):89–96

    Article  Google Scholar 

  • Corradi N (2015) Microsporidia: eukaryotic intracellular parasites shaped by gene loss and horizontal gene transfers. Annu Rev Microbiol 69:167–183

    Article  CAS  PubMed  Google Scholar 

  • Flegel TW (2006) Detection of major penaeid shrimp viruses in Asia, a historical perspective with emphasis on Thailand. Aquaculture 258(1–4):1–33

    Article  Google Scholar 

  • Franzen C, Müller A (1999) Molecular techniques for detection, species differentiation, and phylogenetic analysis of microsporidia. Clin Microbiol Rev. 12(2):243–85.

  • Gunalan B, Soundarapandian P, Anand T, Kotiya AS, Simon NT (2014) Disease occurrence in Litopenaeus vannamei shrimp culture systems in different geographical regions of India. Int J Aquac Fish Sci 10:4

    Google Scholar 

  • Ha NT, Ha DT, Thuy NT, Lien VT (2010) Enterocytozoon hepatopenaei parasitizing on tiger shrimp (Penaeus monodon) infected by white feces culture in Vietnam, has been detected. Agric Rural Dev Sci Technol 12:45–50 (translation from Vietnamese)

    Google Scholar 

  • Itsathitphaisarn O, Jaroenlak P, Sanguanrut P, Salachan PV, Wiredu-Boakye D, Williams BAP, Stantiford GD, Flegel TW, Sritunyalucksana K (2016) A new and improved PCR detection method for Enterocytozoon hepatopenaei (EHP) based on a gene encoding a spore wall protein. https://enaca.org/?id=115&title=new-pcr-detection-method-for-enterocytozoon-hepatopenaei. Accessed Dec 2016

  • Jaroenlak P, Sanguanrut P, Williams BA, Stentiford GD, Flegel TW, Sritunyalucksana K, Itsathitphaisarn O (2016) Nested PCR assay to avoid false positive detection of the microsporidian Enterocytozoon hepatopenaei (EHP) in environmental samples in shrimp farms. PLoS One 11(11):e0166320

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Landgdon JS (1991) Microsporidiosis due to a pleistophorid in arron, Cheraxtenuimanus (Smith), (Decapoda: Parastacidae). J Fish Dis 14:33–44

    Article  Google Scholar 

  • Larsson JR, Ebert D, Manginand KL, Vavra J (1998) Ultrastructural study and description of Flabelliforma magnivora sp. n.(Microspora: Duboscqiidae), a microsporidian parasite of Daphnia magna (Crustacea: Cladocera: Daphniidae). Acta Protozool 1(37):41–52

    Google Scholar 

  • Lightner DV (1996) A handbook of shrimp pathology and diagnosis procedures for diseases of cultured Penaeid shrimp. World Aquaculture Society, Baton Rouge

    Google Scholar 

  • Liu Z, Zhang QL, Wan XY, Huang J (2015) Development of real-time PCR assay for detection of microsporidian Enterocytozoon hepatopenaei and detection in shrimp samples under different growth rates. Fish Sci. https://doi.org/10.11758/yykxjz.20150512003. (in Chinese, English abstract)

  • Liu YM, Qiu L, Sheng AZ, Wan XY, Cheng DY, Huang J (2017) Quantitative detection method of Enterocytozoon hepatopenaei using TaqMan probe real-time PCR. J Invertebr Pathol 151:191–196

    Article  CAS  PubMed  Google Scholar 

  • Pasharawipas T, Flegel TW (1994) A specific DNA probe to identify the intermediate host of a common microsporidian parasite of Penaeus merguiensis and P. monodon. Asian Fish Sci 7:157–167

    Google Scholar 

  • Rajendran KV, Shivam S, Praveena PE, Rajan JJ, Kumar TS, Avunje S, Jagadeesan V, Babu SP, Pande A, Krishnan AN, Alavandi SV (2016) Emergence of Enterocytozoon hepatopenaei (EHP) in farmed Penaeus (Litopenaeus) vannamei in India. Aquaculture 454:272–280

    Article  Google Scholar 

  • Refardt D, Canning EU, Mathis A, Cheney SA, Lafranchi-Tristem NJ, Ebert D (2002) Small subunit ribosomal DNA phylogeny of microsporidia that infect Daphnia (Crustacea: Cladocera). Parasitology 124(4):381–389

    Article  CAS  PubMed  Google Scholar 

  • Sambrook J, Russel DW (2001) Molecular cloning: a laboratory manual. Third ed. 5. CSH Laboratory Press, Cold Spring Harbor, p 62

    Google Scholar 

  • Srinivas D, Venkatrayulu C, Swapna B (2016) Sustainability of exotic shrimp Litopenaeus vannamei (Boone, 1931) farming in coastal Andhra Pradesh, India: problems and issues. Eur J Exp Biol 6(3):80–85

    Google Scholar 

  • Sritunyalucksana K, Sanguanrut P, Salachan PV, Thitamadee S, Flegel TW (2014) Urgent appeal to control spread of the shrimp microsporidian parasite Enterocytozoon hepatopenaei (EHP). NACA Publication. https://enaca.org/publications/health/disease-cards/urgent-appeal-to-control-ehp-3.pdf. Accessed 20 March 2015

  • Stentiford GD, Bateman KS, Longshaw M, Feist SW (2007) Enterospora canceri n. gen., n. sp., intranuclear within the hepatopancreatocytes of the European edible crab Cancer pagurus. Dis Aquat Org 75(1):61–72

    Article  CAS  Google Scholar 

  • Suebsing R, Prombun P, Srisala J, Kiatpathomchai W (2013) Loop-mediated isothermal amplification combined with colorimetric nanogold for detection of the microsporidian Enterocytozoon hepatopenaei in penaeid shrimp. J Appl Microbiol 114(5):1254–1263

    Article  CAS  PubMed  Google Scholar 

  • Tamura K, Stecher G, Peterson D, Filipski A, Kumar S (2013) MEGA6: molecular evolutionary genetics analysis version 6.0. Mol Biol Evol 30(12):2725–2729

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Tang KF, Pantoja CR, Redman RM, Han JE, Tran LH, Lightner DV (2015) Development of in situ hybridization and PCR assays for the detection of Enterocytozoon hepatopenaei (EHP), a microsporidian parasite infecting penaeid shrimp. J Invertebr Pathol 130:37–41

    Article  CAS  PubMed  Google Scholar 

  • Tang KF, Han JE, Aranguren LF, White-Noble B, Schmidt MM, Piamsomboon P, Risdiana E, Hanggono B (2016) Dense populations of the microsporidian Enterocytozoon hepatopenaei (EHP) in feces of Penaeus vannamei exhibiting white feces syndrome and pathways of their transmission to healthy shrimp. J Invertebr Pathol 140:1–7

    Article  PubMed  Google Scholar 

  • Tangprasittipap A, Srisala J, Chouwdee S, Somboon M, Chuchird N, Limsuwan C, Srisuvan T, Flegel TW, Sritunyalucksana K (2013) The microsporidian Enterocytozoon hepatopenaei is not the cause of white feces syndrome in whiteleg shrimp Penaeus (Litopenaeus) vannamei. BMC Vet Res 9(1):139

    Article  PubMed  PubMed Central  Google Scholar 

  • Tourtip S, Wongtripop S, Stentiford GD, Bateman KS, Sriurairatana S, Chavadej J, Sritunyalucksana K, Withyachumnarnkul B (2009) Enterocytozoon hepatopenaei sp. nov.(Microsporida: Enterocytozoonidae), a parasite of the black tiger shrimp Penaeus monodon (Decapoda: Penaeidae): fine structure and phylogenetic relationships. J Invertebr Pathol 102(1):21–29

    Article  CAS  PubMed  Google Scholar 

  • Upadhyay AS (2001) Shrimp and prawn farming development in West Bengal: present status constrains and strategy. Fishing Chimes 20:91–98

    Google Scholar 

Download references

Acknowledgments

The authors are thankful to Dr. Joykrushna Jena, Deputy Director General (Fishery Science), Indian Council of Agricultural Research, New Delhi, for his support and guidance. The authors would like to express their sincere thanks to Mr. Asim Kumar Jana and Mr. Hirak Jyoti Chakraborty for helping in sampling, laboratory assistance and data analysis.

Funding

This work was funded by the National Fisheries Development Board (NFDB) (G/Nat. Surveillance/2013 dated 16.08.2013), Hyderabad under “National Surveillance Program for Aquatic Animal Diseases (NSPAAD)” Project.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to B. K. Behera.

Ethics declarations

Conflict of interest

The authors declare that there is no conflict of interest.

Additional information

Publisher’s note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Behera, B.K., Das, A., Paria, P. et al. Prevalence of microsporidian parasite, Enterocytozoon hepatopenaei in cultured Pacific White shrimp, Litopenaeus vannamei (Boone, 1931) in West Bengal, East Coast of India. Aquacult Int 27, 609–620 (2019). https://doi.org/10.1007/s10499-019-00350-0

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10499-019-00350-0

Keywords

Navigation