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An Improved Microscopic Method for The Rapid Diagnosis of Emerging Microsporidian Parasite, Enterocytozoon hepatopenaei in Shrimp Farms


Affiliations
1 Aquatic Animal Health and Environment Division, ICAR-Central Institute of Brackishwater Aquaculture, 75, Santhome High Road, R.A. Puram, Chennai 600 028, India
 

Penaeus (Litopenaeus) vannamei shrimp samples (hepatopancreas and faeces) collected from grow-out farms were evaluated for the presence of newly emerged microsporidian parasite Enterocytozoon hepatopenaei by concentration techniques using either water–ether sedimentation or Sheather’s sugar solution for spore recovery and subsequent microscopic detection by modified trichrome stain (Ryan-blue method). This improved selective staining protocol can easily resolve the identification of microsporidian spores in hepatopancreatic tubules compared to conventional stains. This method enables differential diagnosis of microsporidian spores by a characteristic staining pattern of pinkish-red, often with a belt-like diagonal stripe seen in the middle of the spore and a halo of unstained area at one end which could be easily distinguished from similarly staining particles or debris.

Keywords

Enterocytozoon hepatopenaei, Microsporidia, Trichrome Staining, Shrimp Aquaculture.
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  • Stentiford, G. D., Feist, S. W., Stone, D. M., Bateman, K. S. and Dunn, A. M., Microsporidia: diverse, dynamic, and emergent pathogens inaquatic systems. Trends Parasitol., 2013, 29, 567– 578.

  • Vavra, J. and Lukes, J., Microsporidia and ‘the art of living together’. Adv. Parasitol., 2013, 82, 253–319.

  • Tourtip, S. et al., 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., 2009, 102, 21–29.

  • Tangprasittipap, A. et al., The microsporidian Enterocytozoon hepatopenaei is not the cause of white feces syndrome in whiteleg shrimp Penaeus (Litopenaeus) vannamei. BMC Vet. Res., 2013, 9, 139–148.

  • NACA, Hepatopancreatic microsporidiosis caused by Enterocytozoon hepatopenaei: disease card, Network of Aquaculture Centres in Asia-Pacific (NACA), Bangkok, Thailand, 2015; www.enaca.org

  • Thitamadee, S. et al., Review of current disease threats for cultivated penaeid shrimp in Asia. Aquaculture, 2016, 452, 69–87.

  • CIBA, Annual Report 2014–15. Central Institute of Brackishwater Aquaculture, Chennai, 2015, p. 96; www.ciba.res.in

  • Rajendran, K. V. et al., Emergence of Enterocytozoon hepatopenaei (EHP) in farmed Penaeus (Litopenaeus) vannamei in India. Aquaculture, 2016, 454, 272–280.

  • Didier, E. S., Vossbrinck, C. R., Stovall, M. E., Green, L. C., Bowers, L., Fredenburg, A. and Didier, P. J., Diagnosis and epidemiology of microsporidia infections in humans. Southeast Asian J. Trop. Med. Public Health (Suppl.), 2004, 35, 65–81.

  • Bukhari, Z. and Smith, H. V., Effect of three concentration techniques on viability of Cryptosporidium parvum oocysts recovered from bovine faeces. J. Clin. Microbiol., 1995, 33, 2592–2595.

  • Dryden, M. W., Payne, P. A., Ridley, R. and Smith, V., Comparison of common fecal flotation techniques for the recovery of parasite eggs and oocysts. Vet. Ther., 2005, 6, 15–28.

  • Ryan, N. J. et al., A new trichrome blue stain for detection of microsporidial species in urine, stool and nasopharyngeal specimens. J. Clin. Microbiol., 1993, 31, 3264–3269.

  • Lom, J., On a new taxonomic character in myxosporidia, as demonstrated in description of two new species of Myxobolus. Folia Parasitol., 1969, 16, 97–103.

  • Bell, T. A. and Lightner, D. V., A Handbook of Normal Penaeid Shrimp Histology, World Aquaculture Society, Baton Rouge, LA, USA, 1988, p. 114.

  • Tang, K. F. J., Pantoja, C. R., Redman, R. M., Han, J. E., Tran, L. H. and Lightner, D. V., Development of in situ hybridization and PCR assays for the detection of Enterocytozoon hepatopenaei (EHP), a microsporidian parasite infecting penaeid shrimp. J. Invertebr. Pathol., 2015, 130, 37–41.

  • Weber, R., Bryan, R. T., Owen, R. L., Wilcox, C. M., Gorelkin, L. and Visvesvara, G. S., Improved light-microscopical detection of microsporidia spores in stool and duodenal aspirates. N. Engl. J. Med., 1992, 326, 161–166.


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  • An Improved Microscopic Method for The Rapid Diagnosis of Emerging Microsporidian Parasite, Enterocytozoon hepatopenaei in Shrimp Farms

Abstract Views: 256  |  PDF Views: 75

Authors

P. Ezhil Praveena
Aquatic Animal Health and Environment Division, ICAR-Central Institute of Brackishwater Aquaculture, 75, Santhome High Road, R.A. Puram, Chennai 600 028, India
T. Bhuvaneswari
Aquatic Animal Health and Environment Division, ICAR-Central Institute of Brackishwater Aquaculture, 75, Santhome High Road, R.A. Puram, Chennai 600 028, India
A. Navaneeth Krishnan
Aquatic Animal Health and Environment Division, ICAR-Central Institute of Brackishwater Aquaculture, 75, Santhome High Road, R.A. Puram, Chennai 600 028, India
V. Jagadeesan
Aquatic Animal Health and Environment Division, ICAR-Central Institute of Brackishwater Aquaculture, 75, Santhome High Road, R.A. Puram, Chennai 600 028, India
J. Joseph Sahaya Rajan
Aquatic Animal Health and Environment Division, ICAR-Central Institute of Brackishwater Aquaculture, 75, Santhome High Road, R.A. Puram, Chennai 600 028, India
K. P. Jithendran
Aquatic Animal Health and Environment Division, ICAR-Central Institute of Brackishwater Aquaculture, 75, Santhome High Road, R.A. Puram, Chennai 600 028, India

Abstract


Penaeus (Litopenaeus) vannamei shrimp samples (hepatopancreas and faeces) collected from grow-out farms were evaluated for the presence of newly emerged microsporidian parasite Enterocytozoon hepatopenaei by concentration techniques using either water–ether sedimentation or Sheather’s sugar solution for spore recovery and subsequent microscopic detection by modified trichrome stain (Ryan-blue method). This improved selective staining protocol can easily resolve the identification of microsporidian spores in hepatopancreatic tubules compared to conventional stains. This method enables differential diagnosis of microsporidian spores by a characteristic staining pattern of pinkish-red, often with a belt-like diagonal stripe seen in the middle of the spore and a halo of unstained area at one end which could be easily distinguished from similarly staining particles or debris.

Keywords


Enterocytozoon hepatopenaei, Microsporidia, Trichrome Staining, Shrimp Aquaculture.

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DOI: https://doi.org/10.18520/cs%2Fv115%2Fi4%2F758-763