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A potential prokaryotic and microsporidian pathobiome that may cause shrimp white feces syndrome (WFS)

Anuphap Prachumwat, Natthinee Munkongwongsiri, Wiraya Eamsaard, Kanokwan Lertsiri, Timothy W. Flegel, Grant D. Stentiford, Kallaya Sritunyalucksana
doi: https://doi.org/10.1101/2021.05.23.445355
Anuphap Prachumwat
1Aquatic Animal Health Research Team, Integrative Aquaculture Biotechnology Research Group, National Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency (NSTDA), Yothi office, Rama VI Rd., Bangkok, Thailand 10400
2Center of Excellence for Shrimp Molecular Biology and Biotechnology (Centex Shrimp), Faculty of Science, Mahidol University, Rama VI Rd., Bangkok, Thailand 10400
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Natthinee Munkongwongsiri
1Aquatic Animal Health Research Team, Integrative Aquaculture Biotechnology Research Group, National Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency (NSTDA), Yothi office, Rama VI Rd., Bangkok, Thailand 10400
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Wiraya Eamsaard
1Aquatic Animal Health Research Team, Integrative Aquaculture Biotechnology Research Group, National Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency (NSTDA), Yothi office, Rama VI Rd., Bangkok, Thailand 10400
2Center of Excellence for Shrimp Molecular Biology and Biotechnology (Centex Shrimp), Faculty of Science, Mahidol University, Rama VI Rd., Bangkok, Thailand 10400
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Kanokwan Lertsiri
1Aquatic Animal Health Research Team, Integrative Aquaculture Biotechnology Research Group, National Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency (NSTDA), Yothi office, Rama VI Rd., Bangkok, Thailand 10400
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Timothy W. Flegel
2Center of Excellence for Shrimp Molecular Biology and Biotechnology (Centex Shrimp), Faculty of Science, Mahidol University, Rama VI Rd., Bangkok, Thailand 10400
3National Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency (NSTDA), Klong Luang, Pathumthani, 12120, Thailand
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Grant D. Stentiford
4International Centre of Excellence for Aquatic Animal Health, Centre for Environment Fisheries and Aquaculture Science (Cefas), Weymouth Laboratory, Weymouth, Dorset DT4 8UB, UK
5Centre for Sustainable Aquaculture Futures, University of Exeter, College of Life and Environmental Sciences, University of Exeter, Exeter EX4 4QD, UK
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Kallaya Sritunyalucksana
1Aquatic Animal Health Research Team, Integrative Aquaculture Biotechnology Research Group, National Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency (NSTDA), Yothi office, Rama VI Rd., Bangkok, Thailand 10400
2Center of Excellence for Shrimp Molecular Biology and Biotechnology (Centex Shrimp), Faculty of Science, Mahidol University, Rama VI Rd., Bangkok, Thailand 10400
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  • For correspondence: kallaya@biotec.or.th
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Abstract

White feces syndrome (WFS) in shrimp cultivation ponds is characterized by the occurrence of shrimp with abnormal, white intestines (midguts) combined with large floating mats of white, shrimp fecal strings. The etiology for WFS is complex, similar to diarrhea in humans. EHP-WFS is a type of WFS characterized by massive quantities of spores from the microsporidian parasite Enterocytozoon hepatopenaei (EHP) together with mixed, unidentified bacteria in the shrimp hepatopancreas, midgut and fecal strings. However, WFS does not always develop in shrimp with severe EHP infections in controlled laboratory challenges. Further, in EHP-WFS outbreak ponds, some shrimp show white midguts (WG) while others in the same pond show grossly normal midguts (NG). We hypothesized that comparison of the microbial flora between WG and NG from the same EHP-WFS pond would reveal probable combinations of microbes significantly associated with EHP-WFS. To test this hypothesis, we selected a pond exhibiting a severe EHP-WFS outbreak in cultivated Penaeus vannamei and used a combination of microscopic and microbial profiling analyses to compare WG and NG samples. By histology, EHP plasmodia and spores were confirmed in the hepatopancreas (HP) and midgut of WG and NG shrimp, but pathological severity and spore quantity was higher in the WG shrimp. In addition, intestinal microbiomes in WG shrimp were less diverse and had higher abundance of bacteria from the genera Vibrio and Propionigenium. Propionigenium quantity in the HP of WG shrimp was significantly higher (P = 1.08e-5) than in NG shrimp (4,506 vs. 3 copies /100 ng DNA, respectively). These findings supported our hypothesis by revealing two candidate bacterial genera that should be tested in combination with EHP as a potential eukaryote-prokaryote pathobiome that causes EHP-WFS in P. vannamei.

Highlights

  • White feces syndrome (WFS) shrimp often harbor the microsporidian Enterocytozoon hepatopenaei (EHP)

  • The hepatopancreas (HP) and midgut of EHP-WFS shrimp had more EHP copies and spores than EHP-non.-WFS shrimp

  • Vibrio spp., Propionigenium sp. and EHP dominated in HP microbiomes of EHP-WFS shrimp

  • Propionigenium copy numbers were uniquely high in the HP of EHP-WFS shrimp

  • EHP-WFS shrimp also showed intestinal microbiomes of reduced diversity but more heterogeneity

Competing Interest Statement

The authors have declared no competing interest.

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
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Posted May 26, 2021.
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A potential prokaryotic and microsporidian pathobiome that may cause shrimp white feces syndrome (WFS)
Anuphap Prachumwat, Natthinee Munkongwongsiri, Wiraya Eamsaard, Kanokwan Lertsiri, Timothy W. Flegel, Grant D. Stentiford, Kallaya Sritunyalucksana
bioRxiv 2021.05.23.445355; doi: https://doi.org/10.1101/2021.05.23.445355
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A potential prokaryotic and microsporidian pathobiome that may cause shrimp white feces syndrome (WFS)
Anuphap Prachumwat, Natthinee Munkongwongsiri, Wiraya Eamsaard, Kanokwan Lertsiri, Timothy W. Flegel, Grant D. Stentiford, Kallaya Sritunyalucksana
bioRxiv 2021.05.23.445355; doi: https://doi.org/10.1101/2021.05.23.445355

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