Munkongwongsiri, N. | bioRxiv Science

bioRxiv2021

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

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. HighlightsO_LIWhite feces syndrome (WFS) shrimp often harbor the microsporidian Enterocytozoon hepatopenaei (EHP) C_LIO_LIThe hepatopancreas (HP) and midgut of EHP-WFS shrimp had more EHP copies and spores than EHP-non.-WFS shrimp C_LIO_LIVibrio spp., Propionigenium sp. and EHP dominated in HP microbiomes of EHP-WFS shrimp C_LIO_LIPropionigenium copy numbers were uniquely high in the HP of EHP-WFS shrimp C_LIO_LIEHP-WFS shrimp also showed intestinal microbiomes of reduced diversity but more heterogeneity C_LI

molecular biology↗

bioRxiv2021

The false mussels (Mytilopsis leucophaeata) can be mechanical carriers of the shrimp microsporidian Enterocytozoon hepatopenaei (EHP)

Enterocytozoon hepatopenaei (EHP) is an obligate intracellular parasite causing hepatopancreatic microsporidiosis (HPM) in cultivated shrimp in Asian countries. One strategy to control EHP is to identify and eliminate biological reservoir(s) in shrimp ponds. Several marine and brackish-water organisms, including false mussels (Mytilopsis) have been reported to test positive for EHP using polymerase chain reaction (PCR) technology. Thus, we tested samples of commonly found Thai false mussel Mytilopsis leucophaeata from 6 EHP-infected shrimp ponds by PCR for the presence of EHP using the spore wall protein (SWP) gene primers. The mussel samples from all 6 ponds were positive. Subsequent bioassays carried out using naive mussels cohabitated with EHP-infected shrimp gave 100% SWP-PCR positive mussels at 20 days. One batch of such PCR-positive mussels was transferred for cohabitation with naive shrimp and gave 37.5% EHP-positive shrimp within 10 days. Tissue analysis of the EHP-PCR-positive mussels using light microscopy, in situ hybridization analysis for the SWP gene and electron microscopy did not confirm EHP infection. In summary, we obtained no evidence that Mytilopsis leucophaeata was itself infected with EHP. However, it was apparently capable carrying infectious spores for some period after ingestion and serving as a mechanical or passive carrier. The results support previous reports warning of the danger of feeding living or fresh bivalves to broodstock shrimp in hatcheries or shrimp in rearing ponds without prior heating or freezing.

zoology↗