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In situ enumeration and localization of the probiotic Pediococcus sp. strain Ab1 in the gut of abalone Haliotis gigantea

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  • Aquaculture
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Abstract

In this study, we established an optimized fluorescence in situ hybridization (FISH) protocol for detecting and enumerating host-derived probiotic Pediococcus sp. strain Ab1 cells in the gut of abalone. Samples were fixed with 4 % paraformaldehyde and hybridized at 40 °C for 3 h using a TAMRA (5-carboxytetramethylrhodamine)-labeled probe (Rpt probe). Strain Ab1 could be clearly discriminated from other control bacteria (Enterococcus, Lactobacillus). Fluorescence signals of Rpt-positive bacteria were easily distinguished from any detectable background noise. Rpt-positive bacteria were detected in both probiotic-supplemented and control abalone; nevertheless, the number of Rpt-positive bacteria in probiotic-supplemented abalone was two orders of magnitude higher than in controls. Furthermore, the mean number of Rpt-positive bacteria (probiotic-supplemented abalone) in the gut-attached section was also two orders of magnitude higher than in the free-living section. Rpt-positive micro-colonies (MCs) were observed by thin-section FISH analysis only in probiotic-supplemented abalone, and these MCs were observed in contact with intestinal epithelial cells. These results indicate that the probiotic Pediococcus sp. strain Ab1 can colonize or adhere to the abalone digestive tract in numbers sufficient for therapeutic effects. The data obtained using our FISH method provide clear and direct evidence of the formation of probiotic colonies in the gut of supplemented abalone.

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Acknowledgments

We thank Mr. Atsushi Hamabe of the Owase Aquaculture Center for kindly providing the abalone samples. This work was supported by the Strategic Young Researcher Overseas Visits Program for Accelerating Brain Circulation of the Japan Society for the Promotion of Science (JSPS).

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Authors and Affiliations

  1. Graduate School of Bioresources, Mie University, Kurimamachiya 1577, Tsu, Mie, 514-8507, Japan

    Reiji Tanaka, Hiroyuki Mitsuya, Mizuki Aoki & Taeko Miyazaki

  2. Research and Development Department, Hokkaido Industrial Technology Center, Kikyo 379, Hakodate, Hokkaido, 041-0801, Japan

    Masashi Ootsubo

  3. Laboratory of Aquaculture and Artemia Reference Center, Ghent University, Rozier 44, 9000, Ghent, Belgium

    Peter Bossier

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  1. Reiji Tanaka
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  2. Hiroyuki Mitsuya
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Correspondence to Reiji Tanaka.

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Tanaka, R., Mitsuya, H., Aoki, M. et al. In situ enumeration and localization of the probiotic Pediococcus sp. strain Ab1 in the gut of abalone Haliotis gigantea . Fish Sci 82, 481–489 (2016). https://doi.org/10.1007/s12562-016-0981-0

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  • DOI: https://doi.org/10.1007/s12562-016-0981-0

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