Abstract
Flavobacterial diseases, caused by multiple members of the Family Flavobacteriaceae, elicit serious losses in wild and farmed fish around the world. Flavobacteria are known to be transmitted horizontally; however, vertical transmission has been suspected but proven only for one fish-pathogenic flavobacterial species (e.g., Flavobacterium psychrophilum). Herein, we report on the isolation and molecular identification of multiple Flavobacterium and Chryseobacterium taxa from the ovarian fluid and eggs of feral Great Lakes Chinook salmon (Oncorhynchus tshawytscha). Identified egg- and ovarian fluid-associated flavobacteria were either well-known flavobacterial fish pathogens (e.g., F. psychrophilum and F. columnare), most similar to emerging fish-associated flavobacteria (e.g., F. spartansii, F. tructae, F. piscis, C. piscium, C. scophthalmum), or were distinct from all other described Chryseobacterium and Flavobacterium spp., as determined by phylogenetic analyses using neighbor-joining, Bayesian, and Maximum Likelihood methodologies. The gamete-associated flavobacteria fell into three groups (e.g., those that were recovered from the ovarian fluid but not eggs; those that were recovered from the ovarian fluid and eggs; and those that were recovered from eggs but not ovarian fluid), a portion of which were recovered from eggs that were surface disinfected with iodophor at the commonly used dose and duration for egg disinfection. Some gamete-associated flavobacteria were also found in renal, splenic, and neurological tissues. Systemic polymicrobial infections comprised of F. psychrophilum and F. columnare were also detected at nearly an 11% prevalence. This study highlights the potential role that sexual products of female Great Lakes Chinook salmon may play in the transmission of fish-associated flavobacteria.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
References
Agbonlahor, D.E., Odugbemi, T.O., and Udofia, P.O. 1983. Differentiation of Gram-positive and Gram-negative bacteria and yeasts using a modification of the “string” test. Am. J. Med. Technol. 49, 177–178.
Anacker, R.L. and Ordal, E.J. 1955. Study of a bacteriophage infecting the myxobacterium Chondrococcus columnaris. J. Bacteriol. 70, 738–741.
Barnes, M.E., Bergmann, D., Stephenson, H., Gabel, M., and Cordes, R.J. 2005. Bacterial numbers from landlocked fall Chinook salmon eyed eggs subjected to various formalin treatments as determined by scanning electron microscopy and bacteriological culture methods. N. Am. J. Aquacult. 67, 23–33.
Brown, L.L., Cox, W.T., and Levine, R.P. 1997. Evidence that the causal agent of bacterial cold-water disease Flavobacterium psychrophilum is transmitted within salmonid eggs. Dis. Aquat. Org. 29, 213–218.
Bullock, G., Hsu, T.C., and Shotts, E.Jr. 1986. Columnaris disease of fishes. Fish Disease Leaflet 72. U.S. Fish and Wildlife Service, Washington DC, USA.
Cai, W., De La Fuente, L., and Arias, C.R. 2013. Biofilm formation by the fish pathogen Flavobacterium columnare: development and parameters affecting surface attachment. Appl. Environ. Microbiol. 79, 5633–5642.
Chalupnicki, M., Dittman, D., Starliper, C.E., and Iwanowicz, D.D. 2015. Efficacy of iodine for disinfection of lake sturgeon eggs from the St. Lawrence River, New York. N. Am. J. Aquacult. 77, 82–89.
Cipriano, R.C. 2015. Bacterial analysis of fertilized eggs of Atlantic salmon from the Penobscot, Naraguagus, and Machias rivers, Maine. J. Aquat. Anim. Health 27, 172–177.
Cipriano, R.C., Novak, B.M., Flint, D.E., and Cutting, D.C. 2001. Reappraisal of the federal fish health recommendation for disinfecting eggs of Atlantic salmon in iodophor. J. Aquat. Anim. Health 13, 320–327.
Clark, S.E., Jude, B.A., Danner, G.R., and Fekete, F.A. 2009. Identification of a multidrug efflux pump in Flavobacterium johnsoniae. Vet. Res. 40, 55.
Cole, J.R., Wang, Q., Fish, J.A., Chai, B., McGarrell, D.M., Sun, Y., Brown, C.T., Porras-Alfaro, A., Kuske, C.R., and Tiedje, J.M. 2014. Ribosomal database project: data and tools for high throughput rRNA analysis. Nucleic Acids Res. 42, D633–642.
Johnson, S.L., Villarroel, M., Rosengrave, P., Carne, A., Kleffmann, T., Lokman, P.M., and Gemmell, N.J. 2014. Proteomic analysis of Chinook salmon (Oncorhynchus tshawytscha) ovarian fluid. PLoS One 9, e104155.
Kim, O.S., Cho, Y.J., Lee, K., Yoon, S.H., Kim, M., Na, H., Park, S.C., Jeon, Y.S., Lee, J.H., Yi, H., et al. 2012. Introducing EzTaxon-e: a prokaryotic 16S rRNA gene sequence database with phylotypes that represent uncultured species. Int. J. Syst. Evol. Microbiol. 62, 716–721.
Kumagai, A., Takahashi, K., Yamaoka, S., and Wakabayashi, H. 1998. Ineffectiveness of iodophore treatment in disinfecting salmonid eggs carrying Cytophaga psychrophila. Fish Pathol. 33, 123–128.
Lindstrom, N.M., Call, D.R., House, M.L., Moffitt, C.M., and Cain, K.D. 2009. A quantitative enzyme-linked immunosorbent assay and filtration-based fluorescent antibody test as potential tools to screen broodstock for infection with Flavobacterium psychrophilum. J. Aquat. Anim. Health 21, 43–56.
Lo, H.H. and Chang, S.M. 2014. Identification, characterization, and biofilm formation of clinical Chryseobacterium gleum isolates. Diagn. Microbiol. Infect. Dis. 79, 298–302.
Loch, T.P. and Faisal, M. 2014. Deciphering the biodiversity of fishpathogenic Flavobacterium spp. recovered from the Great Lakes basin. Dis. Aquat. Org. 112, 45–57.
Loch, T.P. and Faisal, M. 2015a. Emerging flavobacterial infections in fish: A review. J. Adv. Res. 6, 282–300.
Loch, T.P. and Faisal, M. 2015b. Polyphasic characterization reveals the presence of novel fish-associated Chryseobacterium spp. in the Great Lakes of North America. Dis. Aquat. Org. 113, 113–125.
Loch, T.P. and Faisal, M. 2016. Flavobacterium spartansii induces pathological changes and mortality in experimentally challenged Chinook salmon Oncorhynchus tshawytscha (Walbaum). J. Fish Dis. 39, 483–488.
Loch, T.P., Fujimoto, M., Woodiga, S.A., Walker, E.D., Marsh, T.L., and Faisal, M. 2013. Diversity of fish-associated flavobacteria of Michigan. J. Aquat. Anim. Health 25, 149–164.
Long, A., Call, D.R., and Cain, K.D. 2014. Investigation of the link between broodstock infection, vertical transmission, and prevalence of Flavobacterium psychrophilum in eggs and progeny of rainbow trout and coho salmon. J. Aquat. Anim. Health 26, 66–77.
Marchesi, J.R., Sato, T., Weightman, A.J., Martin, T.A., Fry, J.C., Hiom, S.J., and Wade, W.G. 1998. Design and evaluation of useful bacterium-specific PCR primers that amplify genes coding for bacterial 16S rRNA. Appl. Environ. Microbiol. 64, 795–799.
Michel, C., Matte-Tailliez, O., Kerouault, B., and Bernardet, J.F. 2005. Resistance pattern and assessment of phenicol agents’ minimum inhibitory concentration in multiple drug resistant Chryseobacterium isolates from fish and aquatic habitats. J. Appl. Microbiol. 99, 323–332.
Nilsen, H., Sundell, K., Duchaud, E., Nicolas, P., Dalsgaard, I., Madsen, L., Aspan, A., Jansson, E., Colquhoun, D.J., and Wiklund, T. 2014. Multilocus sequence typing identifies epidemic clones of Flavobacterium psychrophilum in Nordic countries. Appl. Environ. Microbiol. 80, 2728–2736.
OIE. 2013. Manual of diagnostic tests for aquatic animals. OIE, Paris, France.
Rambaut, A. 2009. FigTree v1. 3.1 2006-2009. Institute of Evolutionary Biology, University of Edinburgh, UK.
Ronquist, F., Teslenko, M., van der Mark, P., Ayres, D.L., Darling, A., Höhna, S., Larget, B., Liu, L., Suchard, M.A., and Huelsenbeck, J.P. 2012. MrBayes 3.2: efficient Bayesian phylogenetic inference and model choice across a large model space. Syst. Biol. 61, 539–542.
Starliper, C.E. 2011. Bacterial coldwater disease of fishes caused by Flavobacterium psychrophilum. J. Adv. Res. 2, 97–108.
Starliper, C. and Schill, W. 2011. Flavobacterial diseases: columnaris disease, coldwater disease, and bacterial gill disease. In Woo, P.T.K. and Bruno, D.B. (eds.), Fish diseases and disorders, pp. 606–631. CAB International, Oxfordshire, UK.
Sundell, K. and Wiklund, T. 2011. Effect of biofilm formation on antimicrobial tolerance of Flavobacterium psychrophilum. J. Fish Dis. 34, 373–383.
Tamura, K., Stecher, G., Peterson, D., Filipski, A., and Kumar, S. 2013. MEGA6: molecular evolutionary genetics analysis version 6.0. Mol. Biol. Evol. 30, 2725–2729.
Toyama, T., Kita-Tsukamoto, K., and Wakabayashi, H. 1994. Identification of Cytophaga psychrophila by PCR targeted 16S ribosomal RNA. Fish Pathol. 29, 271–275.
USFWS. 2012. Iodophor disinfection of eggs. Handbook of aquatic animal health procedures and protocols, Vol. 3: disinfection, isolation, and quarantine guidelines. United States Fish and Wildlife Service, USA.
Welker, T.L., Shoemaker, C.A., Arias, C.R., and Klesius, P.H. 2005. Transmission and detection of Flavobacterium columnare in channel catfish Ictalurus punctatus. Dis. Aquat. Org. 63, 129–138.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Loch, T.P., Faisal, M. Gamete-associated flavobacteria of the oviparous Chinook salmon (Oncorhynchus tshawytscha) in lakes Michigan and Huron, North America. J Microbiol. 54, 477–486 (2016). https://doi.org/10.1007/s12275-016-5629-3
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s12275-016-5629-3