Adverse Effects of Immobilised Pseudoalteromonas on the Fish Pathogenic Vibrio anguillarum: An In Vitro Study

Journal of Marine Biology, Sep 2016

As a prerequisite for use in marine aquaculture, two immobilisation systems were developed by employing the probiotic bacterium Pseudoalteromonas sp. strain MLms_gA3. Their impact on the survivability of the fish pathogen Vibrio anguillarum was explored. Probiotic bacteria either grown as a biofilm on ceramic tiles or embedded in alginate beads were added to sterile artificial seawater that contained the fish pathogen. While immobilisation on ceramics followed a recently developed protocol, a medium allowing for alginate microencapsulation was newly developed. Anti-Vibrio activities were obtained with both immobilisation systems. The viable cell counts of V. anguillarum constantly decreased within the first two weeks of the treatments evidencing the potential of the immobilisation systems for providing probiotic-based protection against this pathogen.

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Adverse Effects of Immobilised Pseudoalteromonas on the Fish Pathogenic Vibrio anguillarum: An In Vitro Study

Adverse Effects of Immobilised Pseudoalteromonas on the Fish Pathogenic Vibrio anguillarum: An In Vitro Study Wiebke Wesseling,1 Michael Lohmeyer,1 Sabine Wittka,1 Julia Bartels,2 Stephen Kroll,2,3 Christian Soltmann,4 Pia Kegler,5 Andreas Kunzmann,5 Sandra Neumann,6 Burkhard Ramsch,6 Beate Sellner,6 and Friedhelm Meinhardt7 1Mikrobiologisches Labor Dr. Michael Lohmeyer GmbH, Mendelstraße 11, 48149 Münster, Germany 2Advanced Ceramics, University of Bremen, Am Biologischen Garten 2, 28359 Bremen, Germany 3MAPEX Center for Materials and Processes, Bibliothekstraße 1, 28359 Bremen, Germany 4Novelpor UG, Huchtinger Heerstraße 47, 28259 Bremen, Germany 5Leibniz-Center for Tropical Marine Ecology GmbH, Fahrenheitstraße 6, 28359 Bremen, Germany 6AquaCare GmbH & Co. KG, Am Wiesenbusch 11, 45966 Gladbeck, Germany 7Institut für Molekulare Mikrobiologie und Biotechnologie, Westfälische Wilhelms-Universität Münster, Corrensstraße 3, 48149 Münster, Germany Received 7 April 2016; Revised 5 July 2016; Accepted 28 July 2016 Academic Editor: Horst Felbeck Copyright © 2016 Wiebke Wesseling et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Abstract As a prerequisite for use in marine aquaculture, two immobilisation systems were developed by employing the probiotic bacterium Pseudoalteromonas sp. strain MLms_gA3. Their impact on the survivability of the fish pathogen Vibrio anguillarum was explored. Probiotic bacteria either grown as a biofilm on ceramic tiles or embedded in alginate beads were added to sterile artificial seawater that contained the fish pathogen. While immobilisation on ceramics followed a recently developed protocol, a medium allowing for alginate microencapsulation was newly developed. Anti-Vibrio activities were obtained with both immobilisation systems. The viable cell counts of V. anguillarum constantly decreased within the first two weeks of the treatments evidencing the potential of the immobilisation systems for providing probiotic-based protection against this pathogen. 1. Introduction It is not surprising that probiotics gain more and more importance for human and animal nutrition and welfare as well as for aquaculture, even for marine ornamental fish purposes [1]. Antibiotics, the alternative, frequently provoke resistant phenotypes in harmful germs and, thus, their administration must in general be considered as a critical issue (reviewed in [2]). Also, antibiotics should, and customarily are, being administered solely upon emergence of clear signs of microbial diseases, whereas bacteria with “antagonism toward pathogens” [3] that is probiotics can prophylactically exert their protective capacities already from the time of stocking and subsequently during all phases of life. There are promising previous studies from temperate areas, revealing probiotic protection for a number of different fish species, such as Atlantic cod, Atlantic salmon, and turbot [4–9]. The hitherto commercially available probiotic products for use in aquaculture are usually powders or fluids (such as Sanolife® MIC-F (http://shrimpcare.com/newwp/inve-mic-f-probiotic Access: 12th August 2016) or DrTim’s Aquatics® Eco-Balance Probiotic Bacteria for Reef, Nano and Seahorse Aquaria (http://store.drtimsaquatics.com/Eco-Balance-Probiotic-Bacteria-for-Reef-Nano-and-Seahorse-Aquaria_p_389.html Access: 9th July 2015.), resp.), which are readily water soluble and, thus, are immediately available for the fish. It remains, however, at least uncertain whether probiotic planktonic bacteria can withstand intestinal environments, where they encounter low pH values as well as oxygen limitation. Also, there is a risk of being (completely) absorbed within the aquarium’s filter system. Hence, immobilised probiotics, such as Shewanella putrefaciens in alginate beads, were previously tested for oral administration in sole [10]. Moreover, it was proven in vitro that microencapsulation protects bacteria against the action of digestive enzymes and low pH values [11]. Immobilisation is in general considered to give the cells shelter from harmful conditions while the immobilised bacteria are in the stationary growth phase and efficiently produce secondary metabolites [12]. Although such secondary metabolites are known to be generated by free-living bacteria as well, immobilised microorganisms are probably more potent producers [13, 14]. Secretion of bioactive molecules is indeed rather common for bacteria embedded in particles/beads or fixed on solid surfaces [8, 15–18]. A number of bacteria immobilised in extracellular polymer substances (EPS) were seen to detach from the biofilm and upon dispersal into the environmental fluid they became directly fish-available similar to the nonimmobilised (fluid and powder) formulations [19]. There is also the risk that bacterial bioactiv (...truncated)


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Wiebke Wesseling, Michael Lohmeyer, Sabine Wittka, Julia Bartels, Stephen Kroll, Christian Soltmann, Pia Kegler, Andreas Kunzmann, Sandra Neumann, Burkhard Ramsch, Beate Sellner, Friedhelm Meinhardt. Adverse Effects of Immobilised Pseudoalteromonas on the Fish Pathogenic Vibrio anguillarum: An In Vitro Study, Journal of Marine Biology, 2016, 2016, DOI: 10.1155/2016/3683809