Berglin, Mattias

Elwing, Hans

Jonsson, Per

Pansch, Christian

Pinori, Emiliano

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Balanus (Amphibalanus) improvisus in culture  •  Can be found worldwide •  By far the most common barnacle species in the brackish Baltic Sea •  All year round culture at the Sven Lovén Centre for Marine Sciences - Tjärnö (University of Gothenburg, Sweden) provides continuous supply of cyprids: •  Adults (collected in July) are kept in flow-through seawater (25 PSU & 20 °C) fed with Artemia salina ad libitum •  Larvae reared in 20 l buckets at 26 °C and fed diatom algae (Skeletonema marinoi, Chaetoceros calcitrans, Thalassiosira pseudonana) for 6 days •  Cyprids are collected and used in the diverse assays Preliminary Results  •  Preliminary tests with biocide-free panels show high settlement rates and demonstrate that settled barnacles can be monitored after metamorphosis to explore time-dependent post-settlement effect of biocides.  •  This novel flow-cell assay is now used to test for antifouling activity against barnacles, where the biocide effect is triggered when post-settlement barnacles penetrate the coating. ICMCFsg0193a0001 Lab-based all year round anti-fouling bioassay to screen for pre-  and post-settlement biocide activity against barnacles. Christian Pansch1, Per Jonsson1, Emiliano Pinori2, Mattias Berglin2 and Hans Elwing1  1 University of Gothenburg, Sweden, 2 SP Technical Research Institute of Sweden Rationale  •  Current antifouling technologies are mainly based on the continuous release of biocides and consequently discharge into the environment. •  Barnacles are important fouling organisms worldwide - largely increasing the hydrodynamic drag of ships and damaging coatings on underwater surfaces. •  Barnacles are extensively used as a model in antifouling research mostly in static, lab-based systems - however, reliable flow-through test assays for the screening of antifouling paints and its incorporated biocides are rare. •  We have developed a test assays to screen diverse low-release biocide paints and their pre- as well as post-settlement activity against barnacles. •  A minimum of 200 cyprids per flow cell •  Flow-through seawater (25 PSU & 20°C) •  Flow rates from 1.5 to 30 l per h •  Water level adjustable from 1/3 (65 ml) to 2/3 (250 ml) •  Pre-soaking of panels Counts of settlers and survivors every second day •  Feeding with diatom algae (Skeletonema marinoi, Chaetoceros calcitrans, Thalassiosira pseudonana) in 1.5 l beakers  •  Feeding with Artemia salina in 6 l aquaria The Flow Cell Assay  •  Test assay for coatings where antifouling activity is confined to the paint surface and biocides are bound to the paint matrix - antifouling activity exercised only when the fouling organism makes contact with the paint, e.g. at settlement or post-settlement penetration: •  Open flow cells with continuous flow-through seawater prevent accumulation of biocides  •  Design and nylon net ensure the panels to be the only surfaces available for settlement •  Choice or no-choice tests: each flow cell holds two treated or one treated and one control panel (11 x 11 cm) Contact  Christian Pansch  Dept. of Chemistry & Molecular Biology -  Tjärnö, University of Gothenburg, 45296 Strömstad, Sweden  From October 2014 at the GEOMAR - Kiel, Germany  E-Mail: ch.pansch@gmail.com www.leaf-antifouling.eu  GU    LEAF Flow-through seawater  (10/30 l per h, 25 PSU & 20°C) 0	  5	  10	  15	  20	  25	  B	   T	   R	   T	   B	   B	   R	   R	   T	   T	  1	   2	   3	   4	   5	  %	  Se%lement	  Settlement tests Post-settlement mortality tests Figure 1 - Settlement and post-settlement growth of B. improvisus on PMMA (Plexiglas®; T=transparent), LEAF-4.1 paint (R=red) and LEAF-A paint (B=black) without biocides. Figure 2 - Settlement of B. improvisus on PMMA (Plexiglas®) and LEAF-4.1 paint without (Co) and with biocide (Ab & Iv), photo=LEAF-4.1-Co. 0	  5	  10	  15	  20	  25	  PMMA	   Co	   Ab	   Iv	  %	  Se%lement	  Conclusions  This settlement assay produces all-year round reliable results in order to test for large-scale and more realistic anti-settlement activity of biocides against barnacles.  The settlement assay can be easily extended to screen for post-settlement activity of biocides.  It has the potential to be further applied in various other fields of anti-fouling research, in pre- and post-settlement inhibition studies.  This assays is at the moment applied within the European FP7 Collaborative project “LEAF 1”, aiming to develop sustainable antifouling solutions based on a newly discovered “Low Emission AntiFouling” paint concept. adult barnacle Nauplius II Nauplius VI Cypris Acknowledgements  The research leading to these results has received funding from the European Union Seventh Framework Programme (FP7/2007- 2013) under grant agreement n° 314697 (LEAF).  Travel was supported by ONR Award N62909-13-1-C196 and LEAF 500	  µm	  5000	  µm	  Flow cell Panel 

Lab-based all year round anti-fouling bioassay to screen for pre- and post-settlement biocide activity against barnacles

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