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Aims: Microalgae are ubiquitous in aquatic environments, including aquaculture farms, but few studies have delved into their phytoplankton diversity and bioremediation potential. In this study, the cultivable phytoplankton of a rainbow trout freshwater aquaculture farm was isolated, phylogenetically analysed and used to assemble a consortium to polish an aquaculture derived effluent, with low concentrations of ammonium, nitrite and nitrate. Methods and Results: Through standard plating in different selective media, a total of 15 microalgae strains were isolated from sludge from a rotary drum filtering system which removes suspended solids from the water exiting the facility. Based on 18S rRNA gene sequences, isolates were assigned to nine different genera of the Chlorophyta phylum: Asterarcys, Chlorella, Chloroccocum, Chlorosarcinopsis, Coelastrella, Desmodesmus, Micractinium, Parachlorella and Scenedesmus. Species from most of these genera are known to inhabit freshwater systems in Galicia and continental Spain, but the Coelastrella, Asterarcys or Parachlorella genera are not usually present in freshwater streams. In an onsite integrative approach, the capacity of a consortium of native microalgae isolates to grow on aquaculture derived effluents and its nutrient removal capacity were assessed using a raceway pond. After 7 days, removal efficiencies of approximately 99%, 92% and 49% for ammonium, nitrite and nitrate, respectively, were achieved concomitantly with a microalgae biomass increase of ca. 17%. Conclusions: Sludge from the aquaculture filtering system presents a high diversity of microalgae species from the Chlorophyta phylum, whose application in a consortial approach revealed to be efficient to polish aquaculture derived effluents with low nutrient content. Significance and Impact of the Study: The use of native microalgae consortia from aquaculture systems can contribute to the development of efficient treatment systems for low nutrient wastewater, avoiding nutrients release to the environment and promoting water recirculation. This may further strengthen the use of phycoremediation at the industrial scale, as an environment friendly strategy.

The dissolved oxygen (DO) concentration in water streams is one of the most important and critical quality parameters in aquaculture farms. The main objective of this study was to evaluate the potential of two Continuous Flow Granular Reactors, one based on Partial Nitrification-Anammox biomass (Aquammox CFGR) and the other on Microalgae-Bacteria biomass (AquaMab CFGR), for improving dissolved oxygen availability in the recirculation aquaculture systems (RAS). Both reactors treated the extremely low-strength effluents from a freshwater trout farm (1.39 mg NH4+-N/L and 7.7 mg TOC/L). The Aquammox CFGR, removed up to 68% and 100% of ammonium and nitrite, respectively, but the DO concentration in the effluent was below 1 mg O2/L while the anammox activity was not maintained. In the AquaMab CFGR, bioaugmentation of aerobic granules with microalgae was attained, producing an effluent with DO concentrations up to 9 mg O2/L and removed up to 77% and 80% of ammonium and nitrite, respectively, which is expected to reduce the aeration costs in fish farms The authors would like to thank the EU, the Spanish Government (AEI) (PCIN-2017-047) and Fundação para a Ciência e Tecnologia (FCT) (Water JPI/0003/2016) for funding, in the frame of the collaborative international Consortium AQUAVAL financed under the ERA-NET WaterWorks2015 Cofunded Call. This ERA-NET is an integral part of the 2016 Joint Activities developed by the Water Challenges for a Changing World Joint Programme Initiative (Water JPI) and the CDTI (Centro para Desarrollo Tecnológico Industrial, E.P.E., Spain). Authors also thank the Spanish Government (AEI) for funding in the frame of the project TREASURE (CTQ 2017-83225-C2-1-R) and the FCT for funding in the frame of the project UIDB/50016/2020. S. Santorio, A. Val del Rio and A. Mosquera-Corral belong to the Galician Competitive Research Groups (GRC)_ED431C-2021/37 co-funded by FEDER (EU) SI

Lactococcus garvieae, Yersinia ruckeri and Flavobacterium psychrophilum are three of the most important pathogens in worldwide rainbow trout aquaculture (Oncorrhynchus mykiss Walbaum). In this work, a multiplex quantitative PCR (qPCR) method for the simultaneous detection of the three pathogens was developed and tested in a commercial trout farm. Fifty-four spleen and brain samples from a trout farm were analyzed using both multiplex qPCR and classic microbiological methods. When comparing qPCR results and classic plate diagnostic techniques no negative deviations were observed, which resulted in a 100% relative sensitivity of the multiplex qPCR method developed. In addition, efficiency of the qPCR method ranged between 97.5% and 108.8%. The specificity of the combination of primers and probes was successfully tested in 57 target and non-target bacterial strains Hence, the multiplex qPCR method developed in the present study could be used as a reliable diagnostic tool for the detection of L. garvieae, Y. ruckeri and F. psychrophilum in rainbow trout farms, enabling faster diagnostics (only a few hours) and contributing to a quick response and management of bacterial disease outbreaks.

Ammonium and nitrite levels in water are crucial for fish health preservation and growth maintenance in freshwater aquaculture farms, limiting water recirculation. The aim of the present work was the evaluation and comparison of two granular sludge reactors which were operated to treat freshwater aquaculture streams at laboratory-scale: an Aerobic Granular Sludge - Sequencing Batch Reactor (AGS-SBR) and a Continuous Flow Granular Reactor (CFGR). Both units were fed with a synthetic medium mimicking an aquaculture recycling water (1.9–2.9 mg N/L), with low carbon content, and operational temperature varied between 17 and 25 °C. The AGS-SBR, inoculated with mature granules from a full-scale wastewater treatment plant, achieved high carbon and ammonium removal during the 157 operational days. Even at low hydraulic retention time (HRT), varying from 474 to 237 min, ammonium removal efficiencies of approximately 87–100% were observed, with an ammonium removal rate of approximately 14.5 mg NH4+-N/(L⋅d). Partial biomass washout occurred due to the extremely low carbon and nitrogen concentrations in the feeding, which could only support the growth of a small portion of bacteria, but no major changes on the reactor removal performance were observed. The CFGR was inoculated with activated sludge and operated for 98 days. Biomass granulation occurred in 7 days, improving the settling properties due to a high up-flow velocity of 11 m/h and an applied HRT of 5 min. The reactor presented mature granules after 32 days, achieving an average diameter of 1.9 mm at day 63. The CFGR ammonium removal efficiencies were of approximately 10–20%, with ammonium removal rates of 90.0 mg NH4+-N/(L⋅d). The main biological processes taking place in the AGS-SBR were nitrification and heterotrophic growth, while in the CFGR the ammonium removal occurred only by heterotrophic assimilation, with the reactor also presenting complete and partial denitrification, which caused nitrite production. Comparing both systems, the CFGR achieved 6 times higher ammonium removal rates than the AGS-SBR, being suitable for treating extremely high flows. On the other hand, the AGS-SBR removed almost 100% of ammonium content in the wastewater, discharging a better quality effluent, less toxic for the fish but treated lower flows.