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Comparing Methods for Measuring Phytoplankton Biomass in Aquaculture Ponds

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Comparing Methods for Measuring Phytoplankton Biomass in Aquaculture Ponds Vol. 8: 665–673, 2016 AQUACULTURE ENVIRONMENT INTERACTIONS Published December 29 doi: 10.3354/aei00208 Aquacult Environ Interact OPENPEN ACCESSCCESS Comparing methods for measuring phytoplankton biomass in aquaculture ponds Yi-Kuang Wang1,*, Pei-Yu Chen1, Hans-Uwe Dahms2,3, Shinn-Lih Yeh4, Ying-Jer Chiu4 1Department of Ecoscience and Ecotechnology, National University of Tainan, 33, Sec. 2, Shu-Lin St., Tainan City 70005, Taiwan, ROC 2Department of Biomedical Science and Environmental Biology, Kaohsiung Medical University, 100 Shih-Chuan 1st Road, Kaohsiung City 80424, Taiwan, ROC 3Department of Marine Biotechnology and Resources, National Sun Yat-Sen University, No. 70, Lienhai Road, Kaohsiung City 80424, Taiwan, ROC 4Mariculture Research Center, Fisheries Research Institute, the Council of Agriculture, 4 Haipu, Sangu village, Cigu District, Tainan City 72442, Taiwan, ROC ABSTRACT: Algal biomass is important for aquaculture in terms of eutrophication, as a secondary metabolite threat, and as a food source for suspension feeders and benthos. In order to evaluate the applicability of an in situ FluoroProbe instrument, chlorophyll a (chl a) measured with this device was compared to chl a measured by a fluorometer and to algal densities and biovolumes determined by direct counts. Monthly surveys were conducted in 3 types of aquaculture ponds (tilapia, milk fish, and common orient clam) from October 2011 through April 2012 in the Cigu region of Tainan City, Taiwan. Results showed that green algae were the predominant phyto- plankton group, followed by cyanobacteria, euglenoids, and diatoms. The 3 types of aquaculture −3 ponds had wide ranges of chl a, pH, salinity, turbidity, PO4 -P, and suspended solids. Chl a measured by the FluoroProbe had significant regressions with chl a values measured by a fluo- rometer, and with algal densities and biovolumes determined by counting; however, when chl a was >250 µg l−1, the correlation between the FluoroProbe and the fluorometer diminished. The difference in chl a reads between these 2 methods increased when chl a concentration exceeded 200 µg l−1. Of the 4 differently colored algal groups measured by the FluoroProbe, only the green and blue groups had significant regressions with their respective biovolumes, whereas the red and brown groups had no significant regression with their respective biovolumes. Our results show the applicability of the FluoroProbe in algal monitoring of aquaculture ponds, although caution is needed at higher chl a levels. KEY WORDS: FluoroProbe · Phytoplankton · Fluorometer · Algal density · Chlorophyll a INTRODUCTION zooplankton or larvae of fish and invertebrates) or from the benthic environment (e.g. bivalves or by Aquaculture production can be positively or nega- epistrata feeders, such as fish, shrimps, or poly- tively affected by microalgal biomass (Zimba et al. chaetes) (Uddin et al. 2009, Li et al. 2015, Zhang et 2001, Hemaiswarya et al. 2011, Anand et al. 2013, al. 2016). Algae may also cause problems in aqua- Hu et al. 2013). Algal primary production can be culture (Paerl & Tucker 1995, Alonso-Rodríguez & used by suspension feeders in the plankton (such as Páez-Osuna 2003, Casé et al. 2008), as they may © The authors 2016. Open Access under Creative Commons by *Corresponding author: [email protected] Attribution Licence. Use, distribution and reproduction are un - restricted. Authors and original publication must be credited. Publisher: Inter-Research · www.int-res.com 666 Aquacult Environ Interact 8: 665–673, 2016 deteriorate the cultures of farmed organisms by MATERIALS AND METHODS oxygen depletion at night or after death (Dahms 2014), and by producing toxins that threaten both Study area the health of the cultured or ganisms and the health of consumers (Ettoumi et al. 2011, Florczyk et al. The surveyed aquaculture ponds were situated in 2014). Algae may impart a bad smell and taste to the Cigu region of Tainan City in southern Taiwan aquaculture products (Zimba & Grimm 2003, Smith (Fig. 1). We investigated 3 common types of aqua - et al. 2008, Green & Schrader 2015). Due to the culture ponds in this area, where the following prod- abovementioned problems, the measurement of bio- ucts were raised: tilapia (Oreochromis spp.), milk fish mass, density, and diversity of microalgae has Chanos chanos, and common orient clam Meretrix become a critical issue in aquaculture management lusoria (hereafter simply ‘clam’). Each aquaculture (Mischke 2014, Kurten & Barkoh 2016, Pucher et al. type had 3 ponds. A canal, connected to a water 2016). However, algal measurements are often time- source for aquaculture ponds, was also surveyed to consuming and require taxonomic expertise, which expand the range of algal levels. Tilapia ponds had is not practical for farmers. The recent development water depths from 1.8 to 2.5 m, areas from 0.3 to of a fast, easy-operating, in situ measuring instru- 0.7 ha, a density of about 12 000 fish ha−1, and a ment, the FluoroProbe, has improved the aforemen- growth period about 1 yr. Milk fish ponds had water tioned shortcomings of algal measurements (Beutler depths from 0.7 to 1.2 m, areas from 0.45 to 1.2 ha, et al. 2002, Rolland et al. 2010). densities of 7000 fish ha−1, and a growth period of Several studies have applied the submersible Fluoro Probe in reser- voirs and rivers (Leboulanger et al. 2002, Gregor & Maršálek 2004, Gre- gor et al. 2005, Rolland et al. 2010, Catherine et al. 2012). These studies indicated that measurements made with a sub mersible FluoroProbe and the spectro photometer method were similar at low chlorophyll a (chl a) le - vels, where as the FluoroProbe might underestimate chl a at high ambient Fig. 1. Sampling loca- chl a levels (Gregor & Maršálek 2004, tions of aquaculture Rolland et al. 2010). The above stud- ponds in Tainan City, ies were conducted in environments Taiwan with relatively low chl a. To date, no study has tested the FluoroProbe in aquaculture ponds, where relatively high chl a levels can be reached. Therefore, such an evaluation seems timely. The objective of this study was to assess whether the FluoroProbe is applicable to the particular environ- mental conditions of aquaculture ponds, particularly to high densities of planktonic microalgae. Two ap- proa ches were taken: (1) to compare results of the FluoroProbe to that of a fluorometer; and (2) to compare the results of overall and individual algal groups measured by the FluoroProbe to algal density and biovolume meas- urements made by direct counting. Wang et al.: Comparison of phytoplankton measurements 667 less than 1 yr. Clam ponds had a water depth of about algae, algal samples were fixed onto a slide using the 1.1 m, areas from 1.6 to 4.8 ha, and a cultivating syrup method (Stevenson 1984). Subsamples of period of 6 mo to 1 yr. Water in the tilapia and milk phytoplankton of known volumes were added to fish ponds was not changed unless the water quality 0.2 ml of 10% syrup that covered a 22 × 22 mm cover had severely deteriorated, whereas water in the clam slip (Stevenson 1984), and then dried in the dark and ponds was flushed by the tides. sealed onto a slide by nail polish. Algal taxa were identified at 1000× magnification under a compound microscope (Olympus BX51) and then counted at Sampling and sample processing 400×. Biovolumes of algae were measured according to the shape measurements of Hillebrand et al. Samples were taken monthly from October 2011 to (1999). At least 300 natural units were recorded April 2012, when cultured organisms were harves- (Charles et al. 2002). For algal identification, Ding & ted. Field water quality metrics, including water Li (1991), Carmelo (1997), and Yamagishi (1992) temperature, pH, conductivity, and dissolved oxygen were consulted. (DO), were measured with a multiprobe meter (YSI Chl a was analyzed according to the ethanol ex - 556) and a DO meter (YSI DO 200). A Rutt ner sam- traction method of the National Institute of Environ- pler (Hydro-Bios) was used to collect water at depths mental Analysis (NIEA, Taoyuan, Taiwan; Protocol from 30 to 60 cm. Each phytoplankton sample was No. E509.01C). Chl a was concentrated on a filter 900 ml, and each chl a and water chemistry sample paper of 1 µm pore size and 47 mm in diameter was 300 ml. Phytoplankton samples were preserved (Whatman GF/C) by filtering 100 ml of the pond with 5% Lugol’s, whereas chl a and water chemistry water sample. The extraction was done by dissolving samples without preservatives were stored on ice in a chl a on filter paper with 10 ml of 99.5% ethanol in a dark container in the field. centrifuge tube, which was further soaked in a 60°C A FluoroProbe instrument (bbe-Moldaenke) was water bath with periodic shaking for 30 min. After used to measure chl a and 4 algal groups in situ. The extraction, the sample was centrifuged at 3500 × g sensor of the FluoroProbe was lowered to 50 cm be - (13 min). An aliquot from the upper layer of the sam- low the water surface, and values were read every 2 s ple was used for measurements of chl a by a fluoro - (25 times). The data were initially stored within the meter (multipurpose, Turner). FluoroProbe before statistics were applied as de - scribed below. The FluoroProbe commonly measures fluorescence Water chemistry emitted by Photosystem II core pigments at 680 nm as chl a (Beutler et al. 2002). It also uses different Four types of nutrients were measured in the + − fluo rescent wavelengths to separate 4 major algal laboratory: NH4 -N and NO3 -N were analyzed by groups by their absorption maxima (Beutler et al. the NOVA 60 water quality instrument (Merck), − −3 2002). The green group, which is rich in chl a and b, where as NO2 -N and PO4 -P were analyzed by the − includes chlorophytes and euglenophytes, whereas Cintra 20 spectrophotometer (GBC).
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