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1 J. of Aquaculture Vol. 19(4) : , 2006 µ Journal of Aquaculture Korean Aquaculture Society» cyclopoid ƒ Paracyclopina nana,» 1, x * w w, 1 w w œw Grazing of Brackish Water Cyclopoid Copepod Paracyclopina nana on Different Microalgae Species and Concentrations Byeong-Hee Min, Heum-Gi Park 1 and Sung Bum Hur* Department of Aquaculture, Pukyong National University, Busan , Korea 1 Faculty of Marine Bioscience & Technology, Kangnung National University, Gangneung , Korea This study was carried out to investigate grazing of brackish water cyclopoid copepod Paracyclopina nana on four microalgae species (Tetraselmis suecica, Isochrysis galbana, Phaeodactylum tricornutum and Dunaliella tertiolecta) and different food concentrations raging from 5 to 55 ng chl a/ml. The grazing of P. nana was examined by the analysis of decreased number of microalgae and chlorophyll a content in rearing water and pigment content in the gut of P. nana. The maximum content of decreased chlorophyll a and gut pigment in P. nana varied with microalgae species and concentrations. It appeared at the food concentration 30 ng chl a/ml in T. suecica, 40 ng chl a/ml in I. galnaba and D. tertiolecta, and 45 ng chl a/ml in P. tricornutum, respectively. The grazing rate of a P. nana per hour also varied with different microalgae species and concentrations. The maximum grazing rate per hour of P. nana fed T. suecica with 39.3 ng chl a/ml was the highest with 0.63 ng chl a/h, but lowest with 0.52 ng chl a/h. From these results, it can be concluded that T. suecica is the best species among four microalgae species for the mass culture of P. nana and daily optimum food concentration of P. nana is 25~39 ng chl a (approximately 10~ cells). Keywords: Copepod, Grazing, Microalgae, Paracyclopina nana ƒ w š ƒ ù (Nagaraj, 1992). ƒ w ƒ w. ƒ d w ü s j v w y d w ù ü w. ƒ (Dam and Peterson, 1988), (Dagg and Walser, 1987) (Head, 1988; Mayzaud et al., 1998). û ƒ š (Head et al., 1985; Atkinson et al., 1996, Tirelli and Mayzaud, 1999). w» ƒ š (Ohno and Okamura, 1988; Støttrup and Norsker, 1997). ù ƒ w *Corresponding author: hurs@pknu.ac.kr š wš w ƒ w» y w (Lee et al., 2005). Paracyclopina nana ù» xw w ƒ w Artemia nauplius w (Lee, 2004; Lee et al., 2006).» cyclopoid ƒ P. nana w P. nana ww 4 (Tetraselmis suecica, Isochrysis galbana, Phaeodactylum tricornutum, Dunaliella tertiolecta) (Lee et al., 2006; Min et al., 2006) P. nana p w. Paracyclopina nana w 4 (T. suecica KMCC P-9, I. galbana KMCC 310

2 H-2, P. tricornutum KMCC B-45, D. tertiolecta KMCC C-9) w w w w w w. f/2 (Guillard and Ryther, 1962) w, 20±1 C š 1 o 80 µmol m 2 s w. 250 ml ƒ flask w z 1 L ƒ v j, 3 L v j 20 L j x» w. P. nana P. nana » y y(18 psu) 100 µm plankton net w š f 1 w z 500 ml f w 20 o C, 15 psu w. P. nana nauplius 20 L j x» w 120 µm 40 µm w nauplius w z 20 L» ¾ w. Nauplius 28 o C, ¾ 20 o C w š(lee et al., 2006), 15 psu, 2 µmol m 2 s 1, Ÿ» 12L : 12Dw w. T. suecica P. nana cells, cells 1 1z œ w. P. nana w P. nana w» w ü w chlorophyll a w P. nana ü w (gut pigment content) d w. chlorophyll a d f s» g w. 1 L f( 1 L) s w P. nana f 1 /ml w T. suecica, I. galbana, P. tricornutum D. tertiolecta œ w. x w š 3 xw. ml chlorophyll a w 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55 ng w œ w. œ z µm P. nana 100 w z ü w chlorophyll a w P. nana ü w w. wr ƒ 55 ng chl a/ml œ w z (0.5, 1, 1.5, 2, 4, 6, 12, 18, 24 ) P. nana ü w y w. Chlorophyll a d œ 24 z P. nana 90 µm wš, û GF/C filter w z Lorenzen (1967) d w. 100 P. nana 15 ml w 90% acetone 10 ml ƒw w z 4500 rpm 10 w z 24 h þ w w -80 o C w. 665 nm y Ÿ d wš ƒ Paracyclopina nana 311 1N HCl 2 z 665 nm y z Ÿ d w (Lorenzen, 1967). ü w (gut pigment) chlorophyll a phaeopigment w w w. chlorophyll a ( mg m 3 ) phaeopigments ( mg m 3 ) 26.7( a ) v = : y 665 nm Ÿ, 665 a: y z 665 nm Ÿ, v: acetone (ml), V: (liters), l: m w q (cm) wr P. nana (G, ng chl a equiv. ind 1 h 1 ) w. G V ( C 0 C tf ) = N t C 0 x» s, C tf t z x s, V x, N»ü ƒ, f x ƒ m one-way ANOVA test w z Duncan's multiple range test (Duncan, 1955) w s³ (P<0.05) SPSS program (Ver. 10.1) w. P. nana P. nana 24 z ü chlorophyll a w Fig. 1. T. suecica chlorophyll a œ ƒ 30 ng chl a/ml ¾ ƒw ƒ 35~55 ng chl a/ml w. I. galbana D. tertiolecta chlorophyll a 40 ng chl a/ml ¾ ƒw ƒ 45~55 ng chl a/ml w. P. tricornutum 45 ng chl a/ml ¾ ƒw z w w. wr P. nana 24 ü w w Table 1. 4 ƒ ü w ƒw š w z w w. T. suecica 30 ng chl a/ml 35 ng chl a/ml ü w še š, I. galbana D. tertiolecta 40 ng chl a/ml, P. tricornutum 45 ng chl a/ml še. 30 ng chl a/ml ¾ T. suecica V l 26.7( 1.7[ 665 a ] ) v = V l

3 312,», x Fig. 1. Decreased chlorophyll a (mean±sd) in the water of Paracyclopina nana fed different food concentrations of microalgae. ƒ 3 w ü w w ƒ š, P. tricornutum ƒ û w (P<0.05). œ z P. nana ü w y Table 2. 4 x œ z 0.5 ü w še z, 4 ¾ wš z w w w. T. suecica œ P. nana ü w œ z ng pigment, ng pigment k x w w j w (P<0.05), z w. I. galbana D. tertiolecta œ œ z 2 ƒƒ ng pigment T. suecica w ƒ. P. tricornutum œ œ z 6 ¾ x w w w w. ù 12 l 4 x w w. P. nana y Fig. 2. T. suecica œ w P. nana 1 ƒ 39.3 ng chl a/ml 0.63 ng chl a ƒ ùkû ù, ƒ 25 ng chl a/ml l w. I. galbana ƒ 44.7 ng chl a/ml 0.60 ng chl a ind. 1 h 1 ƒ ùkû. P. tricornutum ƒ 44.5 ng chl a/ml, š D. tertiolecta ƒ 49.8 ng chl a/ml, P. nana 1 ƒƒ 0.41 ng, 0.52 ng chl a ƒ ùkû. Table 1. Total pigments in the gut of a Paracyclopina nana fed different food concentrations of microalgae Food concentrations (ng chl a/ml) Tetraselmis suecica Isochrysis galbana Dunaliella tertiolecta Phaeodactylum tricornutum ±0.003 a 0.17±0.007 ab 0.17±0.005 a 0.16±0.005 b ±0.009 a 0.17±0.004 b 0.17±0.009 b 0.16±0.008 b ±0.011 a 0.21±0.014 a 0.22±0.012 a 0.18±0.010 b ±0.011 a 0.24±0.008 b 0.16±0.007 d 0.21±0.013 c ±0.010 a 0.28±0.017 b 0.30±0.012 ab 0.25±0.007 c ±0.018 a 0.31±0.015 c 0.35±0.007 b 0.25±0.009 d ±0.020 a 0.33±0.017 c 0.38±0.016 b 0.31±0.014 c ±0.008 a 0.40±0.013 a 0.41±0.015 a 0.34±0.012 b ±0.011 a 0.38±0.014 a 0.38±0.021 a 0.39±0.019 a ±0.010 a 0.37±0.010 ab 0.37±0.009 ab 0.36±0.017 b ±0.003 a 0.37±0.012 a 0.37±0.008 a 0.35±0.017 b Values in the same row with different superscript letters are significantly different (P<0.05). Table 2. Variations of total pigment content in the gut of Paracyclopina nana fed different microalgal diet during twenty-four hours Hour Tetraselmis suecica Isochrysis galbana Dunaliella tertiolecta Phaeodactylum tricornutum ±0.028 a 0.40±0.013 a 0.44±0.029 a 0.42±0.036 a ±0.029 ab 0.35±0.007 b 0.38±0.014 ab 0.39±0.019 a ±0.007 c 0.31±0.013 b 0.29±0.018 b 0.34±0.006 a ±0.011 b 0.22±0.012 ab 0.21±0.023 b 0.26±0.032 a ±0.003 c 0.20±0.005 b 0.20±0.012 b 0.22±0.012 a ±0.003 b 0.17±0.007 b 0.17±0.005 b 0.19±0.009 a ±0.003 a 0.17±0.007 a 0.17±0.005 a 0.17±0.017 a ±0.003 a 0.17±0.007 a 0.17±0.005 a 0.16±0.007 a ±0.003 a 0.17±0.007 ab 0.17±0.005 a 0.16±0.005 b Values in the same row with different superscript letters are significantly different (P<0.05).

4 Fig. 2. Variations of grazing rate of a Paracyclopina nana per hour, which fed different food concentrations of microalgae. š œ ƒ s w e ƒ w (Corkett and McLaren, 1970; Mayzaud et al., 1998; Knuckey et al., 2005), cyclopoid ƒ Oithona similis f ù w e š w (Sabatini and Kiørboe, 1994). 4 ƒ P. nana ƒw ƒ w w. Calanus hyperboreus C. pacificus Ditylum brightwellii, Thalassiosira fluviatilis ƒ Paracyclopina nana 313 ³ w œ w (Mullin and Brooks, 1963), Frost (1972) x w. cyclopoid ƒ Oithona similisƒ s (Sabatini and Kiørboe, 1994) ew. Calbet and Alcaraz (1997) ƒ» nauplius j w šw. Lee (2004) P. nanaƒ nauplius T. suecica œ ƒw w e ùkù, copepodid 1~ cells/ml» w ù ƒw, w e š w. x œ z ü w y w, P. nana œ z ƒ y wš ú w 4~6 z w w.. w, P. nana y 4~6 d, x w 4 ƒ yw T. suecica, D. tertiolecta, I. galbana, P. tricornutum q. ƒ ƒ û w š (Mullin and Brooks, 1963; Frost, 1972; Huntley, 1981). Acartia tonsa œ 7~150 µgc/l w ƒ š w (Reeve and Walter, 1977). T. suecica, I. galbana, P. tricornutum, š D. tertiolecta w P. nana 1 ƒ œ ƒ ƒƒ 39.3, 44.7, ng chl a/ml ƒƒ 0.63, 0.60, ng chl a ùkù P. nana. Chlorophyll a s w ƒ chlorophyll a s y w ƒƒ , , , cells/ml. š P. nana ü w ƒw ƒ e z ƒ ƒw w. P. nana œ w. P. nana p T. suecica P. nana ƒ z, ng chl a ( cells) w ù œ 25~39 ng chl a ( 10~ cells) q.» cyclopoid ƒ Paracyclopina nana w. œ 24

5 314,», x z P. nana ü w w. T. suecica 30 ng chl a/ml, I. galbana D. tertiolecta 40 ng chl a/ ml, P. tricornutum 45 ng chl a/ml œ w P. nana ü w ƒ. T. suecica, I. galbana, P. tricornutum, š D. tertiolecta w P. nana 1 ƒ œ ƒ ƒƒ 39.3, 44.7, ng chl a/ml ƒƒ 0.63, 0.60, ng chl a ùkû. P. nana ƒ ww œ ƒ û ƒ T. suecica, 1 1 œ 25~39 ng chl a ( 10~ cells)ƒ ƒ q. w ww 2000 p w w w w w tw. š x Atkinson, A., P. Ward and E. J. Murphy, Diel periodicity of subantarctic copepods: relationships between vertical migration, gut fullness and gut evacuation rate. J. Plankton Res., 18, Calbet, A. and M. Alcaraz, Growth and survival rates of early developmental stages of Acartia grani (Copepoda: Calanoida) in relation to food concentration and fluctuations in food supply. Mar. Ecol. Prog. Ser., 147, Corkett, C. J. and I. A. McLaren, Relationships between development rate of eggs and older stages of copepods. J. Mar. Biol. Ass. U. K., 50, Dagg, M. J. and W. E. J. Walser, Ingestion, gut passage, and egestion by the copepod Neocalanus plumchrus in the laboratory and in the the subarctic Pacific Ocean. Limnol. Oceanogr., 32, Dam, H. J. and W. T. Peterson, The effect of temperature on the gut clearance rate constant of planktonic copepods. J. Exp. Mar. Bio. Ecol., 123, Duncan, D. B., Multiple-range and multiple F tests. Biometrics, 11, Frost, B. W., Effects of size and concentration of food particles on the feeding behavior of the marine plankton copeopd Calanus pacificus. Limnol. Oceanogr., 17, Guillard, R. R. L. and J. H. Ryther, Studies on marine planktonic diatoms.. Cyclotella nana Hustedt and Detonula confervacea (Clece) Gran. Can. J. Microbiol., 3, Head, E. J. H., Copepod feeding behaviour and the measurement of grazing rates in vivo and in vitro. Hydrobiologia, 167/168, Head, E. J. H., L. R. Harris and C. Abou Debs, Effect of daylength and food concentration on in situ diurnal feeding rhythms in arctic copepods. Mar. Ecol. Prog. Ser., 24, Huntley, M., Nonselective, nonsaturated feeding by three calanoid copepod species in the Labrador Sea. Limnol. Oceanogr., 26, Knuckey, R. M., G. L. Semmens, R. J. Mayer and M. A. Rimmer, Development of an optimal microalgal diet for the culture of the calanoid copepod Acartia sinjiensis: Effect of algal species and feed concentration on copepod development. Aquaculture, 249, Lee, C. S., P. J. O Bryen and N. H. Marcus, Copepod in aquaculture. Blackwell Publishing, Iowa, 269 pp. Lee, K. W., Mass culture and food value of the cyclopoid copepod Paracyclopina nana Smirnov. Ph.D. thesis, Kangnung National University, 124 pp. Lee, K. W., H. G. Park, S. M. Lee and H. K. Kang, Effects of diets on the growth of the brackish water cyclopoid copepod Paracyclopina nana Smirnov. Aquaculture, 256, Lorenzen, C. J., Determination of chlorophyll and phaeopigments: spectrophotometric equations. Limnol. Oceanogr., 12, 3[ Mayzaud, P., V. Tirelli, J. M. Bernard and O. Roche-Mayzaud, The influence of food quality on the nutritional acclimation of the copepod Acartia clausi. J. Mar. Syst., 15, Min, B. H., H. G. Park,GK. W. Lee and S. B. Hur, Selection of optimum microalgal species for culture of the brackish water cyclopoid copepod Paracyclopina nana. J. Kor. Fish. Soc., 39 (in Press). Mullin, M. M. and E. R. Brooks, Some factors affecting the feeding of marie copepod of the genus Calanus. Limnol. Oceanogr., 8, Nagaraj, M., Combined effects of temperature and salinity on the development of the copepod Eurytemora affinis. Aquaculture, 103, Ohno, A. and Y. Okamura, Propagation of the calanoid copepod, Acartia tsuensis, in outdoor tanks. Aquaculture, 187, Reeve, M. R. and M. A. Walter, Observations on the existense of lower threshold and upper critical food concentrations for the copepod Acartia tonsa Dana. J. Exp. Mar. Biol. Ecol., 29, Sabatini, M. and T. Kiørboe, Egg production, growth and development of the cyclopoid copepod Oithona similis. J. Plankton Res., 16, Støttrup, J, G. and N. H. Norsker, Production and use of copepods in marine fish larviculture. Aquaculture, 155, Tirelli, V. and P. Mayzaud, Gut evacuation rates of Antarctic copepods during austral spring. Polar Biol., 21, š : :

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