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Size distribution in ultraphytoplankton: A comparative analysis of counting methods

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Abstract

Water samples taken at three depth layers from the offshore oligotrophic Cretan Sea were analyzed for ultraphytoplankton size fractionation using different methods: (a) sequential filtration on filters of pore size 5, 1 and 0.2 μm, (b) separate filtration using filters 5 and 0.2 μm as well as 1 and 0.2 μm and (c) direct filtration on 0.2 μm filters after staining of the samples with DAPI. Total abundance of photosynthetic organisms as well as the abundance of different groups such as flagellates and cyanobacteria measured by means of sizing after DAPI staining were significantly higher than those obtained by the other methods. This indicates that although there were no significant differences between the estimates provided by the separate and sequential filtration, both these methods underestimated total abundance by at least 25–50%. The estimates for the size fractions were also found to range from relatively imprecise to completely unreliable depending on the group and the size range. Although size fractionation through direct observation after staining largely depends on the expertise of the observer, this study suggests that it may provide more informative estimates than the other two methods. Although it is difficult to generalize the results of this study in a global context, the paper provides strong indications on the limitations of the sequential and separate methods for size fractionation of photosynthetic organisms and implies that their results are likely to be less accurate than is presently believed.

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References

  • Acosta Pomar, L., Bruni, V., Decembrini, F., Giuffre, G. and Maugeri, T.L.: 1988, ‘Distribution and activity of picophytoplankton in a brackish environment’, Deep-Sea Res. 21, 129–138.

    Google Scholar 

  • Booth, B.C., Lewin, J. and Norris, R.E.: 1982, ‘Nanoplankton species predominant in the subarctic Pacific in May and June 1978’, Deep-Sea Res. 29, 185–200.

    Google Scholar 

  • Booth, B.C.: 1993, ‘Estimating Cell Concentration and Biomass of Autotrophic Plankton Using Microscopy’, in: P. F. Kemp, B. F. Sherr, E. B. Sherr and J. J. Cole (eds), Handbook of Methods in Aquatic Microbial Ecology, Lewis Publishers, Boca Raton, pp. 199–205.

    Google Scholar 

  • Caron, D.A.: 1983, ‘Technique for enumeration of heterotrophic and phototrophic nanoplankton, using epifluorescence microscopy, and comparison with other procedures’, Appl. Environ. Microbiol. 46, 491–498.

    CAS  Google Scholar 

  • Chepurnova, E.A. and Gutveyb, L.G.: 1988, ‘Distribution and size structure of phototrophic bacterioplankton (Cyanobacteria) in the Mediterranean Sea and Atlantic Ocean’, Oceanology 28, 643–647.

    Google Scholar 

  • Chisholm, S.W., Olson, R.J., Zettler, E.R., Goericke, R., Waterbury, J.B. and Welschmeyer, N.A.: 1988, ‘A novel free-living prochlorophyte abundant in the oceanic euphotic zone’, Nature 334, 340–343.

    Google Scholar 

  • Christaki, U., Van Wambeke, F. and Dolan, J.R.: 1999, ‘Nanoflagellates (mixotrophs, heterotrophs and autotrophs) in the oligotrophic eastern Mediterranean: Standing stocks, bacterivory and relationships with bacterial production’, Mar. Ecol. Prog. Ser. 181, 297–307.

    Google Scholar 

  • Davis, P.G., Caron, D.A., Johnson, P.W. and Sieburth, J.McN.: 1985, ‘Phototrophic and apochlorotic components of picoplankton and nanoplankton in the North Atlantic: Geographic, vertical, seasonal and diel distributions’, Mar. Ecol. Prog. Ser. 21, 15–26.

    Google Scholar 

  • DuRand, M.D. and Olson, R.J.: 1996, ‘Conributions of phytoplankton light scattering and cell concentration changes to diel variations in beam attenuation in the equatorial Pacific from flow cytometric measurements of pico-, ultra-and nanoplankton’, Deep-Sea Res. II 43, 891–906.

    Google Scholar 

  • Ferrier-Pagès, C. and Gattuso, J.P.: 1998, ‘Biomass, production and grazing rates of pico-and nanoplankton in coral reef waters (Miyako Island, Japan)’, Microb. Ecol. 35, 46–57.

    Google Scholar 

  • Glover, H.E., Campbell, L. and Prezelin, B.B.: 1986, ‘Contribution of Synechococcus spp. to size-fractioned primary productivity in three water masses in the Northwest Atlantic Ocean’, Mar. Biol. 91, 193–203.

    Google Scholar 

  • Glover, H.E., Prezelin, B., Campbell, L. and Wyman, M.: 1988, ‘Pico-and ultraplankton Sargasso Sea communities: Variability and comparative distributions of Synechococcus spp. and algae’, Mar. Ecol. Prog. Ser. 49, 127–139.

    Google Scholar 

  • Glover, H.E., Smith, A.E. and Shapiro, L.: 1985, ‘Diurnal variations in photosynthetic rates: Comparisons of ultraphytoplankton with a larger phytoplankton size fraction’, J. Plankton Res. 7, 519–535.

    Google Scholar 

  • Goericke, R. and Welschmeyer, N.A.: 1993, ‘The marine prochlorophyte Prochlorococcus contributes significantly to phytoplankton biomass and primary production in the Sargasso Sea’, Deep-Sea Res. 40, 2283–2294.

    Google Scholar 

  • Goldman, J.C. and Dennett, M.R.: 1985, ‘Susceptibility of some marine phytoplankton species to cell breakage during filtration and post-filtration rinsing’, J. Exp. Mar. Biol. Ecol. 86, 47–58.

    Google Scholar 

  • Guillou, L., Jacquet, S., Chrétiennot-Dinet, M.-J. and Vaulot, D.: 2001, ‘Grazing impact of the small heterotrophic flagellates on Prochlorococcus and Synechococcus’, Aquat. Microb. Ecol. 26, 201–207.

    Google Scholar 

  • Hadas, O. and Berman, T.: 1998, ‘Seasonal abundance and vertical distribution of Protozoa (flagellates, ciliates) and bacteria in Lake Kinneret, Israel’, Aquat. Microb. Ecol. 14, 161–170.

    Google Scholar 

  • Hannah, F.J. and Boney, A.D.: 1983, ‘Nanophytoplankton in the Firth of Clyde, Scotland: Seasonal abundance, carbon fixation and species composition’, J. Exp. Mar. Biol. Ecol. 67, 105–147.

    CAS  Google Scholar 

  • Havskum, H. and Hansen, A.S.: 1997, ‘Importance of pigmented and colourless nano-sized protists as grazers on nanoplankton in a phosphate-depleted Norwegian fjord and in enclosures’, Aquat. Microb. Ecol. 12, 139–151.

    Google Scholar 

  • Hobbie, J.E., Daley, R.J. and Jasper, S.: 1977, ‘Use of Nuclepore filters for counting bacteria by fluorescence microscopy’, Appl. Environ. Microbiol. 33, 1225–1228.

    CAS  Google Scholar 

  • Iriarte, A.: 1993, ‘Size-fractionated chlorophyll a biomass and picophytoplankton cell density along a longitudinal axis of a temperate estuary (Southampton Water)’, J. Plankton Res. 15, 485–500.

    Google Scholar 

  • Iriarte, A. and Purdie, D.A.: 1994, ‘Size distribution of chlorophyll a biomass and primary production in a temperate estuary (Southampton Water): The contribution of photosynthetic picoplankton’, Mar. Ecol. Prog. Ser. 115, 283–297.

    CAS  Google Scholar 

  • Jacquet, S., Lennon, J.-F., Marie, D. and Vaulot, D.: 1998, ‘Picoplankton population dynamics in coastal waters of the northwestern Mediterranean Sea’, Limnol. Oceanogr. 43, 1916–1931.

    CAS  Google Scholar 

  • Johnson, P.W. and Sieburth, J.M.: 1979, ‘Chroococcoid cyanobacteria in the sea: A ubiquitous and diverse phototrophic biomass’, Limnol. Oceanogr. 24, 928–935.

    Google Scholar 

  • Joint, I.R., Owens, N.J.P. and Pomroy, A.J.: 1986, ‘Seasonal production of photosynthetic picoplankton and nanoplankton in the Celtic Sea’, Mar. Ecol. Prog. Ser. 28, 251–258.

    Google Scholar 

  • Kirchman, D.L., Rich, J.H. and Barber, R.T.: 1995, ‘Biomass and production of heterotrophic bacteria along 140W in the equatorial Pacific: Effect of temperature on the microbial loop’, Deep Sea Res. 42, 603–619.

    Google Scholar 

  • Kuosa, H.: 1988, ‘Occurrence of autotrophic picoplankton along an open sea—Inner archipelago gradient in the Gulf of Finland, Baltic Sea’, Ophelia 28, 85–93.

    Google Scholar 

  • Kuylenstierna, M. and Karlson, B.: 1994, ‘Seasonality and composition of pico-and nanoplanktonic cyanobacteria and protists in the Skagerrak’, Bot. Mar. 37, 17–33.

    Article  Google Scholar 

  • Li, W.K.W.: 1990, ‘Particles in “particle-free” seawater: Growth of ultraphytoplankton and implications for dilution experiments’, Can. J. Fish Aquat. Sci. 47, 1258–1268.

    Article  Google Scholar 

  • Li, W.K.W., Subba Rao, D.V., Harrison, W.G., Smith, J.C., Cullen, J.J., Irwin, B. and Platt, T.: 1983, ‘Autotrophic picoplankton in the tropical ocean’, Science 219, 292–295.

    CAS  Google Scholar 

  • Li, W.K.W., Zohary, T., Yacobi, Y.Z. and Wood, A.M.: 1993, ‘Ultraphytoplankton in the eastern Mediterranean Sea: Towards deriving phytoplankton biomass from flow cytometric measurements of abundance, fluorescence and light scatter’, Mar. Ecol. Prog. Ser. 102, 79–87.

    Google Scholar 

  • Logan, B.E.: 1993, ‘Theoretical analysis of size distributions determined with screens and filters’, Limnol. Oceanogr. 38, 372–381.

    Article  Google Scholar 

  • Logan, B.E., Passow, U. and Alldredge, A.L.: 1994, ‘Variable retention of diatoms on screens during size separations’, Limnol. Oceanogr. 39, 390–395.

    Google Scholar 

  • Magazzu, G., Bruni, V., Piccione, A., Platt, T., Irwin, B. and Rao, D.V.S.: 1987, ‘Picoplankton: Contribution to phytoplankton production in the strait of Messina’, PSZNI: Mar. Ecol. 8, 21–31.

    CAS  Google Scholar 

  • Malone, T.C.: 1980, ‘Algal Size’, in: I. Morris (ed.), The Physiological Ecology of Phytoplankton, Blackwell Scientific Publications, London, pp. 1–625.

    Google Scholar 

  • Marshall, H.G.: 1995, ‘Autotrophic picoplankton distribution and abundance in the Chesapeake Bay, U.S.A’, Mar. Nat. 4, 33–42.

    Google Scholar 

  • Maugeri, T.L., Acosta Pomar, M.L.C., Bruni, V. and Salomone, L.: 1992, ‘Picoplankton and picophytoplankton in the Ligurian Sea and straits of Messina (Mediterranean Sea)’, Bot. Mar. 35, 493–502.

    Article  Google Scholar 

  • Murphy, L.S. and Haugen, E.M.: 1985, ‘The distribution and abundance of phototrophic ultraplankton in the North Atlantic’, Limnol. Oceanogr. 30, 47–58.

    Google Scholar 

  • Partensky, F., Blanchot, J., Lantoine, F., Neveux, J. and Marie, D.: 1996, ‘Vertical structure of picophytoplankton at different trophic sites of the tropical northeastern Atlantic Ocean’, Deep-Sea Res. I 43, 1191–1213.

    CAS  Google Scholar 

  • Platt, T., Rao, D.V.S. and Irwin, B.: 1983, ‘Photosynthesis of picoplankton in the oligotrophic ocean’, Nature 300, 702–704.

    Google Scholar 

  • Porter, K.G. and Feig, Y.S.: 1980, ‘The use of DAPI for identifying and counting aquatic microflora’, Limnol. Oceanogr. 25, 943–948.

    Google Scholar 

  • Quinones, R.A.: 1994, ‘A comment on the use of allometry in the study of pelagic ecosystem processes’, Sci. Mar. 58, 11–16.

    Google Scholar 

  • Reynolds, N.: 1973, ‘The estimation of the abundance of ultraplankton’, Br. Phycol. J. 8, 135–146.

    Google Scholar 

  • Sanders, R.W., Berninger, U.G., Lim, E.L., Kemp, P.F. and Caron, D.A.: 2000, ‘Heterotrophic and mixotrophic nanoplankton predation on picoplankton in the Sargasso Sea and Georges Bank’, Mar. Ecol. Prog. Ser. 192, 103–118.

    Google Scholar 

  • Sheldon, R.W. and Sutcliffe, W.H. Jr.: 1969, ‘Retention of marine particles by screens and filters’, Limnol. Oceanogr. 14, 441–444.

    Google Scholar 

  • Sheldon, R.W.: 1972, ‘Size separation of marine seston by membrane and glass-fiber filters’, Limnol. Oceanogr. 17, 494–498.

    Google Scholar 

  • Shimada, A., Hasegawa, T., Umeda, I., Kadoya, N. and Maruyama, T.: 1993, ‘Spatial mesoscale patterns of West Pacific picophytoplankton as analyzed by flow cytometry: Their contribution to subsurface chlorophyll maxima’, Mar. Biol. 115, 209–215.

    CAS  Google Scholar 

  • Sieburth, J. McN., Smetacek, V. and Lenz, J.: 1978, ‘Pelagic ecosystem structure: Heterotrophic compartments of the plankton and their relationship to plankton size fractions’, Limnol. Oceanogr. 23, 1256–1263.

    Google Scholar 

  • Stockner, J. G., Klut, M. E. and Cochlan, W. P.: 1990, ‘Leaky filters: A warning to aquatic ecologists’, Can. J. Fish Aquat. Sci. 47, 16–23.

    Article  Google Scholar 

  • Utermöhl, H.: 1958, ‘Zur Vervollkommnung der quantitativen Phytoplankton-methodik’, Ecol. Monogr. 57, 61–88.

    Google Scholar 

  • Vaulot, D., Partensky, F., Neveux, J., Mantoura, R. F. C. and Llewellyn, C.: 1990, ‘Winter presence of prochlorophytes in surface waters of the north-western Mediterranean Sea’, Limnol. Oceanogr. 35, 1156–1164.

    Article  Google Scholar 

  • Waterbury, J.B., Watson, S.W., Guillard, R.R.L. and Brand, L.E.: 1979, ‘Widespread occurrence of a unicellular, marine, planktonic cyanobacterium’, Nature 277, 293–294.

    Google Scholar 

  • Williams, R.B.: 1964, ‘Division rates of salt marsh diatoms in relation to salinity and cell size’, Ecology 45, 877–880.

    Google Scholar 

  • Zaika, V.Y.: 1986, ‘Vertical distribution of autotrophic picoplankton in the Indian Ocean and the Mediterranean Sea’, Oceanology 26, 209–212.

    Google Scholar 

  • Zaika, V.E. and Yashin, V.A.: 1984, ‘Lumenescing picosuspension (0.2–2 μm) in oligotrophic waters of the Mediterranean and Black seas’, Dokl. Biol. Sci. 275, 262–264.

    Google Scholar 

  • Zimmermann, R. and Meyer-Reil, L.A.: 1974, A new method for fluorescence staining of bacterial populations on membrane filters’, Kieler Meeresforsch. 30, 24–27.

    Google Scholar 

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Pitta, P., Karakassis, I. Size distribution in ultraphytoplankton: A comparative analysis of counting methods. Environ Monit Assess 102, 85–101 (2005). https://doi.org/10.1007/s10661-005-2690-5

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