Molecular Ecology of Aquatic Communities pp 265-277 | Cite as
RNA—DNA ratio and other nucleic acid-based indicators for growth and condition of marine fishes
Abstract
Nucleic acid analysis has provided useful tools to study recent growth and mortality of young fishes and their responses to environmental variability. The ratio of RNA-DNA (R/D) has been shown to respond to changes in feeding conditions and growth after periods as short as 1-3 days in a variety of fish species. The earliest studies used primarily UV-based methods, but most investigators now use more sensitive, fluorometric dye-binding assays to estimate RNA and DNA in individual larvae. These newer methods are very sensitive to procedural details and choice of standards. Analytical methods, normalization and calibration procedures to optimize information obtained from nucleic acid analysis are discussed. We present examples illustrating the technique’s utility, and problems encountered when applying nucleic acid-based indices to fish larvae and early juveniles. The wide use of R/D analysis in studies of fish early life stages, together with a proliferation of analytical methods, demands a major intercalibration exercise.
Key words
nucleic acids fish larvae RNA/DNA condition growthPreview
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References
- Bergeron, J. P., 1997. Nucleic acids in ichthyoplankton ecology: a review, with emphasis on recent advances for new perspectives. J. Fish Biol. 51 (Suppl. A): 284–302.CrossRefGoogle Scholar
- Bergeron, J. P., M. Boulhic & R. Galois, 1991. Effet de la privation de nourriture sur la teneur en ADN de la larve de sole (Solea solea L.) ICES J. mar. Sci. 48: 127–134.Google Scholar
- Bergeron, J. P., J. Person-Le Ruyet & C. Koutsikopoulos, 1997. Use of carbon rather than dry weight to assess the DNA content and nutritional condition index in sole larvae. ICES J. mar. Sci. 54: 148–151.CrossRefGoogle Scholar
- Bertram, D. F., T. I. Miller & W. C. Leggett, 1996. Individual variation in growth and development during the early life stages of winter flounder, Pleuronectes americanus. Fish. Bull. U.S.A. 95: 1–10.Google Scholar
- Brodie, S., J. Giron & S. A. Latt, 1975. Estimation of accessibility of DNA in chromatin from fluorescence measurements of electronic excitation energy transfer. Nature 253: 470–471.CrossRefPubMedGoogle Scholar
- Buckley, L. I., 1979. Relationships between RNA-DNA ratio, prey density and growth rate in cod (Gadus morhua L.) larvae. J. Fish Res. Bd Can. 36: 1497–1502.CrossRefGoogle Scholar
- Buckley, L. J., 1980. Changes in RNA, DNA, and protein content during ontogenesis of the winter flounder Pseudopleuronectes americanus) and the effect of starvation. Fish. Bull. U.S.A. 77: 703–708Google Scholar
- Buckley, L. J., 1982. Effects of temperature on growth and biochemical composition of larval winter flounder, Pseudopleuronectes americanus. Mar. Ecol. Progr. Ser. 8: 181–186.CrossRefGoogle Scholar
- Buckley, L. J., 1984. RNA-DNA ratio: an index of larval fish growth in the sea. Mar. Biol. 80: 291–298.CrossRefGoogle Scholar
- Buckley, L. J. & F. J. Bulow, 1987. Techniques for estimation of RNA, DNA, and protein in fish. In Summerfelt R. C. and G. E. Hall, (eds), Age and Growth of Fish. Iowa State University Press, Ames, Iowa. 345–354.Google Scholar
- Buckley, L. J. & R. G. Lough, 1987. Recent growth, biochemical composition, and prey field of larval haddock (Melanogrammus aeglefinus) and Atlantic cod Gadus morhua) on Georges Bank. Can. J. Fish. aquat. Sci. 44: 14–25.CrossRefGoogle Scholar
- Buckley, L. J., S. I. Turner, T. A. Halavik, A. S. Smigielski, S. M. Drew, & G. C. Laurence, 1984. Effects of temperature and food availability on growth, survival and RNA-DNA ratio of larval sand lance Ammodytes americanus). Mar. Ecol. Progr. Ser. 15: 91–97CrossRefGoogle Scholar
- Buckley, L. J., A. S. Smigielski, T. A. Halavik, B. R. Burns & G. C. Laurence, 1993. Growth and survival of the larvae of three species of temperate marine fishes at discrete prey densities. II. Cod (Gadus morhua), winter flounder (Pseudopleuronectes americanus), and silver hake (Merluccius bilinearus). In Walther, B. T. & H. I. Fyhn, (eds), Physiological and Biochemical Aspects of Fish Development. Univ. of Bergen, Norway.Google Scholar
- Bulow, F. J., 1970. RNA-DNA ratios as indicators of recent growth rates of a fish. J. Fish Res. Bd Can. 27: 2343–2349.CrossRefGoogle Scholar
- Bulow, F. J., 1987. RNA-DNA ratios as indicators of growth in fish. In R. C. Summerfelt and G.E. Hall (eds), The Age and Growth of Fish. Iowa State University Press, Ames, Iowa. 45–64.Google Scholar
- Caldarone, E. M. & L. J. Buckley, 1991. Quantitation of DNA and RNA in crude tissue extracts by flow injection analysis. Anal. Biochem. 199: 137–141.CrossRefPubMedGoogle Scholar
- Canino, M. F. & E. M. Caldarone], 1995. Modification and comparison of two fluorometric techniques for determining nucleic acid contents of fish larvae. Fish. Bull. U.S.A. 93: 158–Google Scholar
- Canino, M. F., K. M. Bailey & L. S. Incze, 1991. Temporal and geographic differences in feeding and nutritional condition of walleye pollock larvae Theragra chalcogramma in Shelikof Strait, Gulf of Alaska. Mar. Ecol. Prog. Ser. 79: 27–35.CrossRefGoogle Scholar
- Chicharo, M. A., L. Chicharo, L. Valdes, E. Lopez-Jamar & P. Re, 1998. Estimation of starvation and diel variation of the RNA/DNA ratios in field-caught Sardina pilchardus larvae off the north of Spain. Mar. Ecol. Prog. Ser. 164: 273–283.CrossRefGoogle Scholar
- Clemmesen, C., 1987. Laboratory studies on RNA/DNA ratios of starved and fed herring (Clupea harengus) and turbot (Scoph-thalmus maximus) larvae. J. Cons. perm. int. Explor. Mer. 43: 122–1Google Scholar
- Clemmesen, C., 1988. An RNA and DNA fluorescence technique to evaluate the nutritional condition of individual marine fish larvae. Meeresforch. 32: 134–143Google Scholar
- Clemmesen, C., 1989. RNA/DNA ratios of laboratory-reared and wild herring larvae determined with a highly sensitive fluorescence method. J. Fish Biol. 35 (Suppl. A): 331–333.CrossRefGoogle Scholar
- Clemmesen, C., 1993. Improvements in the fluorimetric determination of the RNA and DNA content in individual marine fish larvae. Mar. Ecol. Prog. Ser. 100: 177–183.CrossRefGoogle Scholar
- Clemmesen, C., 1994. Importance and limits of RNA/DNA ratios as a measure of nutritional condition in fish larvae. Proc. International Workshop: Survival Strategies in Early Life Stages of Marine Resources. 67–82. Yokohama, Japan. 11-14 October 1994.Google Scholar
- Clemmesen, C., 1996. Importance and limits of RNA/DNA ratios as a measure of nutritional condition in fish larvae. In Y. Watanabe, Y. Yamashita & Y. Ooseki (eds), Survival Strategies in Early Life Stages of Marine Resources. A.A. Balkema. Rotterdam: 67–82.Google Scholar
- Clemmesen, C. & T. Doan, 1996. Does otolith structure reflect the nutritional condition of a fish larva? Comparison of otolith structure and biochemical index (RNA/DNA ratio) determined on cod larvae. Mar. Ecol. Prog. Ser. 138: 33–39.CrossRefGoogle Scholar
- Clemmesen, C., R. Sanchez & C. Wongtschowski, 1997. A regional comparison of the nutritional condition of S.W. Atlantic anchovy larvae, Engraulis anchoita, based on RNA/DNA ratios. Arch. Fish. Mar. Res. 45: 17–43.Google Scholar
- Ferron, A. & W. C. Leggett, 1994. An Appraisal of Condition Measures for Marine Fish Larvae. Adv. Mar. Biol. 30: 217–303.CrossRefGoogle Scholar
- Folkvord, A. & E. Moksness, 1995. RNA/DNA ratios and growth of herring larvae. Mar. Ecol. Prog. Ser. 121: 311–312.CrossRefGoogle Scholar
- Folkvord, A., L. Ystanes, L. Johannessen & E. Moksness, 1996. RNA:DNA ratios and growth of herring (Clupea harengus) larvae reared in mesocosms. Mar. Biol. 126: 591–602.CrossRefGoogle Scholar
- Foster, A. R., D. F. Houlihan, S. J. Hall, & L. J. Burren. 1992. The effects of temperature acclimation on protein synthesis rates and nucleic acid content of juvenile cod (Gadus morhua L.). Can. J. Zool. 70:2095–2102.CrossRefGoogle Scholar
- Fry, J. E. J., 1971. The effects of environmental factors on the physiology of fish. In Hoar, W. S. & D. J. Randall (eds), Fish Physiology, Academic Press, New York, 6: 1–98.Google Scholar
- Goolish, E. M., M. G. Barron & I. R. Adelman, 1984. Ther-moclimatory response of nucleic acid and protein content of carp muscle tissue: influence of growth rate and relationship to glycine uptake by scales. Can. J. Zool. 62: 2164–2170.CrossRefGoogle Scholar
- Gronkjaer, P., C. Clemmesenn & M. St. John, 1997. Nutritional condition and vertical distribution of Baltic cod larvae. J. Fish Biol. 51 (Suppl. A): 352–369.CrossRefGoogle Scholar
- Hewitt, R. P., G. H. Theilacker & N. C. H. Lo, 1985. Causes of mortality in young jack mackerel. Mar. Ecol. Prog. Ser. 26: 1–10.CrossRefGoogle Scholar
- Houde, E. D., 1987. Fish Early Life Dynamics and Recruitment Variability. Amer. Fish. Society. Symp. 2: 17–29.Google Scholar
- Houlihan, D. F., S. J. Hall, C. Gray & B. S. Noble, 1988. Growth rates and protein turnover in Atlantic cod, Gadus morhua. Can. J. Fish. aquat. Sci. 45: 951–964.CrossRefGoogle Scholar
- Hovenkamp, F. & J. Witte, 1991. Growth, otolith growth and RNA/DNA ratios of larval plaice Pleuronectes platessa in the North Sea 1987–1989. Mar. Ecol. Prog. Ser. 70: 105–116.CrossRefGoogle Scholar
- Jennings, S., 1991. The effects of capture, net retention and preservation upon lengths of larval and juvenile bass, Dicentrarchus labrax (L.). J. Fish Biol. 38: 349–357.CrossRefGoogle Scholar
- Koban, M., A. A. Yup, L. B. Agellon & D. A. Powers. Molecular adaptation to environmental temperature: heat-shock response of the eurythermal teleost Fundulus heteroclitus. Molec. mar. Biol. Biotechnol. 1: 1–17.Google Scholar
- Lough, R. G., E. M. Caldarone, T. K. Rotunno, E. A. Broughton, B. R. Burns & L. J. Buckley, 1996. Vertical distribution of cod and haddock eggs and larvae feeding and condition in stratified and mixed waters on southern Georges Bank May 1992. Deep Sea Research 43: 1875–1904.CrossRefGoogle Scholar
- Love, R. M., 1970. The Chemical Biology of Fishes. Academic Press, New York.Google Scholar
- Malloy, K. D. & T. E. Targett, 1994. The use of RNA:DNA ratios to predict growth limitation of juvenile summer flounder (Paralich-thys dentatus) from Delaware and North Carolina estuaries. Mar. Biol. 118:367–375.CrossRefGoogle Scholar
- Martin, F. D. & D. A. Wright, 1987. Nutritional state analysis and its use in predicting striped bass recruitment: laboratory calibration. Am. Fish. Soc. Symp. 2: 109–114.Google Scholar
- Mathers, E. M., D. F. Houlihan & M. J. Cunningham, 1992. Nucleic acid concentrations and enzyme activities as correlates of growth rate of the saithe Pollachius virens: growth-rate estimates of open-sea fish. Mar. Biol. 112: 363–369.CrossRefGoogle Scholar
- Mathers, E. M., D. F. Houlihan & L. J. Burren, 1994. RNA, DNA, and protein concentrations in fed and starved herring Clupea harengus larvae. Mar. Ecol. Prog. Ser. 107: 223–231.CrossRefGoogle Scholar
- McGurk M. & M. W. Kusser, 1992. Comparison of three methods of measuring rRNA and DNA concentrations of individual Pacificherring, Clupea pallasi, larvae. Can. J. Fish. aquat. Sci. 49: 967–974.CrossRefGoogle Scholar
- McNamara, P. T. & L. J. Buckley, 1994. Identification and characterization of metallothionein cDNA from mRNA transcripts induced by starvation in Atlantic cod (Gadus morhua). Molec. Mar. Biol. Biotechnol. 3: 252–2Google Scholar
- Morgan, A. R., D. H. Evans, J. S. Lee & D. E. Pulleyblallnk, 1979. Review: Ethidium fluorescence assay. Part II: Enzymatic studies and DNA-protein interactions. Nucleic Acids Res. 7: 571–594.Google Scholar
- Ong, T. L., R. Armstrong & L. Buckley. Diel study of RNA productions associated with starvation in Atlantic cod Gadus morhua larvae. (In prep.).Google Scholar
- Raae, A. J., I. Opstad, P. Kvenseth & B. T. Walther, 1988. RNA, DNA and protein during early development in feeding and starved cod (Gadus morhua L.) larvae. Aquaculture 73: 247–259.CrossRefGoogle Scholar
- Radtke, R. L., 1989. Larval fish age, growth, and body shrinkage: information available from otoliths. Can. J. Fish. aquat. Sci. 46: 1884–1894.CrossRefGoogle Scholar
- Richard, P., J.-P. Bergeron, M. Boulhic, R. Galois & J. P.-L. Ruyet, 1991. Effect of starvation on RNA, DNA and protein content of laboratory-reared larvae and juveniles of Solea solea. Mar. Ecol. Prog. Ser. 72: 69–77.CrossRefGoogle Scholar
- Robinson, S. M. C. & D. M. Ware, 1988. Ontogenetic development of growth rates in larval Pacific herring, Culpea harengus pallasi, measured with RNA-DNA ratios in the Strait of Georgia, British Columbia. Can. J. Fish. aquat. Sci. 45: 1422–1429.CrossRefGoogle Scholar
- Rooker, J. R. & G. J. Holt, 1996. Application of RNA:DNA ratios to evaluate the condition and growth of larval and juvenile red drum (Sciaenops ocellatus). Mar. Freshwat. Res. 47: 283–2CrossRefGoogle Scholar
- Sclafani, M., G. Sterling & W. C. Leggett. 1997. Osmoregulation, nutritional effects and buoyancy of marine larval fish: a bioas-say for assessing density changes during the earliest life-history stages. Mar. Biol. 129: 1–9CrossRefGoogle Scholar
- Steel, R. G. D. & J. H. Tome, 1960. Principles and Procedures of Statistics, McGraw-Hall, 481 pp.Google Scholar
- Suthers, I. M., J. J. Cleary, S. C. Battaglene & R. Evans, 1996. Relative RNA content as a measure of condition in larval and juvenile fish. Mar. Freshwater Res. 47: 301–307.CrossRefGoogle Scholar
- Theilacker, G. H., 1980. Changes in body measurements of larval northern anchovy, Engraulis mordax, and other fishes due to handling and preservation. Fish. Bull. U.S.A. 78: 685–692.Google Scholar
- Theilacker, G. H., K. M. Bailey, M. F. Canino & S. M. Porter, 1996. Variations in larval walleye pollock feeding and condition: a synthesis. Fisheries Oceanography 5 (Suppl. 1): 112–123.CrossRefGoogle Scholar
- Wagner, M., L. J. Buckley, A. Durbin and T. Durbin, 1998. RNA:DNA ratios as indicators of nutritional condition in the copepod Calanus finmarchicus. Marine Ecol. Prog. Ser. 162: 173–181.CrossRefGoogle Scholar
- Westerman, M. E. & G. J. Holt, 1988. The RNA-DNA ratio: measurement of nucleic acids in larval Sciaenops ocellatus. Contrib. Mar. Sci. 30: 117–124.Google Scholar
- Westerman, M. E. & G. J. Holt, 1994. RNA:DNA ratio during the critical period and early larval growth of the red drum Sciaenops ocellatus. Mar. Biol. (Berlin) 121: 1–9.CrossRefGoogle Scholar
- Wilder, I. B. & J. G. Stanley, 1983. RNA-DNA ratio as an index to growth in salmonid fishes in the laboratory and in streams contaminated by carbaryl. J. Fish Biol. 22: 165–172.CrossRefGoogle Scholar
- Wright, D. A. & F. D. Martin, 1985. The effect of starvation on RNA:DNA and growth of larval striped bass, Morone saxatilis. J. Fish Biol. 27: 479–485.CrossRefGoogle Scholar
- Young, V. R. 1970. The role of skeletal and cardiac muscle in the regulation of protein metabolism. In Munro H.H. (ed), Mammalian protein metabolism, Academic Press, London. 585–674.Google Scholar