Abstract
Annual killifish development is unique compared to other teleosts and is characterized by the dispersion and subsequent reaggregation of pre-embryonic blastomeres and the occurrence of embryonic diapause. Austrofundulus limnaeus is an excellent species to use for studies of development and embryonic diapause in annual killifish. A. limnaeus has a high fecundity, reproduces readily in a laboratory environment, and has a relatively long laboratory life span compared to many other species of annual killifish. Methods are presented for rearing A. limnaeus in the laboratory with an emphasis on collecting and incubating large numbers of embryos for biochemical and physiological studies. Females produce an average of 29 eggs during a two to four hour spawning. Egg quality (% fertilization and survival) and egg production (eggs female-1) are affected by the number of days between spawning events. Percent fertilization of eggs and survival of embryos decreases as the interval between spawning increases from two to eight days. The number of fertile embryos produced per female remains relatively constant as a function of spawning interval. Fertilization rates may be maintained at high levels by replacing aged males (1.5 years old) with younger males. An embryo medium was formulated to mimic the natural waters inhabited by A. limnaeus. The developmental rate and survival of embryos in the embryo medium was essentially equivalent when compared to Yamamoto's fish saline solution.
Similar content being viewed by others
References cited
Bailey, R.G. 1972. Observations on the biology of Nothobranchius guentheri (Pfeffer) (Cyprinodontidae), an annual fish from the coastal region of east Africa. The African Journal of Tropical Hydrobiology and Fisheries 2: 33–43.
Balon, E.K. 1990. Epigenesis of an epigeneticist: the development of some alternative concepts on the early ontogeny and evolution of fishes. Guelph Ichthyol. Rev. 1: 1–48.
Baroiller, J.F., D. Chourrout, A. Fostier & B. Jalabert. 1995. Temperature and sex chromosomes govern sex ratios of the mouthbrooding cichlid fish Oreochromis niloticus. J. Exp. Zool. 273: 216–223.
Boulekbache, H., J. Bastin, M. Andriamihaja, B. Lefebvre & C. Joly. 1989. Ageing of fish oocytes: effects on adenylic nucleotides content, energy charge and viability of carp embryo. Comp. Biochem. Physiol. 93B: 471–476.
Cameron, I.L. & K.E. Hunter. 1984. Regulation of the permeability of the medaka fish embryo chorion by exogenous sodium and calcium ions. J. Exp. Zool. 231: 447–454.
Carter, C.A. & J.P. Wourms. 1990. An ultrastructural analysis of the dispersed cell phase during development of the annual fish, Cynolebias. J. Morphol. 204: 209–225.
Carter, C.A. & J.P. Wourms. 1991. Cell behavior during early development in the South American annual fishes of the genus Cynolebias. J. Morphol. 210: 247–266.
Conover, D.O., D.A. van Hoorhees & A. Ehtisham. 1992. Sex ratio selection and the evolution of environmental sex determination in laboratory populations of Menidia menidia. Evolution 46: 1722–1730.
Craik, J.C.A. & S.M. Harvey. 1984. Egg quality in rainbow trout: the relation between egg viability, selected aspects of egg composition, and time of stripping. Aquaculture 40: 115–134.
Denucé, J.M. 1989. Developmental changes in annual fish embryos kept in the proximity of adult fishes. Acta. Embryol. Morphol. Exper. 10: 87–94.
Inglima, K., A. Perlmutter & J. Markofsky. 1981. Reversible stage-specific embryonic inhibition mediated by the presence of adults in the annual fish Nothobranchius guentheri. J. Exp. Zool. 215: 23–33.
Kinne, O. & E.M. Kinne. 1962. Rates of development in embryos of a cyprinodont fish exposed to different temperature-salinity-oxygen combinations. Can. J. Zool. 40: 231–253.
Kroll, W. 1984. Morphological and behavioral embryology and spontaneous diapause in the african killifish, Aphyosemion gardneri. Env. Biol. Fish. 11: 21–28.
Levels, P.J. & J.M. Denucé. 1988. Intrinsic variability in the frequency of embryonic diapauses of the annual fish Nothobranchius korthausae, regulated by light: dark cycle and temperature. Env. Biol. Fish. 22: 211–223.
Levels, P.J., L.J.H. Van Tits & J.M. Denucé. 1986. The effect of the presence of adult fishes, gonad homogenates, and embryo homogenates on the dispersion-reaggregation phase during early embryonic development of the annual fish Nothobranchius korthausae. J. Exp. Zool. 240: 259–264.
Lilyestrom, C. & D. Taphorn. 1982. El control biologico de mosquitos mediante peces en la cuenca del Lago de Maracaibo, Venezuela. Informe Tecnico del Vice — Rectorado de Produccion Agricola 2: 1–29.
Markofsky, J. & J.R. Matias. 1977a. Effects of light: dark cycles and temperature on embryonic diapause in the East African annual fish, Nothobranchius guentheri. Chronobiologia 4: 130–131.
Markofsky, J. & J.R. Matias. 1977b. The effects of temperature and season of collection on the onset and duration of diapause in embryos of the annual fish Nothobranchius guentheri. J. Exp. Zool. 202: 49–56.
Markofsky, J., J.R. Matias, K. Inglima, J.H. Vogelman & N. Orentreich. 1979. The variable effects of ambient and artificial light: dark cycles on embryonic diapause in a laboratory population of the annual fish Nothobranchius guentheri. J. Exp. Biol. 83: 203–215.
Markofsky, J. & A. Perlmutter. 1972. Age at sexual maturity and its relationship to longivity in the male annual cyprinodont fish, Nothobranchius guentheri. Exp. Geront. 7: 131–135.
Matias, J.R. & J. Markofsky. 1978. The survival of embryos of the annual fish Nothobranchius guentheri exposed to temperature extremes and the subsequent effects on embryonic diapause. J. Exp. Zool. 204: 219–228.
Matias, J.R. 1982. Embryonic diapause in annual fishes: evaporative water loss and survival. Experientia 38: 1315–1317.
Matias, J.R. 1983. The effect of exposure to gaseous ammonia on the duration of diapause II in the embryos of the annual fish, Nothobranchius guentheri. Experientia 39: 1148–1150.
Matias, J.R. 1984. The stage-dependent resistance of the chorion to external chemical damage and its relationship to embryonic diapause in the annual fish, Nothobranchius guentheri. Experientia 40: 753–754.
Mollah, M.F.A. & E.S.P. Tan. 1983. Viability of catfish (Clarias macrocephalus, Gunther) eggs fertilized at varying post-ovulation times. J. Fish Biol. 22: 563–566.
Myers, G.S. 1952. Annual fishes. Aquarium Journal 23: 125–141.
Nico, L.G. & J.E. Thomerson. 1989. Ecology, food habits and spatial interactions of Orinoco Basin annual killifish. Acta Biol. Venez. 12: 106–120.
Peters, N. 1963. Embryonale Anpassungen oviparer Zahnkarpfen aus periodisch austrocknenden Gewassern. Int. Rev. ges. Hydrobiol. 48: 257–313.
Podrabsky, J.E., T. Hrbek & S.C. Hand. 1998. Physical and chemical characteristics of ephemeral pond habitats in the Maracaibo Basin and Llanos region of Venezuela. Hydrobiologia 362: 67–78.
Strüssmann, C.A., S. Moriyama, E.F. Hanke, J.C.C. Cota & F. Takashima. 1996. Evidence of thermolabile sex determination in pejerrey. J. Fish Biol. 48: 643–651.
Taphorn, D.C. & J.E. Thomerson. 1978. A revision of the South American cyprinodont fishes of the genera Rachovia and Austrofundulus, with the description of a new genus. Acta Biol. Venez. 9: 377–452.
Trinkaus, J.P. 1967. Fundulus. pp. 113–121. In: F.H. Wilt & N.K. Wessells (ed.) Methods in Developmental Biology, Thomas Y. Crowell Co., New York.
Walford, R.L. & R.K. Liu. 1965. Husbandry, life span, and growth rate of the annual fish, Cynolebias adloffi E. Ahl. Exp. Geront. 1: 161–171.
Westerfield, M. 1995. The zebrafish book. A guide for the laboratory use of zebrafish (Danio rerio), edition 3. University of Oregon Press, Eugene. 378 pp.
Wourms, J.P. 1964. Comparative observations on the early embryology of Nothobranchius taeniopygus (Hilgendorf) and Aplocheilichthys pumilis (Boulenger) with special reference to the problem of naturally occurring embryonic diapause in teleost fishes. pp. 68–73. In: East African Freshwater Fisheries Research Organization, Annual Report for 1964, Jinja.
Wourms, J.P. 1967. Annual fishes. pp. 123–137. In: F.H. Wilt & N.K. Wessells (ed.) Methods in Developmental Biology, Thomas Y. Crowell Co., New York.
Wourms, J.P. 1972a. Developmental biology of annual fishes I. Stages in the normal development of Austrofundulus myersi Dahl. J. Exp. Zool. 182: 143–168.
Wourms, J.P. 1972b. The developmental biology of annual fishes II. Naturally occuring dispersion and reaggregation of blastomeres during the development of annual fish eggs. J. Exp. Zool. 182: 169–200.
Wourms, J.P. 1972c. The developmental biology of annual fishes III. Pre-embryonic and embryonic diapause of variable duration in the eggs of annual fishes. J. Exp. Zool. 182: 389–414.
Zar, J.H. 1996. Biostatistical analysis, 3rd ed. Prentice-Hall, Upper Saddle River. 662 pp.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Podrabsky, J.E. Husbandry of the Annual Killifish Austrofundulus limnaeus with Special Emphasis on the Collection and Rearing of Embryos. Environmental Biology of Fishes 54, 421–431 (1999). https://doi.org/10.1023/A:1007598320759
Issue Date:
DOI: https://doi.org/10.1023/A:1007598320759