CHAPTER III

PHYSIOLOGICAL AND
BIOCHEMICAL ASPECTS OF
ARTEMIA ECOLOGY

JAMES S. CLEGG
Molecular and Cellular Biology and Bodega Marine Laboratory
University of California (Davis)
Bodega Bay, CA 94923
USA

CLIVE N.A. TROTMAN

Department of Biochemistry
University of Otago, Box 56
Dunedin, New Zealand

1. Introduction

This chapter considers life cycle-dependent biochemical and physiological

adaptations critical to the survival of Artemia in nature. Thus, the encysted
gastrula embryo ('cyst') is arguably the most resistant of all animal life history
stages to extremes of environmental stress, while the motile stages are among
the best osmoregulators in the animal kingdom. These and other adaptations
are emphasized not only because they are essential to the success of brine
shrimp, enabling them to cope with the many challenges presented by their
harsh environment, but because this animal is a useful model system for the
study of a variety of general biological processes. We hope our coverage
will encourage others to study this remarkable organism.
A number of books (Persoone et al. 1980; Decleir et al. 1987; MacRae et
al. 1989; Warner et al. 1989; Browne et al. 1991) and reviews (Clegg, 1986a,b;
Hand and Hardewig, 1996; Guppy and Withers, 1999) have been published
that treat these aspects to varying degrees. As a result, the reference list is
not exhaustive, particularly for many of the earlier publications and for bio-
chemical or physiological studies that are not easily related to the ecology
of Artemia, the major focus of this chapter (completed in November 2000).
We apologize to authors of those papers, and to those whose work we over-
looked inadvertently.
Finally, we note that almost all of the research to be considered here involves
populations of a single species, Artemia jranciscana, mainly from the South
San Francisco Bay of California and the Great Salt Lake, Utah, USA. Unless

Th. J. Abatzopoulos et al. (eds.), Artemia: Basic and Applied Biology, 129-170.
© 2002 Kluwer Academic Publishers.
130 JAMES S. CLEGG AND CLIVE N.A. TROTMAN

stated otherwise, the coverage refers to this species. Whether generalizations

developed here apply to other Artemia species remains to be seen.

2. The Two Paths of Development

Whether bisexual or parthenogenetic, development of a given clutch of eggs

follows one of two paths: ovoviviparous, resulting in nauplius larvae, or
oviparous resulting in encysted gastrula embryos (cysts) that enter diapause,
a state of obligate dormancy, released into their, usually hypersaline, envi-
ronment. Diapause is terminated by environmental cues which differ in detail
for the various species and even in geographically separated populations of the
same species (Lavens and Sorgeloos, 1987; Drinkwater and Clegg, 1991).
Embryos released from diapause resume development under permissive con-
ditions of oxygen availability, water content and temperature. The embryo's
inner embryonic cuticle is apparently impermeable to non-volatile solutes
(De Chaffoy et al. 1978; Clegg and Conte, 1980) and, as we shall see, this
relative independence from the environment has substantial adaptive signifi-
cance in terms of inorganic ion homeostasis. Larvae produced from cysts
appear to be morphologically the same as those produced ovoviviparously;
however, significant biochemical differences exist (Liang and MacRae, 1999).
It is certainly true that newly hatched larvae have adaptive repertoires very
different from the encysted embryos, which is not surprising since the envi-
ronmental challenges they face are so different.
These two paths involve adaptive features related in an obvious way to their
ecological setting. Although exceptions exist, ovoviviparity is commonly taken
when conditions are favourable (such as low or moderate salinity, high oxygen
tension, and abundance of food) whereas diapause cysts often result when these
and other conditions are not as favourable. This scenario is a common one
in organisms that experience wide fluctuations in environmental parameters
and are capable of producing either active or dormant stages.
For the sections that follow it may be useful to refer to the structure of
encysted embryos, one of which is illustrated in Figure 1. This cyst, repre-
sentative of many examined, had been dried to terminate diapause, then fully
re-hydrated (at 2-4 °C to suppress metabolism) before being prepared for
electron microscopy by conventional methods (Clegg et al. 2000a). To our
knowledge the only detailed electron microscopic description of diapause
embryos is the doctoral dissertation of Jardel (1986), although Criel (1991a)
describes a few of their ultrastructural features. The early study by Morris
(1968) on electron microscopy of dried (activated) cysts revealed what one
might expect for cells that are able to desiccate reversibly: although dense
and highly compact, ultrastructural integrity was retained and most cellular
components were easily recognized, although some with modification. Later
we consider some possible mechanisms underlying the retention of ultra-
structural integrity in the desiccated embryo. Other ultrastructural observations