Journalof IndustrialMicrobiology(1996) 17, 477-489

9 1996 Societyfor IndustrialMicrobiology016%4146/96/$12.00

Biodiversity and application of microalgae

FB Metting Jr

Battelle Pacific Northwest National Laboratory, MSIN P7-54, PO Box 999, 902 Battelle Boulevard, Richland, Washington
99352, USA

The algae are a polyphyletic, artificial assemblage of 02-evolving, photosynthetic organisms (and secondarily non-
photosynthetic evolutionary descendants) that includes seaweeds (macroalgae) and a highly diverse group of micro-
organisms known as microalgae. Phycology, the study of algae, developed historically as a discipline focused on
the morphological, physiological and ecological similarities of the subject organisms, including the prokaryotic blue-
green algae (cyanobacteria) and prochlorophytes. Eukaryotic algal groups represent at least five distinct evolution-
ary lineages, some of which include protists traditionally recognized as fungi and protozoa. Ubiquitous in marine,
freshwater and terrestrial habitats and possessing broad biochemical diversity, the number of algal species has
been estimated at between one and ten million, most of which are microalgae. The implied biochemical diversity is
the basis for many biotechnological and industrial applications.

Keywords: microalgae; cyanobacteria; biodiversity; applied phycology

Introduction sequencing, flagellar hairs and roots, plastid and mitochon-

drial structure, the mitotic apparatus) has combined to cre-
The phylogenetic diversity of the algae is very broad and
ate an exciting, dynamic field of inquiry. Biodiversity
is reflected in an equally wide range of metabolisms and
implies that groups of organisms are differentiated by some
biochemical properties. This paper considers the phylogeny
measure of the extent to which their gene pools are separ-
(evolutionary relatedness), classification (taxonomy), ecol-
ated and how this is expressed phenotypically. For purposes
ogy and distribution, biochemical diversity and industrial
of classification and for understanding biological and evol-
application of eukaryotic and prokaryotic microalgae in an
utionary relatedness (phylogeny), the species is the funda-
attempt to illustrate the biodiversity of this assemblage of
mental unit for classifying groups of organisms. Thus, in
largely microscopic phototrophs. The volume of infor-
order to assess biodiversity, it is important to consider the
mation on these subjects is more than sufficient for a num-
species concept as it is applied to the algae. With the
ber of textbooks and this article is by no means believed
increasing availability of molecular information (ie protein
or intended to be authoritative or complete. For in depth
and nucleic acid sequence data), there has been a movement
information, interested readers are encouraged to consult
toward reconciliation of taxonomic and phylogenetic
the literature cited in the text and Bold and Wynne [10]
approaches. This is leading to classification systems that
and van den Hoek et al [93] on algal biology, systematics,
reflect some biological reality, such as the degree to which
and classification, Cox [26] on photosynthetic flagellates.
groups of populations are genetically similar, with impli-
Margulis et al [59] and O'Kelly [70] for perspectives on
cations for evolutionary history and speciation. The results
phylogeny and classification, Andersen [2], Norton et al
of ongoing molecular level interrogation of cultured and
[68], and Radmer [77] on biodiversity, Darley [28], Rogers
natural microbial populations is leading to completely new
and Gallon [85] and Stewart [91] on physiology and bio-
frameworks for categorization at the higher taxonomic lev-
chemistry, and Akatsuka [l], Borowitzka and Borowitzka
els. For an in depth treatment of the interrelationships
[14], and Lembi and Waaland [55] on commercial appli-
among phylogeny and classification based on current
cations. Much of our fundamental knowledge of microalgae
debate on the species concept applied to algae, see Manhart
is based on research with isolates cultured in the laboratory.
and McCourt [58].
For a discussion of culturing and a listing of culture collec-
Historically, species of microalgae were recognized on
tions throughout the world, see Andersen [3].
the basis of phenotypic properties, such as whole organism
morphology, cellular anatomy and ultrastructure, and
Phylogeny and classification metabolism and physiology and were described and catego-
rized according to the International Code of Botanical
Our understanding and interpretation of the phylogeny of
Nomenclature. More recently, it has been recommended by
algae and related organisms has evolved dramatically in
some bacteriologists that the taxonomy of prokaryotic blue-
recent years. Molecular and ultrastructural evidence of
green algae (cyanobacteria or cyanophytes*) and prochloro-
evolutionarily conserved features (eg ribosomal RNA gene
phytes be treated under the International Code of
Nomenclature of Bacteria, with the botanical system serv-
Correspondence: FB Metting Jr, Battelle PacificNorthwest National Lab-
oratory, MSIN P7-54, PO Box 999, 902 Battelle Boulevard, Richland,
Washington 99352, USA *In this article, the terms blue-green algae, cyanophytes,and cyanobac-
Received 19 March 1996; accepted 30 August 1996 teria are used interchangeably.
 Biodiversity and application of microalgae
FB Metting Jr
478
ing as the baseline [21]. However, very few species have lar coccoid, colonial, and filamentous forms with or without
been described under the bacteriological code and most branching or differentiation of specialized cells, such as
researchers agree to the pressing needs for resolution of akinetes (spores) and heterocysts (anaerobic N2-fixing
this issue. Geitler [32] continues to be used as the baseline cells). Blue-greens are thought to be the evolutionary origin
authority for the identification and preliminary classi- of plastids within eukaryotic algae and land plants (an event
fication of genera and putative species of blue-green algae. that probably occurred more than once) [80] and respon-
Species are placed in Orders that are defined on the basis sible for oxygenation of the atmosphere early in our earth's
of morphology and contrasting modes of cell division. history. Thylakoids (photosynthetic membranes) are not
Because of the dynamic nature of research and debate in arranged in stacks, in contrast to prochlorophytes and most
the phylogeny of eukaryotic microorganisms and the cur- other algae. Pigmentation includes chlorophyll a and d,
rent uncertainty about placement of a number of groups, blue and red phycobilins, /3-carotene, and several xantho-
and also for continuity with research in ecology and distri- phylls. The storage product, cyanophycean starch (an c~-(1-
bution, biochemical diversity and applied phycology, this 4)-linked glucan), is formed as granules between the thy-
article will rely on a classification scheme based on a tra- lakoids [93]. Blue-green algae produce at least 50 com-
ditional approach. Historically the major groups of algae pounds which are toxic to vertebrates in addition to anti-
are classified into divisions (equivalent to the zoological biotics and other secondary metabolites [19].
phylum) on the basis of pigmentation, the chemical nature Prochlorophytes are prokaryotic microalgae in the same
of the photosynthetic storage product, thylakoid organiza- bacterial division and class as the blue-green algae. They
tion and other ultrastructural features of the chloroplast, the are distinct from the blue-greens by possessing both of the
chemistry and structure of the cell wall, the number, chlorophylls found in green algae, euglenoids and land
arrangement and ultrastructure of flagella (if any), and the plants (a and b or divinyl derivatives). Discovered less than
occurrence of any special features. Ten of the divisions 30 years ago as extracellular symbionts within subtropical
recognized by van den Hoek et al [93], representing vir- ascidians (sea squirts), but now known to include free-liv-
tually all known algae, are listed in Table 1. It is now ing freshwater and marine plankton, the prochlorophytes
agreed that the algae are not a cohesive, natural assemblage may themselves be polyphyletic in origin [72].
of organisms. Depending on how the data for problematic
groups are interpreted, algae are included in as many as
seven [2], eight [70] or more eukaryotic and one or more
Glaucophytes
The eukaryotic division Glaucophyta includes three genera
eubacterial lines within the domains Eukarya and Bacteria
of biflagellate unicellular microalgae whose chloroplasts
(sensu Woese et al [99]). At this level, many people agree
strongly resemble coccoid blue-green algae in that the thy-
that eukaryotic algae for which sufficient data are available
lakoids are not stacked and are surrounded by a thin pep-
represent a minimum of four or five phyletic assemblages:
tidoglycan layer (a wall typical of Gram-negative bacteria).
green plants (including green algae and land plants), red
Also like blue-greens, glaucophytes have chlorophyll a and
algae, euglenoids, chromophyte algae, and dinoflagellates
phycobilins, although the range of xanthophylls present is
(which are either included with the chromophytes or con-
smaller. They are rare freshwater inhabitants [93].
sidered to be an independent lineage). The diversity among
these major lineages is illustrated in Figure 1, where it can
be seen that algae are phylogenetically more diverse than Red algae
either land plants or animals. Groups for which no clear Red algae (Rhodophyta) include seaweeds and microalgae
consensus has yet emerged because of paucity of data or distinct from other algae by having the following combi-
opposing interpretation of what evidence is available nation of properties: (1) the lack of any flagellate stages;
include the glaucophytes, prymnesiophytes, chloroacrachni- (2) the presence of accessory phycobilin pigments (shared
ophytes and cryptophytes (Table 1). Among the algae, fos- with cyanobacteria, glaucophytes, and cryptomonads);
sil records show the blue-green lineage arising early in the (3) chloroplasts with singly-occurring thylakoids;
Precambrian. The red and green lineages date from the mid- (4) ftoridean starch (an c~-(1-4-glucan) as the photosynth-
to late-Precambrian. Brown algae are first seen in the Paleo- ate; and (5)oogamy in species with sexual reproduction
zoic while most other lineages date from the early to late [93]. Microalgal species have not been shown to reproduce
Mesozoic [3]. sexually. Chlorophyll a is the only chlorophyll. In addition
to phycobilins (phycoerythrin imparts the red color), rhodo-
Blue-green algae and prochlorophytes phytes have /3-carotene, zeaxanthin and, as a group, five
All blue-green algae and prochlorophytes are Gram-nega- other known xanthophylls. Two classes are usually recog-
tive eubacteria classified by bacteriologists as the class nized on the basis of whether the plant body is relatively
Oxyphotobacteria in the eubacterial division Gracilicutes simple (Bangiophyceae, includes microalgae) or complex
[22,67]. Species are described on the basis of morphology, (Floridophyceae) together with a number of anatomical
although phylogenetic analysis based on molecular data has characteristics and attributes of the life history [93]. Red
revealed that some morphologically cohesive assemblages, seaweeds are distributed worldwide along coastlines. In
such as the non-heterocystous filamentous 'oscillatorian' addition to marine habitats, red microalgae occur in fresh-
algae, include phylogenetically distinct groups suggesting water, on soil, and as endosymbionts within tropical fora-
that some species and genera may require extensive taxo- minifera (protozoa). One marine gastropod harbors a red
nomic revision [33,97]. Among the approximately 2500 algal chloroplast as a photosynthetic quasi-organelle.
described species of blue-green algae are included unicellu- Nearly all of the approximately 5500 species are seaweeds.
 Biodiversity and application of microalgae
FB MettingJr
479
Table 1 Biodiversity, distribution, and estimated numbers of algal species

Divisiona Distinguishing Major groups a Distribution of microscopic species Estimated number

(common features t' (common name of species
name) and/or features) Marine Fresh- Terrestrial Symbiotic (microalgae and
water seaweeds)~

Cyanophyta Chlorophyll a,d: phycobilins: Chroococcales + + + + ~125 described

(blue-green /3-carotene; zeaxanthin, (Unicellular cocci and
algae) echinenone, canthaxanthin, rods; binary fission and
myxoxanthophyll, budding)
oscillaxanthin, 3 minor
xanthophylls; phycobilins;
prokaryotic; Gram-negative
cell walls
Pleurocapsales + + - - ~35 described
(Unicellular and
aggregate-forming;
multiple fission)
Oscillatoriales + + + - 1000
(Filamentous; binary
fission; no specialized
cells)
Nostocales + + + + iooo
(Filamentous; binary
fission; specialized cells;
heterocysts, akinetes)
Stigonematales - + + - ~35 genera
(Filamentous; branched;
cell division in 3 planes;
specialized cells;
heterocysts, akinetes and
motile, gliding
hormonogia)
Prochlophyta Chlorophyll a,b; /3-carotene; Prochlorales + ? ? + <10 described
(prochlorophytes) several xanthophylls; (Coccoid and
phycobilins, prokaryotic; filamentous; binary
Gram-negative cell walls fission)
Glaucophyta Chlorophyll a,b; phycobilins; 3 described
(glaucophytes) /3-carotene; zeaxanthin, /3-
cryptoxanthin; starch;
cellulose wall
Rhodophyta Chlorophyll a; c~-, /3- Bangiophycidae + + + + 5500-20 000 (both
(red algae) carotenes; phycobilins; Floridophycidae + classes)
zeaxanthin, 5 minor (seaweeds only)
xanthophylls; ftoridean starch;
cellulose and other cell wall
materials
Cryptophyta Chlorophyll a,c2; ee-, /3-, e- + + ? ? 1200
(cryptophytes) carotenes; phycobilins;
alloxanthin, zeaxanthin, c~-1,4
glucan starch; exterior organic
periplast
Chlorophyta Chlorophyll a,b; oe-, /3-, "y- Micromonadophyceae + + ? ? 500
(green algae) carotenes; zeaxanthin, lutein, (prasinophytes)
violaxanthin, neoxanthin;
several minor xanthophylls;
starch; diverse cell wall
materials, incl cellulose
Charophyceae + + + ? 20 500
(stoneworts, filamentous
microalgae)
Ulvophyceae + + + ? 3000
(seaweeds, filamentous
microalgae)
Pleurastrophyceae + + + + 6 genera
Chlorophyceae + + + + 10 000-100 000
 Biodiversity and application of microalgae
FB Metting Jr
480
Table 1 Continued

Division a Distinguishing Major groups ~ Distribution of microscopic species Estimated number

(common features b (common name of species
name) and/or features) Marine Fresh- Terrestrial Symbiotic (microalgae and
water seaweeds)C

Euglenophyta Chlorophyll a,b; /3-W- + + + ? 2000

(euglenoids) carotenes; neoxanthin,
diadinoxanthin, diatoxanthin;
paramylon; wall-like organic
pellicle internal to cell
membrane
Chlorarachnio- Chlorophyll a,b; unknown + - - + 2 genera, each with 1
phytes carotenoids; amoeboid with species
zoospores
PyiThophytad Chlorophyll a,c; /3-carotene; + + ? + 3500-11000
(dinoflagellates) diadinoxanthin, diatoxanthin,
fucoxantbin, peridinin, 5
minor xanthophylls; a-l,4
glucan starch; cellulose walls
Chromophyta Chlorophyll a,c,,c2,Cz; o~-, ,6- Chrysophyceae (golden- + + ? ? 3400
(heterokont and e-carotenes; several major brown algae)
algae) ~ and minor xanthophylls;
chrysolaminarin, other
glucans, oils; variety of cell
wall materials
B acill ariophyceae + + + ? 100 000-10 000 000
(diatoms)
Xantbophyceae (yellow- + + + ? 2000
green algae)
Eustigmatopbyceae + + + 9 1000-10 000
(eustigmatophytes)
Raphidophyceae + + ? ? 100
(raphidophytes,
cbloromonads)
Prymnesiophyceae + + ? ? 2000
(baptophytes or
prymnesiophytes,
including
coccolithophorids)
Dictyochophyceae + . . . . 15
(silicoflagellates)
Phaeophyceae (brown + + + ? 2000
seaweeds, no
microalgae)

aSensu Castenholtz and Waterbury [22] for cyanophyte orders, Lewin [56] for prochlorophyte orders, and van den Hoek et al [93] for all eukaryotic
divisions and classes.
bNot all classes within a division include taxa displaying all the distinguishing features. In particular, classes of chromophyte algae vary considerably
in pigmentation and nature of the photosynthate [9].
CFrom Andersen [2] and Norton et al [68].
dSome dinoflagellates include endosymbiotic brown algae so that while most species have peridinin as the principal xanthophyll, others have fucoxantbin.
This may represent multiple endosymbiotic events in the evolutionary past [70].
~Some authors recognize additional classes of chromophyte algae, including the Synurophyceae and Pelagophyceae [2,88].

Ctyptophytes [23] allies them with the chromophyte algae. Starch is

Cryptophytes (Cryptophyta) are a small group of mostly stored as distinct granules and some species also store
unicellular biflagellate microalgae with a broad collection oils [93].
of pigments, including chlorophylls a and c2 and phycobil-
ins, and a single chloroplast with a thylakoid structure inter- Green algae
mediate between the red algae and other plants. The cell is The green algae (Chlorophyta) comprise one of the largest
enclosed by a stiff proteinaceous periplast made of rec- in terms of number of species, most widely distributed, and
tangular or polygonal plates. Ultrastructural properties of morphologically diverse groups of algae. Land plants prob-
the chloroplast and unique cellular inclusions place crypto- ably evolved directly from green algae and share many bio-
monads apart from other algae, although Cavalier-Smith chemical, metabolic and ultrastructural features. Most chlo-
 Biodiversity and application of microalgae
FB Metting Jr

481
by prymesiophytes, a class of chromophyte algae. Euglen-
oids do not have cell walls, but rather possess a protein-
aceous pellicle internal to the cell membrane. Osmotrophic
and phagotrophic facultative and obligate heterotrophs are
common. All euglenoids are microscopic and include
mostly uninucleate and unicellular flagellates, although col-
onial species are common [93].

Chromophyto algae
The chromophyte algae are a highly diverse group, most of
which share a common storage product (chrysolaminarin, a
/3-1inked glucan) and a preponderance of carotenoids over
chlorophylls that results in all groups having golden,
golden-brown, brown or yellow-green rather than the grass-
green, blue-green or red coloration typical of other major
algal divisions. Five of the many chromophyte classes
(Table 1) feature chlorophylls a, Ca, and c2. Eustigmato-
phytes have only chlorophyll a. Fats and oils are stored by
many chromophyte algae and prymnesiophytes make para-
mylon, found elsewhere only in euglenoids. Ultrastructural
and molecular sequence comparisons over the past decade
have revealed that chromophytes may be closely related to
certain fungi and protozoa as well and it was recently pro-
posed to place them with these non-photosynthetic relatives
into a single kingdom with the common name strameno-
Figure 1 Depiction of the phylogenetic relatedness of some groups of piles, reflecting the common presence of tripartite (bipartite
algae. Lengths of line segments are proportional to evolutionary distance for the prymnesiophytes) mastigonemes, or hairs on one
based on analysis of ribosomal RNA gene sequences (modified from
Radmer and Parker [78]).
flagellum [73]. This division includes ecologically and mor-
phologically distinctive microalgat classes such as the dia-
toms, coccolithophorids, and silicoflagellates [93].
rophytes store starch and the group as a whole includes a
wide diversity in the number and arrangements of chloro- Dinotlagellatos
plasts and flagella associated with individual cells. Thy- The dinoflagellates (Pyrrhophyta) are a diverse assemblage
lakoids are tightly stacked in bundles of from two to six of coccoid, filamentous, palmelloid, and amoeboid microal-
lamellae within plastids bound by two membranes, in com- gae within which the evolutionary acquisition of plastids
mon with red algae and glaucophytes. Green algae include and phototrophy has occurred more than once [70].
unicellular, colonial, filamentous and pseudoparenchyma- Together with photosynthesis, a range of heterotrophic
tons uni- or multinucleate forms and large numbers of both nutritional patterns is represented, with many saprophytic,
seaweeds and microscopic species have been described. phagotrophic, symbiotic and parasitic species having been
Major taxonomic categories (Classes) listed in Table 1 are described. Dinoflagellates share the presence of chloro-
delineated largely on the basis of ultrastructural details of phylls a, ca, and Cz and some carotenoids with chromophyte
flagellate cells and mechanisms of cell division and so are algae, but also have unique carotenoids and a cellulose
thought to accurately reflect phylogeny within the wall. Most dinoflagellates have two uniquely positioned
division [60,93]. flagella. Ultrastructural aspects of flagellar and nuclear
organization are also unique to this group. Dinoflagellates
Chlorarachniophytes are important freshwater and marine inhabitants [93].
This rare group includes species in two genera that appear
to be some evolutionalT combination of an amoeboid, plas-
modial, phagotrophic host with an eukaryotic chlorophyll
Ecology and distribution
a and b containing endosymbiont derived from a euglenoid As the predominant component of the marine and fresh-
or chlorophyte [70]. They are described from association water plankton, microalgae are primarily responsible for the
with marine siphonous green algae [93]. 40-50% of total global photosynthetic primary production
contributed by all algae [41]. Included in aquatic habitats
Euglonoids are microplankton (20-200/xm), nanoplankton (2-20/zm)
Euglenoids (Euglenophyta) share pigmentation with the and picoplankton (0.2-2 p,m) [90]. In recent years it has
prochlorophytes, green algae, and land plants, but possess become evident that much or most primary production is
unique cellular and biochemical features that, together with contributed by cells < 3 - 5 / x m in diameter, known as ultra-
ultrastructural and molecular evidence, places these micro- plankton, that together with similar-sized heterotrophic
organisms closer to trypanosomes than to any algal group plankton constitute as much as 80% of the pelagic biomass
[70]. The storage product is paramylon, a/3-1,3 linked glu- [2,89]. While the contribution by pelagic, coastal, and
can, located freely in the cytoplasm that is also produced freshwater communities is relatively well understood, the
 Biodiversity and application of microalgae
FB Metting Jr

482
significance of terrestrial microalgae is not known with any microalgae in the order Porphyridiales. The genera Rhod-
degree of precision, but they may be important on a local ella and Rhodosorus include marine phytoplankton species.
or regional scale because of their predominance on exten- They are regular inhabitants at some locations, such as
sive areas of exposed land surface in arid and semi-arid Norwegian fjords. Under favorable circumstances, the
steppes and deserts [63]. release of male gametes by red seaweeds makes a signifi-
Phytoplankton reside throughout the photic zone from cant ephemeral contribution to coastal benthic phytoplank-
the surface layer (neuston) to 250 m or more in depth in ton communities [92]. Porphyridium inhabits freshwater
some clear oligotrophic waters and depending on latitude. and terrestrial ecosystems [93].
Andersen [2] calculated that as many as 3.6 x 1025 individ-
ual phytoplankters may inhabit the world's oceans at any Cryptophytes
given time and are responsible for an annual primary pro- Members of this group are sporadically present in fresh-
duction of over 5 x 1013 kg, constituting the base of the water and marine habitats. Some species of Chroomonas
marine food web. As a consequence of diurnal, seasonal, exhibit broad tolerance to salt concentrations, living in estu-
vertical, and geographic variation in nutrient availability, aries and salt marshes [92]. One species is symbiotic within
temperature, light and other factors, the distribution and the marine ciliate Mesodinium.
metabolic activities of groups of microalgae may be con-
siderably more heterogeneous than is currently understood. Green algae
The interpretation of data from flow cytometry, extraction Green algae are ubiquitous in freshwater habitats, where
and analysis of community-level nucleic acids, phospho- they commonly dominate phytoplankton assemblages, and
lipid and other biochemical assays, remote sensing, and tra- are abundant in terrestrial environments as well. The green
ditional microscopy has resulted in some generalizations algae range in size from microscopic to macroscopic in all
regarding the distribution of phytoplankton. These are cer- of these environments. Predominantly photosynthetic, there
tain to be refined as more surveys across geographic and are nonetheless numerous examples of facultative and obli-
temporal gradients are reported and as our knowledge gate heterotrophs [93].
improves about the distribution and significance of viable Representative genera that are widely distributed in mar-
but non-culturable species, believed by many people to ine environments include mostly uniflagellate (eg Pedino-
include the bulk of microbial biodiversity. The discussion monas, Micromonas) and biflagellate prasinophytes (eg
of the occurrence and general distribution patterns of the Mamiella, Mantoniella) and chlorophytes (eg Pyrami-
major groups of marine microalgae that follows is largely monas, Tetraselmis). Various coccoid chlorophytes less fre-
taken from Thomsen's 1986 survey [92] in which he indi- quently documented (eg Chlorella, Nanochloris,
cated that the depth of information about the different algal Halochlorococcum) may also prove ultimately to have
groups is highly variable and that creation of a systematic worldwide distribution and importance. Dunaliella, a uni-
framework for presenting or interpreting the current data is cellular biflagellate phytoplankter found in extremely saline
difficult at best. Thus, the following treatment is meant only habitats, such as seaside rock pools and saline lakes, includ-
to provide an appreciation for the ecological diversity of ing the Dead Sea and Great Salt Lake, is a focus of efforts
microalgae with the degree of attention paid to each group for commercial production of/3-carotene [15].
in no way reflecting its perceived or real importance in Green microalgae are abundant and diverse in freshwater
the biosphere. habitats. For example, unicellular (eg Chlamydomonas,
Phacotus) and colonial (eg Volvox, Pyrobotrys) flagellates,
Blue-green algae and prochlorophytes coccoid non-flagellate species (eg ChlorelIa, Pediastrum,
Cyanobacteria are ubiquitous in marine, freshwater, and ter- Ankistrodesmus, Scenedesmus), and filamentous forms (eg
restrial environments as free-living populations and endo- Chaetophora, Oedogonium) are ubiquitous in ponds, lakes,
symbionts of marine animals, lichens, bryophytes, and and streams at most latitudes. Many green microalgae are
cycad roots. In the past two decades, ultraplanktonic cyano- important as 'weeds' in lakes, canals, and other waterways
bacteria (eg Synechococcus) have been shown to contribute [54]. Most of these are mat-forming (eg Spirogyra,
as much or more to oceanic primary production as any other Hydrodictyon) or attached (eg Cladophora, Ulothrix,
single group of microalgae [31,95]. In addition, filamentous Stigeoclonium) filamentous species [10,93].
cyanobacteria such as Anabaena, Nostoc, Microcoleus, Green microalgae also occur in terrestrial habitats,
Oscillatoria and Mastigocladus are commonly the domi- including on and in soil and rocks. The most commonly
nant component of microbial mats in brackish waters, hot encountered are coccoid (eg Chlorococcum), palmelloid--
springs, on semi-arid and arid soils, and in rice paddies aggregates of cells invested by a common mucilage--(eg
[20,62,84]. Prochlorophytes (eg Prochloron) were first Palmella, Gloeococcus), sarconoid--packets of 4, 8, 16 or
described two decades ago from symbiotic association with 32 cells--(eg Tetracystis, Chlorosarcina), and filamentous
marine didemnids and holothurians and are now known to (eg Stichococcus, Klebshormidium) genera [62]. Trentepho-
also exist as free-living components of the pelagic nano- lia inhabits tree trunks and other subaerial surfaces while
plankton where their distribution and importance in this other green microalgae are symbiotic, such as Trebouxia,
habitat is becoming clearer [25]. a common lichen phycobiont.

Red algae Euglenoids

Consisting mostly of seaweeds categorized into two major Euglena, Trachelomonas, Colacium and other euglenoid
botanical classes, the red algae include genera of free-living genera include many common freshwater and edaphic spec-
 Biodiversityand applicationof microalgae
FB MettingJr
483
ies. This group is less common in marine environments, known to bloom in eutrophic waters. Coccolithophores
although species in the genus Eutreptia occasionally form have been the subject of more investigations of geographic
very dense blooms in coastal waters. Most euglenoids are distribution in the oceans than most other major groups of
facultatively heterotrophic and/or phagotrophic. Colorless, algae. Subtropical and tropical marine waters support
phagotrophic species (eg Peranema) are also common [93]. diverse communities that include Emiliana, Discosphaera,
Rhabdosphaera and many other genera [98]. Coccolithus
Chromophyte algae and Emiliana are two coccolithophore genera common in
Chromophyte microalgae are highly diverse and important open oceans at arctic latitudes. Members of the genus Chry-
components of most marine and freshwater habitats. With sochromulina are common in all marine environments
the exception of Pelagococus, which has been cultured where they often dominate and can form dense populations
from across the north Pacific, golden-brown microalgae even in dilute brackish waters. Prymnesiophytes are
(Chrysophyceae: eg Ochromonas, Chromulina), although uncommon in freshwater and unknown from terrestrial
often encountered, are not often abundant in marine habitats.
environments. Other golden-browns are, however, known
to be important freshwater phytoplankton that commonly Dinoflagellates
form blooms in lakes and ponds. Synura, for example, is a Dinoflagellates are often important components of the
nuisance alga responsible for imparting undesirable odors microplankton of freshwater and marine habitats in which
and taste to drinking water from lakes and reservoirs. they are found. Dinophysis, Gymnodinium, Amphidinium,
Golden-brown microalgae have not been reported from soil. Peridinium, Ceratium and Prorocentrum all include photo-
In contrast, silicoflagellates (eg Dichtyota) are very com- synthetic flagellate species found at most latitudes in fresh
mon in the oceans, where they often form dense blooms. and marine bodies of water [92]. Dinoflagellates also
These microalgae are uncommon in freshwater and not include many specialized species, including endo- and ecto-
known from terrestrial habitats. Pedinella is another pelagic parasites of marine animals (eg Blastodinium, Haplozoon)
genus whose abundance under ice can color the seawater and the photosynthetic partners of many corals known as
yellow [921. zooxanthellae [93]. Dinoflagellates include species capable
Diatoms (Bacillariophyceae) are ubiquitous in marine, of bioluminescence (eg Gonyaulax, Pyrocystis) and those
freshwater and terrestrial environments and probably responsible for 'red tides' (see section on Bioactive com-
include the greatest number of extant species (up to ten pounds, below).
million) of any group of microalgae [49,86]. Diatoms are
categorized into three classes on the basis of having bilat-
Biochemical diversity
eral or radial symmetry. Bacillariophyceae include bilater-
ally symmetrical species with raphes (an opening or fissure The phylogenetic breadth of microalgae is reflected in an
in the valve). Common genera include Navicula, Nitzschia, equally broad biochemical diversity of pigments, photosyn-
and Phaeodactylum. Diatoms displaying bilateral symmetry thetic storage products, cell walls and mucilages, fatty acids
but without raphes are in the class Fragilariophyceae (eg and lipids, oils, sterols and hydrocarbons, and bioactive
Diatoma, Synedra, Fragilaria). Radially symmetrical dia- compounds, including secondary metabolites. Much of the
toms are in the class Coscinodiscophyceae (eg Chaeto- material to follow is from contributed chapters to volumes
ceros, Biddulphia, Thalassiosira). Diatoms are mostly uni- edited by Borowitzka and Borowitzka [14], Rogers and
cellular, although filamentous species are also abundant and Gallon [85], and Stewart [91].
diverse. These microalgae commonly dominate planktonic
communities at all latitudes, particularly in and under ice, Pigments
in temperate coastal and pelagic environments, on mud Chlorophylls, carotenoids, and phycobilins comprise the
flats, in still or moving freshwater, and on some soils [86]. range of pigments produced in various combinations by
Yellow-green microalgae (Xanthophyceae: eg Vau- photosynthetic microalgae. Chlorophyll a is present in all
cheria, Bumilleria) are primarily known from freshwater algae and land plants, including the prokaryotic cyanobac-
environments. Unicellular, aggregate and filamentous xan- teria and prochlorophytes. Other chlorophylls (b, cl, c2, d)
thophytes are also common on soil. The eustigmatophytes are accessory light-harvesting molecules whose distribution
Nanochloris and Nanochloropsis (Enstigmatophyceae) among algal groups is used, in part, for taxonomic purposes
have been discovered over the past two decades to be at the division level (Table 1). Chlorophylls absorb blue
important contributors to primary production in estuaries, and red light and can range up to 2% or more of cellular
salt marshes, and brackish seas and are present, but less dry weight. Together with carotenoids and phycobilins, the
abundant, in the open ocean [3]. synthesis, content, and abundance of chlorophylls is depen-
Prymnesiophytes, also known as haptophytes dent on light, nutrition and other environmental factors
(Prymnesiophyceae or Haptophyceae) are microalgae that [61,82].
commonly dominate the marine phytoplankton in numbers Carotenoids are tetraterpenes comprised of eight
and biomass (eg Phaeocystis). They are sometimes dis- branched 5-C isoprenoid units. The variety of carotenoids
tinguished at the species level by the pattern of organic in algae is far greater than for land plants, with more than
scales and, in the case of the coccolithophorids, CaCO3 40 carotenes and xanthophylls having been isolated and
deposits on the cell surface. The biodiversity of prymnesi- characterized [9]. Several algal groups derive their primary
ophytes is greatest in warm oligotrophic regions, such as coloration from the mix of carotenoids they produce, for
the Red Sea and Gulf of California, although they are also example the yellow-green, golden, golden-brown and
 Biodiversity and application of microalgae
FB Metting Jr
484
brown coloration found within the chromophyte algae. /3 in euglenoids and to the outside in cryptophytes. Cyanobac-
carotene is chemically the most simple and is found in all teria and prochlorophytes have Gram-negative bacterial
algae and other plants. Hypersaline species of Dunaliella walls composed of c~- and e-diamino-pimelic acid and glu-
accumulate /3-carotene up to 14% of total dry weight. cosamine (peptidoglycan). Many or most green algae and
Astaxanthin is another important carotenoid that is pro- dinoflagellates have cellulose walls. Other chlorophyte
duced in large concentrations by green microalgae, includ- walls are constructed from hydroxy-proline glucosides, xyl-
ing Haematococcus and others that inhabit snow and ice ans, or mannans; some are calicified [93].
(eg Chlamydomonas spp). Many carotenes and xantho- Diatoms construct elaborately decorated walls of silicon
phylls are commonly restricted to one or a few algal groups. (frustules) made of two identical halves (valves) that fit
For example, lutein is shared by chlorophytes and land together like the lid and bottom of a petri dish. Devoid
plants, myxoxanthin and myxoxanthophyll are character- of walls, silicoflagellates instead produce siliceous internal
istic of cyanobacteria, as are peridinin for dinoflagellates and/or external skeletons, often with spiny projections,
and fucoxanthin for brown algae and diatoms. around and through which the protoplast moves. Prymnesi-
Phycobilins are water-soluble accessory pigments pro- ophytes, although without a rigid wall, are covered by
duced by cyanobacteria, glaucophytes, red algae, and superficial organic scales visible only with an electron
cryptophytes. As many as three different phycobilins can microscope. These scales are calcified in the coccolithopho-
be produced by an individual organism. In some algae, they rids [93].
modulate a metabolic process called chromatic adaption in Microalgal representatives in all of the divisions produce
which one form or another is preferentially synthesized a variety of extracellular polysaccharide mucilages. Much
according to the quality of available light [34]. Phycobilins smaller quantities are usually produced during active
are extracted from red and blue-green algae for use as flu- growth and cell division as compared to stationary phase.
orescent markers in cell biology and to impart color to food Mucilages can be responsible for aggregation of plankton,
and cosmetic products [12]. contribution to biofilm formation by attached species, and
desiccation tolerance of terrestrial forms and can possess
Photosynthetic storage products chelating and flocculant properties. The best-studied muci-
Microalgae synthesize a wide variety of photosynthetic lages are produced by cyanobacteria [4], diatoms and other
storage products (photosynthates), the chemical nature of chromophyte algae [42], chlorophytes [79], and red
which is also used as a criterion for taxonomic treatment microalgae [94]. Cell wall materials from brown and red
at the division level. Photosynthates include c~-(1-4)-linked seaweeds form the basis of a large marine hydrocolloid
glucans (starches), /3-(l-3)-glucans, fructans, low molecu- industry [77].
lar weight carbohydrates, and fats and oils. Starches pro-
duced by various groups include floridean starch Lipids, oils, sterols and fatty acids
(amylopectin subunits) by red algae and myxophycean The total oil and fat content of microalgae ranges from 1%
starch (amylopectin or glycogen-like subunits) by blue- to 70% of the dry weight and tends to be inversely pro-
green algae. Some green algae form a cross-linked amyl- portional to the rate of growth with greater accumulations
ose-amylopectin starch that is essentially similar to starch during stationary phase [11]. The percent of total lipid as
in land plants. Others store fructosans (inulin-like fructose neutral lipid, glycolipid, and phospholipid also varies
oligosaccharides). Cryptophytes and dinoftagellates also widely among and within groups of microalgae [11,50].
store c~-(1-4)-linked glucans. Chrysophytes store oils or Some species manufacture hydrocarbons. An example is
chrysophycean starch, a water-soluble /3-(1-3) glucopyr- Botryococcus, a chlorophyte capable of accumulating up to
anoside. Other chromophyte algae store various hydro- 90% of its colonial dry weight as a mixture of ten hydro-
carbons and oils (see below). Paramylon, composed solely carbon compounds in globules occluded among cells
of /3-(l-3)-linked D-glucose, is unique to euglenoids and embedded in a colonial matrix. Hydrocarbons up to C-37
prymnesiophytes [93]. in size from different strains of B. braunii include straight
Many microalgae in different divisions store monosacch- chain alkadienes, branched triterpenoids (botryococcenes),
arides (eg glucose, galactose, many others), disaccharides and the tetraterpenoid lycopadiene [69].
(ie sucrose, trehalose, maltose), glycerol glycosides, and A wide range of common and rare sterols are also
polyols (eg mannitol). Low molecular weight compounds synthesized by microalgae, including, for example,
are also accumulated by microalgae for osmoregulation. cholesterols (cyanobacteria, rhodophytes), chondrillasterol
Examples include sugars (eg Ochromonas), free glycerol (chlorophytes, euglenoids), clinoasterol (xanthophytes),
(eg Dunaliella and various zooxanthellae) and proline dinosterol (dinoflagellates), ergosterol (chlorophytes,
(some Chlorella spp). Industrial processes for glycerol [7] rhodophytes, euglenoids), epibrassicasterol (diatoms), pori-
and proline [53] production were developed, but not com- ferasterol (chlorophytes, chrysophytes), and sitosterol
mercialized. (cyanobacteria, chlorophytes, xanthophytes) [37,75].
Because of increased awareness of the possible health
Cell walls and extracellular mucilages aspects of including w-3 fatty acids in the diet (see Appli-
Most microalgae form rigid cell walls. Exceptions include cations, below), the manufacture and content of fatty acids
numerous 'naked' microalgae, such as Dunaliella, some by microalgae is the subject of a good deal of ongoing
dinoftagellates and chrysophytes, and the motile spores of research and development [44]. Microalgal lipids are
various groups. Cryptomonads and euglenoids have distinc- mostly esters of glycerol and fatty acids with a chain length
tive proteinaceous pellicles, internal to the cell membrane of C~4 to C22 and may be saturated or unsaturated. Cyano-
 Biodiversity and application of microalgae
FB MettingJr f%~
485
bacteria tend to have large amounts of polyunsaturated fatty 1950s when productivity and yield were first studied in
acids while eukaryotic microalgae contain, in addition, a mass culture [18]. Early interest was focused on exploiting
wide range of saturated and monosaturated fatty acids with the fact that optimal culture conditions result in theoretical
fatty acid profiles widely variable among taxa. Specific conversion of sunlight to biomass at yields of eight to ten
fatty acid profiles vary widely among algal groups, with times the most productive natural or agricultural plant com-
age and growth stage, and with environmental conditions. munities. In practice, sustained commercial mass culture
There are, however, some generalizations. For example, results in productivities of about 15 dry g cm-2 day -1 (54
linolenic acid (C18:3) is common in green algae, whereas tons ha i year1). More recently, microalgae have been tar-
diatoms contain palmitic (C16:o), hexadecenoic (C~6:~) and geted as a source of bioactive compounds and pharmaceut-
C-2o polyenoic acids. Red microalgae have high contents icals, specialty chemicals, health foods, aquaculture feeds,
of arachidonic acid (C2o:4) as well as palmitic, oleic (C~8:1) and for waste treatment and agriculture [1,14,55,77].
and linoleic acids. Chrysophytes contain significant quan-
tities of highly unsaturated C 18:4and C2e:6 acids in addition Microalgal mass culture
to unsaturated C~6:o and C2o:o compounds, fL3 fatty acids Microalgal biomass is produced in engineered facilities
are produced by a variety of microalgae. Eicosapentaenoic whose fundamental design and infrastructure depend on the
(Czo:5) and docosahexaenoic (C22:6) acids are produced by growth requirements of the microalgae of interest and the
some species of green and red microalgae, cryptophytes, nature and value of the final product. Two principal pro-
dinoflagellates, prymnesiophytes and diatoms. Certain duction methods are the photobioreactor and the open pond.
species have also been shown to synthesize unusual fatty Photobioreactors are lit fermentors of variable sophisti-
acids such as, for example myristic (C~4:o) in the diatom cation that are closed to the environment in order to prevent
Fragilaria, lignoceric (C24:o) in the premnesiophyte lso- chemical or biological contamination, including parasitism,
chrysis, gamma linolenic (C 18:3) in the golden-brown alga predation, and the need to control competition from
Ochromonas, and arachidonic (C20:4) in the red microalga unwanted microalgae ('weeds') when the desired species is
Porphyridium [24]. otherwise uncompetitive. Only products with high value
such as reagent grade phycobilins or isotopically-labeled
Bioactive compounds research compounds are produced in closed systems.
Microalgae produce a wide array of compounds with bio- Industrial scale production of microalgal biomass for the
logical activity including antibiotics, algicides, other toxins, health food market, for animal feed ingredients, and for
pharmaceutically-active compounds, and plant growth wastewater treatment relies on engineered ponds open to
regulators [13,65]. Antibiotic substances produced by the environment (Figure 2). Maintenance of single species
microalgae include certain fatty acids, bromophenols, tan- (unialgal) cultures in open ponds relies on the competitive
nins, terpenoids, polysaccharides, alcohols, and other com- ability of the microalga of interest. Thus, Spirulina nat-
pounds [76]. Most antibiotics are known from chlorophytes urally predominates in highly alkaline water, as does Duna-
and golden-browns, prymnesiophytes, diatoms, cyanobac- liella in saline culture. Wastewater treatment systems that
teria, and dinoflagellates. Activity has been demonstrated rely on microalgae for O2 production are dominated by
against bacteria, fungi, protozoa and other microalgae. For chlorophytes with high specific growth rates that tolerate
example, the cyanobacterium Scytonema hofmanii is known environments with high concentrations of dissolved and
to excrete a ,/-lactone algicidal to a broad range of poten- particulate organic compounds (eg Ankistrodesmus, Chlor-
tially competing microalgae in freshwater environments ella, Scenedesmus). Engineering design considerations are
[36]. considered in detail by Jassby [48], Oswald [71], and Rich-
Dinoflagellates, cyanobacteria and other microalgae pro- mond and Becker [83].
duce compounds toxic to invertebrate and vertebrate ani-
mals, including the neurotoxic agents of diarrhetic and Pharmaceuticals
paralytic shellfish poisoning (red tides) and ciguaterra fish By virtue of their larger size and accessibility, seaweeds
poisoning, an endemic disease in human populations that have until recently attracted more attention than microalgae
consume reef-inhabiting fish in the South Pacific. A number as sources of natural products, including identification of
of icthyotoxins are also produced by dinollagellates. At novel compounds for pharmaceutical development [45,46].
least twelve toxins are produced by species of Protogony- Screening of marine algae for antitumor activity began in
aulax alone [47,65]. In very small quantities a saxitoxin the 1970s. Early discoveries included tubercidin, a hetero-
has been used in microsurgical procedures and as an exper- cyclic nitrogen compound from the cyanobacterium Toly-
imental treatment for near-sightedness. Neurotoxins (eg pothrix byssoidea, shown to have in vitro activity against
anatoxin, aphanatoxin) produced by the cyanobacteria Ana- P-388 lymphocytic leukemia. Another compound, an 17
baena flos-aquae, Aphanizomenon flos-aquae, and hepato- aspinarigenase from the green microalga Chlamydomonas,
toxins (eg microcystin) from Microcystis aeruginosa in inhibits growth of lymphosarcoma in mice. Marine blue-
temperate freshwater have been implicated in poisonings green algae have been one target of a more recent Natural
and occasional death of cattle and other livestock [19,38]. Cancer Institute screening program aimed at anticancer and
antiviral (anti-HIV) activity that has identified a number of
compounds with potential for drug development, such as
Applications dibromoaplysiatoxin from Lyngbya majuscula. Approxi-
Microalgae have been the subject of applied research for mately 10% of 600 cultures from numerous genera of mar-
their commercial and industrial potential since the early ine blue-greens showed some activity in reducing cyto-
 Riodiversity and application of microalgae
FB Metting Jr
486
As the base of the marine food chain, microalgae are
responsible for manufacture of the straight-chain polyunsat-
urated fatty acids associated with lessened incidence of
heart disease in humans whose diets are rich in w-3 fatty
acids from consumption of cold water fishes. Prymnesi-
ophytes, prasinophytes, diatoms, and golden-brown
microalgae have been investigated as potential commercial
sources of the fatty acids of interest. Eicosapentaenoic acid
(EPA: C2o:5) and docosahexaenoic acid (DHA: C22:6) from
microalgae are available commercially from at least two
companies who market them as additives to various pro-
ducts, including infant formulas [43].
Microalgae have been investigated as sources of polysac-
charides, lipids, oils and hydrocarbons [11] and are used
for production of high value research biochemicals such
as phycobilins for fluorescent cell labeling [35] and stable
isotopically-labeled fatty acids, proteins and other mol-
ecules, and deuterated lubricants [27,51]. Phycocyanin, a
blue phycobilin from Spirulina is used to color food and
cosmetic products in Japan [12].

Health foods
The market niche for microalgal products within the inter-
national market for 'health foods' is conservatively esti-
mated at $20-25 million and is by far the largest commer-
cial application of microalgal culture. Currently, the largest
industrial operations product Spirulina biomass in outdoor
ponds in Mexico, Hawaii, California, and Thailand for for-
mulation into health food products and poultry feed sup-
plements [6]. Worldwide annual Spirulina production may
Figure 2 Spirulina biomass is produced in 5000 m 2 ponds at Earthrise be as much as 1500 dry tons (Yoshmichi Ota, personal
Farms, Calipatria, CA for formulation and sale as health food supplements
and poultry feed ingredients. Individual ponds are agitated by paddle-
communication). On a smaller scale, Chlorella
wheels (courtesy of Earthrise Farms). The total area is 150000 m 2. (Chlorophyceae) is also produced for consumption as a
health food [81].
Nutritional research has focused on Spirulina as a source
pathic effects normally associated with cultured cell lines of protein based on standard measures of utilization, includ-
[74]. Microalgae, including cyanobacteria and colorless ing protein content, digestibility, and biological value. As
(apochlorotic) variants of diatoms, may also be potential a source of useable protein, Spirulina is comparable to meat
sources of antiviral sulfolipids (eg sulfoquinosovyl and dairy products, superior to plant materials, but inferior
diglyceride) [40]. to poultry and fish. Its value as a source of protein quality
is compromised by low levels of sulfur-containing essential
Specialty chemicals amino acids, although it is comparable to other plant and
Markets for the production of high value chemicals and single-cell protein sources and can be modified by strain
food supplements from microalgal biomass have grown selection and modification of culture conditions, such as
considerably over the past decade. Microalgae have been providing high sulfate levels. Spirulina also contains
investigated for the production of vitamins and vitamin pre- unusually high concentrations of vitamins A and B12 [48].
cursors, including L-ascorbic acid (vitamin C), riboflavin, Products sold as variously formulated tablets and powder
and c~-,/3-, and y-tocopherol for food and cosmetic formu- in Europe, North America, and Asia also have putative
lations and mariculture [16,17,29]. Commercially viable therapeutic effects (eg antiviral, enhanced immune
heterotrophic production of ascorbic acid by Chlorella was response, radioprotection) and other claims that lack rigor-
reported by Running et al [87]. Carotenoids, such as /3- ous clinical documentation [5]. Mention has already been
carotene from halophilic Dunaliella and astaxanthin from made of commercial efforts to produce vitamins, caroteno-
Haematococcus and other chlorophytes, euglenoids and red ids, and (o-3 fatty acids in various microalgae for formu-
microalgae are also targets for commercialization. /3-caro- lation into traditional and health food products (see pre-
tene (pro-vitamin A) is extracted on a commercial scale vious section).
from Dunaliella mass-cultured in israel and Australia for
formulation into food supplements for human consumption Aquaculture feeds
[15]. Astaxanthin is responsible for the pink coloration of Mass culture of microalgae as feed for molluscs (clams and
cultured salmonid fishes and other seafoods, but is not as oysters), crustaceans (shrimp), and fish is an important
yet commercially competitive with synthetic products in the component of the mariculture industry [8,30]. Microalgal
aquaculture industry [8]. feeds are particularly valuable for seafood species with fas-
 Biodiversityand applicationof microalgae
FB Metting Jr
487
tidious dietary requirements that cannot be met by formu- to Gerald Cysewski, Cyanotech Corporation, Kailua-Kona,
lation with traditional agricultural commodity products, HI and Yoshimichi Ota, Earthrise Farms, Calipatria, CA
such as corn, soybeans, and fish and food processing by- for insights regarding commercial Spirulina and Dunaliella
products. In these cases, microalgae commonly provide markets: to Mr Ota for Figure 2; and to Sharon Lepel and
essential amino acids, fatty acids or other unidentified Sue Carver (Pacific Northwest National Laboratory),
growth factor requirements or are used to provide caroten- respectively, for secretarial and library assistance.
oids for coloration of the final product. In nearly all cases,
the microalgae are produced at the aquaculture facility, fre-
quently by simple fertilization of incoming seawater, and References
fed directly to the animals as dilute, living cultures [8]. 1 Akatsuka I (ed). 1990. Introductionto Applied Phycology. SPB Aca-
demic Publishing,The Hague.
Waste treatment 2 Andersen RA. 1992. Diversity of eukaryotic algae. Biodiversityand
Microalgae are principal or otherwise important biological Conservation 1: 267-292.
3 AndersenRA. 1996. Algae. In: MaintainingCultures for Biotechnol-
components of various systems for treating municipal and ogy and Industry(Hunter-CeveraJC and A Belt, eds), pp 29-64, Aca-
industrial effluents. Oxygen from photosynthesis is the pri- demic Press, San Diego.
mary microalgal contribution to treatment of municipal 4 Bar-Or Y and M Shilo. 1988. Unique characteristics and exopolysac-
wastewaters in open ponds or impoundments. The most charides of benthic cyanobacteria of potential importance for their
common designs include facultative ponds, which are mass cultivation.In: Polysaccharidesfrom Microalgae: A New Agro-
industry (Ramus J and MC Jones, eds), pp 26-32, InternationalWork-
deeper and support surface growth of microalgae, and high- shop Proceedings, Duke UniversityMarine Laboratory, Beaufort.
rate oxidation ponds which are specifically engineered to 5 Belay A, Y Ota, K Miyakawa and H Shimamatsu. 1993. Current
encourage maximum oxygenation by microalgae. High-rate knowledge on potentialhealth benefits of Spirulina. J Appl Phycol 5:
ponds are shallow and depend on mechanical mixing to 235-241.
6 Belay A, Y Ota, K Miyakawa and H Shimamatsu. 1994. Production
maximize algal production and removal of biochemical of high quality Spirulina at Earthrise Farms. In: Algal Biotechnology
oxygen demand [71]. In both configurations, microalgal in the Asia-PacificRegion (Phang SM et al, eds), pp 92-102, Univer-
communities are commonly dominated by green microalgae sity of Malaya, Kuala Lumpur.
(eg Ankistrodesmus, Chlorella, Scenedesmus). Biomass 7 Ben-AmotzA and M Avron. 1980. Glycerol,/3-caroteneand dry algal
from high-rate oxidation ponds can be harvested for use as meal production by commercial cultivation of Dunaliella. In: Algae
Biomass. Productionand Use (Shelef G and CJ Soeder, eds), pp 603-
animal feed and the concept has been demonstrated as a 610, Elsevier, Amsterdam.
component of integrated approaches to recycling of live- 8 BenemannJR. 1992, Microalgae aquaculture feeds. J Appl Phycol 4:
stock wastes [57]. Non-living microalgal biomass has also 233-245.
been subject to investigation and early-stage commercializ- 9 BjornlandT and S Liaanen-Jensen.1989. Distributionpattern of carot-
enoids in relation to chromophytephylogenyand systematics. In: The
ation for its potential in recovery of metals and radio- Chromophyte Algae: Problems and Perspectives (Green JC, BSC
nuclides from aqueous wastestreams as an alternative to Leadbetter and WL Diver, eds), pp 37-60, ClarendonPress, Oxford.
ion exchange resins [39]. Additionally, research has been 10 Bold HC and MJ Wynne. 1985. Introductionto the Algae. 2nd edn.
targeted at the use of microalgal systems for mitigation of Prentice-Hall, Englewood Cliffs.
waste heat [96] and CO= [52] from industrial processes. 11 Borowitzka MA. 1988. Fats, oils and hydrocarbons. In: Micro-algal
Biotechnology(MA and LJ Borowitzka, eds), pp 257-287, Cambridge
UniversityPress, Cambridge, UK.
Biofertilizers 12 BorowitzkaMA. 1988. Vitaminsand fine chemicalsfrom micro-algae.
Microalgae are employed in agriculture as biofertilizers and In: Micro-algalBiotechnology(MA and LJ Borowitzka, eds), pp 153-
soil conditioners [64]. Biofertilizers consist of N=-fixing 196, Cambridge UniversityPress, Cambridge, UK.
13 BorowitzkaMA. 1988. Microalgae as sources of pharmaceuticals and
cyanobacteria in lowland tropical rice cultivation. Filamen- other biologicallyactive compounds. J Appl Phycol 7: 3-15.
tous species of Anabaena, Nostoc, Aulosira, Tolypothrix 14 BorowitzkaMA and LJ Borowitzka (eds). 1988. Micro-algalBiotech-
and Scytonema are common in China, India and elsewhere nology. Cambridge UniversityPress, Cambridge.
in Asia, where they can provide 20 or more kg N ha -1 15 BorowitzkaMA and LJ Borowitzka. 1988, Dunaliella. In: Micro-algal
year -I, sufficient for one third or more of the requirements Biotechnology (MA and LJ Borowitzka, eds), pp 27-58, Cambridge
University Press, Cambridge, UK.
of traditional rice cultivars [84]. In China, Vietnam and 16 Brown MR and CL Farmer. 1994. Riboflavincontent of six species
Africa, Anabaena azollae, symbiotic in leaf cavities of the of microalgae in mariculture. J Appl Phycol 6: 61-65.
water fern Azolla is used as a green manure or companion 17 Brown Mr and KA Miller. 1992. The ascorbic acid content of eleven
crop in rice production [66]. On a small scale, palmelloid species of microalgae used in maricnlture, J Appl Phycol 4: 205-212.
18 Burlew JS (ed). 1953. Algal Culture from Laboratory to Pilot Plant.
(mucilage-producing) microalgae in the genus Chlamydo- Carnegie Institute, Washington,DC.
monas (Chlorophyceae) have been used as soil-condition- 19 Carmichael WW. 1992. Cyanobacteria secondary metabolites--the
ing agents for erosion control of pivot-irrigated soils in cyanotoxins. J Appl Bacteriol 72: 445-459.
North America [64]. 20 CastenholzRW. 1969. Thermophilicblue-greenalgae and the thermal
environment.Bacteriol Rev 33: 476-504.
21 CastenholzRW. 1992. Species useage, concept, and evolution in the
Acknowledgements cyanobacteria (blue-greenalgae). J Phycol 28: 737-745.
22 Castenholz RW and JB Waterbury. 1989. Oxygenic photosynthetic
Thanks are due to Robert Andersen, Center for Culture of bacteria. Group I. Cyanobacteria.In: Bergey's Manual of Systematic
Marine Phytoplankton, Bigelow Laboratory for Marine Bacteriology, Vol 3 (Staley JT et al, eds), pp 1710-1798, Williams
and Wilkens, Baltimore, Maryland.
Sciences, West Boothbay Harbor, ME, for sharing preprints 23 Cavalier-SmithT. 1989. The kingdom Chromista. In: The Chromo-
and references to the literature on algal diversity and for phyte Algae: Problems and Perspectives (Green JC, BSC Leadbetter
providing a thorough review of the preliminary manuscript; and WL Diver, eds), pp 381-407, ClarendonPress, Oxford.
~ Biodiversityand applicationof microalgae
FBMettingJr
488 24 Chen H, SE Bingham, V Chantler, B Pritchard and DJ Kyle. 1990. 49 John DM. 1994. Biodiversity and conservation: an algal perspective.
~3C-labeled fatty acids from microalgae. Dev lnd Microbiol 31:57 64. The Phycologist 38: 3-15.
25 Chishom SW, SL Frankel, R Goericke, RJ Olson, B Palenik, JB Water- 50 Kates M. 1987. Lipids of diatoms and of halophilic species, in: The
bury, L West-Johnsrud and ER Zettler. 1992. Prochlorococcus mar- Metabolism, Structure, and Function of Plant Lipids (Stumpf PK, JB
inus nov gen sp: an oxyphototrophic marine prokaryote containing Mudd and WD Nes, eds), pp 613-621, Plenum Press, New York.
divinyl chlorophyll a and b. Archive fur Mikrobiologie 157: 297-300. 51 Kyle DJ. 1989. Biodeuteration: a novel method for the production of
26 Cox ER (ed). 1980. Phytoflagellates. Developments in Marine Biology deuterated lipids. Lubrication Engineering 45:355 359.
2: Elsevier/North Holland, New York. 52 Laws EA and JL Beming. 1991. A study of the energetics and econo-
27 Cox J, H Chen, C Kabacoff, J Singer, S Hoeksema and D Kyle. 1989. mics of microalgal mass culture with the marine chlorophyte Tetra-
The production of 2H-, 13C-, and ~SN-labelled biochemicals using selmis suecica. Implications for use of power stack gases. Biotechnol
microalgae. In: Stable Isotopes in Pediatric, Nutritional, and Metabolic Bioeng 37: 936-947.
Research (Chapman T , ed), Intercept Ltd Press, Andover. 53 Leavitt R. 1986. Osmotic regulation in Chlorella sp 580 as a mech-
28 Darley WM. 1982. Algal Biology: a Physiological Approach. anism for the production of iJproline. Beihefte zur Nova Hedwigia.
Blackwell Scientific Publications, Oxford. Heft 83: 139-141.
29 De Roeck-Holtzman Y, I Quere and C Claire. 1991. Vitamin analysis 54 Lembi CA, SW O'Neil and DF Spencer. 1988. Algae as weeds: econ-
of five planktonic microalgae and one macroalga. J Appl Phycol 3: omic impact, ecology, and management alternatives. In: Algae and
259-264. Human Affairs (Lembi CA and JR Waaland, eds), pp 455M81, Cam-
30 DePauw N and G Persoone. 1988. Micro-algae for aquaculture. In: bridge University Press, Cambridge.
Micro-algal Biotechnology (MA and LA Borowitzka, eds), pp 197 55 Lembi CA and JR Waaland (eds). Algae and Human Affairs. Cam-
221, Cambridge University Press, Cambridge. bridge University Press, Cambridge.
31 Fogg GE. 1987. Marine planktonic cyanobacteria. In: The Cyanobac- 56 Lewin RA. 1989. Group II. Order Prochlorales. In: Bergey's Manual
teria (Fay P and C Van Baalen, eds), pp 393-414, Elsevier, Amster- of Systematic Bacteriology, Vol 3 (Staley JT et al, eds), pp 1799-
dam. 1806, Williams and Wilkens, Baltimore.
32 Geitler L. 1932. Cyanophyceae. In: L. Rabenhorst's Kryptogammen- 57 Lincoln EP and JFK Earle. 1990. Wastewater treatment with microal-
flora. Reprinted in 1985 by Koeltz Scientific Books, Koenigstein. gae. In: Introduction to Applied Phycology. (Akatsuka I, ed), pp 429
33 Giovannoni SJ, S Turner, GJ Olsen, S Barns, DJ Lane and NR Pace. 446, SPB Academic Publishing, The Hague.
1988. Evolutionary relationships among cyanobacteria and green chlo- 58 Manhart JR and RM McCourt. 1992. Molecular data and species con-
roplasts. J Bacteriol 170: 3584-3592. cepts in the algae. J Phycol 28: 730-737.
34 Glazer AN. 1987. Phycobilisomes: assembly and attachment, in: The 59 Margulis L, JO Corliss, M Melkonian and DJ Chapman (eds). 1990.
Cyanobacteria (Fay P and C Van Baalen, eds), pp 69-94, Elsevier, Handbook of Protoctista. Jones and Bartlett, Boston.
Amsterdam. 60 Mattox KR and KD Stewart. 1984. Classification of the green algae:
35 Glazer AN. 1994. Phycobiliproteins--a family of valuable, widely a concept based on comparative cytology. In: Systematics of the Green
used fluorophores. J Appl Phycol 6: 105-112. Algae (Irvine DEG and DM John, eds), pp 27-72, Academic Press,
36 Gleason FK, J Porwoll, JL Flippen-Anderson and C George. 1986. X- London.
ray structure determination of the naturally occurring isomer of cyano- 61 Meeks JC. 1974. Chlorophylls. In: Algal Physiology and Biochemistry
bacterin. J Org Chem 51:1615 1616. (Stewart SDP, ed), pp 161-175, Botanical Monographs 10, University
37 Goodwin TW. 1974. Sterols. In: Algal Physiology and Biochemistry of California Press, Berkeley.
(Stewart WDP ed), pp 266-280, Botanical Monographs 10, University 62 Metting FB. 1981. The systematics and ecology of soil algae. Botan
of California Press, Berkeley. Rev 47: I95-312.
38 Gorham PR and WW Carmichael. 1988. Hazards of freshwater blue- 63 Metting FB. 1991. Biological surface features of semi-arid lands and
green algae. In: Algae and Human Affairs (Lembi CA and JR Waa- deserts. In: Semi-Arid Lands and Deserts. Soil Resource and Recla-
land, eds), pp 403-432, Cambridge University Press, Cambridge. mation (Skujins J, ed), pp 257-293, Marcel Dekker, New York.
39 Greene B and GW Bedell. 1990. Algal gels or immobilized algae for 64 Metting FB. 1990. Microalgae applications in agriculture. Dev lnd
metal recovery, in: Introduction to Applied Phycology (Akatsuka I, Microbioi 31: 265-270.
ed), pp 137-150, SPB Academic Publishing, The Hague. 65 Metting FB and JW Pyne. 1986. Biologically active compounds from
40 Gustafson KR, JH Cardellina, RW Fuller, OW Weislow, RF Kiser, microalgae. Enzyme Microb Technol 8: 386-394.
KM Snader, GML Patterson and MR Boyd. 1989. AIDS-antiviral sup 66 Metting FB, WR Rayburn and PA Reynaud. 1988. Algae and agricul-
folipids from cyanobacteria (blue-green algae). J Nail Canc Inst 81: ture. In: Algae and Human Affairs (Lembi CA and JR Waaland, eds),
1252-1258. pp 335-370, Cambridge University Press, Cambridge.
41 Harlin MM and WM Darley. 1988. The algae: an overview. In: Algae 67 Murray RGE. 1989. The higher taxa, or, a place for everything...?
and Human Affairs (Lembi CA and RA Waaland, eds), pp 3-27, Cam- In: Bergey's Manual of Systematic Bacteriology, Vol 3 (Staley JT et
bridge University Press, Cambridge. al, eds), pp 1631-I634, Williams and Wilkens, Baltimore.
42 Hellebust JA. 1988. Polysaccharides produced by chromophyte 68 Norton TA, RA Andersen and M Melkonian. 1996. Algal biodiversity.
microalgae, lu: Polysaccharides from Microalgae: A New Agroindus- Phycologia 35:308 326.
try (Ramus J and MC Jones, eds), pp 13-19, International Workshop 69 Okada S, M Murakami and Y Yamaguchi. 1995. Hydrocarbon compo-
Proceedings, Duke University Marine Laboratory, Beaufort, North sition of newly isolated strains of Botryococcus braunii. J Appl Phycol
Carolina. 7: 555-559.
43 Hodgson J. 1996. Heliosynthese takes on Martek infant formula mar- 70 O'Kelley CJ. 1993. Relationships of eukaryotic algal groups to other
ket. Nature Biotechnol 14: 700. protists. In: Ultrastructure of Microalgae (Beruer T, ed), pp 269-293,
44 Hoeksema SD, PW Behrens, R Gladue, KL Arnett, MS Cole, JM CRC Press, Boca Raton.
Rutten and DJ Kyle. 1989. An EPA-containing oil from microalgae 71 Oswald WJ. 1988. Micro-algae and waste-water treatment. In: Micro-
in culture, in: Health Effects of Fish and Fish Oils (Chandra RK, ed), algal Biotechnology (MA and LJ Borowitzka, eds), pp 305-328, Cam-
pp 337-347, ARTS Biomedical Publishers, St John's, Newfoundland. bridge University Press, Cambridge.
45 Hoppe HA, T Levring and Y Tanaka (eds). 1979. Marine Algae in 72 Palenik B and R Haselkorn. 1992. Multiple evolutionary origins of
Pharmaceutical Sciences, Vol 1. de Gruyter, Berlin. prochlorophytes, the chlorophyll-b containing prokaryotes. Nature
46 Hoppe HA and T Levring (eds). Marine Algae in Pharmaceutical 355: 265-267.
Sciences, Vol 2. de Gruyter, Berlin. 73 Patterson DL. Stramenopiles: cfiromophytes from a protistan perspec-
47 Ikawa M and JJ Sasner. 1990. The chemistry and physiology of algal tive. In: The Chromophyte Algae: Problems and Perspectives (Green,
toxins. In: Introduction to Applied Phycology. I. (Akatsuka 1, ed), JC, BSC Leadbetter and WL Diver, eds), pp 357-379, Clarendon
pp 27 65, SPB Academic Publishing, The Hague. Press, Oxford.
48 Jassby A. t988. Spirulina: a model for microalgae as human food. In: 74 Patterson GML, KK Baker, CL Baldwin, CM Bolis, FR Caplan, LK
Algae and Human Affairs (Lembi CA and JR Waaland, eds), pp 149- Larsen, IA Levine, RE Moore, CS Nelson, KD Tschappat, GD Tuang,
179, Cambridge University Press, Cambridge. MR Boyd, JH Cardellina II, RP Collins, KR Gustafson, KM Snader,
 Biodiversity and application of rnicroalgae
FB Metting Jr

OW Weislow and RA Lewin. 1993. Antiviral activity of cultured blue- 87 Running JA, RH Huss and PT Olsen. 1994. Heterotrophic production
489
green algae (Cyanophyta). J Phycol 29: 125-130. of ascorbic acid by microalgae. J Appl Phycol 6: 99-104.
75 Patterson GW. 1987. Sterol synthesis and distribution and algal phy- 88 Saunders GW, D Potter, MP Paskind and RA Andersen. 1995. Cladis-
logeny. In: The Metabolism, Structure, and Function of Plant Lipids tic analyses of combined traditional and molecular data sets reveal an
(Stumpf PK, JB Mudd and WD Nes, eds), pp 631-636, Plenmn Press, algal lineage. Proc Natl Acad Sci USA 92: 244-248.
New York. 89 Shapiro LP and RRL Guillard. 1986. Physiology and ecology of the
76 Pesando D. 1990. Antibacterial and antifungal activities of marine marine eukaryotic ultraplankton. Can Bull Fish Aquat Sci 214: 371-
algae. In: Introduction to Applied Phycology (Akatsuka l, ed), pp 3 - 389.
26, SPB Academic Publishing, The Hague. 90 Sieburth J McN, V Smetacek and J Lang. 1978. Pelagic ecosystem
77 Radmer RJ. 1996. Algal diversity and commercial algal products. structure: heterotrophic compartments of the plankton and their
BioScience 46: 263-270. relation to plankton size fractions. Limnol Oceanogr 23: 1256-1263.
78 Radmer RJ and BC Parker. 1994. Commercial applications of algae: 91 Stewart WDP (ed). 1974. Algal Physiology and Biochemistry. Botan-
opportunities and constraints. J Appl Phycol 6: 93-98. ical Monographs 10. University of California Press, Berkeley.
79 Rayburn WR. 1988. Polysaccharides of chloropbytes. In: Polysac- 92 Thomsen HA. 1986. A survey of the smallest eukaryotic organisms
charides from Microalgae: A New Agroindustry (Ramus J and MC of the marine phytoplaukton. Can Bull Fish Aquat Sci 214: 121-158.
Jones, eds), pp 20-25, International Workshop Proceedings, Duke Uni- 93 van den Hoek C, D Mann and HM Jahns. 1994. An Introduction to
versity Marine Laboratory, Beaufort. Phycology. Cambridge University Press, Cambridge.
80 Reith M. 1995. Molecular biology of rhodophyte and chromophyte 94 Vonshak A, 1988. Porpbyridium. In: Micro-algal Biotechnology (MA
plastids. Annu Rev Plant Physiol Plant Mol Biol 46: 549-575. and LJ Borowitzka, eds), pp 85-134, Cambridge University Press,
81 Richmond A. 1986. Microalgae of economic potential. In: Handbook Cambridge.
of Microalgal Mass Culture (Richmond A, ed), pp 199-243, CRC 95 Waterbury JB, SW Watson, RRL Guillard and LE Brand. 1979. Wide-
Press, Boca Raton. spread occurrence of a unicellular marine, planktonic cyanobacterium.
82 Richmond A. 1986, Cell response to environmental factors. In: Hand- Nature 277: 293-294.
book of Microalgal Mass Culture (Richmond A, ed), pp 69-99, CRC 96 Wilde EA, JR Benemann, JC Weissman and DW Tillett. 1991. Culti-
Press, Boca Raton. vation of algae and nutrient removal in a waste heat utilization process.
83 Richmond A and EW Becker. 1986. Technological aspects of mass J Appl Phycol 3: 159-167.
cultivation--a general outline. In: Handbook of Microalgal Mass Cul- 97 Wilmotte A and S Golubic. 1991. Morphological and genetic criteria
ture (Richmond A, ed), pp 245-263, CRC Press, Boca Raton. in the taxonomy of Cyanophyta/Cyanobacteria. Algol Stud 64: 1-24.
84 Roger PA and SA Kulasooriya. 1980. Blue-Green Algae and Rice. 98 Winter A, RW Jordan and PH Roth. 1994. Biogeography of living
International Rice Research Institute, Los Bafios, The Philippines. coccolithophores in ocean waters. In: Coccolithophores (Winter A and
85 Rogers LJ and JR Gallon (eds). 1988. Biochemistry of the Algae and WG Siesser, eds), pp 161-176, Cambridge University Press, Cam-
Cyanobacteria. Oxford Scientific Publications, Clarendon Press, bridge.
Oxford. 99 Woese CR, O Kandler and ML Wheelis. 1990. Towards a natural sys-
86 Round FE, RM Crawford and DG Mann. 1990. The Diatoms. Cam- tem of organisms: proposal for the domains Archaea, Bacteria, and
bridge University Press, Cambridge. Eukarya. Proc Natl Acad Sci USA 87: 4576-4579.