Biotecnologia de Microalgas (1988 R) - (De La Noue and de Pauw) - The Potential of Microalgal Biotechnology A Review of Production and Uses of Microalgae

Biotech. Adv. Vol. 6, pp.
725-770, 1988
Printed in Great Britain. All Rights Reserved
0734-9750/88 $0.00 + .50

Copyright (~) 1988 Pergamon Press plc
THE POTENTIAL OF MICROALGAL

BIOTECHNOLOGY: A REVIEW OF PRODUCTION AND
USES OF MICROALGAE
JOEL DE LA NOUE' and NIELS DE PAUW
de Recherche en Recyclage Biologique, Universit~ Laval, Quebec,
Canada G1K 7P4
Laboratory f o r Mariculture, State University of Ghent, Ghent, Belgium
Groupe
ABSTRACT
An overview of the various aspects, promises and limitations of microalgal
biotechnology is presented. The factors of importance in microalgal cultivation
as well as the culture systems are briefly described. Microalgal biomasses can
f u l f i l the nutritional requirements of aquatic larvae and organisms. The
biochemical composition of algae can be improved by the manipulation of culture
conditions.
The nutritive value of the microalgal biomasses for humanand animal
consumption is also commented upon as well as some socio-economical aspects.

Among the sources of required nutrients (N, P), wastewaters and manures can
upgraded as culture media for microalgae the safety of which has to be evaluated.
Harvesting of the biomass is one of the bottlenecks. The various techniques,
physical, physico-chemical and biological are outlined and their f e a s i b i l i t y and
economic interest examined. Microalgal biomasses can be submitted to various
technological transformations. Various processes are reviewed in the light of
their effects on safety and nutritional value. The possible extraction of fine
chemicals and the preparation of protein concentrates is also reported on. The
various uses of microalgae lead to a possible competition, to be evaluated,
between systems for the production of food, energy and chemicals. The review
f i n a l l y covers the application of genetic manipulation to microalgae.
725
J, DE LA NOUE a n d N. DE PAUW
726
KEYWORDS
Microalgae - Solar biotechnology - Culture conditions
safety
Harvesting
Nutritional
value
Microbiological
Technical transformations - Fine
chemicals - Feeds/Foods - Genetic manipulations IMPORTANCE AND POTENTIAL OF MICROALGAE

Microalgae, as well as bacteria and yeasts, belong to those promising
microorganisms which deserve attention within the vast array of traditional as
well as
new biotechnologies. Along with basic research, applied algology has
been developing rapidly over the last 40 years, starting in Germany and extended
in the United States, Japan, Israel, I t a l y , with the aim of producing single cell
protein (SCP} and f a t (see Burlew, 1953).
As a result, already in the 1960s, the
commercial production of Chlorella in Japan and Taiwan as a novel health food

item was a success (Kawaguchi, 1980; Soong, 1980).
In the 1950s the idea of
using microalgae for wastewater treatment was launched and in the 1960s interest
grew in developing e x t r a t e r r e s t r i a l l i f e support systems. In the 1970s attention
went to the production of microalgal biomasses for fuel and f e r t i l i z e r s .
A new
trend in the 1980s is to use microalgae as a source of commonand fine chemicals.

Furthermore, microalgae have since the 1940s been playing a role of increasing
importance in
aquaculture.
For historical reviews on applied algology, the
reader is referred to Goldman (1979a) and Soeder (1980, 1986).

An important feature of microalgal systems is t h e i r v e r s a t i l i t y , making i t
possible to l i n k different applications within the same process, for example
wastewater treatment and production of
food, feed and chemicals.
Another
attractive characteristic of microalgae, in comparison with other microorganisms,

is t h e i r photosynthetic capability to convert solar energy into valuable biomass
with an interesting biochemical composition.
important role in solar biotechnology.
As such, microalgae could play an
Hereby, annual yields of 25 T and more,
even up to 200 T dry weight algae per hectare have been forwarded (Dubinsky e_tt
al..., 1978; Goldman, 1979a; Shelef and Soeder, 1980; Soeder, 1980; Santillan,
1982; Richmond, 1986c). Moreover, more than 60% of the dry weight can be made up
by protein.
Under favourable conditions, microalgal cultures can produce up to
20 to 35 times more protein than soybean and more than 50 times more than rice,
wheat or maize for the same area (Switzer, 1982; C i f e r r i , 1983).
Of particular
importance is also that microalgae can be grown yearound and harvested on a

continuous basis and be cultured on marginal lands in arid regions of the world,
u t i l i z i n g waters unsuitable for conventional agriculture (Thomas, 1983; Ciferri
and Tiboni, 1985; Gauthier et al_~.., 1985; Hall, 1986; Richmond, 1986c). Algal
MICROALGAL BIOTECHNOLOGY
727
cultures also have a lower water consumption than that required by traditional
cultivars (Heussler e_t_tal___:., 1978c). I f one considers that the water used for
algal cultures can be used afterwards for i r r i g a t i o n , algal cultures are even
more advantageous.
Although there is yet no real breakthrough in this f i e l d , due to a number
of fundamental drawbacks (Benemann and Weissman, 1984; Benemann et.t al_._~., 1986;
Soeder, 1986), interest in applied algology has never decreased i f we look at the
recent appearance of several important books on this matter: "Algae Biomass"
(Shelef and Soeder, 1980), "Biotechnology and Exploitation of Microalgae" (Becker
and Venkataraman, 1982), "CRC Handbook of Microalgal Mass Culture" (Richmond,
1986a), "Algal Biomass Technologies" (Barclay and Mclntosh, 1986), "Algal
Biotechnology (Stadler e_t_tal___~., 1988) and "Microalgal Biotechnology'" (Borowitzka
and Borowitzka, 1988). In connection one can mention the establishment in 1980
of an International Association for Applied Algology (IAAA) on ,the occasion of
the International Conference on Microalgae Production in T r u j i l l o , Peru (G.
Shelef, person, commun.) and the "French Association for Applied Algology",
active since 1982 (R. Fox, person, commun.). Also a newsletter called "Applied
Phycology Forum", edited by W.R. Barclay in the US, is distributed worldwide
since 1984 (Barclay, 1984-1985).
PRODUCTIONOF MICROALGAE
Figure l schematically shows the major pathways followed in microalgal
production and u t i l i z a t i o n of the biomasses for different purposes. Production
of microalgae f i r s t involves the cultivation, followed in most cases by harvesting and processing of the algae (Soeder, 1980; Becker and Venkataraman, 1982).
From a systematic point of view, the microalgal group includes several
thousand species belonging to two major groups: the prokaryotes including
blue-green algae (cyanobacteria) and the eukaryotes, including a.o. green algae
(Chlorophyta), red algae (Rhodophyta), and diatoms (Bacillariophyta). Of these,
only 30-40 species have been considered for mass cultivation and only few are
presently of real commercial importance (Borowitzka, 1988a; Richmond, 1986c). To
these belong representatives of the genera Chlorella, Scenedesmus, (green algae),
Spirulina (a blue green alga) and a number of phytoflagellate and diatom species
which are used as live food in larval mariculture (De Pauw and Persoone, 1988).
Other algae of commercial interest in the future could be Dunaliella (a green
flagellate), Porph~ridium (a red alga) and Botrococcus (a green alga) (Richmond,
1986c). Expressed in quantities, the world production of microalgae (mainly
Spirulina and Chlorella) only amounts to about one thousand tons per year
[. DE LA NOUE and N. DE PAUW
728
(~I
CULTIVATION POND
m , c c c ~ , ~ 0m;~ n o l m . r a ~c~
ccMP~
F-~F-~
Figure I .
Flow diagram of algal mass cultivation systems (from Becker and

Venkataraman, 1982).
(Kawaguchi, 1980; Soong, 1980; Ciferri and Tiboni, 1985) but is expected to
increase markedly in the near future (Venkataramanand Becker, 1988).
Determinants of Algal Growth
Major factors
of
temperature, nutrients,
importance in
the
pH and agitation
cultivation
process are
light,
(Becker and Venkataraman, 1982).
Although maximum growth rates are achieved in conditions of l i g h t saturation,

maximum yields w i l l be obtained only when l i g h t is the limiting nutrient since
production is proportional to solar energy conversion efficiency.
Light
limitation is established as a function of irradiance b y adapting the areal
density of the culture and thus the algal concentration. In continuous cultures
this is achieved by changing the detention time or dilution rate of the culture
(Goldman, lgTgb). The maximal theoretical conversion efficiency for total l i g h t
energy has been established at 6.6% (Shelef and Soeder, 1980). However,
sustained efficiencies of conversion of total incident radiation are more l i k e l y
to be within the range of l to 2%, or expressed in PHAR (Photosynthetic Active
Radiation) about 2 to 4.5% (Benemann eta)..., 1977; Goldman, 1979b; Shelef and
Soeder, 1980; Soeder, 1980; Pouliot and de la NoUe, 1985). Translated into daily
yields, these conversion efficiencies correspond to less than lO g up to 30 g and
in exceptional cases even 50 g dry weight per m2 (Goldman, 1980). Although under
729
l i g h t limiting conditions, the effect of temperature is lessened with regard to

the growth rate, too low temperatures may also become limiting especially during
the daytime or in winter(Toerien and Grobbelaar, 1980; Vonshak e.t_t a l . , 1982;
Bedell, 1985). For this reason heating up of the cultures may be beneficial
mainly in areas with a lot of incident radiation. Optimal temperatures for most
species range between 15 and 30C.
Several kinetic as well as empirical models predicting algal productivities
as a function of irradiance and/or temperature have been developed (Goldman,
1979b; M~rkl, 1980; Toerien and Grobbelaar, 1980; Hill and Lincoln, 1981; Grobbelaar et al_~., 1984). Other models also take nutrients (Shelef, 1981, in Grobbelaar et a l . , 1984) and mixing into account (Erickson and Lee, 1986). In open air
cultures, yields are practically linearly correlated with the incident radiation
(Paelinck, 1978, in De Pauwand Van Vaerenbergh, 1983; Castillo e_t_tal_~., 1980).
For optimal growth, the culture must be provided with nutrients in adequate
amounts (Borowitzka, 1988b). These include several macronutrients such as
carbon, nitrogen and phosphorus, sulfur, potassium, (for diatoms also silicon)
and a number of trace elements like minerals (e.g. Co, Mo, Mn) and several
vitamins (e.g. Bl2, thiamin). Besides quantities, the right proportions among
nutrients (e.g. N:P; N:Si) are also important. Basic information on algal
nutrition is given by Kaplan et al. (1986). With regard to the source of carbon,
apart from some algae like Spirulina which are capable of using bicarbonate at
alkaline pH values (Ciferri, 1983), most photo-autotrophically growing algae
prefer to u t i l i z e free CO2 (Heussler et a l . , 1978a; Richmond et a l . , 1982; Kaplan
e_t_t al.__~., 1986). Yields may, however, be increased two- or threefold by using
organic sources of carbon like glucose or acetate (mixotrophic growth) which of
course adds substantially to the production costs (Soong, 1980).
Present attention
is
also directed
towards growing microalgae
heterotrophically in the complete absence of light of which certain species (e.g.
Chlorogonium elongatum and Chlorella p~renoidosa) are capable (Kawaguchi, 1980;
Becker and Venkataraman, 1982; Kreuzberg e t a l . , 1985; Kaplan et a l . , 1986).
This process could also be exploited under conditions of low illumination where
glucose can replace l i g h t as the energy source (Folmann et a l . , 1978). The type
of C-source is related to the pH (Richmond, 1986b). This factor substantially
controls the bio-availability of nutrients. A too high pH for example w i l l ,
through the photosynthetic a c t i v i t y , make the free carbon dioxide unavailable to
most algae. Phosphorus, too, be may precipitated and ammonia nitrogen is also
stripped to the air (De Pauw and Van Vaerenbergh, 1983). For this reason,
addition of free carbon dioxide is beneficial to algal growth (Becker and
730
J. DE LA NOUE and N. DE PAUW
Venkataraman, 1982). Moreover,inorganic carbon levels play an important role in

affecting individual species' growth rates (Novak and Brune, 1985).
avoid
Agitation is also very important to microalgal cultivation, not at least to

sedimentation,
photoinhibition, nutrient
limitation and thermal
stratification but also to increase the l i g h t conversion efficiency (M~rkl, 1980;

Persoone e_t_ta l . , 1980; Richmond, 1986d). Since mixing requires energy, however,
the most economical regime w i l l have to be sought for each cultivation system,
for which the algal output is maximal (e.g. De Pauw et a l . , 1983; de la NoUe e.t_t
al~, 1984). Even i f the above conditions are optimally f u l f i l l e d , a number of
biological problems can and w i l l arise in the mass cultivation of microalgae.
These include contamination, grazing, diseases, premature collapse and lack of
species control (Shelef and Soeder, 1980; Becker and Venkataraman, 1982; De Pauw
e_t_tal~, 1984; Richmond, 1986d). Control measures for avoiding contamination by
bacteria and other algal species are s t e r i l i z a t i o n and u l t r a f i l t r a t i o n of the
culture medium (Ukeles, 1976). Grazing by protozoans and diseases like fungi can
eventually be treated chemically (Heussler et a l . , 1978b; Becker and
Venkataraman, 1982; De Pauw, pers. commun.; Becker, 1986). Larger zooplankters
and insects can be removed mechanically by screening (De Pauw et a l . ,
1983;
Becker, 1986), eradicated chemically (Loosanoff et a l . , 1957) or by changing the

culture conditions (Lincoln et a l . , 1983; SchlUtter and Groenweg, 1981).
Premature collapse of pure algae cultures during upscaling can be avoided by
establishing balanced growth conditions. A protocol for defining such conditions
has been presented by Pruder (1981). Control over the species composition of
large scale cultures on the other hand can be obtained to a certain extent by
proper operational management (Azov et a l . , 1980; Richmondet a l . , 1982; Vonshak
et a l . , 1982; De Pauwet a l . , 1983).
Cultivation Systems
Depending on the purpose of the mass production, the technology employed
may vary from a state close to agriculture to elaborate biotechnology (Soeder,
1980).
With regard to the origin of nutrients involved in the cultivation, a
distinction can be made between clean water and wastewater-based production
processes (Soeder, 1980). In the former case s t r i c t l y defined media are usually
employed in which bacteria are of no significant metabolic importance; in the
l a t t e r case (Fig. 2) less defined media such as sewage and manure are used in
which mixed cultures of bacteria and microalgae are active. While for small
scale culturing one usually relies on media which are complex and expensive, in
MICROALGAL B1OTECHNOLOGY
Figure 2.
LIGHT
731
The role of microalgae in the purification of wastewater by biological

recycling.
large scale culturing one is often forced to use cheaper media composed of
inorganic and organic agricultural f e r t i l i z e r s for reasons of cost. A l o t of
research is
still
necessary to
develop adequate recipes
for
large scale
enrichment. Problems relate to the methodology for distributing the nutrients

into the whole water mass, the dosage and frequency of enrichment, the nature of
the f e r t i l i z e r s , their price and local a v a i l a b i l i t y (De Pauwe_.ttal_~L., 1984).
As far as size and location are concerned, a distinction can be made betwen
indoor and outdoor (open-air) culture systems. Indoor systems serve for
culturing pure algal strains to be used in aquaculture or as inocula for starting
up larger (outdoor) cultures for example. Culture vessels include test-tubes,
Erlenmeyers, flasks, carboys, plastic bags, aquaria, troughs, tanks, raceways and
even vertical columns (Ukeles, 1976; De Pauw, 1981; Fox, J., 1983; Vonshak,
1986).
The algal reactors may be provided with a r t i f i c i a l l i g h t in or outside
the culture medium. In some cases i t may be profitable to work in greenhouses

for heating and to exploit natural l i g h t (Ukeles, 1980).
Water treatment w i l l usually involve s t e r i l i z a t i o n and/or advanced
f i l t r a t i o n while mixing is usually done by bubbling air through the medium,
enriched or not with CO2 (Ukeles, 1976). However, mechanical agitation of the
732
J. DE LA N O U E a n d N. DE P A U W
algal culture with a CO2-air mixture in a closed atmosphere over the culture is
as good (de la NoUe et a l . , 1984). Detailed information on laboratory techniques
for the cultivation of microalgae (culture media, isolation and purification,
maintainance of stocks, inoculation, culture devices) can be found in Ukeles
(1976), Fox, J. (1983), Borowitzka (1988b) and Vonshak (1986). Numerous systems
for large scale outdoor cultivation of microalgae have also been developed
(Goldman, 1979a; Becker and Venkataraman, 1982; Fox, R., 1983).
According to
Richmond and Becker (1986),
two basic
systems can be
distinguished: horizontal turbulent flow created by various types of devices or

inclined baffled surfaces (cascades) where the algal suspension is constantly
pumped back from the lower to the upper part of the unit.
The horizontal ponds
may be circular or consist of simple or meandering oblong raceways. Sloped

cultivation units on the other hand, may be meandering and b u i l t on the ground
(Heussler, 1985) or as a roof structure to cover a greenhouse (Setlik et a l . ,
1970).
Details on pond design and construction
are given by Dodd (1986).
Depending on whether or not the algal biomasses have to be harvested and

concentrated, the culture systems w i l l be shallow, 30 cm or less (Becker and
Venkataraman, 1982; Dodd, 1986; Richmond, 1986c) or deep, 0.5 m and more
(Persoone et a l . , 1980; De Pauw et a l . , 1983). Dependingon the species to be
cultured, fresh, brackish, alkaline, brine or seawater is utilized. The source
of water can be natural or made up a r t i f i c i a l l y . In particular cases i t may be
advantageous to use saline groundwater (Scura etal_~., 1979; Goldstein, 1986) or
nutrient-rich a r t i f i c i a l upwelling water from the deep sea (Roels et a l . , 1976).
One of the major factors influencing the design is the need for adequate
mixing and addition of carbon dioxide. Devices for agitating the culture medium
include paddlewheel(s), rotating spokeswheel arms, propellors, jetpumps, a i r - l i f t
pumps, air injection, mixing boards or even manual, or recirculation of the
medium by pumping on a sloped (baffled) surface (Goldman, 1979a; Becker and
Venkataraman, 1982; Richmond and Becker, 1986; Soeder, 1986). Attention has to
be paid to possible stress or damage to the algae by too intensive mixing
(Bronnenmeier and M~rkl, 1980). For the addition of CO2, specific devices have
also been developed (Heussler e t a l . , 1978a; Dodd, 1986; Richmond and Becker,
1986). So far, however, e f f i c i e n t addition of CO2 remains an unsolved problem.
Suggestions have been made to replace expensive pure CO2 by flue gases or even
wastes (Pruder, 1983).
For the maintenance of optimal culture temperatures, greenhouses (Ukeles,
1976; Richmond, 1986d) or even solar collectors like transparent polyethylene
tubes in which the algae are circulated (Materassi et a l . , 1980) can be used to
733
good effect.
I t has also been shown that greenhouse technology could make
yearound algal production feasible in northern climates with low temperatures but
sufficient solar irradiance (Pouliot and de la NoUe, 1985). Recently proposals
have also been made to u t i l i z e geothermal water (Bedell, 1985; Goldstein, 1986).
With regard to the sophistication of the cultivation technology, the process may
be semi-natural, without inoculation of algae, and a natural bottom, (e.g. the
cultivation of Spirulina at Sosa Texcoco) or a r t i f i c i a l , with inoculation of
precultured algae and lining of the bottom (e.g. Spirulina cultivation in Taiwan)
(Ciferri and Tiboni, 1985).
Finally, i t must be mentioned that the cultivation process can be
batchwise, semi-continuous or continuous (Vonshak, 1986).
Harvestin9 of microalgae
One of the major bottle-necks, limiting the further expansion of most
microalgal biomass applications, is the cost-effective harvesting (Table l ) .
Except for a few larger algae like Spirulina, which can be easily recovered by
simple gravity f i l t r a t i o n and inexpensive microstraining (Becker and
Venkataraman, 1982), this is not at all the case for most species which are
indeed small in size (less than 20 ~m). For this reason, many efforts have been
devoted to the development of suitable technologies for harvesting these small
particles (Mohn, 1980; Richmond and Becker, 1986). Though technically solved,
the handicap s t i l l remains the incompatibility between the efficiency of the
proposed methods and their cost-effectiveness (Benemanne_t_tal~, 1980).
Table I.
Advantages and disadvantages of different harvesting methods

(modified from Benemann et_tal.~., 1977, 1979 in De Pauwand Van
Vaerenbergh, 1983).
Method
Reliability
Cost energy
requirement
Centrifugation
Chemoflocculation
Sandfiltration
Ultrafiltration
Microstraining
Bioflocculation
good
good
fair
good
poor
poor
high
high
low
high
low
low
Quality for
bioconversion
good
poor
poor
good
poor
good
The most successful techniques are centrifugation, f i l t r a t i o n and

flocculation (Mehn, 1980). In practice, a combination of techniques is often
used to preconcentrate and/or concentrate.the algae. For preconcentrating the
734
algae, chemical flocculation eventually followed by flotation is commonpractice

in many operations (Becker and Venkataraman, 1982; De Pauw and Van Vaerenberg,
1983; Lavoie et a l . , 1984; Richmond and Becker, 1986). However, a number of
problems with the use of flocculants, including toxicity and carcinogenicity,
have been recognized (Dodd, 1979). This aspect is certainly important when the
objective is to use the algal biomasses as food or feed. A promising non-toxic
flocculant in this regard could be chitosan which is a natural product derived
from chitin, avail able worldwide (Nigam et a l . , 1980; Becker and Venkataraman,
1982). Presently in its disfavour, is its high price. Process optimization and
recycling of the flocculant, however, could increase the cost-effectiveness of
chitosan (Lavoie and de la NoUe, 1983; Lavoie et a l . , 1984; Morales et a l . ,
1985).
To reduce the high inorganic flocculants demand, the use of polymers or
ozone treatment prior to the flocculation process has also been recently proposed
(Shelef et a l . , 1986).
Another important step forward would be to get control of the process of
bioflocculation (autoflocculation) of microalgae without addition of chemicals
(Benemanne__t_tal~., 1980). Bioflocculation is the formation of cellular aggregates
by means of exocellular polymers. Research is in progress to unravel the
mechanism of this phenomenon(Sukenik and Shelef, 1984; Lavoie, 1985; Sukenike_.tt
a l . , 1985). Bioflocculation can also be enhanced by continuous mixing of the
cultures, promoting the succession of readily settling self-flocculating species
or inhibiting photosynthesis in standing populations (Lincoln and Koopman, 1986).
Recently the great potential of tangential flow f i l t r a t i o n for concentrating
marine microalgae has also been demonstrated (Welsh e_t_ta l . , 1985). The equipment
is, however, s t i l l expensive. Finally, for harvesting Dunaliella, several
procedures have been proposed which exploit the h i g h salinity-dependent
physiological and behavioural characteristics of this species (Borowitzka and
Borowitzka, 1988). Another method is exploiting the salinity-dependent
hydrophobicity of the Dunaliella cell membrane (Curtain and Snook, 1983).
UTILIZATION OF MICROALGAE
MicroalBae for Aquaculture
Microalgae are one of the live foods which are essential in aquaculture for
hatchery rearing of bivalve molluscs and peneid shrimp as well as the culturing
of several zooplankters (rotifers, cladocerans, brine shrimp, copepods) which are
themselves live food organisms for larvae of marine fish and curstaceans (De Pauw
and Pruder, 1986). Numerous, more or less sophisticated systems, have been
developed for culturing some 40 algal species to feed these larvae and
735
zooplankton organisms (Ukeles, 1976; Watson, 1979; De Pauw, 1981; De Pauw and
Pruder, 1986; De Pauw and Persoone, 1988). In particular cases where pure
nutrient-rich well water or deep sea water is available, i t has been shown that
large scale production of monospecific algal cultures to feed oysters and clams
up to market size would be feasible (e.g. Roels et a l . , 1976; Scura e_tt al_~.,
1979). Also, i t has been demonstratedthat closed-cycle rearing of the American
oyster, from the larva up to the market size, is possible (Pruder and Greenhaugh,
1978)o However, in the latter case, the economic feasibility can be put in
question. For this reason, when large quantities of microalgae are needed, the
alternative for pure algae cultures may consist of bloom induction of natural
phytoplankton (De Pauw, 1981; De Pauwand De Leenheer, 1985).
De Pauwet al. (1983) demonstratedthat with proper operational management,
i t is possible to steer the composition of the natural assemblages towards
species suited to the consumers. Basedon that principle, an industrial model
for nursery culturing of bivalve molluscs has been worked out (Claus et a l . ,
1984). The cultivation of microalgae none the less requiring specific skills,
the harvest and eventually the storage of algae grown at latitudes with ample
sunshine would represent a major breakthrough in aquaculture hatchery and nursery
operations (De Pauw et a l . , 1984). Apart from the constraints of economical
harvesting, however, are the processing and storage of the algal harvest and the
(re)treatment of the stored algae to make these acceptable again to the
consumers. Morespecifically, microalgae entangled in the matrix of a flocculant
are of too large particle size to be ingested by most filter-feeders, and
techniques for "declustering" such algal masses need to be developed (COST,
1983).
Apart from technical and economic problems involved with the mass
production of microalgae, the major problems in aquaculture are nutritionrelated (De Pauwe_t_tal_~., 1984). On the one hand, there is the lack of knowledge
of the nutritional requirements of the microalga consumers (molluscs,
crustaceans, fish) which are d i f f i c u l t to assess, and on the other hand, there is
the biochemical composition of the microalgaewhich is determinedby the culture
conditions (Webb and Chu, 1982). Of particular importance here is the presence
of essential fatty acids and the degree of fatty acid unsaturation. These
quantities can be modified by changing the culture conditions (Samson, 1980;
Enright et_t al.__~., 1986b). Of importance, for example, are light, temperature,
N-source, N:P ratio, etc (Sansregret, 1986). The same is true of the content of
amino acids and carbohydrates which are also of importance (Enright e_tt al__~.,
1986a; Terry e_t_tal_~., 1983; Terry, 1986).
736
Microal~ae for Wastewater Treatment

The increasing deterioration of our environment and the need for energy and
food is
forcing us to explore the f e a s i b i l i t y of wastewater recycling and
resource recovery.
Within this context, bio-treatment with microalgae is
particularly attractive because of their photosynthetic capabilities, converting
solar energy into useful biomasses and incorporating nutrients such as nitrogen
and phosphorus causing eutrophication (Fig. 2). This fascinating idea launched
some t h i r t y years ago in the U.S. by Oswald and Gotaas (1957) has since been
intensively tested in many countries (see examples in Goldman, 1979a; Shelef and
Soeder, 1980; De Pauw and Van Vaerenbergh, 1983). Thereby, emphasis may be put
on wastewater treatment and/or algal biomass production.
Depending on the
options taken, deep oxidation ponds or shallow high rate oxidation or algal ponds
(HIROP, HRAP) are used for this purpose. For a review on the variables playing a
role in the design and operation of microalgal wastewater treatment systems we
refer
to
Azov and Shelef (1982), De Pauw and Van Vaerenbergh (1983) and
Abeliovich (1986). Processes involved in the removal of nutrients are

precipitation, stripping and (luxury) uptake by algal biomass. The efficiency of
removal of a particular nutrient (for example N or P) w i l l also dependon whether
or not these nutrients are limiting in the wastewater to be treated. In this
regard i t was shown by de la No~e et al. (1980) that a two-phase culture system
with preconditioning of the algae could increase the nitrogen uptake of the
starved algae. In practice, with adequate stirring more than 90% nitrogen and/or
phosphorus can be removed (e.g. De Pauw e__t_tal__~.., 1978; Lincoln and H i l l , 1980;
Shelef e_t_tal__~., 1980; Martin et a l . , 1985a,b).
Provided sufficient solar irradiance and space are available, (non-toxic)
wastewaters of various origin and nature (municipal, industrial, agricultural,
aquacultural) may be treated with microalgal systems. Particularly promising
seem to be the combined treatment and upgrading of animal manures from
bio-industries with algal biomass production possibly in combination with biogas
production (Chung e_t_tal___~.,1978; Taganaidese__t_tal___~., 197g; Lincoln and H i l l , 1980;
Groeneweg e_t.tal___~.,1980; De Pauwet a l . , 1980a; Pieterse and Le Roux, 1980; Soong,
1980; Duerr, 1985; Martin e_t_tal~, 1985b). Light and temperature being the major
factors determining yield, wastewater treatment with algae will be particularly
suited for application in tropical and subtropical countries
(Soeder, 1984).
Light often being the limiting factor in Northern climates during winter time,
the use of a r t i f i c i a l illumination to increase the performance of wastewater
treatment in Quebec, Canada, has also been considered (Pouliot and de la NoUe,
1985). However, the cost of such a process is presently prohibitive, i f not
forever.
I t is also important to stress that in the treatment process, a large
737
array of microalgal species developing in different aquatic environments (e.g.

fresh-,
brackish-,
sea-,
alkaline-, brine-water)
adapted to
specific
high
nutrient loads can be utilized (De Pauwand Van Vaerenbergh, 1983).

To be effective in wastewater treatment, the algal biomasses must be efficiently
removed. Different pathways may be followed to this end. Usually, harvesting
includes a solid-liquid separation followed by dewatering and drying (Morainee_t_t
al_._~., 1980; Richmond and Becker, 1986). Although harvesting is not presently
cost-effective, i t has been shown that under tropical and subtropical conditions
the cost-benefit of microalgal wastewater treatment processes may compare
favourably with classical wastewater treatment systems such as activated sludge
systems. Particularly in favour of microalgal wastewater treatment systems is
the fact that the harvested algal biomasses can be upgraded in numerous ways, for
example to animal feed, and that no sludges have to be handled (Shelef e_t_ta l . ,
1978).
Wastewater grown microalgae may be used not only as a supplement in animal
feed (Chung e_t_tal.__~., 1978; Lincoln and H i l l , 1980; Sandbank and Hepher, 1980;
Soong, 1980; Saxena e_t_tal_~., 1983) but also in aquaculture for feeding fish,
molluscs and crustaceans (see several examples in Grobbelaar et a l . , 1981; De
Pauw and Van Vaerenbergh, 1983 and de la NoUeet a l . , 1986), and as a source of
energy, fuel, f e r t i l i z e r s and chemicals (Benemann e_t_t al___~., 1977). Indirect
harvesting of the microalgae by filterfeeders through a r t i f i c i a l aquatic food
chains including zooplankton, bivalve molluscs or fish, has also been a promising
and realistic alternative (e.g. Dinges, 1974; Goldmanand Ryther, 1976; De Pauw
et a l . , 1980a,b; Pieterse and Le Roux, 1980; Edwards, 1980; Groeneweg and
Schl~tter, 1981; Tarifeno-Silva e_t_tal...~_., Ig82a; Proulx and de la NoOe, 1985a,b).
Depending on the destination of the algal biomass, different criteria will
have to be taken into account with regard to possible contaminants such as heavy
metals, pesticide residues, pathogenic bacteria and viruses. However, the few
results already available indicate that fear concerning potential risk might be
excessive (Yannai e_t_ta l . , 1980; Tarifeno-Silva et a l . , 1982b; de la NoOee__t_ta l . ,
1986; Gauthier et a l . , 1985; Becker, 1986). Pathogensand bacteria eventually
remaining will be eliminated from the algal biomass at the processing stage. In
this context, microalgal production could also be used as a trap for toxic ions
and molecules (Aaronsone_t_tal_._~., 1980).
Since the
land-space requirements of microalgal wastewater treatment
systems are substantial (De Pauw and Van Vaerenbergh, 1983) efforts are being
made to develop wastewater treatment systems based on the use of
738
J. DE L A N O U E a n d N. DE P A U W
hyperconcentrated algal cultures.

T h i s process, called "activated algae"
(McKinney et a l . , 1971), is highly e f f i c i e n t in removing N and P within very
short periods of time, e.g. less than one hour (Lavoie and de la No~e, 1985).
Concentrated algal cell suspensions experiencing severe light limitation appear
to be more promising for tertiary treatment than for actual biomass production
purposes. The design of workable systems, not only with free microalgae, but
also with flocculated or algae immobilized for example in carrageenanbeads, has
also been shown possible (de la NoUeet a l . , 1983; Chevalier and de la NoUe,
1985a,b; 1986). Also, recycling part of the algae produced, to operate at the
shortest possible retention time, could increase system performance (de la No~e
and Ni Eidhin, 1988).
The challenge will be to make these systems not only
reliable but also economically competitive with conventional wastewater treatment

systems. I t could well be that the f i r s t applications to effluent treatment w i l l
occur in the f i e l d of toxic metal removal from industrial effluents.
Some
promising systems are currently under investigation (see, for example, Kosaric
and Ngcakani, 1988).
Microalgae as HumanFood
Many efforts have been made to promote microalgae as food directly for man.
This idea has been even more supported by the discovery that several native human
populations, at Lake Chad in Africa and Lake Tezcoco in Mexico amongothers, have
subsisted p a r t i a l l y on the nutritional qualities of the naturally occuring
blue-green alga Spirulina (Durand-Chastell, 1980; Ciferri, 1983; Bourges, 1986).
Representatives of this prokaryotic alga are indeed very rich in protein (Table
2) and except for some deficiency of sulphur-containing amino acids (methionine,
cysteine), have a f a i r l y balanced composition (Table 3) comparing favourably with
egg and milk protein. The same is true of essential fatty acid content
(Santillan, 1982; C i f e r r i , 1983; Bourges, 1986). The nutritional quality of
Spirulina protein is even superior to that of soybean (Jaya et a l . , 1980).
Table 2.
Basic chemical composition of the microalgae, Scenedesmusobliquus

and Spirulina maxima, as compared to soya bean (whole seed) and
wheat (whole gra~-~. Modified after Soeder (1980).
Components
Crude protein
Water
Li pi ds
Carbohydrates
Crude fibre
Ash components
Scenedesmus
50-60
4-8
12-I 4
l O-I 7
3-I0
6-I0
Spirulina
56-62
I0
2-3
16-I 8
0.I-0.9
6.4-9.0
Soya seed
34-40
7-10
16-20
19-35
3-5
4-5
Wheat
13.4-13.5
12.8-13.5
2. 1-2.4
78.6-80.5
2.1-2.4
1.6-2.8
M1CROALGAL BIOTECHNOLOGY
Table 3.
739
Essential amino acid composition of Scenedesmus species comparedto the

FAO pattern (g/16 gN).
Amino acid
VAL
LEU
ILEU
PHE+TYR
LYS
MET+CYS
TRY
THR
S. acutusI
S__~.obliquus2
4.7
7.0
3.1
6.0
4.6
3.2
1.7
4.9
FAO3
5.7
8.3
4.1
lO.l
5.9
2.9
n.d.
8.6
5.0
7.0
4.0
6.0
5.5
3.5
l.O
4.0
l A r t i f i c i a l medium (Becker et a l . , 1976).

2 Urban wastewater (Proulx a ~ d T l a NoUe, 1985b).
3 FAO/WHO(1973).
Furthermore, representatives of eukaryotic microalgae such as Scenedesmus,
Chlorella and Coelastrum have more or less the same nutritional characteristics
(EI-Fouly et_tal__~., 1985). Microalgae are also rich in vitamins and other growth
factors. For these reasons, several research groups have been endeavoring to
establish production units for t h e s e algae in tropical or subtropical
(developing) countries like Peru, Thailand, India, and Egypt but also in the USA
and Israel (Soeder, 1980; Castillo e_t_tal__~., 1980; Becker and Venkataraman, 1982;
El-Fouly e_t_ta l . ,
1985; Richmond, 1986c).
In most of these projects, serious
attention has been paid to the nutritional quality and the possible toxicological
effects of the algae (Payer et a l . , Ig80; de la NoUe et al.__~., 1986). The tests
involved humans as well as animals (Pabst, 1978; Becker, 1980; Ciferri, 1983).
Extensive testing has indicated that microalgae are a valuable and safe source of
protein. Testing with malnourished children also showed promise (Gross et a l . ,
1978). The high nucleic acid content, however, limits the admissible daily
consumption to about 5% of the human requirements (Becker, 1986). Based on a
recommended maximum daily supply of 2 g nucleic acids from SCP per adult
(FAO/WHO, 1973, in Becker, 1986), 46 g of dry Spirulina or 15 g Scenedemuswould
pose no problems (Becker, 1978a, 1986; Soeder, 1980; Bourges, 1986). Some
sensitivity reactions have been reported with Chlorella in humans but no major
adverse effects have been reported (Shubert and Larsen, 1985; see also Scrimshaw,
1986).
I t is also of importance that the d i g e s t i b i l i t y and the nutritive value of
microalgae are influenced by the processing technology used (Becker, Ig80~
Venkataraman et a l . , 1980; Becker and Venkataraman, 1982). I t has been shown
that
d i g e s t i b i l i t y especially of
chlorococcalean algae
could
be markedly
740
J. DE LA NOUE a n d N. DE PAUW
increased by cracking the cell wall with appropriate treatments such as drum
drying, spray drying, etc (Mitsuda e t a l . , 1977a,b; Becker, 1980). However,the
high cost of
production including
the harvesting and processing, and the
potential consumer nonacceptance (determined psyche- culturally, poor sensorial

properties) are the major obstacles to a definitive breakthrough. Direct
consumption is therefore presently limited to the use of expensive "health" food
sold for US $ 60 per kg and more on the international market (Bourges, 1986;
Richmond, 1986). Total production amounts to about one thousand tons per year
mainly involving Spirulina from Mexico and the U.S.A. (Ciferri and Tiboni, 1985)
and Chlorella from Japan and Taiwan (Kawaguchi, 1980; Soong, 1980).
In contrast
with the health food cultus (Hills, 1980; Switzer, 1982), a different philosophy
is followed by several other scientists who endeavor to exploit inexpensive
algoculture systems to combat malnutrition in developing countries (Fox, R.,
1980, 1983, 1985; Olguin and Vigueras, 1981; Becker and Venkataraman, 1982). The
integrated village health and energy systems involve cultivation of Spirulina
along with the production of biogas and compost.
Micro~lgae as Animal Feed
As in the case of human food, microalgae have also been successfully used
as an animal feed ingredient.
Feeding experiments with rats, mice, poultry,
pigs, sheep and carp, demonstrated unequivocally that microalgal meals produced
from various strains or species of Chlorella, Scenedesmus and Spirulina are
valuable protein sources lacking any acute toxicity (Becker, 1980, 1986). As
already mentioned is the utilization of wastewater grown microalgal biomasses for
which hygienical criteria are not so stringent as for human food particularly
promising (see examples given in section on "Microalgae for wastewater
treatment"). Primary and secondary toxicological testing of sewage grown algal
biomasses with regard to heavy metals demonstrated the likelihood that routine
use will turn out to be toxicologically safe (Becker, 1980, 1986). Moreover
these biomasses are a by-product of wastewater treatment and thus costcompetitive in with conventional feeds (Shelef et a l . , 1978). In contrast,
though the potential is there, the cost of pure microalgal biomasses like
Chlorella and Spirulina presently prohibits their extensive application in
aquaculture (De Pauwetal____~., 1984).
Microalgae as a Source of Energy
At pilot scale i t has been demonstrated that microalgal biomasses can be
converted by fermentation into energy-rich products such as methanegas, alcohol,
or liquid fuel as vegetable oils or hydrocarbon (Benemannet a l . , 1977, 1986;
Mitsui,
1980; Cohen, 1986).
741
A promising alga in this
regard seems to be
Botrxococcus braunii for the production of long-chain hydrocarbons (Gudin et a l . ,

1983; Benemann and Weismann, 1984; Richmond, 1986c).
Though technically
feasible, a major drawback to practical application is that enormous algal plants

are neededto ensure even a small percentage of our energy requirements. Concepts
have been developed for the exploitation of algal energy farms up to lO0 square
miles in size in which unreclaimable water and wastewater nutrients could be used
(Oswald et a l . , 1977). The microalgal fuel economics and engineering of such
systems are described by Goldman and Ryther (1977) and Benemannet al. (1986).
Aside from the fact that no one is presently running algal farms larger than a
few thousand square meters, major constraints to further development are the
economical harvesting of the algal biomasses and the lack of control over
unwanted algal consumers like zooplankters. Moreover,in the present situation,
methane from microalgal biomasses is not at all competitive with conventional
energy sources (Dubinsk.v e__t_tal__=,1978). Other problems are related to the actual
methane fermentation of the algal biomass (Benemannet a l . , 1977).
Many algal
species can also be induced to produce hydrogen through
biophotolysis. Cultures of nitrogen-fixing heterocystous (e.g. Anabaena) as well

as non-heterocystous blue-green algae (e.g. Spirulina, Oscillatoria) can be used
for this purpose (Benemann and Weissman, 1976; Hallenbeck e_t_tal.__~., 1978; Mitsui,
1980; Kumazawa and Mitsui,
1982, Karube et al~,
1986).
A team from the
university of Miami (Mitsui and co-workers) demonstratedthat a particular strain

of marine blue-green non-heterocystous alga (Oscillatoria) in a chamber 20 feet
square and three feet deep, could produce enough hydrogen to yield lO00
kilowatt-hours of e l e c t r i c i t y per month. Technical as well as economic
constraints, however, presently restrict the practical application of these
systems. Immobilization of microalgae or cyanobacteria by entrapment in various
matrices (polyurethane and polyvinyl foams, for example) might prove to be an
interesting solution to the problem of production of fuels, energy and chemicals
(Hall, 1988).
Microal~ae as Fertilizers
A promising idea gaining more and more interest is the production of easily
harvestable nitrogen-fixing blue-green algae in conjunction with wastewater
treatment which could be converted into organic nitrogen f e r t i l i z e r (Benemann,
1979; Li, 1981; Padhy, 1985; Venkataraman, 1986). I t has also been shown that
unconsumed microalgae in a wastewater-fish production system could be directly
upgraded agriculturally for the production of maize (Edwards et al~, 1981). This
could be particularly rewarding in tropical countries where microalgal wastewater
742
treatment systems could be easily applied. Presently, several companiesinvolved

in algal biotechnology, are exploring the potential of blue-green algae as
agricultural f e r t i l i z e r s and soil conditioners (Applied Phycology Forum, 1985;
Metting, 1985). According to Curtin (1985) one company in the US has already
started producing and selling f e r t i l i z e r s composedof blue-green algae, which are
competitive with conventional agricultural fertilizers.
One kg of algae could
replace 60 kg of conventional nitrogen f e r t i l i z e r .

Microal~ae as a source of commonand fine chemicals
Our modern industrial world depends heavily upon petroleum and its
derivatives to obtain a vast array of useful chemicals. Production of microalgae
in arid and sunny parts of the world may be a solution to the economic and
material stress raised by the needs of the industry. I t has been suggested that
those who will benefit f i r s t from commercial marine biotechnology will be the
producers of sugars and polysaccharides, pharmaceuticals, dyes, bioflocculants,
pigments, vitamins, lipids, o i l , etc. One of the most convincing examples is
that of phycobiliproteins which are fluorescent dyes used in certain immunoassays
and cell separation and worth $75/mg (Curtin, 1985). Two categories of products
can be obtained f~om microalgae (Gudin e_t_tal_~., 1983): endocellular substances
that act as osmoregulators in the cell (glycerol, sorbitol, mannitol, etc) or do
not (starch, amylase, amylopectin, glycogen) and exocellular products, mainly
polysaccharides (glucan, mannan, chitan), hydrocarbon or polyacrylates.
Obviously, the latter can be recovered more easily, and usually without
destruction of the cells.
The many products of interest find applications in the chemical, food,
and pharmaceutical industries and medecine (Tables 4 and 5). For reviews on
chemicals and products from microalgae, the reader is referred to Aaronson et
al. (1980), Benemannand Weissman (1984), Borowitzka (lg88a), Cohen (1986). The
diversity of exo-polysaccharides produced by microalgae is impressive and
undoubtedly represents considerable potential for the food industry as gums,
thickeners, gelling agents and stabilizers and many other diverse uses (Weiner e._t_t
al_~., 1985). A carrageenan-like polysaccharidehas been extracted from the marine
microalga Porph~ridium cultivated in outdoor saltwater ponds (Curtin, 1985) by
Israeli workers (Arad et a l . , 1986). I t has been shown by Weissman and Benemann
(1980) that polysaccharide productivity can be markedly enhanced by nitrogen
starvation. T h i s alga is currently one of the most promising, and efficient
culture systems have been designed (Gudin e_t_ta l . , 1983).
Table 4.
Application of microalgae as a source of chemicals.
Chemicals
Proteins
Lipids
Pigments
Vitamins
Carbohydrates
Pharmaceuticals
Table 5.
743
Example
Protein concentrates
Glycerides (glycerol)
B-carotene, Phytol
Phycobiliproteins
Biotin
Mannitol, Sorbitol
Polysaccharides
Sterols
Chlorella extract
Toxins
Application
Food and feed industries
Fuels, food additives
Precursor vitamins
Dyes, cosmetics
Vitamin-rich meals
A r t i f i c i a l sweeteners
Viscosifiers
Steroid hormones
Antibiotics
Anti-parasitic
Valuable products from microalgae.
Source
Product
Anabaena flos-aquae
Protein
Chlorella sp.
Phaeodactxlum tricornutum
Dunaliella tertiolecta
Asteromonas 9racilis
Microphytes
Lipids
Reference
Molton et a l . , 1980
Molton et a l . , 1980
Sansregr-6t~T986
Starch (insoluble)Williams et a l . , 1978
Glycerol
Dubinskye-t-aT., 1978
Ben-Amotz~ta-a-T., 1982
Agar-Agar
Bonin, 1983
Carrageenan
Chlam~domonas agloeformis
Polysaccharides Moltonet a l . , 1980
Porphxridium cruentum
Gudin et--a.l~,Ig83
Bacillariophyceae d~atoms)
Chitan
Gudin ~ a l . , 1983
Botrxococcus braunii
Hydrocarbons
Gudin et-aT., 1983
Phaeocxstis p ~ i
Polyacrylates
G u d i n ~ a T . , 1983
Scenedesmus acutus
Pigments
Partali e_t_tal___~., 1985
Spirulina
Phycocyanin
Tel-or et a l . , 1980
Dunaliella, Spirulina
g-carotene
Tel-or e't'aT., 1980
Chlorophyll a
Cyanobacteria
Phycobiliprotein Curtin, 1985
Spirulina
Enzymes
Tel-or et a l . , 1980
Ferredoxin
Ferredoxin-NADP reductase
Cytochromes
Ribulose bi-phosphate
Carboxylase
Although the i n i t i a l investments appear high, even prohibitive, the cost of

producing glycerol from Dunaliella could be competitive with that of
petrochemical methods (Ben-Amotz and Avron, 1980). The method is subject to
patent applications and pilot-scale production has been undertaken in Israel
where long-term productivities of 4.5 g of glycerol/m2.day in 3.5 M NaCl have
744
been measured (Ben-Amotz et a l . , 1982). An intracellular glycerol content of up

to 4.4 molar in cell has been reported for D. v i r i d i s (Williams et al._.~., 1978) and
7 molar with D. salina (Borowitzka, 1981). This is about 50% of the dry weight.
Cell concentrations of above l g/L can, however, be d i f f i c u l t to obtain since
optimal growth conditions do not necessarily maximize glycerol production
(Ben-Amotz e_~_tal___~., 1982). Dunaliella exhibits the added advantage of having no
rigid cell wall, thereby f a c i l i t a t i n g extraction.
By bacterial fermentation, the
algal-glycerol mixtures can be converted into neutral solvents (Nakas et al___~.,

1986).
I t is, however, the B-carotene-producing capacity of Dunaliella bardawil
(Ben-Amotz and Avron, 1980, 1983) which has stimulated a large commercial
investment, i.e. $2.5 million by Koor Foods of Israel, over the last 6 years
(Weiner, 1985). About 5 to 9% of the dry biomass may be B-carotene. The same
incentive
has
been triggering
large
companies like
Roche Products and
Wesfarmers to sponsor a scientific team in Australia to develop an industrial

production
unit
for
B-carotene from Dunaliella (Borowitzka e_tt al~,
1986a,b).
In the USSR, attention has also been paid to the mass production of
Dunaliella (Massyuk, 1966). Within the group of
the
carotenoid
1984,
pigments,
xanthophylls are also particularly useful in pigmenting chicken skin, tissue

and egg yolks (Benemann and Weissman, 19@4). Other pigments derived from
microalgae and receiving attention f r o m industrial researchers are the
phycocyanins (blue pigments) making up the photosynthetic apparatus of
Spirulina and valuable as dyes in food or cosmetics (Ciferri, 1983; Sasson,
1983) or even in the jeans industry (Bionov, 1986, personnal commun.). Even
i f the production cost is high ($15/mg) production under a r t i f i c i a l l i g h t is
retained (Curtin, 1985). Other potential products from microalgae are phytol
and a wide range of sterols.
Phytol is a suitable precursor for the synthesis
of several vitamins (A, E, K, K2) and B-carotene while sterols can be used as
substrates for the synthesis of steroid hormones (Borowitzka, 1988a).
Algae as a source of edible oils have begun to receive attention, in
spite of fierce competition among traditional vegetal oils and a slowly
expanding market. Other non-food markets may be suitable for algal oils
because of their chemical diversity and their high l i p i d content, which can be
comprised between l and 85% on a dry weight basis (Shifrin, 1984) depending
upon the species and the culture conditions.
For example, Phaeodact~lum
tricornutum is a marine diatom that contains some highly unsaturated longchain acids, some of which could be used as substitutes of drying oils in paints
and lacquers (Molton e_~_tal__~.,1980). These authors made someeconomic projections
on
process incorporating
manure hydrolysis
and
algal
culture
for
745
oil and feed protein production and calculatea a conservative simple return on
investment of 21%.
I f microalgae are used for oil production, the production cost based on
very conservative figures for capital and operating costs would be within the
range of $0.25-2.00/gai ($0.07-0.75/L), which is quite encouraging (Shifrin,
1984). Certainly one of the factors which has aroused interest is the enormous
productivity possible - up to 25 and more tons/ha.yr requiring however a huge
capital investment. The possibility of coupling production to sewage
treatment may satisfy economic requirements (Dubinsky e_t_tal_~., 1978; Shelef e_t_t
al__~., 1978).
The oil
content of many microalgae is in the range of 40-70%
(Ratledge and Boulton, 1985) and includes a large proportion of neutral lipids
with fatty acids in the C12-22 range (Shifrin, 1984). Species of Chlorella
appear to offer promise as a source of edible oils (Ratledge and Boulton, 1985)
while Botr~ococcus braunii excretes oils (unsaponifiable lipids) and carotenoids
which may have various industrial uses (Gudin et a l . , 1983; Wolf, 1986). Much
research remains to be done both on the production of the oils by the algae in
question, and on their recovery and chemical characterization (Benemann and
Weissman, 1984).
Another area of potential development for microalgae is the extraction
of pharmaceuticals (Table 6).
Despite some d i f f i c u l t i e s , such as relative
inaccessibility and lack of characterization, lower product-yield than usual
sources, complexity of purification, the search for pharmaceuticals from algae
and marine organisms is under way and some very interesting biologically
active
products have been found (Wright, 1984; Borowitzka, 1988a; Cohen,
1986).
For example, a l i p i d antioxidant function may be responsible for the
purported action of B-carotene as an anti-cancer agent (Burton and Ingold,

1984). Extracts from Chlorella and Scenedesmushave been shown to have in vitro
antibacterial activity (Reichelt and Borowitzka, 1984) and stimulate the growth
and yield of yeasts and other microorganisms (Fingerhut e_t_tal~, 1984). This
property finds application in the fermentation industry to improve the growth of
lactic acid bacteria (Soong, 1980). Extracts of Scenedesmus and Spirulina could
also serve as a replacement for serum (Kumamoto, 1984). Certain compounds
extracted from Porpt~vridium could play a role in commercial abalone culture,
inducing the settlement and metamorphosisof the larvae (Morse and Morse, 1984).
Other valuable compounds of interest for the food industry which may be
produced from microalgae include other osmoregulatory substances such as
sorbitol, mannitol, and cyclohexanetetrol, bio-emulsifiers and various lowmolecular weight metabolites, e.g. aminoacids.
The latter may be released
74G
Table 6.
Pharmaceuticalsfrom microalgae.
Source
Product
Reference
Amphiridiumcarterae
(dinoflagellate}
8-2 blocker
Baker et a l . , 1985
Nitzchia ovalis (diatom)
Antibacterial
Maksimova et a l . , 1984
Antioedema
Bronchodilator
Polysynaptic
Blocker
Anticonvulsant
Hypotensive
Baker e_~tal__.~., 1985
Cyanobacteria
(blue-green algae)
Antiinflammatory
Norton and Wells, 1982
Rivularia firma
Analgesic
Anaesthetics
Antiallergic
Norton and Wells, 1982
Scxtonema hofmani
Cyanobacterin (antibiotic)
Wood et a l . , 1982
Lyngbya majuscula
Malyngolide (antibiotic)
Antineoplastic
Cardillo et a l . , 1981
Wright, 1984
Lyngbya lutea
Antiamphetamine
Baker e_t_tal~, 1985
Gomphosphaeria aponina
Aponin (lysis of dinofl.)
Lem and Glick, 1985
Spirulina
Serum replacer
Lem and Glick, 19~5
Toxins (saxitoxin)
Curtin, 1985
Brevetoxin
Wright, 1984
Cosmetics
Bonin, 1983
Dinoflagellates
Ptxchodiscus brevis
Microphytes
from immobilized cyanobacteria (Synechocystis) by osmotic shock (Reed et a l . ,

1986). I t has long been known that some microalgae, such as Spirulina, are
among the richest known sources for vitamins, especially Bl2 (Lem and Glick,
1985). The proportion of an alga's vitamin production excreted to the medium
may be quite high and could be enhanced by manipulating the growth conditions
(Borowitzka, 1988a). Although the studies done with immobilized microalgae
or cyanobacteria are recent (see Rao and Hall, 1984) progress has been recently
made with carrageenan for Scenedesmus (Chevalier and de la NoUe, 1985a),
calcium alginate (Martinez e_~_ta l . , 1987) or chitosan (Proulx and de la NoUe,
1988) for Phormidium., or polyurethane for Porph~ridium (Gudin e_.t_tal___:., 1983).
One interesting possibility, at least on theoretical grounds, is that of the
747
co-immobilization of algae with bacteria, the former producing oxygen for the
l a t t e r which produce the product of interest for the food industry (Chevalier and
de la No~e, 1986).
PROCESSINGOF MICROALGALBIOMASSES
As previously shown, microalgal biomasses can be used as such in animal
feeding, aquaculture or for human consumption, despite low d i g e s t i b i l i t y in
some cases or deficiency in some amino acids, such as sulphur-containing ones.
In some cases, however, processing of the biomasses might be desirable or
necessary either to improve the nutritional quality or to allow the
conservation of the biomass for later use. I f one wishes to extract specific
substances or added value products, additional processing steps are obviously
required.
Processin9 Methods and their Effect on the Safet~ and Nutritional Value of
Algal Biomasses
Sometimes, i t
is possible to obtain strains of microalgae which possess
characteristics that simplify their u t i l i z a t i o n . For example, a strain of

Chlorella vulgaris (CCAP no 211-la) lacks the resistant cell wall component
sporopollenin (Strain e t
al~_., 1984). This
strain
has a relatively high
d i g e s t i b i l i t y (81.7%) without any treatment a n d , therefore, no expensive

processing to increase its d i g e s t i b i l i t y is required. In most cases, however,
i t is necessary to break cell walls either by applying physical, physicochemical or enzymatic treatments. Breaking cell walls is not an easy matter.
For example, the breaking pressure (by gas decompression technique) is 95
atmospheres for Chlam,vdomonas as compared to only 30 atmospheres for cultured
cells of carrot in suspension (Carpita, 1985).
Drying the biomasses is one of the most common treatments applied but
degradation may account for some 30% of the total production cost (Lin, 1985).
The choice of technique used for dehydrating algae affects the appearance, the
texture, the nutritional value and the d i g e s t i b i l i t y of the final product.
Spray-drying appears to be more useful t h a n freeze-drying since toxic
substances are more effectively destroyed (Lin, 1985). The resulting powder
is harder, an important characteristic for the production of hard tablets.
Although these processes are expensive, their overall evaluation shows that
they are superior to drum-drying (Lin, 1985).
For Spirulina, thermal
treatment has been reported to be effective for detoxification (Voronkova et
al___~., 1983).
748
With respect to
true protein content only, drum-drying, deamination,
irradiation and autoclaving appear to be equivalent in the case of Ooc~stis (Lee

et al.__~., 1982). Rats fed with autoclaved algae showed a lower Net Protein Ratio
(NPR) and a lower protein digestibility than rats fed algae treated by other
methods (Becker, 1980; Lee e_~_t a l . , 1982). Improved digestibility after
gamma-irradiation may be expected since this treatment brings about cellulose
depolymerization (Campbell e_~_tal___~., 1986). Studies done in vivo with rats fed
Chlorella have shown that treatment processes give different results.
For
dried cells, digestibility is 60%, and rises to 73% after heating to lO0C for
30 minutes and to 80% for broken cells (Fujiwara-Arasaki, 1984). Considering
the
h i g h cost of
most processing techniques, i t
appears that
the most
economical may be to evaporate water from algae by sun-drying on sand beds

(Lincoln
and Koopman, 1986; Becker and Venkataraman, 1982; Richmond and
Becker, 1986). I t is clear that the choice of the technique is a matter of

balance between cost, intended use and changes in the characteristics of the
processed biomass. In the case of algae intended for human consumption,
concern has been expressed that their color might hinder consumer acceptance.
I t has been shown that decolorization may be achieved by photolysis
(15 000 lux, fluorescent lamp, lO h). Moreover, this treatment resulted in
the removal of the unpleasant odour of the blue-green alga used, Anabaena
flos-aquae (Choi and Markakis, 1981). Enzymatic treatments are s t i l l not
widely used for microalgae. By analogy with what has been done with marine
macrophytes (Fujiwara-Arasaki, 1984), one can expect that treatment with
pepsin, pancreatin and pronase might lead to increased digestibility of
alkali-soluble proteins.
Another aspect of
biomass treatment
is
that
of
supplementation.
Microalgae are known to be deficient in sulphur-containing amino acids

(Becker, 1978b; Ben-Amotz and Avron, 1980; Saxena et a l . , 1983). Supplementation of sun dried Spirulina platensis biomasses with 0.2% methionine has
been shown to significantly improve the biological value and the net protein
utilization with the same digestibility (Narasimha e_t_tal~, 1982). I t would
also be possible to improve the nutritional value of S__~. platensis by
compensating for the low content in methionine through the use of mutants that
have higher intracellular pools of this amino acid (Ciferri, 1983). I t is
suggested that with appropriate genetic enhancement, microalgae producing
desirable amino acids in sufficiently high concentrations could be produced
(Borowitzka, 1988a).
749
Preparation of ProteinConcentrates
Algal biomasses are generally high in protein content, values up to 60%
or more being reported for Spirulina for example (Santillan, 1982; Ciferri and
Tiboni, 1985; Bourges, 1986). This has prompted some investigators to explore
the possibility of preparing protein concentrates that exhibit good functional
properties such as water and fat absorption, emulsification and foaming
capacities, and foam stability. In general, these functional properties for
Spirulina protein concentrate are better than or similar to those of soybean
meal, except for foam stability (Anasuya Devi and Venkataraman, 1984).
Preparation of protein concentrates from blue-green algae such as Anabaena
flos-aquae has been thought to be a means of overcoming its low digestibility
due to cell walls, its unattractive color and its strange flavor (Choi and
Markakis, 1981). Treating the biomass with 3N HCI at 95C for lO minutes and
neutralizing afterwards has been found to be an effective way o f obtaining a
solution containing 80% of the cell nitrogen (Choi and Markakis, 1981).
I t is
likely that enz~nnic treatments could be applied to such concentrates to

prepare partially hydrolyzed fractions.
Unless such fractions yield highly
priced products, i t is unlikely that algal biomasses may constitute a
competitive protein source. However, as the price of traditional sources such
as fish or soybean meal continues to rise, microalgal SCP used as a protein
supplement may become increasingly attractive especially when produced as a
by-product of wastewater treatments (Aaronson and Dubinsky, 1982; Pouliot and
de la NoUe, 1985). Protein isolated from algae shows a much higher
digestibility than entire cells (Choi and Markakis, 1981), a result that is
not surprising.
In vitro digestion of Chlorella proteins with trypsin, for
example, gave digestibilities of 45% for dried cells, 69% for frozen cells,
71% for broken cells and 86% for extracted proteins (Ishii et a l . , 1974;
Mitsuda et a l . , 1977a, b).
COMPETITION BETWEENDIFFERENT INDUSTRIESFOR THE USE OF ALGALBIOMASSES
I f commercial s c a l e production of microalgal biomasses is to be
i n i t i a l l y limited to existing installations, e.g. sewage treatment f a c i l i t i e s
or to their extension as tertiary treatment systems, i t is conceivable that
supply may f a l l short of demand, given all the possible applications already
described. The price of the biomass produced will have to increase before the
number of production sites can increase substantially. T h i s could arise
through other uses for microalgae to which biotechnology might contribute.
Possibilities of low-to-medium value-added products include biosorption of
metals, with a wide range of applications (Darnall et a l . , 1986) ammonia
750
production in soil (Padhy, 1985), nitrogen f e r t i l i z a t i o n of rice fields (Baker e t

al___:., 1985) and energy production through methanization of biomasses or hydrogen
production (Karube e t al__~., 1986), a capacity shared with many nitrogen-fixing
bacteria and also exhibited by some green algae, e.g. Chlam~domonas or
Scenedesmus under anaerobic conditions. Also to be considered is the production
of organic acids under anaerobiosis.
It
has been estimated that 2.35 million Km2 could be used for farming
marine biomass in U.S. coastal waters not currently used for the production of
human food, animal feed or other non-energy products (Snow et a l . , 1978;
Chynoweth and Srivastava, 1980). Although there is s t i l l
some debate about
the future of solar biotechnology, some private industries are entering the
field.
This is the case of Cyanotech Corporation in California which is
supposed to begin this year the commercial distribution of algal-based food
products, vitamin supplement and nitrogen-fixing f e r t i l i z e r at one-third the
cost of conventional nitrogen f e r t i l i z e r (Curtin, 1985). The demand for
high-added-value specialty items such as isotopically labelled metabolites and
intermediates for research on serum replacement for mammalian tissue culture
derived from Spirulina extract (Lem and Glick, 1985) is not expected to
impinge heavily on the available algal raw material production capacity.
One interesting use of algae has been proposed by Oswald (1980), namely
the use of some agricultural wastes, such as bagasses, ensiled with algae and
fermented under appropriate conditions to provide feed for ruminants. Such
production systems would require only 0.02 ha to produce or recycle a l l of the
high quality protein required by one person, i.e. 5 x 306 ha for the USA for
example. Land for other purposes such as grain or forest production or energy
products would be released. Although there have been arguments about the
possible competition between algal culture and valuable farm lands, i t can be
proposed that the former could be conducted in marginal areas (for example
arid zones) and that ingenuity in converting wasteland to productive land
through algal cultures is all that is required (Shifrin, 1984).
APPLICATION OF RECENTBIOTECHNOLOGICALMETHODSTO THE DEVELOPMENTOF
MICROALGAL BIOMASS-BASEDPRODUCTS
According to Ciferri and Tiboni (1985) and Venkataramanand Becker (1988),
i t is conceivable that strain improvement by standard methods utilized in
industrial microbiology or the techniques of genetic engineering may lead to the
isolation of strains endowedwith more favorable characteristics such as faster
751
growth rate, capacity to grow at suboptimal temperatures, higher yields or higher

content of n u t r i t i o n a l l y important components. However, basic genetic research
has been slow to reach algae and studies on them are relatively few (Lemieuxet
al___~., 1985; Necas, 1985; Turmel e_t_tal__~., 1986; Chauvat e_t_tal_._~., 1987; Lee and Tan,
1987), since they do not appear to offer obvious advantagesover the favourites,
i . e . E , c o l i , Bacillus sp. or Saccharomxceseither as a vehicle of expression for
genes of higher organisms or as a model for the genetics of higher eukaryotes.
However, the possibility of inserting new gene sequences in small linear DNA
demonstrated in Chlamxdomonas (Turmel et a l . , 1986) might open interesting
perspectives, even more promising than the use of plasmids (G. Bellemare person.
comm.).
For the study of microalgal genetics the prokaryotic cyanobacteria offer

several advantages. Most can be grown on defined medium, with generation
times as short as five hours and can be handled and grown on,agar medium as
colonies. Transformation does occur, and to-date several shuttle vectors for
cyanobacteria can replicate stably in E. coli
Glick, 1985).
(Baker e_t_tal__:., 1985; Lem and
Algal pond culture may already be considered for a wider range of

products. Extension of the latest genetic techniques to the eukaryotic
microalga Dunaliella is
considered a high
priority
(Baker e_t_t al___~., 1985).
Approaches to the genetics of this alga have been presented by Simon and
Latorella (1986). Use of genetic alterations in cyanobacteria for maximizing
H2 production have been recently demonstrated (Spiller and Shanmugam, 1986).
Spirulina, a tropical species with optimum growth temperature around 30C, has
been manipulated genetically and a strain has been developed by Japanese
workers which grows at 4C (Anon., 1984). This might open vast possibilities
for culture systems under temperate or even cold climatic conditions.
Strain-specific differences in the chemical composition with respect to
l i g h t intensity have been demonstrated for Phaeodactxlumtricornutum (Terry et
al._~., 1983). Genetic studies are also required in order to transfer the
capacity for the production of algal polysaccharides to faster growing
bacteria or to transfer t r a i t s such as pesticide or herbicide resistance into
nitrogen- fixing blue-green algae in rice paddies (Erickson et a l . , 1984)o
Genes coding for the production of polysaccharides can be amplified by
inclusion of s m a l l extrachromosomal elements and genetic manipulations,
including transfer to bacteria, according to one report (Weiner et al_._~., 1985).
Other properties of algae to be exploited, pending genetic characterization
752
(Gallagher, 1986), include the a b i l i t y of certain species such as Chlorella to

grow under extreme conditions of s a l i n i t y and acioity and the super- sensitivity
of some strains of C_.~.saccharophila to the toxic heavy metal cadmium (Kessler,
1985). These are only a few examples of possible interesting applications of
genetic manipulations with microalgae. A much more complete treatment of the
subject will be found in the recent review contribution of Craig et al. (1988).
ACKNOWLEDGMENTS
The authors wish to thank deeply D. Proulx and S. Davids for their help in
preparing this paper, D. Ni Eidhin for revising i t and G. Gagnonfor the typing.
Financial help was provided by FCAR-Equipe grant, NSERC and the minist~re des
Relations Internationales du Quebec.
REFERENCES
I.
S. Aaronson and Z. Dubinsky, Mass production of microalgae, Experientia, 38,
2.
36-40 (1982).
S. Aaronson, T. Berner and Z. Dubinsky, Microalgae as a source of chemicals
and natural products, in A l g a e Biomass, Shelef and Soeder (Eds),
Elsevier/North-Holland Biomedical Press, Amsterdam, 575-601 (1980).
3.
4.
A. Abeliovich, Algae in wastewater oxidation ponds, in CRC Handbook of

Microalgal Mass Culture, Richmond (Ed), CRC Press Inc., Boca Raton, Florida,
331-338 (1986).
M. Anasuya Devi and L.V. Venkataraman, Functional properties of protein
products of mass cultivated blue-green alga Spirulina platensis, J.
5.
Sci., 49, 24-27 (1984).

Anonymous,Spirulina et g~nie g~n~tique, Biofutur (21), 12-13 (1984).
6.
7.
Applied Algology Forum, ~ (2), I I (1985).

S. Arad, M. Adda and E. Cohen, Effect
of
Food
growth conditions on the
production of the cell-wall polysaccharide in Porphxridium, in Algal

Biomass Technologies, Barclay and Mclntosh (Eds), Nova Hedwigia Beih. 83,
8.
J. Cramer, Berlin, 75-78 (1986).

Y. Azov and G. Shelef, Operation of high-rate oxidation ponds: theory and
experiments, Water Res., 16, I153-I160 (1982).
9.
Y. Azov, G. Shelef, R. Moraine and A. Levi, Controlling algal genera in high

rate wastewater oxidation ponds, in Algae Biomass, Shelef and Soeder (Eds),
lO.
J.T. Baker, J.L. Reichelt and D.C. Sutton, Pharmacologically active and
related marine microbial products, in Comprehensive Biotechnology,
Moo-Young (Ed), PergamonPress, Vol. 3, 225-230 (1985).
II.
12.
13.
14.
15.
16.
17.
18.
19.
20.
21.
22.
23.
24.
753
W.R. Barclay (Ed), Applied phycology forum. A Newsletter for algal

biotechnologies, P.O.Box II157, Boulder, Colorado 80301, USA (1984-85).
W.R. Barclay and R.P. Mclntosh (Eds), Algal Biomass Technologies, An
Interdisciplinary Perspective, Nova Hedwigia Beih. 83, J. Cramer, Berlin,
Stuttgart (1986).
E.W. Becker, The legislative background for utilization of microalgae and
other types of single cell protein, Arch. Hydrobiol. Beih. Ergebn. Limnol.,
] ] , 56 (1978a).
E.W. Becker, Major results of the Indo-German algal project~ Arch.
Hydrobiol./Beih. Ergebn. Limnol., II,23-40 (1978b).
E.W. Becker, Comparative toxicological studies with algae in India, Thailand
and Peru, in Algae Biomass, Shelef and Soeder (Eds), Elsevier/North-Holland
Biomedical Press, Amsterdam, 767-786 (1980).
E.W. Becker, Nutritional properties of microalgae: potential and
constraints, in CRC Handbook of Microalgal Mass Culture, Richmond (Ed), CRC
Press Inc., Boca Raton, Florida, 339-419 (1986).
E.W. Becker, L.V. Venkataraman and P.M. Khanum, Digestibility coefficient
and biological value of the proteins of the alga "Scenedesmus acutus
processed by different methods, Nut. Rep. Intern., 14, 457-466 (1976).
E.W. Becker and L.V. Venkataraman, Biotechnology and exploitation of algae.
The Indian approach, Deutsche Ges. techn. Zusammenarbeit (GTZ), GmbH
Eschborn, Fedl. Republic Germany (1982).
G.W. Bedell, Stimulation of commercial algal biomass production by the use
of geothermal water for temperature control, Biotechnol. Bioeng., 27,
I063-I066 (1985).
A. Ben-Amotz and M. Avron, Glycerol, beta-carotene and dry algal meal
production by commercial cultivation of Dunaliella, in Algae Biomass,
Shelef and Soeder (Eds), Elsevier/North-Holland Biomedial Press, Amsterdam,
603-610 (1980).
A. Ben-Amotz and M. Avron, On the factors which determine massive
beta-carotene accumulated in the alga Dunaliella bardawil, Plant Physiol.,
72, 593-597 (1983).
A. Ben-Amotz, I. Sussman and M. Avron, Glycerol production by Dunaliella,
Experientia, 38, 52 (1982).
J . R . Benemann, Production of nitrogen f e r t i l i z e r with nitrogen fixing
blue-green algae, Enz. Microb. Technol., ~, 83-90 (1979).
J.R. Benemann and J.C. Weissman, Biophotolysis: problemsand prospects, in
Microbial Energy Conversion, Schlegel and Barnea (Eds), Erich G~Itze, K.G.
G~ttingen, Germany, 413-426 (1976).
754
25.
26.
27.
28.
29.
30.
31.
32.
33.
34.
35.
J.R.
Benemann and J.C.
Weissman, Chemicals f r o m microalgae,
in
Bioconversion Systems, Wise (Ed), CRC Press, Boca Raton,Flori~a, 59-70
(1984).
J.R. Benemann, J.C. Weissman, R.P. Goebel and D.C. Augenstein, Microalgae
fuel economics, in Algal Biomass Technologies, Barclay and Mclntosh (Eds),
Nova Hedwigia Beih. 83, J. Cramer, Berlin, 176-191 (1986).
J.R. Benemann, J.C. Weissman, B.L. Koopman and W.J. Oswald, Energy
production by microbial photosynthesis, Nature, 268,19-23 (1977).
J.R. Benemann, B. Koopman, J. Weissman, D. Eisenberg and R. Goebel,
Development of microalgae harvesting and high-rate pond technologies in
California, in Algae Biomass, Shelef and Soeder (Eds), Elsevier/NorthHolland Biomedical Press, Amsterdam, 457-495 (1980).
D.J. Bonin, Les microalgues. Biomasse Actualit~s, NO special 3 (suppl. ~ NO
12), les v~g~taux aquatiques, 22-23 (1983).
L.J. Borowitzka, Solute accumulation and regulation of cell water a c t i v i t y ,
in The Physiology and Biochemistry of Draught Resistance in Plants, Paleg
and Aspinall (Eds), Academic Press, Sydney, 97-130 (1981).
M.A. Borowitzka, Vitamins and fine chemicals from microalgae, in Microalgal
Biotechnology, Borowitzka and Borowitzka (Eds), Cambridge Univ. Press,
Cambridge, 153-196 (1988a).
M.A. Borowitzka, Algal media and sources of algal cultures, in Microalgal
Biotechnology, Borowitzka and Borowitzka (Eds), Cambridge Univ. Press,
Cambridge, 456-465 (1988b).
M.A. Borowitzka and L.J. Borowitzka, Microalgal Biotechnology, Cambridge
Univ. Press, Cambridge (1988).
L.J. Borowitzka, M.A. Borowitzka and T. Moulton, The mass culture of
Dunaliella salina for fine chemicals: from laboratory to p i l o t plant,
Hydrobiologia, l l 6 / l l 7 , I15-134 (1984).
L.J. Borowitzka, T.P. Moulton and M.A. Borowitzka, Salinity and the
commercial production of Beta-carotene from Dunaliella salina, in Algal
Biomass Technologies. Barclay and Mclntosh (Eds), Nova Hedwigia Beih. 83,
J. Cramer, Berlin, 224-229 (1986a).
36.
L.J. Borowitzka, T.P. Moulton, N. Sammy and D.J. Vincent, The mass culture
of Dunaliella salina for beta-carotene:
from p i l o t plant to production
plant, 12th Int. Seaweed Symposium, Sao Paulo, Brazil, July 27-August l
(1986).
37.
H.
Bourges,
Spirulina,
a food product?,
in
Internat.
Symp. "Food &
Biotechnology", de la NoUe, Goulet and Amiot (Eds), Centre de recherche en

n u t r i t i o n , Universit~ Laval, Quebec, 251-266 (1986).
38.
755
R. Bronnenmeier, and H. M~rkl, Hydrodynamic stress capacity of

microorganisms (experiments with green and blue green algae), 3rd Int. Conf.
on Production and Use of Microalgae, T r u j i l l o , Peru, Oct. 26-31, (Ig80).

J.S. Burlew (Ed), Algal culture from laboratory to p i l o t plant, Public. No.
600, Carnegie Institute, Washington, D.C. (1953).
40. G.W. Burton and K.U. Ingold, Beta-carotene:
an unusual type of l i p i d
antioxidant, Science, 224, 569-573 (1984).
41. G.L. Campbell, H.L. Classen and G.M. Ballance, Gamma irradiation treatment
of cereal grains for chick diets, J. Nutr., l l 6 , 560-569 (1986).
42. G. Cardillo, M. Orena, G. Porzi and S. Sandri, Synthesis of malyngolide, an
antibiotic from the marine blue-green alga Lyngbya ma~uscula Gomont, J.
39.
43.
44.
45.
46.
47.
48.
49.
50.
51.
52.
53.
Org. Chem., 46, 2439-2442 (1981).

N.C. Carpita, Tensile strength of cell walls of l i v i n g cells,
Plant.
Physiol., 79, 485-488 (1985).

J. Castillo, F. Merino and P. Heussler, Production and ecological
implications of algae mass culture under peruvian conditions, in Algae
Biomass, Shelef and Soeder (Eds), Elsevier/North-Holland Biomedical Press,
Amsterdam, 123-134 (1980).
F. Chauvat, J. Labarre et F. Ferrino, Developments of genes cloning systems
in the unicellular cyanobacterium Synechocystis 680382, Recent Progress in
Algal Biotechnology, Villeneuve d'Ascq, Sept. 14-17, Cl (1987).
P. Chevalier and J. de la NoUe, Wastewater nutrient removal with microalgae
immobilized in carrageenan, Enz. Microb. Technol., ~, 621-624 (]985a).
P. Chevalier and J. de la NoQe, Efficiency of immobilized hyperconcentrated
algae for ammonium and orthophosphate removal from wastewaters, Biotechnol.
Letters, ~, 395-400 (1985b).
P. Chevalier and J. de la NoUe, Alpha-amylase production by Bacillus
s u b t i l i s co-immobilized in carrageenan with the microalga, Scenedesmus
obliquus, Internat. Symp. "Food and Biotechnology", Quebec, Aug. 19-22,
Abst., 44 (1986).
Y.R. Choi and P. Markakis, Blue-green algae as a source of proteins, Food
Chem., ~, 239-247 (1981).
Po, Chung, W.G. Pond, J.M. Kingsbury, E.F. Walker, Jr. and L. Krook,
Production and n u t r i t i v e value of Arthrospira platensis, a spiral
blue-green alga grown on swine wastes, J. Anim. Sci., 47, 319-330 (1978).
D.P. Chynoweth, and V.J. Srivastava, Methane production from marine
biomass, in Proc. Intern. Symp., Biogas. Microalgae and Livestock Wastes,
Chung (Ed), Taipei, Taiwan, Sept. 15-17 (1980).
O. C i f e r r i , Spirulina, the edible microorganism, Microbiol. Reviews, 47,
551-578 (1983).
O. Ciferri and O. Tiboni, The biochemistry and industrial potential of
Spirulina, Ann. Rev. Microbiol., 39, 503-526 (1985).
756
54.
C. Claus, W.P. Davidse, N. De Pauwand A.C. Drinkwaard, Oesternurseries, in

Maricultuur achter de Oosterscheldekring, Interimrapport 1984, Projectgroep
MARIOS, Uitgave Directie van de Visserijen, Ministerie van Landbouw en
Visserij in samenwerking met de Deltadienst van Rijkswaterstaat,
55.
s'Gravenhage, 44-87 (1984).

Z. Cohen, Products from microalgae, in Handbookof Microalgal Mass Culture,
56.
Richmond (Ed), CRC Press Inc., Boca Raton, Florida, 421-454 (1986).
COST, Proposal for a Research Programme on Selected Research in
Aquaculture, COST project 41, Rep. of the Mgmt Comm, COST Project 46 on
57.
Mariculture to COSTSenior Officials Comm., COST/226/83, Brussels (1983).

R. Craig, B.Y. Reichelt and J.L. Reichelt, Genetic engineering of
microalgae, in Micro-algal Biotechnology, Borowitzka and Borowitzka (Eds),
58.
Cambridge University Press, Cambridge, 415-455 (1988).

C.C. Curtain and H. Snook, Method for harvesting algae, P.C.T./AV82/ 00165
(8 October 1982), US Serial No 511 135 (7 June 1983) (1983).
59.
M.E. Curtin, Chemicals from the sea, Biotechnology, ~,34-37 (1985).
60.
D.W. Darnall,
B.
Greene, M.T. Menz, J.M. Hosea, R.A. McPherson, J.
Sheeddon, and D.M. Alexander, Selective recovery of gold and other metal
61.
ions from an algal biomass, Environ. Sci. Technol., 20, 206-208 (1986).
J. de la NoOe, and D. Ni Eidhin, Improved performance of intensive semicontinuous cultures of Scenedemus by biomass recirculation, Biotechnol.
Bioeng., 31, 397-406 (1988).
62.
J. de la No~e, A. Lavoie and P. Walsh, Hyperintensive wastewater tertiary

treatment by flocculated activated algal sludge, Proc. Internat. Conf.
Commercial Applications and Implications of Biotechnology, London,
I005-I015 (1983).
63. J. de la NoUe, L. Cloutier-Mantha, P. Walsh and G. Picard, Influence of

agitation and aeration modes on biomass production by Oocystis sp. grown on
wastewaters, Biomass, 4_, 43-58 (1984).
64.
J. de la No~e, G.A. Picard, J.C. Piette and C. Kirouac, Utilisation de

l'algue Oocxstis pour le traitement t e r t i a i r e des eaux us~es. I I .
Effet du
conditionnement pr~alable des cellules en cyclostat sur la vitesse de prise

en charge de l'azote lors d'incubations de longue dur~e, Water Res., 14,
Ill5-1130 (1980).
65.
J. de la NoUe, D. Proulx, R. Guay, Y. Pouliot and J. Turcotte, Algal

biomass production from wastewaters and swine manure: nutritional and
safety aspects, in Microbial Biomass Protein, Moo-Young and Gregory (Eds),
Elsevier Applied Science, London, 141-145 (1986).
66.
N. De Pauw, Use and production of microalgae as food for nursery bivalves,

in Nursery Culturing of Bivalve Molluscs, Claus, De Pauw and Jaspers (Eds),
Europ. Maricult. Soc., Special Public. 7, Bredene, Belgium, 35-69 (1981).
757
67.
N. De Pauw and L. De Leenheer, Outdoor mass production of marine microalgae
68.
Limnol., 20, 139-145 (1985).

N. De Pauw and G. Persoone, Microalgae for aquaculture, in Microalgal
69.
Cambridge, 197-221 (1988).

N. De Pauw and G. Pruder, Use and production of microalgae as food in
for nursery culturing of bivalve molluscs, Arch. Hydrobiol. Beih. Ergebn.
Biotechnology, Borowitzka and Borowitzka (Eds), Cambridge University Press,
aquaculture,
in Realism in Aquaculture: Achievements, Constraints,
Perspectives, B i l i o , Rosenthal and Sindermann (Eds), Europ. Aquacult. Soc.,
70.
Bredene, Belgium, 77-I06 (1986).

N. De Pauwand E. Van Vaerenbergh, Microalgal wastewater treatment systems:
potentials and limits, in Phytodepuration and the Employmentof the Biomass
Produced, Ghetti (Ed), Centro Ric. Produz. Animali, Reggio Emilia, I t a l y ,
71.
211-287 (1983).
N. De Pauw, E. Bruggeman and G. Persoone, Research on the tertiary
treatment of swine manure by mass culturing of algae, Mitt.
72.
Verein. Limnol., 21, 490-506 (1978).

N. De Pauw, J. Morales and G. Persoone, Mass culture of microalgae in
aquaculture systems: progress and constraints,
73.
Internat.
Hydrobiologia, l l 6 / l l T ,
121-134 (1984).
N. De Pauw, J. Verboven and C. Claus, Large scale microalgae production for
nursery rearing of marine bivalves, Aquac. Engin., 2, 27-47 (1983).
74.
N. De Pauw, H. Verlet and L. De Leenheer, Heated and unheated outdoor

cultures of marine algae with animal manure, in Algae Biomass, Shelef and
Soeder (Eds), Elsevier/North-Holland Biomedical Press, Amsterdam, 315-341
75.
(1980a).
N. De Pauw, L. De Leenheer Jr., P. Laureys, J. Morales and J. Reartes,
Cultures d'algues et d'invert~br~s sur d~chets agricoles,
Pisciculture en Etang, Billard (Ed), I.N.R.A., P u b l . Paris,
( l g8Ob).
76.
in La
189-214
R. Dinges, The a v a i l a b i l i t y of Daphnia for water quality improvement and as

an animal food source, in Proc. Wastewater Use in the Production of Food
and Fiber, EPA 660/2-74-041, US Environmental Protection Agency,
Washington, DC, 142-161 (1974).
77.
J.C.
Dodd, Algae production
and harvesting
f r o m animal wastewater,
Agricult. Wastes, ! , 23-27 (1979).

78.
J . C . Dodd, Elements of pond design and construction, in

Microalgal Mass Culture, Richmond (Ed), CRC Press Inc.,
Florida, 265-283 (1986).
Handbook of
Boca Raton,
758
79.
80.
81.
Z. Dubinsky, T. Berner and S. Aaronson, Potential of large-scale algal

culture for biomass and l i p i d production in arid lands, Biotechnol. Bioeng.
Symp., (8), 51-68 (1978).
E. Duerr, Production of biomass for use in animal feeds: current research
at the Oceanic Institute, Hawaii, Appl. Phycol. Forum, ~, 6-7 (1985).
H. Durand-Chastel, Production and use of Spirulina in Mexico, in Algae
Amsterdam, 51-64 (1980).

P. Edwards, A review of recycling organic wastes into fish with emphasis on
the tropics, Aquaculture, 21, 261-279 (1980).
83. P. Edwards, O.-A. Sinchumpasak, V.K. Labhsetwar and M. Tabucanon, The
harvest of microalgae from the effluent of a sewage fed high rate
stabilization pond by Tilapia nilotica. Part 3: Maize cultivation
experiment, bacteriological studies and economic assessment, Aquaculture,
82.
84.
85.
86.
87.
88.
89.
90.
91.
23, 149-170 (1981).

M. EI-Fouly, F.E. Abdalla, F.K. EI-Baz and F.H. Mohn, Experience with algae
production within the Egypt-German Microalgae project, Arch. Hydrobiol.
Beih. Ergebn. Limnol., 20, 9-15 (1985).
C.T. Enright, G.F. Newkirk, J.S. Craigie and J.D. Castell, Evaluation of
phytoplankton as diets for juvenile Ostrea edulis L., J. Exp. Mar. Biol.
Ecol., 96, 1-13 (1986a).
C.T. Enright, G.F. Newkirk, J.S. Craigie and J.D. Castell, Growth of
juvenile Ostrea edulis L. fed Chaetoceros 9racilis SchUtt of varied
chemical composition, J. Exp. Mar. Biol. Ecol., 96, 15-26 (1986b).
J.M. Erickson, M. Ralsire, P. Bennoun, P. Delepelaire, B. Diner and J.D.
Rochaix, Herbicide resistance in Chlamxdomonas reinhardtii results from a
mutation in the chloroplast gene for the 32-kilodalton protein of
photosystem I I , Experientia, 8_11,3617-3621 (1984).
L.E. Erickson and H.Y. Lee, Process analysis and design of algal growth
systems, in Algal Biomass Technologies, Barclay and Mclntosh (Eds), Nova
Hediwgia Beih. 83, J. Cramer, Berlin, 197-206 (1986).
FAO/WHO, Energy and protein requirement. Food and Agriculture Organization
of United Nations, Food Nutrition Meeting Report, Series NO 53, Rome
(1973).
U. Fingerhut, L.E. Webb and C.J. Soeder, Increased yields of Rhizobium
japonicum by an extract of the green alga, Scenedesmus obliquus (276-3a),
Appl. Microbiol. Biotechnol., 19, 358-360 (1984).
H. Folmann, H. Markl and D. Vortmeyer, Continuous production of microalgae
under mixotrophic conditions, Germ. Chem. Eng., ~, 335-339 (1978).
92.
93.
94.
95.
96.
97.
98.
99.
lO0.
lOl.
I02.
I03.
J.M. Fox, Intensive algal culture techniques,

Mariculture, Crustacean Aquaculture, McVey (Ed),
759
in CRC Handbook of
CRC Press Inc., Boca
Raton, Florida, 15-41 (1983).

R.D. Fox, An integrated village health system, in Proc. Nat. Works. Algal
Systems. Indian Soc. Biotechnol., Shri A.M.M. Murugappa Chettiar Res.
Centre, Photosynthesis and Energy Div., Tharamani, Madras, 135- 138 (1980).
R.D. Fox, Algoculture, Ph.D. Thesis, Univ. of Strasbourg, France (1983).
R.D. Fox, Applied Phycol. Forum, ~, 8 (1985).
T. Fujiwara-Arasaki, In vitro d i g e s t i b i l i t y of algal proteins, Jap. J.
Phycol., 32, 293-299 (1984).
J. Gallagher, Population genetics of microalgae, in Algal Biomass
Technologies, Barclay and Mclntosh (Eds), Nova Hedwigia Beih. 83, J.
Cramer, Berlin, 6-14 (1986).
J. Gauthier, P. Talbot, G. Lessard, Y. Pouliot and J. de la NoOe, Epuration
du l i s i e r de porc et production de Spiruline. Rapport, Compagnie Lallemand
et Universit~ Laval (1985).
J. Goldman, Outdoor algal mass cultures. I. Applications, Water Res., 13,
1-19 (1979a).
J. Goldman, Outdoor algal mass cultures. I I .
Photosynthetic yield
limitations, Water Res., L3, I19-136 (1979b).
J. Goldman, Physiological aspects in algal mass cultures, in Algae Biomass,
Shelef and Soeder (Eds), Elsevier/North-Holland Biomedical Press, Amsterdam,
343-359 (1980).
J.C. Goldman and J.H. Ryther, Waste reclamation in an integrated food chain
system, in Biological Control of Water Pollution, Tourbier and Pierson
(Eds), Univ. of Pennsylvania Press, Philadelphia, 197-214 (1976).
J.C. Goldman and J.H. Ryther, Mass production of algae: bioengineering
aspects, Biological Solar Energy Conversion, Mitsui et al. (Eds), Academic
Press, New York, 367-378 (1977).
I04. B. Goldstein, Saline groundwater aquaculture I I . The growth of Crassostrea

spp. in the saline groundwater of new Mexico, Abstract, World Mariculture
Soc. Annual Meeting, Jan. 19-23 (1986).
I05. J.U. Grobbelaar, C.J. Soeder and D.F. Toerien, Wastewater for aquaculture,
Univ. of Orange Free State, Publ., Ser. C, (3), 1-215 (198I).
I06. J. Grobbelaar, C.J. Soeder and E. Stengel, Modelling algal productivity and
oxygen production in large outdoor cultures, Kernforschungsanlage J~lich
GmbH. I n s t i t . f~r Biotechnologie JUlich-Spez, 282, W-Germany (1984).
I07. J. Groeneweg and M. SchlOtter, Mass production of freshwater rotifers on
liquid wastes. I I . Mass production of Brachionus rubens Ehrenberg 1938 in
the effluent of high-rate algal ponds used for the treatment of piggery
waste, Aquaculture, 25, 25-33 (1981).
76o
I08. J. Groeneweg, B. Klein, F.H. Mohn, K.H. Runkel and E. Stengel, First
results of outdoor treatment of pig manure with algal bacterial systems, in
Algae Biomass, Shelef and Soeder (Eds), Elsevier/North-Holland Biomedical
Press, 255-264 (1980).
109. R. Gross, U. Gross, A. Ramirez, K. Cuadra, C. Collazos and W. Feldheim,
Nutritional tests with green alga Scenedesmus with healthy and malnourished
children, Arch. Hydrobio1. Beih. Ergebn. Limnol., I I , 161 (1978).
llO. C. Gudin, A. Bernard, D. Chaumont and C. Th~penier, Produits chimiques a
partir de microalgues, in La biomasse source d'interm~diaires industriels,
Heslot et al. (Eds), Technique et documentation Lavoisier, Paris, 275-289
(1983).
I l l . D.O. Hall, The production of biomass: a challenge to our society, in CRC
Handbook of Microbial mass culture, Richmond (Ed). CRC Press Inc., Boca
Raton, Florida, 1-24 (1986).
I12. D.O. Hall, Immobilized photosynthetic system for the production of fuels
and chemicals, 8th International Biotechnology Symposium, Paris, July
17-22, Abstract Book, 34 (1988).
l l 3 . P.C. Hallenbeck, L.V. Kochian, J.C. Weissman and J.R. Benemann, Solar
energy conversion with hydrogen-producing cultures of the blue-green algae
Anabaena c~lindrica, Biotechnol. Bioengin. Symp., (8), 283-297 (1978).
If4. P. Heussler, Aspects of sloped pond engineering, Arch. Hydrobiol. Beih.
Ergebn. Limno1., 20, 71 (1985).
I15. P. Heussler, J. Castillo and F. Merino, Parasite problems in the outdoor
cultivation of Scenedesmus, Arch. Hydrobiol. Beih. Ergebn. Limnol., I I ,
223-227 (1978a).
l l 6 . P. Heussler, J. Castillo, S. Merino and V. Vasquez, Improvements in pond
construction and CO2 supply for the mass production of microalgae, Arch.
Hydrobiol., I I , 254 (1978b).
l l 7 . P. Heussler, J. Castillo and F. Merino, Ecological balance of algal
cultures in arid climates: major results of the Peruvian-German Microalgae
Project at T r u j i l l o , Arch. Hydrobiol. Beih., I I , 17-22 (1978c).
If8. D.T. Hill and E.P. Lincoln, Development and validation of a comprehensive
model of large-scale production of microalgae, Agricultural Wastes, ~,
43-64 (Ig81).
If9. C. H i l l s , The secrets of Spirulina, Medical discoveries of Japanese
Doctors., Univ. of the Trees Press, Boulder Creek, Calif. (Ig80).
12O. T. I s h i i , M. Kandachi and M. Kamedaka, Nutritional value of Chlorella
proteins, J. Jap. Soc. Food Nutr., 27, 103-108 (1974).
121. V.T. Jaya, M.L. Scarino and A.M. Spadoni, Caratteristiche n u t r i t i o n a l i in
viso di Spirulina maxima, in Prospettive delle Coltura di Spirulina in
I t a l i a , Materassi (Ed). Acad. dei Geografili, Firenze, 195 (Ig8O).
761
122. D. Kaplan, A.E. Richmond, Z. Dubinsky and S. Aaronson, Algal n u t r i t i o n , in

Handbook of Microalgal Mass Culture, Richmond (Ed), CRC Press Inc., Boca
Raton, Florida, 147-198 (1986).
123. I. Karube, H. Ikemoto, K. Kajiwawa, E. Tamiya and H. Matsuoka,
Photochemical energy conversion using immobilized blue-green algae, J.
Biotechnol., 4, 73-80 (1986).
124. K. Kawaguchi, Microalgae production systems in Asia, in Algae Biomass,
Shelef and Soeder (Eds), Elsevier/North-Holland Biomedical Press, Amsterdam,
25-33 (1980).
125. E. Kessler, An extremely cadmium-sensitive strain of Chlorella, Experientia,
41, 16 (1985).
126. N. Kosaric and Z.T. Ngcakani, Floculated algae slurry upflow bioreactor for
toxic industrial treatment, 8th International Biotechnology Symposium,
Paris, July 17-22, Abstract Book, 232 (1988).
127. K. Kreuzberg, G. Reznicezk and G. Kl~ck, Properties of algal biomass
production and the parameters determining i t s fermentative degradation,
Third Conf. Energy from Biomass, March 25-29, Venice, I t a l y (1985).
128. S. Kumamoto, U.S. patent 4, 468, 460 (1984).
129. S. Kumazawa and A. Mitsui, Hydrogen metabolism of photosynthetic bacteria
and algae, in CRC Handbook of Biosolar Resources, Vol. I , Part I , Zaborsky
(Ed), CRC Press Inc., Boca Raton, Florida (1982).
130. A. Lavoie, Traitement t e r t i a i r e des eaux us~es par Scenedesmus obliquus:
influence de l ' ~ t a t physiologique sur la prise en charge des nutriments et
la r~colte des biomasses produites, Th~se Ph.D., Universit~ Laval, Quebec
(1985).
131. A. Lavoie and d. de la NoOe, Harvesting microalgae with chitosan, J. World
Maricult. Soc., 14, 685-694 (1983).
132. A. Lavoie and J. de la No~e, Hyperconcentrated cultures of Scenedesmus
obliquus, A new approach for wastewater biological t e r t i a r y treatment?,
Water Res., 19, 1437-1442 (1985).
133. A. Lavoie, J. de la No~e and J.B. S~rodes, R~cup~ration de microalgues en
eaux us~es: ~tude comparative de divers agents floculants, Can. J. Cir.
Eng., l l , 266-272 (1984).
134. B.H. Lee, G.A. Picard and G. Goulet, Effects of processing methods on the
nutritive value and d i g e s t i b i l i t y of Ooc~stis alga in rats, Nut. Rep.
Intern., 25, 417-429 (1982).
135. Y.K. Lee and H.M. Tan, Genetic recombination through protoplast fusion in
algae, Recent Progress in Algal Biotechnology, Villeneuve d'Ascq, Sept.
14-17, C2 (1987).
762
136. N.W. Lem and B.R. Glick, Biotechnological uses of cyanobacteria, Biotech.
Adv., ~, 195-208 (1985).
137. C. Lemieux, M. Turmel, R.W. Lee and G. Bellemare, A 21 kilobase-pair
deletion-addition difference in the inverted repeat sequence of chloroplast
DNA from Chlamydomonasengauretos and C. moewusii, Plant Molecul. Biol., 5,
77-84 (1985).
138. S. Li, Nitrogen-fixing blue-green algae, a source of b i o f e r t i l i z e r , in Proc.
Joint China-U.S. Phycology Symp., 479 (1981).
139. L.P. Lin, Microstructure of spray-dried and freeze-dried microalgal powders,
Food Microst., 4, 341-348 (1985).
140. E.P. Lincoln and D.T. H i l l , An integrated microalgae system, in Algae
Amsterdam, 229-244 (1980).
141. E.P. Lincoln and B.
Koopman, Bioflocculation of
microalgae in
mass
cultures, in Algal Biomass Technologies, Barclay and Mclntosh (Eds), Nova

Hedwigia Beih. 83, J. Cramer, Berlin, 207-211 (1986).
142. E.P. Lincoln, T.W. Hall and B. Koopman,Zooplankton control in mass algal
cultures, Aquaculture, 32, 331-337 (1983).
143. V.L. Loosanoff, J.E. Hanks and A.E. Ganaros, Control of certain forms of
zooplankton in mass algal cultures, Science, 125, I092-I093 (1957).
144. I.V. Maksimova, 0.0. Malakovskaya, E.G. Pryadil'Shchikova, Antibacterial
activity in diatoms. I. Lipids of Nitzschia ovalis and their antibacterial
a c t i v i t y , Fiziol. Rast. (Mosc.), 31, 944-950 (In Russian) (1984).
145. H. M~rkl, Modelling of algal production systems, in Algae Biomass, Shelef
and Soeder (Eds), Elsevier/North-Holland Biomedical Press, Amsterdam,
361-383 (1983).
146. C. Martin, J. de la No~eand G. Picard, Intensive cultivation of freshwater
microalgae on aerated pig manure, Biomass, ~, 245-259 (1985a).
147. C. Martin, G. Picard and J. de la No~e, Epuration biologique du l i s i e r de
porc par production de biomasses d'algues unicellulaires, Mircen J. Appl.
Microbiol. Biotechnol., I I , 173-184 (1985b).
148. A. Martinez, A. Alana, M.J. Llama and J.L. Serra, Sustained photoproduction
of ammonia from nitrate or n i t r i t e by free and immobilized cells of
Phormidium laminosum, in Inorganic Nigrogen Metabolism, Ullrich et al.
(Eds), Springer-Verlag, Berlin, 220-222 (1987).
149. N.P. Massuyk, Large-scale culture of the carotene-bearing alga Dunaliella
salina Teod, British Library Leading Div. Translating Progr. RTS 4107
(1966).
763
150. R. Materassi, C. Paoletti, W. Balloni and G. Florenzano, Some

considerations on the production of l i p i d substances by microalgae and
cyanobacteria, in Algae Biomass, Shelef and Soeder (Eds), Elsevier/
North-Holland Biomedical Press, Amsterdam, 619-626 (1980).
151. R.E. McKinney, E.C. McGriff, R.J. Sherwood, N.V. Wahbeh and D.W. Newport,
Ahead: activated Algae?, Water and Wastes Engin., 8, 51-52 (1971).
152. B. Metting, Agronomic uses of algae, Appl. Phycol. Forum, ~, II (1985).
153. H. Mitsuda, M. Higuchi, A. Yamamoto and K. Nakahima, Protein concentrate
from the blue-green algae, its nutritive value, J. Jap. Soc. Food and
Nutr., 30, 23-28 (1977a).
154. H. Mitsuda, Y. Nishikawa, M. Higuchi, K. Nakahima and F. Kawai, Effect of
the breaking of Chlorella cells on the d i g e s t i b i l i t y of Chlorella proteins,
J. Jap. Soc. Food and Nutr., 30, 93-98 (1977b).
155. A. Mitsui, Saltwater based biological solar energy conversion for fuel,
chemicals, f e r t i l i z e r , food and feed, in Proc. Bio-energy '80. Bio-energy
Council, Washington, D.C., 486-491 (1980).
156. H. Mohn, Experiences and strategies in the recovery of biomass from mass
cultures of microalgae, in Algae Biomass, Shelef and Soeder (Eds),
Elsevier/ North-Holland Biomedical Press, Amsterdam, 547-571 (1980).
157. P.M. Molton, G.R. Cysewski and D.B. Anderson, Algae grown on animal wastes
as a source of industrial o i l s , polysaccharides and animal feed, in
Proceed. Intern. Symp. Biogas, Microalgae and Livestock Wastes, Chung (Ed),
Taipei, Taiwan, Sept. 15-17, 381-387 (1980).
158. R. Moraine, G. Shelef, E. Sandbank, Z. Bar-Moshe and L. Shvartzburd,
Recovery of sewage-grown algae: flocculation, flotation and centrifugation
techniques, in Algae Biomass, Shelef and Soeder (Eds), Elsevier/NorthHolland Biomedical Press, Amsterdam, 531-545 (1980).
159. J. Morales, J. de la NoUe and G. Picard, Harvesting marine microalgae
species by chitosan flocculation, Aquacult. Engin., ~, 257-270 (1985).
160. A.N.C. Morse and D.E. Morse, GABA-mimetic molecules f r o m Porph~ra
(Rhodophyta) induce metamorphosis of Haliotis (Gastropoda) larvae,
Hydrobiologia, l l 6 / l l 7 , 155-158 (1984).
161. J. Nakas, M. Schaedle, D.W. Whitely and S.W. Tanenbaum, Bioconversion of
algal biomass to neutral solvents, in Algal Biomass Technologies, Barclay
and Mclntosh (Eds), Nova Hedwigia Beih. 83, J. Cramer, Berlin, 171-175
(1986).
162. D.L.R. Narasimha, G . S . Venkataraman, S . K . Duggal and B.O. Eggum,
Nutritional quality of the blue-green alga Spirulina platensis Geitler, J.
Sci. Food A g r i c u l t . , 33, 456-460 (1982).
163. J. Necas, Genetic aspects of the biotechnology of freshwater algae and
blue-green algae, Biologicke Listy, 50, 98-122 (In Czechoslovakian) (1985).
764
164. B.P. Nigam, P.K. Ramanathanand L.V. Venkataraman, Application of chitosan

as a flocculant for the cultures of the green alga: Scenedesmus acutus,
Arch. Hydrobiol., 88, 378-387 (1980).
165. R.S. Norton and R.J. Wells, A series of polybrominated bi-indoles from the
marine blue-green alga Rivularia firma. Application of 13C NMR Spin-lattice
relaxation data and 13C-'H coupling constants to structure elucidation, J.
Am. Chem. Soc., I04, 3628-3635 (1982).
166. J.T. Novak and D.E. Brune, Inorganic carbon limited growth kinetics of some
freshwater algae, Water Res., I_~9, 215-225 (1985).
167. E.J. Olguin and J.M. Vigueras, Unconventional food production at the
village level in a desert area of Mexico, Proc. 2nd Wld. Congr. Chem. Eng.,
Montreal, Canada, 4-9 October (1981).
168. W.J. Oswald, Integrated systems for community waste management, in Proceed.
Internat.
Symp. Biogas, Microalgae and Livestock
Wastes, Chung (Ed),
Taipei, Taiwan, Sept. 15-17 (1980).

169. W.J. Oswald and H.B. Gotaas, Photosynthesis in sewage treatment, Trans. Am.
Soc. Civ. Eng., 122, 73-I05 (1957).
170. W.J. Oswald, J.R. Benemann and B.L. Koopman, Production of biomass from
freshwater aquatic systems - concepts of large scale bioconversion systems
using microalgae, Proc. Fuels from Biomass Symp., Univ. of I l l . , Champaign,
I l l , 59-81 (1977).
171. W. Pabst, Nutritional evaluation of nonsewagemicroalgae by the rat balance
method, Arch. Hydrobiol. Beih. Ergebn. Limnol., l l , 65-70 (1978).
172. R.N. Padhy, Cyanobacteria employed as f e r t i l i z e r s and waste disposers,
Nature, 31__]7,475-476 (1985).
173. V. Partali, Y. Olsen, P. Foss and S. Liaaen-Jensen, Carotenoids in food
chain studies. I. Zooplankton (Daphnia magna) response to a unialgal
(Scenedesmus acutus) carotenoid diet, to spinach, and to yeast diets
supplemented with individual carotenoids, Comp. Biochem. Physiol., 82B,
767-772 (1985).
174. H.D. Payer, W. Pabst and K.H. Runkel, Review of the nutritional
and
toxicological properties of the green alga Scenedesmusobliquus as a single

cell protein, in Algae Biomass, Shelef and Soeder (Eds), Elsevier/NorthHolland Biomedical Press, Amsterdam, 787-797 (1980).
175. G. Persoone, J. Morales, H. Verlet and N. De Pauw, A i r - l i f t pumps and the
effect of mixing on algal growth, in Algae Biomass, Shelef and Soeder
(Eds), Elsevier/North-Holland Biomedical Press, Amsterdam, 505-522 (1980).
176. A.J.H. Pieterse and J. Le Roux, Algal culture using wastewater, in Symp.
on Aquaculture in Wastewater, Nov. 1980, CSIR, Pretoria (1980).
765
177. Y. Pouliot and J. de la NoUe, Mise au point d'une installation-pilote

d'(puration tertiaire des eaux us(es par production de microalgues, Rev.
Fran~. Sci. de l'Eau, ~, 207-222 (1985).
178. D. Proulx and J. de la Noue, Harvesting Daphnia magna grown on urban
tertiarily-treated effluents, Water Res., 19, 1319-1324 (1985a).
179. D. Proulx and J. de la NoUe, Growth of Daphnia magna grown on urban
wastewaters tertiarily-treated with Scenedesmus sp., Aquacult. Engin., ~,
93-111 (I 9BBb).
180. D. Proulx and J. de la No~e, Removal of macronutrients from wastewaters by
immobilized microalgae, in Bioreactor Immobilized Enzymes and Cells
Fundamentals and Applications, Moo-Young (Ed.), Elsevier Applied Science,
London, 301-310 (1988).
181. G.D. Pruder, Aquatic production systems: algae, in Nursery Culturing of
Bivalve Molluscs, Claus et al. (Eds), Eur. Maricult. Soc. Spec. Publ. 7,
Bredene, Belgium, 219-226 (1981).
182. G. Pruder, Biological control of gas exchange in intensive aquatic
production systems, I.E.E.E, 1002-I004 (1983).
183. G.D. Pruder and E.E. Greenhaugh, Bivalve molluscs rearing process, United
States Patent 4,080,930. March 28 (1978).
184. K.K. Rao and D.O. Hall, Photosynthetic production of fuels and chemicals in
immobilized systems, Trends in Biotechnology, ~, 124-129 (1984).
185. C. Ratledge and C . A . Boulton, Fats and oils,
in Comprehensive
biotechnology, Vol. l , Moo-Young (Ed), Pergamon Press, 983-1003 (1986).
186. R.H. Reed, S.R.C. Warr, N.W. Kerby and W.D.P. Stewart, Osmotic
shock-induced release of low-molecular weight metabolites from free- living
and immobilized cyanobacteria, Enz. Microb. Technol., 8, lOl-104 (1986).
187. J.L. Reichelt and M.A. Borowitzka, Antimicrobial activity from marine
algae: results of a large-scale screening programme, Hydrobiologia,
l l 6 / l l 7 , 158-168 (1984).
188. A. Richmond (Ed), CRC Handbook of microalgal mass culture, CRC Press Inc.,
Boca Raton, Florida (1986a).
189. A. Richmond, Cell response to environmental factors, in CRC Handbook of
Microalgal Mass Culture, Richmond (Ed), CRC Press Inc., Boca Raton,
Florida, 69-99 (1986b).
190. A. Richmond, Microalgae of economic potential, in CRC
Florida, 199-243 (1986c).
191. A. Richmond, Outdoor mass cultures of microalgae, in CRC
Florida, 285-329 (1986d).
Handbook of
Boca Raton,
Handbook of
Boca Raton,
766
J. DE LA NOUEand N. DEPAUW
192. A. Richmond and E.W. Becker, Technological aspects of mass cultivation - a

general outline, in CRC Handbookof Microalgal Mass Culture, Richmond (Ed),
CRC Press Inc., Boca Raton, Florida, 245- 263 (1986).
193. A. Richmond, S. Karg and S. Boussiba, Effects of bicarbonate and carbon
dioxide on
the
competition
between Chlorella vul9aris
and Spirulina
platensis, Plant Cell Physiol., 23, 1411-1417 (1982).

194. O.A. Roels, K.C. Haines and J.B. Sunderlin, The potential yield
of
a r t i f i c i a l upwelling mariculture, in Proc. lOth Eur. Symp. Mar. Biol.,

Persoone and Jaspers (Eds), Universa Press, Wetteren, Belgium, 381-390
(1976).
195. R. Samson, Caract~ristiques
physiologiques de la croissance et de la
production de polysaccharides chez neuf esp~ces d'algues endog~nes isol~es

des effluents du traitement secondaire de l'usine d'~puration de
Valcartier, Th~se M.Sc., Univ. Laval, Quebec (1980).
196. E. Sandbank and B. Hepher, Microalgae grown in wastewater as an ingredient
in the diet of warmwater fish, in Algae Biomass, Shelef and Soeder (Eds),
197. J.L. Sansregret, Culture de Phaeodact1umtricornutum sur milieux enrichis,
Th~se M.Sc., Universit~ Laval, Quebec (1986).
198. C. Santillan, Mass production of Spirulina, Experientia, 38, 40-43 (1982).
199. A.
Sasson, Les biotechnologies, d~fis et promesses, Unesco, 134- 219
(1983).
200. P.N. Saxena, M.R. Ahmad, R. Shyam and D.V. Amla, Cultivation of Spirulina
in sewage for poultry feed, Experientia, 39, I077-I083 (1983).
201. M. SchlUtter and J. Groeneweg, Mass production of freshwater rotifers on
liquid wastes. I. The influence of some environmental factors on population
growth of Brachionus rubens, Aquaculture, 25, 17-24 (1981).
202. N.S. Scrimshaw, Nutritional and tolerance considerations in the feeding of
single cell protein, in Proc. Internat. Symp. "Food and Biotechnology", de
la No~e et al. (Eds), Centre de recherche en nutrition, Universit~ Laval,
Quebec, 197-214 (1986).
203. E.D. Scura, A.M. Kuljis, R.H. York Jr. and R.S. Le Goff, The commercial
production of oysters in an intensive raceway system, in lOth Annual
Meeting World Maricult. Soc., Avault (Ed), Louisiana St. Univ., Baton
Rouge, Louisiana, 624-630 (1979).
204. I. Setlik, S. Veladimir and I. Malek, Dual purpose open circulation units
for large scale culture of algae in temperate zones. I. Basic design
considerations and scheme of p i l o t plant, Algal. Stud. (Trebon, CSSR), ~,
II (1970).
205. G. Shelef and C.J. Soeder (Eds), Algae Biomass, Production and Use,
Elsevier/North-Holland Biomedical Press, Amsterdam (1980).
767
206. G. Shelef, G. Oron and R. Moraine, Economic aspects of microalgae

production on sewage, Arch. Hydrobiol. Beih. Ergebn. Limnol., I I , 281-294
(1978).
207. G. Shelef, A. Sukenik and M. Green, Separation and harvesting of marine
microalgal biomass, in Algal Biomass Technologies, Barclay and McIntosh
(Eds), Nova Hedwigia Beih. 83, J. Cramer, Berlin, 245- 251 (1986).
208. G. Shelef, Y. Azov, R. Moraine and G. Oron, Algal mass production as an
integral part of a wastewater treatment and reclamation system, in Algae
Amsterdam, 163-189 (1980).
209. N.W. Shifrin, Oils from microalgae, in Biotechnology for the oils and fats
industry, Ratledge et al. (Eds), American Oil Chemist's Soc., 145-162
(1984).
210. L.E. Shubert and B.D. Larsen, Nutritional value of Spirulina and Chlorella
for human consumption, Appl. Phycol. Forum, &, 12 (1985).
211. R. Simon and A.H. Latorella, Approaches to the genetics of Dunaliella, in
Algal Biomass Technologies, Barclay and Mclntosh (Eds), Nova Hedwigia Beih.
83, J. Cramer, Berlin, 15-20 (1986).
212. J.T. Snow, L.E. Piper, S.A. Lupton and G.R. Stegen, A comparative
213.
214.
215.
216.
assessment of marine biomass materials, in EPRI/GRI Workshop on biomass

resources and conversion, Electric Power Research Institute, Palo Alto, CA,
II-33 (1978).
C.J. Soeder, Massive cultivation of microalgae: results and prospects,
Hydrobiologia, 72, 197-209 (1980).
C.J. Soeder, Aquatic bioconversion of excrements in ponds, in Animals as
waste converters, Ketelaar and lwema (Eds), PUDOC, Wageningen, 130-136
(1984).
C.J. Soeder, An historical outline of applied algology, in Handbook of
Microalgal Mass Culture. Richmond (Ed.), CRC Press Inc., Boca Raton,
Florida, 25-41 (1986).
P. Soong, Production and development of Chlorella and Spirulina in Taiwan,
in Algae Biomass, Shelef and Soeder (Eds), Elsevier/North- Holland
Biomedical Press, Amsterdam, 97-I13 (1980).
217. H. Spiller and K.R. Shanmugam, Genetic modification of Anabaena variabilis

for enhancement of H2 evolution, in Algal Biomass Technologies, Barclay and
McIntosh (Eds), Nova Hedwigia Beih 83, J. Cramer, Berlin, 3-5 (1986).
218. T. Stadler, J. Mollion, M.-C. Verdus, Y. Karamanos, H. Morvan and D.
Christiaen (Eds), Algal Biotechnology, Elsevier Applied Science Publ.,
Barking, England, 1988.
219. A. Sukenik and G. Shelef, Algal autoflocculation
verification and
proposed mechanism, Biotechnol. Bioengin., 26, 142-147 (1984).
768
220. A. Sukenik, W. SchrOder, J. Lauer, G. Shelef and C . J . Soeder,

Coprecipitation of microalgal biomass with calcium and phosphate ions,
Water Res., 19, 127-129 (1985).
221. J.J. Strain, H . J . Fellowfield and M.K. Garrett, The composition and
nutritive value of Chlorella vul~aris grown in laboratory scale and p i l o t
plant culture, Proceed. Nutr. Soc., 43, 92A (1984).
222. L. Switzer, Spirulina, the whole food revolution, Proteus Corporation,
Bantam Books, Toronto (1982).
223. E.P. Taganaides, K.C. Chau and B.Y. Lee, Animal waste managements and
utilization in Singapore, Agricult. Wastes, ~, 129-141 (1979).
224. E. Tarifeno-Silva, L.Y. Kawasaki, D.P. Yu, M.S. Gordon and D.J. Chapman,
Aquacultural approaches to recycling of dissolved nutrients in secondarily
treated domestic wastewaters. I f . Biological productivity of a r t i f i c i a l
food chains, Wat. Res., I_6_6,51-57 (1982a).
225. E. Tarifeno-Silva, L.Y. Kawasaki, D.P. Yu, M.S. Gordon and D.J. Chapman,
Aquaculture approaches to recycling of dissolved nutrients in secondarily
treated domestic wastewaters. I I I . Uptake of dissolved heavy metals by
a r t i f i c i a l food chains, Wat. Res., I_6_6,59-65 (1982b).
226. O. Tel-Or, S. Boussiba and A.E. Richmond, Products and chemicals from
Spirulina platensis, in Algae Biomass, Shelef and Soeder (Eds), Elsevier/
North-Holland, Biomedical Press, Amsterdam, 611-613 (1980).
227. K.L. Terry, The diel periodicity of algal physiological processes:
implications for the optimization of culture yield, in Algal Biomass
Technologies, Barclay and Mclntosh (Eds), Nova Hedwigia Beih 83, J. Cramer,
Berlin, 95-99 (1986).
228. K.L. Terry, J. Hi rata and E.A. Laws, Light - limited growth of two strains
of
the marine diatom Phaeodact~lum tricornutum Bohlin:
chemical
composition, carbon partitioning and the diel periodicity of physiological
processes, J. Exp. Mar. Biol. Ecol., 68, 209-227 (1983).
229. W.H. Thomas, D.N.R. Seibert, M. Alden, P. Eldridge, A. Neori and S. Gaines,
Microalgae from desert saline waters as potential biomass producers, Prog.
in Solar Energy, 6, 143-145 (1983).
230. D.F. Toerien, Growth kinetic prediction of algal growth in mass cultivation
systems, in Wastewater for Aquaculture, Grobbelaar et al. (Eds). U.O.F.S.
Publ. Series C, (3), 168-172 (198).
231. D.F. Toerien and J.U. Grobbelaar, Algal mass cultivation experiments in
South Africa, in Algae Biomass, Shelef and Soeder (Eds), Elsevier/NorthHolland Biomedical Press, Amsterdam, 73-80 (1980).
232. M. Turmel, G. Bellemare, R.W. Lee and C. Lemieux, A linear DNA molecule of
5.9 kilobase pairs is highly homologous to the chloroplast DNA in the green
alga Chlamydomonas moewusii, Plant Molecul. Biol., 6, 313-319 (1986).
769
233. R. Ukeles, 4. Cultivation of plants, 4.1. Unicellular plants, in Marine

Ecology, Kinne (Ed), John Wiley and Sons, London, New York, 367-466 (1976).
234. R. Ukeles, American experience in the mass culture of microalgae for feeding
larvae of the American oyster, Crassostrea virginica, in Algae biomass,
Shelef and Soeder ( E d s ) , Elsevier/North-Holland
Biomedical Press,
Amsterdam, 287-306 (1980).
235. L.V. Venkataraman, Blue-green algae as b i o f e r t i l i z e r , in CRC Handbook of
Microalgal Mass Culture, Richmond (Ed), CRC Press Inc., Boca Raton,
Florida, 455-471 (1986).
236. L.V. Venkataraman, B.P. Nigam and P.K. Ramanathan, Rural oriented fresh
water cultivation and production of algae in India, in Algae Biomass,
Shelef and Soeder ( E d s ) , Elsevier/North-Holland
Biomedical Press,
Amsterdam, 81-95 (1980).
237. L.V. Venkataraman and E.W. Becker, Production of food, feed, biochemicals
and bioenergy from microalgae, 8th International Biotechnology Symposium,
Paris, July 17-22, Abstract Book, 53 (1988).
238. A. Vonshak, Laboratory techniques for the cultivation of microalgae, in CRC
Handbook of Microalgal Mass Culture, Richmond (Ed), CRC Press Inc., Boca
Raton, Florida, I17-145 (1986).
239. A. Vonshak, A. Abeliovich, S. Boussiba, S. Arad and A. Richmond,
Production of Spirulina biomass: effects of environmental factors and
population density, Biomass, ~, 175-185 (1982).
240. S.S. Voronkova, O.N. Al'Bitshaya, N.P. Sadkova, S.V. Rogozhin, S.A.
Pavlova, M.K. Isaev, I.A. Abakumova and V.I. Fofanov, Relative nutritional
value harmlessness and physicochemical properties of Spirulina platensis
proteins, Scl'skohozyaistennaya Biologiya (In Russian), 4, 63-67 (1983).
241. A.S. Watson, Aquaculture and algae culture, Process and production, Noyes
Data Corporation, Park Ridge, N.J. (1979).
242. K.L. Webb and F.L. Chu, Phytoplankton as a food source for bivalve larvae,
in Biochemical and Physiological Approaches to Shellfish Nutrition, Pruder
et al. (Eds), World Maricult. Soc. Spec. Publ. 2, Baton Rouge, Louisiana,
272-291 (1982).
243. J.
Weiner, Marine biotech in the Negev desert,
Biotechnology, ~, 41
(1985).
244. R.M. Weiner, R.R. Colwell, R.J. Jarman, D.C. Stein, C.E. Somerville and
D.B. Bonar, Applications of biotechnology to the production, recovery and
use of marine polysaccharides, Biotechnology, ~, 899-902 (1985).
245. J.C. Weissman and J.R. Benemann, Polysaccharide production by microalgae.
Phase I. Final report to National Science Foundation, NSF/RA- 800567
(1980).
770
J. DE LA NOUE a n d N. DE PAUW
246. D.J. Welsh, F.J. Petrovits, R.A. Kraus, L.M. Wuethrich, C.A. Withstandley
and K.N. Wilson, An evaluation of tangential flow f i l t r a t i o n of microalgal
cell harvesting, Aquacultural Res. Corp., Denis, Mass. 02638, Final project
report NSF, Washington (1985).
247. L.A. Williams, E.L. Foo, A.S. Foo, I. KUhn and C.G. Heden, Solar
bioconversion systems based on algal glycerol production, Biotechnol.
Bioengineer. Symp., (8), I15-130 (1978).
248. F.R. Wolf, Physiological and metabolic studies on a branched hydrocarbon
producing strain of Botr~ococcus braunii, in Algal Biomass Technologies,
Barclay and Mclntosh (Eds), Nova Hedwigia Beih. 83, J. Cramer, Berlin,
160-170 (1986).
249. J.M. Wood, F.K. Gleason, C.P. Mason, J. Pignatello and R. Carlson,
A c t i v i t y of a chlorine-containing a n t i b i o t i c isolated from a freshwater
cyanobacteria, Fed. Proc. Am. Soc. Exper. Biol.-CISTI, 4:640, Abst, No 2163
(1982).
250. J.L.C. Wright, Biologically active marine metabolites:
some recent
examples, Proceed. Nova Scotia Inst. Sci., 34, 133-161 (1984).
251. S. Yannai, S. Mokaday, K. Sachs, B. Kantorowitz and Z. Berk, Certain
contaminants in algae and in animals fed algae containing diets and
secondary t o x i c i t y of the algae, in Algae Biomass, Shelef and Soeder (Eds).

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Biotech. Adv. Vol. 6, pp.

0734-9750/88 $0.00 + .50

THE POTENTIAL OF MICROALGAL

The nutritive value of the microalgal biomasses for humanand animal

consumption is also commented upon as well as some socio-economical aspects.

Technical transformations - Fine

chemicals - Feeds/Foods - Genetic manipulations IMPORTANCE AND POTENTIAL OF MICROALGAE

new biotechnologies. Along with basic research, applied algology has

As a result, already in the 1960s, the

commercial production of Chlorella in Japan and Taiwan as a novel health food

In the 1950s the idea of

trend in the 1980s is to use microalgae as a source of commonand fine chemicals.

For historical reviews on applied algology, the

reader is referred to Goldman (1979a) and Soeder (1980, 1986).

food, feed and chemicals.

attractive characteristic of microalgae, in comparison with other microorganisms,

As such, microalgae could play an

Hereby, annual yields of 25 T and more,

Under favourable conditions, microalgal cultures can produce up to

importance is also that microalgae can be grown yearound and harvested on a

[. DE LA NOUE and N. DE PAUW

Flow diagram of algal mass cultivation systems (from Becker and

(Becker and Venkataraman, 1982).

Although maximum growth rates are achieved in conditions of l i g h t saturation,

l i g h t limiting conditions, the effect of temperature is lessened with regard to

J. DE LA NOUE and N. DE PAUW

Venkataraman, 1982). Moreover,inorganic carbon levels play an important role in

Agitation is also very important to microalgal cultivation, not at least to

stratification but also to increase the l i g h t conversion efficiency (M~rkl, 1980;

Becker, 1986), eradicated chemically (Loosanoff et a l . , 1957) or by changing the

The role of microalgae in the purification of wastewater by biological

develop adequate recipes

enrichment. Problems relate to the methodology for distributing the nutrients

The algal reactors may be provided with a r t i f i c i a l l i g h t in or outside

the culture medium. In some cases i t may be profitable to work in greenhouses

Richmond and Becker (1986),

distinguished: horizontal turbulent flow created by various types of devices or

The horizontal ponds

may be circular or consist of simple or meandering oblong raceways. Sloped

Details on pond design and construction

are given by Dodd (1986).

Depending on whether or not the algal biomasses have to be harvested and

Advantages and disadvantages of different harvesting methods

The most successful techniques are centrifugation, f i l t r a t i o n and

J. DE LA NOUE and N. DE PAUW

algae, chemical flocculation eventually followed by flotation is commonpractice

To reduce the high inorganic flocculants demand, the use of polymers or

J. DE LA NOUE and N. DE PAUW

Microal~ae for Wastewater Treatment

forcing us to explore the f e a s i b i l i t y of wastewater recycling and

Abeliovich (1986). Processes involved in the removal of nutrients are

I t is also important to stress that in the treatment process, a large

array of microalgal species developing in different aquatic environments (e.g.

nutrient loads can be utilized (De Pauwand Van Vaerenbergh, 1983).

land-space requirements of microalgal wastewater treatment

hyperconcentrated algal cultures.

The challenge will be to make these systems not only

reliable but also economically competitive with conventional wastewater treatment

Basic chemical composition of the microalgae, Scenedesmusobliquus

Essential amino acid composition of Scenedesmus species comparedto the

l A r t i f i c i a l medium (Becker et a l . , 1976).

1985; Richmond, 1986c).

In most of these projects, serious