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Copyright i 1978 American Society for Microbiology Printed in U.S.A.
Algal Toxins
MIKE COLLINS
Civil Engineering Department, University ofMissouri, Columbia, Missouri 65211
INTRODUCTION .............................. 725
CHLOROPHYCOPHYTA ............................... 725
725
Caulerpa spp.725.....................................................
Cheatomorpha m inm 726
Uwva pp ....................... 726
CYANOPHYCOPHYTA ..................... 727
Micrcyatis aen ...................................................... 727
Ochrdnw8
nas pp............................... 734
INTRODUCTION CHLOROPHYCOPHYTA
This is a literature review of the toxins pro- The Chlorophycophyta have been associated
duced by algae. For this paper, only toxicity to with toxicity only in rare instances. Aside from
multicellular organisms was considered. This the three genera discussed below, Prescott's
eliminated a large number of publications deal- book The Algae: a Review (122) lists Chlorella
ing with antibacterial and antiviral substances and Scenedesmus as death-inducing algae, but
released by algae. Because the paper addresses information on these species is rare.
those toxins produced by algae, the phenomenon
of bioaccumulation of environmental contami- Caulerpa spp.
nants is not discussed. Also not discussed is the The marine benthic green alga Caulerpa is
one pathogenic algal genus, Prototheca, because responsible for the production of two toxic sub-
there is no indication of a toxin being released stances, namely, caulerpicin and caulerpin (4,38,
(review by Sudman [165]). 39). Both of these compounds have demon-
What are discussed are a wide variety of toxins strated toxicity in mice. They were originally
produced by five phyla of algae: Chlorophyco- isolated from C. racemosa but were also identi-
phyta (green algae), Cyanophycophyta (blue- fied in C. sertulariodes, C. lentillifera, and C.
green algae [cyanobactena]), Chrysophyco- lamourouxii. It is interesting to note that Cau-
phyta (diatoms, yellow-green and golden algae), lerpa is probably the most popular edible alga
Pyrrhophycophyta (dinoflagellates), and Rho- in the Philippines but becomes toxic during the
dophycophyta (red algae). The types of mole- rainy months. The toxicity is believed to derive
cules involved are diverse, going from simple from the agitation of the plant during the rainy
ammonia to complicated polypeptides and poly- season (37).
saccharides. The physiological effects are also The infrared spectrum of caulerpicin indicated
varied, ranging from the acute toxicity of para- that this compound was a long-chain saturated
lytic shellfish poison of Gonyaulax, leading to hydroxy amide. Aguilar-Santos and Doty (4)
death in a short period of time, to the chronic hypothesized from the spectral data that the
toxicity of carrageenans from red algae, which structure was as follows:
induce carcinogenic and ulcerative tissue
changes over long periods of time. Perhaps the CH20H
only link among this wide variety of toxin is I
that each is produced by some form of alga. CH3-(CHI2)i3-- CH-NH-CO(CH2) - CH3
725
726 COLLINS MICROBIOL. REV.
(n = 23, 24, 25). Mass spectral information led TABLE 1. Ichthyotoxic and hemolytic activities of
them to believe that the actual substance was a fatty acids occurring in C. minima (from reference
mixture of these homologous molecules. This 45)
structure has not received full confirmation Hemo- Avg death timeb
(144). The human physiological symptoms as- lytic ac-
sociated with caulerpicin ingestion include Fatty acid tivitya
(saponin 5 mg/100 ml 1 mg/100 ml
numbness and a cold sensation of the extremi- units/mg)
ties, rapid and difficult breathing, slight depres- 1.37 11 min (5) 44.6 min (5)
sion, and eventually loss of balance. Depending 8:0
9:0 0.73 12 min (5) 34.2 min (5)
on the dose, the effects are usually gone within 10:0 0.49 13 min (5) 6.6 h (4)
a couple of hours to a day. 12:0 0.52 16 min (5) 20.2 min (5)
Caulerpin was found to be a heterocyclic, red 14:0 2.48 9.4 h (2) (0)
substance after it was crystallized from ether 16:0 3.01 (0) (0)
extracts of the alga. It is a pyrazine derivative. 16:1 7.17 1.2 h (5) 2.4 h (5)
Both spectral analysis and degradation reactions 18:0 0.35 15.2 h (3) (0)
were used to determine the hypothesized struc- 18:1 4.52 2.5 h (5) 1.9 h (2)
ture of dimethyl 6,13-dihydrodibenzo[ b, i] phen- 18:2 9.49 2.8 h (1) (0)
azine-5,12-dicarboxylate (Fig. 1) (144). 20:0 0.26 14.3 h (1) (0)
Estimated by the method of Oshima et al. (104)
H
+
+H;
HzN HN
/
HO + OH
0 CH
FIG. 2. Anatoxin A (from Huber [63]). FIG. 3. Saxitoxin (from Shimizu [155]).
730 COLLINS MICROBIOL. REV.
soluble (46) and dialyzable (72). They are also of the nerve fibers (46). This mechanism is the
very hygroscopic and resistant to crystallization same as those described for saxitoxin (95) and
(72). Hydrogenation generally does not destroy tetrodotoxin, a powerful toxin from the puffer
their toxicities, although hydrogenation of saxi- fish (61, 73, 102). However, Gentile (45) notes
toxin does destroy its toxicity (142). that Aphanizomenon toxin blocks calcium-de-
Aphanizomenon toxin was originally classi- pendent action potentials, which are found in
fied as another VFDF, even though it differed certain invertebrate muscles, but tetrodotoxin
chemically from the Anabaena toxin, because it does not.
also acts rapidly (137). When injected i.p., the
toxins usually act within 1 to 2 min, and the Toxic Marine Cyanophycophyta
latent period is seldom more than 5 min (47, As a result of the readily observable deaths of
137). The minimum lethal dose for the Aphan- waterfowl, cattle, and fish, the toxicity of fresh-
izomenon toxin has been reported by Gentile water blue-green algae has received more atten-
(46) to be from 0.05 to 0.10 mg/kg in mice. It is tion then the toxicity of marine blue-greens.
significant to note that this minimum lethal dose However, it seems well documented that the
is less than that of either Microcystis toxin or marine forms produce toxic compounds. The
Anabaena toxin (47, 137). well-documented cases of marine blue-green-al-
The symptoms and cause of death, when in- gal toxins have all been found in the family
- - -- Longitudinal sulcus
. - -- Flagellum
--.-----
Hypotheca
Flagellum
Most of toxic dinoflagellates are marine orga- about 1,500,000 cells per ml, caused fish mortal-
nisms, and they are generally within the range ity in 15 to 25 min (85). The culture supernatant
of 40 to 60 Sn in diameter. Blooms are usually from A. carteri was also found to be ichthyotoxic
dependent upon an annual cycle of environmen- (65). Ikawa and Taylor (65) hypothesized that
tal parameters to produce the right temperature, the toxin was choline-like, and, consequently,
salinity, illumination, pH, and concentrations of they assayed A. carteri cells and found that 0.36
trace elements and growth factors. Generally, to 0.41% of the dry weight was choline chloride
toxic species are found greater than 300 latitude equivalent. This assay included pharmacologi-
(139). cally inactive forms of choline as well as phar-
Recently, there has been work to substantiate macologically active forms. For this reason, they
the hypothesis that ciguatera (a type of fish assayed Amphidinium cells which were proc-
poisoning) is due to a toxic dinoflagellate (95). A essed by a different method to find the fraction
single investigation has concluded that Diplos- of the total amount of choline-like molecules
alis sp. nov. is very likely to be the cause of that were in the water-insoluble, inactive form.
ciguatera (189). It was found that 0.07 to 0.10% of the dry weight
was in the inactive form of cholines, which
Peridinium polonicum means that 0.26 to 0.34% of the dry weight was
There has been evidence to incriminate the in the active form of the choline-like compounds.
By combining the use of assay procedures and
Aluterus schoepfi ............ Orange filefish Mycteroperca bonaci ......... Black grouper
Anchoa mitcheUi ............ Bay anchovy Myrophis punctatus ... Speckled worm eel
Archosargus probatocephalus Sheepshead Mystriophis intertinctus Spotted spoon-nose eel
Arius felis Sea catfish Ogcocephalus vespertilio .... Longnose batfish
Astroscopus y-graccum Southem stargazer Oligoplites saurus .. ...... Leatherjacket
Bagre marina ................ Gaff-topsail catfish Ophichthys gomesi ....... Shrimp eel
BairdieUa chrysura ........... Silver perch Ophichthys ocellatus .... Pale-spotted eel
Balistes capriscus ............ Gray tiggerfish Ophidion holbrooki Bank cusk-eel
Balistes vetula ............... Queen triggerfish Opisthonema oglinum ........ Atlantic thread herring
Bascanichthys teres .......... Sooty eel Opsanus tau Oyster toadfish
Brevoortia patronus .......... Gulf menhaden Orthopristis chrysopterus .... Pigfish
Calamusp Porgy family Paralichthys albiguttus Gulf flounder
.......
Caranx crysos ................ Blue runner Peprilus alepidotus .......... Harvest fish
Caranx hippos ............... Crevalle jack Peprilus burti....... Gulf butterfish
Centropomus undecimalis ..... Snook Peprilus triacanthus ... Butterfish
Chaetodipterus faber ......... Atlantic spadefish Pogonias cromis ..
.. .. Black drum
Chilomycterus schoepfi ........ Striped burrfish Pomacanthus arcuatus Gray angelfish
Menidia beryUina ........... Tidewater silverside Limulus polyphemus ......... Horseshoe crab
.
nature of the toxic compound have combined to dated because of its instability. Kim and Padilla
yield a variety of results. The recent use of high- (77) have found that the original toxin is broken
pressure liquid chromatography for the analysis down to a more stable toxic component, as well
of biological compounds (24, 180) has led to the as to other stable but nontoxic components. The
conclusion that there is actually only a single purified toxin was found to be unstable when
ichthyotoxic component (77). The chemical stored in the dark at 4°C in the dry state. G.
structure of this component has not been eluci- Padilla believes that the toxic molecule has a
738 COLLINS MICROBIOL. REV.
molecular weight of approximately 400 and that The hemolytic fraction of the toxin does not
there is no sulfur or phosphorus in the toxin and exhibit any lethal activity (2).
very little nitrogen, if any (personal conversa-
tion). G. breve produces a separate hemolytic Gonyaulax spp.
component which has been separated from the There are six different species of the marine
ichthyotoxic component (78, 109, 115). dinoflagellate Gonyaulax which have been doc-
The physiological action of the G. breve toxin umented as toxic species. The three species G.
is as much of an enigma as the chemical struc- catenella, G. acatenella, and G. tamarensis
ture of this compound. The dominant hypothesis have been shown to cause paralytic shellfish
as to the mode of action of G. breve toxin is that poisoning, whereas the species G. monilata and
it acts as a depolarizing agent, similar to the G. polygramma have been related to ichthy-
toxin from G. veneficum (1, 164). Depolarizing otoxic incidents. G. catenella is found on the
agents block nerve transmission and are respon- northern Pacific coasts of North America and
sible for decreasing or eliminating the resting Japan, G. acatenella is found on the Pacific
membrane potentials. These agents can also re- coast of North America, and G. tamarensis is
duce the abilities of the nerves and muscles to found off the North American Atlantic coast and
transmit action potentials. G. breve toxin has in the North Sea. G. polygramma has been
been responsible for reducing the resting mem- associated with red tides and has caused fish
brane potentials and the amplitudes of action toxicity off Massachusetts and oyster deaths off
phon. In the siphon, the rate of toxin release or On a cellular level, the physiological action of
destruction is low, so these clams may remain Gonyaulax toxins is the blocking of sodium mol-
toxic for 1 year or more after the environment is ecules that would passively cross excited mem-
cleared of the Gonyaulax (139). branes, essentially preventing action potentials.
Most chemical isolations have been done on The toxins have a negligible effect on potassium
G. catenella and G. tamarensis. It was found and chloride permeabilities (49, 101, 102). This
that the poisons extracted from clams, mussels, effectively blocks axonal and muscular conduc-
and G. catenella were chemically approximately tion while having a minimal effect on the neu-
the same (143). G. tamarensis is considered a romuscular synapse (74). This type of block
more severe practical problem, because the red occurs without depolarization.
tides of this alga occur more frequently and The pharmacological activity of the Gonyau-
cover a larger area than the tides of G. catenella lax toxins includes potent central and peripheral
(48). Recent work with thin-layer and high-pres- neurotoxic actions. Central nervous system ac-
sure liquid chromatography and electrophoresis tions include cardiovascular and respiratory ef-
has indicated that the toxic constituents of these fects, whereas peripheral nervous system actions
two algal species are relatively similar (105). are effects at the neuromuscular synapses and
There have been seven separate toxins isolated sensory nerve endings (151). Schantz (139) de-
from G. tamarensis: saxitoxin, gonyautoxin 1, scribes the human symptoms of paralytic shell-
gonyautoxin 2, gonyautoxin 3, gonyautoxin 4, fish poisoning as follows.
CH20H CH20OSO3
O I 0 0
`\O~
0 OH OH oSo3 i n
_
A
CH20H
I
In