Marine Biology (2003) 143: 1013–1027
DOI 10.1007/s00227-003-1147-z
J. I. Carreto Æ N. G. Montoya Æ H. R. Benavides
R. Guerrero Æ M. O. Carignan
Characterization of spring phytoplankton communities
in the Rı́o de La Plata maritime front using pigment
signatures and cell microscopy
Received: 20 November 2002 / Accepted: 14 May 2003 / Published online: 16 July 2003

Springer-Verlag 2003
Abstract The surface abundance and species composi-
tion of phytoplankton communities were studied in a
section across the continental shelf between the Rı́o de
La Plata and the oceanic waters of the Subtropical
Convergence, during late spring (November 1999). Al-
gal communities were examined using light microscopy
and HPLC-derived (high-performance liquid chroma-
tography) pigment concentrations. The CHEMTAX
program was used to estimate the chlorophyll a (chl a)
biomass of different algal classes. The inclusion of the
most abundant members of the chl c pigment family
(chl c1, chl c2, chl c3 and chl c2 monogalactosyldiacyl-
glyceride esters) in the pigment matrix improved the
CHEMTAX interpretation of field data. Using this
novel approach four haptophyte populations were
distinguished across the studied section, even though
they had qualitatively similar pigment signatures,
although one subtype lacked 19¢-hexanoyloxyfucoxan-
thin (Hex-Fuco). Five different phytoplankton assem-
blages, spatially segregated by the prevailing
environmental conditions, were distinguished during
the studied period. All of them showed a complex
community structure, formed by a background of
small-sized cells such as cyanobacteria, cryptophytes,
haptophytes and prasinophyceans, on which diatom,
cryptophyte or some haptophyte blooms were over-
lapped. In the estuarine assemblage, where maximum
chl a concentrations where found, diatoms were always
the dominant group (30–60% of total chl a), but
cryptophytes (10–40%), prasinophyceans (2–20%) and
dinoflagellates (2–12%) were also relevant. In the
coastal assemblage diatoms were also the dominant
group (35–45%), but haptophytes lacking Hex-Fuco
Communicated by O. Kinne, Oldendorf/Luhe
J. I. Carreto (&) Æ N. G. Montoya Æ H. R. Benavides
R. Guerrero Æ M. O. Carignan
Instituto Nacional de lnvestigación y Desarrollo Pesquero
(INIDEP), Paseo Victoria Ocampo no. 1,
7600 Mar del Plata, Argentina
E-mail: jcarreto@inidep.edu.ar
were subdominant (20–35%). The continental shelf
assemblage showed an almost exclusive dominance
(90%) of haptophytes resembling the coccolithophorid
E. huxleyi. Another haptophyte (Phaeocystis sp.) was
dominant (75–85%) in the Malvinas Current assem-
blage. The Brazil Current assemblage was characterized
by the codominance of cyanobacteria (45%) and ha-
ptophytes (35%). These results are discussed in rela-
tionship to the complex hydrographic features of the
area.
Introduction
The Rı́o de La Plata drains the second largest basin of
South America. It flows into the Atlantic Ocean with a
total discharge of 20,000–25,000 m3 s)1 (Framiñan and
Brown 1996). The main characteristics of the Rı́o de La
Plata estuary are its large spatial scale and the occur-
rence of a quasi-permanent salt wedge regime gener-
ating a border system (Carreto et al. 1986; Guerrero
et al. 1997; Mianzan et al. 2001). Tidal currents are
typically below 45 cm s)1 (Framiñan et al. 1999), and
the estuary therefore becomes a typically two-layer
system with strong vertical stratification that gradually
weakens seaward (Mianzan et al. 2001). Although the
main flow of the Rı́o de La Plata is expected to have a
NNE direction along the Uruguayan coastline, its tra-
jectory, forced by the dominant winds, fluctuated sea-
sonally. Its influence may be detected, in spring and
summer, in a southward direction along the coastal
region of Argentina (Guerrero et al. 1997) and/or along
the mild-shelf regime (Carreto et al. 1995). Occasionally
its area of influence may extend to the continental
slope, where these estuarine waters contact the warm,
subtropical Brazil Current water (Lusquiños and Val-
des 1971; Provost et al. 1995), and the nutrient-rich
waters of the Malvinas flow (Negri et al. 1992), or it
may be found between both currents. These waters can
be carried offshore by the return flux of the Malvinas
1014
Current (Provost et al. 1995) in an area referred to as
the Subtropical Confluence. The time and intensity of
the occurrence of these events may be dependent on the
cyclical excursions of the main currents along the
continental slope, on the volume discharge of the river
(Provost et al. 1995) and on the meteorological con-
ditions (Lusquiños and Valdes 1971; Carreto et al.
1986; Glorioso and Flather 1995; Guerrero et al. 1997).
The strong contrast in various ocean properties
(e.g. salinity, sea surface temperature and nutrient
concentration), together with complex mesoscale
patterns, make this area an important biogeographic
boundary between phytoplankton communities of
brackish and oceanic waters of subtropical and sub-
Antarctic origin. However, relatively few phytoplank-
ton studies have been carried out in this area since
Hentschel’s (1932) early data. Phytoplankton taxonomy
and biogeography of dinoflagellates, diatoms and sili-
coflagellates have been previously studied by Balech
(1949, 1971, 1986), by Frenguelli and Orlando (1959),
Müller-Melchers (1959) and Lange (1985), respectively.
Negri et al. (1988) and Bayssé et al. (1986) have studied
relative phytoplankton abundance and community
structure of net samples and their relationship with
environmental factors. Gayoso (1995) and Gayoso and
Podestá (1996) reported on the phytoplankton abun-
dance, although cells <5 lm in diameter were not in-
cluded in their analyses. The scenario resulting from
these previous studies displayed a region where robust
diatoms, armored dinoflagellates, calcareous cocco-
lithoporids and Phaeocystis (Prymnesiophyceae) colo-
nies are the most frequent forms. The recognition of
the importance of delicate flagellates and minute coc-
coid forms has come only recently, after more appro-
priate techniques for studying the picoplankton were
applied in this area (Silva and Negri 2000).
Picoplankton and many of the smaller nanoplankton
cells are difficult to study. Their small size and the ab-
sence of clearly discernible features often preclude
identification by light microscopy, even to the class level,
and many are too fragile to be adequately preserved
(Jeffrey and Vesk 1997). Fluorescence microscopy and
flow cytometry have proven to be invaluable for the
enumeration of cyanobacteria and prochlorophytes, but
the identification of the less distinctive cells remains a
problem.
The characteristic pigment composition of phyto-
plankton groups and the development of modern of
high-performance liquid chromatography (HPLC)
methods has favored the present use of pigment analysis
in phytoplankton community research (Gieskes and
Kraay 1983; Bidigare et al. 1990; Wright et al. 1996;
Schlüter et al. 2000). Pigment chemo-taxonomy has been
especially useful in samples from oceanic areas (Veldhuis
and Kraay 1990; Letelier et al. 1993; Peeken 1997), and
proved to be valuable for the characterization of small-
sized phytoplankton in coastal marine areas (Schlüter
et al. 2000) and estuaries (Roy et al. 1996). The signature
role of the pigments has been summarized in various
papers, although the interpretation of HPLC field data is
not always a straightforward task (see Jeffrey et al. 1997).
Recent developments in HPLC technology using C18
polymeric and C8 monomeric HPLC columns, and
pyridine as modifier (Garrido and Zapata 1997; Jeffrey et
al. 1999; Zapata et al. 2000), have uncovered additional
chlorophyll c (chl c) pigments and acyloxyfucoxanthins
(Garrido and Zapata 1998; Egeland et al. 2000; Garrido
et al. 2000; Zapata et al. 2001). The improved resolution
of HPLC methodology and the potential utility of these
pigments as biological markers (Zapata et al. 2001) could
facilitate the interpretation of field data.
Several possibilities to estimate quantitatively the
various phytoplankton groups from marked pigment
concentrations have been developed (Gieskes et al.
1988; Everitt et al. 1990; Letelier et al. 1993). A po-
tential source of errors in these approaches was that
pigment ratios might change as a consequence of long-
term changes in light climate and nutrient availability
(Goericke and Montoya 1998; Mackey et al. 1998; van
Leeuwe and Stefels 1998). Mackey et al. (1996) intro-
duced the CHEMTAX program to obtain biomass
estimations of algal groups that share common
pigments, taking into account the physiological and
genetic variation in pigment ratios. Nevertheless, inter-
species (Jeffrey and Vesk 1997; Zapata et al. 2001) and
inter-clones (Stolte et al. 2000) variation in pigment:chl
a ratios, was also considerable. These findings indicated
that the ratios chosen for the applications of the
CHEMTAX calculations should, if at all possible,
reflect the dominant phytoplankton species present in
the sample (Schlüter et al. 2000).
In the present work, the structure of surface phyto-
plankton communities was studied during November
1999 in a section across the Argentine continental shelf
between the Rı́o de la Plata and the oceanic waters of the
Subtropical Convergence. Microscopic enumeration of
phytoplankton species, HPLC-derived pigment concen-
trations and the CHEMTAX software were used for the
first time in the area to evaluate the distribution and
abundance of the different phytoplankton groups. The
spatial distribution of the different phytoplankton
communities and their dominant species were analyzed
in relation to the complex environmental features of the
region.
Materials and methods
Sample collection
Sampling procedures were performed from 6 to 11 November 1999,
on board the R.V. ‘‘Cap. E. Canepa’’ (Fig. 1a). Vertical profiles for
temperature, salinity and fluorescence were obtained with a CTD-
SBE 911 equipped with a Seapoint fluorometer. CTD data and in
vivo fluorescence measurements were calibrated and reduced to
1-m bins. Light penetration was measured with a PUV 500 (Bio-
spherical Instruments) underwater radiometer. Surface water
samples (0 m depth) were collected with a clean plastic bucket, and
discrete sampling of the water column (5 m depth) for CTD cali-
bration was performed with a Niskin bottle.

Fig. 1 a AVHRR sea surface temperature mapped image from 10
November 1999, showing location of occupied stations (MC
Malvinas Current; BC Brazil Current; RP Rı́o de La Plata).
b SeaWiFS level 3 standard mapped image for chlorophyll, 6–12
November 1999 composite
Remote data on SST and ocean color
Sea surface temperature (SST) field was derived from data collected
by the Advanced Very High Resolution Radiometer (AVHRR)
installed on the NOAA 11 polar satellite and processed by the
Argentine National Space Research Commission (CONAE) using a
multi-channel algorithm. Chlorophyll surface field was derived
from data collected by SeaWiFS installed on the Orb View-2 polar
satellite and processed to level 3 (standard mapped image) by
CONAE.
Microscopic analysis
Surface water subsamples (0 m depth) for phytoplankton identifi-
cation and cell counts were obtained from five selected stations and
fixed with neutral Lugol’s iodine. Sample volumes of 50 and 100 ml
were settled. Phytoplankton species were identified and enumerated
with an Olympus IX-70 inverted microscope using bright-field
optics and ·200, ·400, ·600 and ·1,000 (oil) magnifications. Algal
cells were identified to species level when possible, except for the
1015
small and fragile ones for which only the genus or alga group was
recorded. Unicellular cyanobacteria were observed by epi-fluores-
cence microscopy, and could only be reliably quantified in the clear
water of the Brazil Current.
Pigment analysis
Surface seawater samples of 2–4 l (0 m depth) were filtered through
Whatman GF/F glass fiber filters. Filters were deep-frozen imme-
diately and stored in liquid nitrogen to prevent pigment modifica-
tion before the analysis. Pigments were extracted with 100%
methanol and sonicated (Vibra Cell, Sonic and Materials) at 0C.
The extracts were filtered through GF/F filters to remove cell
debris. Water was added to the extract immediately before injection
to obtain an 80% methanol dilution (Jeffrey et al. 1997). Sample
solution aliquots were automatically injected into an HPLC system
Shimadzu LC 10 AC. For pigment elution we use the method of
Garrido and Zapata (1997), slightly modified. A C18 Vydac 201
TP 54, 250·4.6 mm i.d. column was used, protected with an Opti-
Guard C18 guard column. The mixing chamber and column were
thermostated at 27C with a CTO-10 AC (Shimadzu) column oven.
Eluent A was a 45:35:20 mixture of methanol, acetonitrile and
aqueous solution (0.25 M pyridine, pH adjusted to 5.3 with acetic
acid), and eluent B was always acetone. Peak detection was carried
out using a diode array detector (SPD-M10Avp) and a spectro-
fluorometer (FR-10Axl). Pigments were identified by their reten-
tion time, and absorption spectra were obtained from the on-line
diode array detector. High purity standards were provided by VKI
(The International Agency for 14C Determination, Denmark) or
isolated from cultures of Emiliania huxleyi (clone CCMP370) and
Alexandrium tamarense (clone MDQ1096). The HPLC system was
calibrated using genuine standards from VKI or the extinction
coefficients reported by Jeffrey et al. (1997) for prepared standards.
Novel pigments were quantified considering the extinction coeffi-
cients for the most similar documented chromophore (e.g. chl c3
and non-polar chl c was measured as chl c2 equivalents); 4-keto-
hexanoyloxyfucoxanthin (4-k-Hex-Fuco) and fucoxanthin (Fuco)
could not be well resolved, probably because of the high volume
injection employed. Nevertheless, the diode array of the spectros-
copy analysis indicated that Fuco was the dominant pigment in all
samples. Unfortunately, the major carotenoid of Chlorophyceae,
lutein (Lut), coeluted with diatoxanthin (Diato), and they were not
quantified.
Data processing
The chl a of phytoplankton groups at each sampled station were
calculated using CHEMTAX, a matrix factorization program
running under MATLAB (Mackey et al. 1996). All diagnostic
pigments detected, as well as those ambiguous markers indicative
of only a few groups, were included in the program MATRIX.
Chl c1, chl c2, chl c3 and chl c2 monogalactosyldiacylglyceride es-
ters were also included, as the HPLC method used is very useful to
separate the members of this pigment family. Prochlophytes were
not included in the data matrix, as divinyl chl a and b could not be
separated from monovinyl chl a and b with this method. The light-
protecting carotenoids of the xanthophyll cycle, diadinoxanthin
(Diadino) and diatoxanthin (Diato), b,-carotene (b,-Car), b,b-
carotene (b,b-Car) and other pigments only detected in trace
amounts were not included (Mackey et al. 1996).
A matrix using initial pigment:chl a ratios derived from the
literature was constructed (Wright et al. 1996; Jeffrey and Vesk
1997; Peeken 1997; Mackey et al. 1998; Schlüter et al. 2000; Stolte
et al. 2000; Zapata et al. 2001), since data on pigment ratios for the
dominant phytoplankton species in the area were not available.
Although the original data must be previously split into homoge-
neous groups, the samples were all analyzed together since the
small number of samples adversely affected the accuracy of the
CHEMTAX calculations (Mackey et al. 1996). However, four
1016

categories were considered in our calculations because of the high

Table 1 Pigment to chlorophyll a ratio matrix after fitting by the CHEMTAX program used in the calculation of algal group compositions (E1, Emiliania; C2, Chrysochromulina; for

Chl a

1.000
1.000
1.000
1.000
1.000
1.000
1.000
1.000
1.000
1.000
1.000
complexity observed within the Haptophyta, having nine recog-
nized subgroups (Jeffrey and Vesk 1997; Jeffrey and Anderson
2000). The inclusion of four categories accommodated the natural

non-polarchl c
split of the haptophytes between the different ecosystems studied.

Chl b Chl c2–MGDG Chl c2–MGDG Unknown

These categories were delineated on the basis of: (1) identification
of pigment assemblages and their relation to the hydrography and

0.000
0.000
0.000
0.000
0.075
0.000
0.000
0.000
0.000
0.000
0.000
(2) the microscopic analysis of representative samples from each
pigment assemblage. The taxonomic groups considered and the
pigment ratios obtained from CHEMTAX calculations are shown
in Table 1.

C2-type
Results

0.000
0.000
0.000
0.000
0.000
0.188
0.000
0.000
0.000
0.000
0.000
Hydrography

The satellite-derived SST data on 10 November (Fig. 1a)

provided useful scenery to understand some of the main

E1-type

0.000
0.000
0.000
0.082
0.000
0.000
0.000
0.000
0.000
0.000
0.000
hydrographic features of the area. The Brazil Current
separated from the shelf-break at 36.5S, where it
encountered a narrow filament of cooler waters flowing

0.000
0.000
0.000
0.000
0.000
0.000
0.000
0.230
1.156
0.000
0.000
along the upper shelf-break between 39S and the lati-
tude of the Brazil Current separation. The Malvinas

0.000
0.000
0.000
0.000
0.000
0.000
0.000
0.010
0.000
0.000
1.115
Current separation is not spatially coincident with the

Zea
separation of the Brazil Current, showing coherence

0.000
0.000
0.000
0.000
0.000
0.000
0.700
0.000
0.000
0.000
0.000
with the results of Olson et al. (1988), who observed a

Chl c2 Viola Allo

difference of 3 in latitude in the separation of both
currents. A band of intermediate temperatures occupies

0.000
0.000
0.000
0.000
0.000
0.000
0.000
0.099
0.061
0.000
0.000
the area between the two strong thermal fronts. This
transition zone seems to be filled by an eddy of sub-

0.208
0.320
0.247
0.220
0.282
0.220
0.174
0.000
0.000
0.470
0.000
tropical waters wrapped between the cold filament
flowing along the upper shelf-break and the southward
flow of pure Brazil waters. The contact between the 0.000
0.000
0.000
0.000
0.000
0.000
0.000
0.000
0.372
0.000
0.000
Chl c1 Chl c3 Pras

Malvinas and Brazil Currents occurred south of the

anticyclonic eddy, where a filament of pure Malvinas
waters turns offshore. On the shelf, the satellite-derived
0.000
0.000
0.296
0.230
0.211
0.240
0.000
0.000
0.000
0.240
0.000
SST data showed the occurrence of two well-defined
thermal fronts: (1) a shelf-break front separating shelf
0.178
0.000
0.000
0.000
0.000
0.000
0.000
0.000
0.000
0.000
0.000
and Malvinas waters (up to 3830¢S) and (2) a front
between coastal waters and Rı́o de La Plata outflow
waters.
0.000
0.000
0.000
0.000
0.000
0.000
0.000
0.060
0.062
0.000
0.000
But-fuco Perid Fuco Hex-fuco Neo

The temperature and salinity diagram for occupied

CTD stations, as shown in Fig. 2, reveals five different
water masses: Rı́o de la Plata estuarine (A), coastal (B),
0.000
0.000
0.000
1.000
0.740
0.714
0.000
0.000
0.000
0.000
0.000
0.863
0.000
0.465
0.100
0.480
0.354
0.000
0.000
0.000
0.166
0.000
pigment abbreviations see Fig. 6 legend)

0.000
0.800
0.000
0.000
0.000
0.000
0.000
0.000
0.000
0.000
0.000
0.000
0.000
0.000
0.020
0.050
0.000
0.000
0.000
0.000
1.330
0.000
Prasinophyceans
Haptophytes D

Chlorophyceans
Haptophytes C
Haptophytes B

Haptophytes E

Pelagophyceans
Dinoflagellates

Cyanobacteria
Cryptophytes

Fig. 2 Temperature–salinity and density (rt) distribution for

Diatoms

occupied CTD stations. Five water masses are identified: Rı́o de

la Plata estuarine (plus sign, A), coastal (circle, B), continental shelf
(triangle, C), upper Malvinas (square, D) and Brazil (cross, E)

continental shelf (C), upper Malvinas waters (D) and
Brazil (E). Rı́o de la Plata estuarine waters were char-
acterized by a salinity between 0 and 29 psu and tem-
peratures above 16C. This stratified system is
associated with the head of the estuary (stations 1–6),
where a layer of highly diluted water overlies salty
coastal water. Vertical mixing of estuarine and conti-
nental shelf waters forms this coastal water (stations 7–
11). The influence of the Rı́o de la Plata extends offshore
up to station 13 (280 km away for the head), capping
continental shelf waters, with coastal water of 30 psu
(upper 15 m). Continental shelf waters are characterized
by a salinity between 33.3 and 33.6 psu and tempera-
tures ranging from 13C to 9.4C; these waters are
constrained to depths below 20 m (stations 12–13).
Upper Malvinas waters, lying over the continental shelf
break (stations 14–15), are characterized by a salinity of
33.8 psu and a temperature range from 6.3C to 10.3C.
Brazil waters have a salinity >34.5 psu and tempera-
tures >16C (stations 16–19). At some stations of the
Brazil waters and throughout the anticyclonic eddy de-
scribed above, a variable layer of low-salinity water
coming from the continental shelf system capped the
surface waters.
Structure of phytoplankton communities
Distribution of chl a
The SeaWiFS (Fig. 1b) imagery provided valuable
information on the overall distribution of near-surface
phytoplankton biomass in the region and its relation
with hydrography. The lowest chl a values were ob-
served in both the warm water of the Brazil Current,
where phytoplankton is nutrient limited, and in the
nutrient rich but highly turbulent waters of Malvinas
Current. In the transition zone between these boundary
currents, the chlorophyll concentration was enhanced,
especially near the shelf-break, where filaments of high
chlorophyll concentrations were found (Gayoso and
Podestá 1996; Brandini et al. 2000). These filaments
appear to be related to a band of high chlorophyll
concentrations present along the west side of the frontal
region separating the Malvinas Current from shelf wa-
ters (Carreto et al. 1986, 1995). Except for this frontal
region, the over-shelf chlorophyll concentration was
slight and similar to that observed in the transition zone.
Another noteworthy region was the Rı́o de La Plata
estuary, where an extremely high chlorophyll signal was
found. However, chl a surface measured ‘‘in situ’’ of the
estuary stations (1–6) was much lower than that ob-
tained from SeaWiFS (Fig. 3), and did not vary signifi-
cantly with depth, as observed in the vertical profiles of
station 2, representative of this sector (Fig. 4a). There-
fore, the optical signal produced by the dissolved or-
ganic matter and suspended sediments in the area was
likely to be stronger than the phytoplankton signal
1017
Fig. 3 Chlorophyll a concentration (lg l)1) at the surface (0 m
depth) of occupied stations along the section, as determined by
HPLC
(Framiñan and Brown 1996). Chl a surface concentra-
tion was low in the coastal area near the estuary (sta-
tions 7–11) (Fig. 3), but increased gradually with depth
(Fig. 4b). Except for the estuarine area, the highest
surface chlorophyll concentration (2.14 lg l)1) was
measured in the over-shelf stations 12 and 13, where
5.0 lg l)1 maximum was observed at 10–15 m depth,
at the base of the mixed layer (Fig. 4c). The high chl a
values in the photic zone of these stations appeared to be
related to the previously mentioned high chl a concen-
tration along the west side of the shelf-break front re-
trieved by the SeaWiFS imagery. In agreement with
SeaWiFS data, surface chl a concentrations measured in
the domain of Malvinas waters near the shelf-break
(stations 14–15) were lower than those in the frontal
region (Fig. 3). Vertical distribution was rather homo-
geneous at these stations (Fig. 4d). In contrast, the
minimum surface chl a values (Fig. 3) measured ‘‘in
situ’’ in the oceanic region dominated by the subtropical
water of the Brazil Current (stations 16–19) were
accompanied by a maximum (1.8 lg l)1) at the base of
the photic zone (Fig. 4e).
Pigment profiles and microscopy data
Table 2 lists the microalgal pigments detected in
increasing elution order, spectral characteristics of pig-
ment in the mobile phase are also included. The chro-
matograms obtained (Fig. 5) from five selected stations
showed the elution patterns of a range of chlorophyll
and carotenoid pigments characteristic of the phyto-
plankton communities that prevail in the different
studied ecosystems. The species composition of the
phytoplankton communities estimated by cell counts, in
these selected stations are presented in Table 3.
Phytoplankton communities
Three main phytoplankton assemblages, closely related
to the hydrography of the area, were differentiated on
the basis of the dominant carotenoid: (1) estuarine and
coastal communities, (2) continental shelf and Malvinas
communities and (3) the Brazil Current community
(Fig. 6a).
1018
Fig. 4 Vertical profiles of temperature (C), salinity (psu) and
chlorophyll a (lg l)1), from five selected representative stations of:
a estuarine (station 2), b coastal (station 7), c continental shelf
(station 12), d Malvinas Current (station 15) and e Brazil Current
waters (station 16). Data derived from continuous CTD-fluoro-
meter readings
Estuarine and coastal communities
Fuco was the dominant carotenoid in the Rı́o de La
Plata estuarine and coastal area. Microscopic analysis
(Table 3) confirmed that several diatom species were
dominant in this phytoplankton assemblage, explaining
the observed high relative Fuco concentration. This
community was separated into estuarine (A) and
coastal (B) assemblages on the basis of pigment com-
plements and microscopy data.
The relative concentration of the unequivocal
Cryptophyceae marker alloxanthin (Allo) was high in
the estuarine sector, but sharply dropped in coastal
waters (Fig. 6b). Microscopy showed that free-living
cryptophytes (Cryptomonas sp.) were more abundant in
estuarine than in coastal waters, explaining the observed
distribution of Allo. Pigments associated with the green
algae (Chlorophyceae and Prasinophyceae) also revealed
two different phytoplankton assemblages (Fig. 6b). The
amount of chl b with respect to chl a was higher in
assemblage B. Prasinoxanthin (Pras), an unequivocal
marker for some prasinophyceans (Micromonadophy-
ceae), was also found in high proportions in this
assemblage, strongly correlated with the chl b concen-
tration. Consequently, most chl b arises from prasino-
phyceans. Microscopy data were coincident with
pigment evidence, as the prasinophyceans Micromonas
sp. and Nephroselmis sp. were frequent in assemblage B,
but were absent in assemblage A. The relative low
abundance of Pras in assemblage A suggests that chl b
arises from chlorophyceans or prasinophyceans lacking
Pras. Unfortunately, Lut, the major carotenoid of
Chlorophyceae, coeluted with Diato and was not
quantified. The microscopic examination of these sam-
ples showed the prasinophyte Pyramimonas sp. as the
most abundant species, indicating that most of the chl b
arises from prasinophyceans lacking Pras in assem-
blage A.
Except for the estuarine waters (Fig. 6c), the
unequivocal marker for dinoflagellates, peridinin (Pe-
rid), was in relatively low concentrations in the overall
area. The abundance of Perid in assemblage A appeared
to be related to some autotrophic dinoflagellates, espe-
cially Prorocentrum minimum and Scrippsiella trocoidea
(Table 3). Although dinoflagellate cell numbers were
also important in assemblage B, the phytoplankton
community showed a more diverse dinoflagellate
assemblage, including some heterotrophic or phago-
trophic species such as Noctilluca scintillans.
According to the dominant diatom flora, chl c3 was
detected only in trace amounts in assemblage A
(Fig. 6d), but a sharp increase in the chl c3:chl a ratio
was observed in B; however, Hex-Fuco was rarely
observed in this assemblage (Fig. 6a). Moreover, as in
assemblage A, diatoms prevailed and no haptophytes
were found by light microscopy. Some clones of the
diatoms Thalassionema nitzschioides and Rhizosolenia
setigera (identified in our samples) are known to con-
tain chl c3 (Stauber and Jeffrey 1988). However, as
diatom species composition was similar in both assem-
blages, the relative increase of chl c3 detected in
assemblage B was not related to a change in the diatom
flora. Another possibility was that some small or delicate
haptophytes with Fuco, such as the main carotenoid,
would have been present (Jeffrey and Vesk 1997). The
detection of chl c2–monogaloctosyldiacylglyceride ester
 1019

Table 2 Peak identification

table. Wavelengths given in Peak no. Pigment Retention Maxima in eluant
parentheses denote shoulders (min) (nm)
(MGDG monogalactosyldiacyl-
glyceride) 0 (Solvent front) 2.52
1 Peridininol 4.73 480
2 Chlorophyllide a 5.68 430 581 663
3 Peridinin 9.17 473
4 19¢-Butanoyloxyfucoxanthin 9.85 447 469
5 4-keto-19¢-Hexanoyloxyfucoxanthin 10.89 447 470
6 Fucoxanthin 11.11 449
6b MgDVP:Mg-38-divinyl-pheoporphyrin 11.15 439 577 628
a5 monomethyl estera
7 19¢-Hexanoyloxyfucoxanthin 12.03 447 469
8 9¢-cis-Neoxanthin 12.52 413 438 465
9 Chlorophyll c1 13.10 448 581 631
10 Monovinyl–chlorophyll c3 13.32 449 584 628
11 Chlorophyll c3 13.75 457 588 628
12 Prasinoxanthin 14.50 455
13 Chlorophyll c2 14.98 452 583 633
14 Violaxanthin 15.25 416 440 470
15 Dinoxanthin 15.76 418 441 470
16 Unknown carotenoid 17.69 (424) 444 471
17 Diadinoxanthin 18.87 (422) 447 476
18 Unknown carotenoid 20.52 (422) 444 470
19 Alloxanthin 21.90 (426) 452 482
20 Diatoxanthin 22.55 (426) 453 481
21 Zeaxanthin 23.20 (426) 453 478
22 Unknown chlorophyll c 23.29 454 586 633
23 Chlorophyll b 26.11 462 599 648
24 Crocoxanthin-like 27.36 (422) 447 476
25 Chlorophyll a allomer 29.32 430 615 664
26 Chlorophyll a 29.68 431 616 663
27 Chlorophyll a epimer 30.39 430 615 663
28 Unknown non-polar chlorophyll c 32.41 455 584 633
29 Chlorophyll c2–MGDG Emiliania-type 33.36 455 584 633
30 bb-Carotene 34.59 (426) 454 481
a 31 Chlorophyll c2–MGDG Chrysochromulyna-type 35.23 455 584 633
Quantified by fluorescence

from Emiliania huxleyi (chl c2–MGDG Emiliania-type,
Garrido et al. 2000) and chl c2–monogaloctosyldiacyl-
glyceride from Chrysochromulina polylepis (chl c2-
MGDG Chrysochromulina-type, Zapata et al. 2001) as
associated trace pigments (Fig. 5b) supported this
assumption.
Continental shelf and Malvinas communities
Hex-Fuco was the dominant carotenoid in continental
shelf and Malvinas waters. Herein, the relative abundance
of the associated pigments also indicated at least two
spatially segregated pigment associations (Fig. 5c, d).
The relative proportion of Fuco to Hex-Fuco in the
continental shelf assemblage (C) was lower than that
observed in the Malvinas assemblage (D) (Fig. 6a). In
addition, although the two assemblages presented sev-
eral non-polar types of chl c (Fig. 5c, d), a different
proportion of these compounds was evident (Fig. 6e).
In assemblage C the HPLC retention time of the main
compound (rt=33.36 min) was coincident with that of
the principal non-polar chl c from E. huxleyi (chl c2–
MGDG Emiliania-type) (Garrido et al. 2000). The
retention time of the major non-polar chl c in assem-
blage D was slightly slower (rt=32.41 min) and similar
to that observed for the minor non-polar chl c (un-
known non-polar chl c) from E. huxleyi (clo-
ne CCMP370). A compound with the same retention
time and spectral properties (Table 2) was also present
in assemblage C, although at a trace level. Also, trace
amounts of a pigment with a retention time
(rt=35.23 min) similar to that observed for chl c2–
MGDG Chrysochromulina-type (Zapata et al. 2001),
were present in both assemblages. Another unknown
chl c–like derivative of intermediate polarity
(rt=23.29 min) was observed only in assemblage D
(Fig. 5f).
Microscopy showed (Table 3) that haptophytes were
the dominant group in both phytoplankton assem-
blages. However, E. huxleyi was the dominant species
in assemblage C and Phaeocystis sp. was dominant in
assemblage D. Some other unidentified Coccosphaerales
species were also present in assemblage D. Microscopy
showed that some diatom species were present in low
abundances in both assemblages, although chl c1 was
not detected in our chromatograms (Fig. 5c, d).
Microscopy also showed that the Prasinophyceae
Micromonas sp. was present in both assemblages as a
minor component, explaining the small amounts of
chl b and Pras detected in these samples (Fig. 5c, d).
1020

Fig. 5 HPLC chromatograms

of pigment extracts from
surface samples (0 m depth) of
five selected representative
stations: a estuarine (station 2),
b coastal (station 7), c
continental shelf (station 12),
d and f Malvinas Current
(station 15) and e Brazil
Current waters (station 16).
Detection was by absorbance at
436 nm (a, b, c, d and e) and by
fluorescence at Ex440/Em650 nm
(f); peak identification as in
Table 2

Brazil Current community electron microscopy (Andersen et al. 1993). However, as

Zea was the principal carotenoid in the community
corresponding to the subtropical waters of the Brazil
Current. Microscopy showed that picoplanktonic cy-
anobacteria (Synechococcus spp.) were the most abun-
dant cells in the phytoplankton assemblage. The relative
proportions of chl c3, Fuco, But-Fuco and Hex-Fuco
were also high (Fig. 5e), indicating that haptophytes
and/or pelagophyceans were abundant. Chl c2–MGDG
Chrysochromulina-type (Zapata et al. 2001) was the main
non-polar chl c, although several non-polar types of
chl c, with similar retention times to those observed in
assemblages C and D, were present in trace levels
(Fig. 5e). The pigment profile of the dominant hapto-
phytes in assemblage E was different from that observed
for E. huxleyi (C) and Phaeocystis sp. populations (D).
Microscopy showed that, although E. huxleyi was also
present, some unidentified coccolithophorid species were
the dominant haptophytes in this assemblage (Table 3).
Pelagophyceans were not observed by light microscopy,
as these small cells can only be properly identified by
the ratio of But-Fuco to total fucoxanthins was signifi-
cantly higher than that observed for the other assem-
blages (Fig. 5e) pelagophyceans were expected to be
present. Despite the error arising from the quantification
of low pigment amounts associated with green algae, a
significant relation between chl b and Pras was observed.
On the other hand, Lut was only present in trace
amounts and then was not quantified. The evidence
indicated that, in this assemblage, chl b stemmed from
prasinophyceans containing Pras. However, the Prasin-
ophyceae lacking Pras, Pyramimonas sp., was the only
green alga observed by microscopy (Table 3). Another
uncertainty is related to the HPLC method, as divinyl
chl a and b could not be separated from monovinyl chl a
and b. Hence, the presence of picoplanktonic prochlo-
rophytes could not be totally excluded. The unequivocal
Cryptophyceae marker Allo was present in low amounts
in the samples from this assemblage (Fig. 6b), but this
group was not detected by microscopy; cryptophytes
probably could not withstand the fixation process
(Gieskes and Kraay 1983).
 1021

Table 3 Abundance (cells l)1) of

major species in various algal Phytoplankton assemblage Estuarine Coastal Cont. shelf Malvinas Brazil
groups at the surface (0 m Stn 2 Stn 7 Stn 12 Stn 15 Stn 16
depth) of five representative
stations (stations 2, 7, 12, 15, 16) CHROMOPHYTA
Bacillariophyceae
Thalassionema nitzschioides 21,600 5,820 900 – –
Actinocyclus ehrembergii 3,840 1,060 240 1,227 –
Thalassiosira decipiens 960 460 – – –
Thalassiosira sp. 320 – – – –
Coscinodiscus radiatus 80 – 20 – –
Pleurosigma normanii 80 80 – – –
Rhizosolenia setigera 80 – – – –
Paralia sulcata – 180 – –
Dinophyceae
Noctiluca scintillans – 40 100 – –
Prorocentrum minimum 720 – – – –
Prorocentrum compressum – 40 – – –
Protoperidinium conicum 480 60 – – –
Protoperidinium depressum – 20 – – –
Scrippsiella trocoidea 320 – 20 – –
Dinophysis caudata 80 – – –
Ceratium tripos – 180 380 – –
Ceratium fusus – 20 – – –
Ceratium candelabrum – 40 – –
Dictyochophyceae
Dictyocha fibula – – 51 – –
Prymnesiophyceae
Phaeocystis sp. – – 10,000 1,811,596 –
O. coccosphaerales – – – 50,307 18,750
Emiliania huxleyi – – 20,000 – 1,250
Cryptophyceae
Cryptomonas sp. 38,400 7,500 – – –
CHLOROPHYTA
Chlorophyceae
Scenedesmus sp. 320 – – – –
Prasinophyceae
Pyramimonas sp. 19200 – – – 6,250
Micromonas sp. – 7500 10,000 1,227 –
Nephroselmis sp. – 2500 – – –
CYANOPHYTA
O. Nostocales (n/quantified) Frequent Scarce – – –
O. chroococcales – – – – 768,750

CHEMTAX: abundance of algal groups
The total and relative contribution of the various groups
of phytoplankton to chl a, calculated by CHEMTAX
are shown in Figs. 7 and 8, respectively.
As expected from pigment results, diatoms were al-
ways the dominant group (30–60% of total chl a) in the
phytoplankton community of the estuary assemblage
(A), where a maximum chl a concentration was found.
In terms of chl a calculated by CHEMTAX, crypto-
phytes were the subdominant group (10–40% of total
chl a). In decreasing order, chlorophyceans (2–20% of
total chl a) and dinoflagellates (2–12% of total chl a)
were also relevant. The contribution of other groups
such as cyanobacteria (<10% of total chl a) and Pras-
containing prasinophyceans (<5% of total chl a) was
small.
Total phytoplankton biomass was very low in the
coastal assemblage (B), but the phytoplankton com-
munity was more diverse. The pigment proportions
detected with CHEMTAX showed that diatoms
prevailed (35–45% of total chl a), as in the estuary com-
munity. In terms of chl a calculated by CHEMTAX,
haptophyte B was the subdominant group (20–35% of
total chl a). Cyanobacteria (5–15% of total chl a) and
prasinophyceans (2–12% of total chl a) were also rel-
evant in decreasing order. The contribution of other
groups was small (chlorophyceans, prasinophyceans
lacking Pras, cryptophytes, and other haptophyte
types).
The algal group composition obtained by CHEM-
TAX for the continental shelf area (C) showed an
almost total dominance (90% of total chl a) of ha-
ptophytes C, probably represented by the coccolitho-
phorid E. huxley. The contribution of other groups was
very low, with cyanobacteria the most important of
these groups (about 5% of total chl a).
A high dominance (75–85% of total chl a) of ha-
ptophytes D was detected by CHEMTAX in the
Malvinas Current assemblage (D), mainly represented
by the flagellated cells of Phaeocystis sp. However,
some differences between the two stations (14 and 15)
1022
Fig. 6 Relative abundance of major pigments across the transect
(station 1–19) grouped in separate panels: a fucoxanthin (Fuco),
19¢-hexanoyloxyfucoxanthin (Hex-Fuco), 19¢-butanoyloxyfucoxan-
thin (But-Fuco), zeaxanthin (Zea); b alloxanthin (Allo), chloro-
phyll b (chl b), prasinoxanthin (Pras); c chlorophyllide a (Chlide
a), peridinin (Perid), phaeophytin a (Phytin a); d chlorophyll c1
(Chl c1), chlorophyll c3 (Chl c3), chlorophyll c2 (Chl c2); e chloro-
phyll c2–monogalactosyldiacylglyceride (Chl c2–MGDG) Emilia-
nia-type, chlorophyll c2–MGDG Chrysochromulina-type, unknown
non-polar chlorophyll c
can be expected after considering the subdominant
groups. Haptophyte E was the subdominant group
(15% of total chl a) at the oceanic station (15), whereas
diatoms were subdominant (5% of total chl a) on the
west side of the shelf-break (station 14). The contri-
bution of each of the other detected groups was <5%.
The algal group composition detected by CHEM-
TAX in the Brazil Current assemblage (E) showed the
codominance of cyanobacteria (45%) and haptophytes
E (35%). The contribution of prasinophyceans, cryp-
tophytes and pelagophyceans was similar, each group
representing about 10% of the total chl a.
Fig. 7 Concentration of chl a (lg l)1) in the different phytoplank-
ton groups across the transect (station 1–19) estimated by
interpretation of pigment HPLC data using the CHEMTAX
program, grouped in separate panels: a diatoms, cryptophytes,
dinoflagellates and chlorophyceans; b haptophytes B, hapto-
phytes C, haptophytes D and haptophytes E; c pelagophyceans,
cyanobacteria and prasinophyceans
Discussion
As in most studies, the lack of local known pigment ratios
was a limitation for the application of CHEMTAX
analysis. Nevertheless, our pigment ratio estimations are
consistent with those obtained by previous authors. The
CHEMTAX program assumes that each taxonomic cat-
egory has a consistent set of pigment ratios through all
samples. However, some changes in both, the pigment
ratios within a given algae species and the different species
composition of an algae group, would be expected along
the section. Our results showed that the inclusion in the
pigment matrix of the most abundant member of the chl c
pigment family (chl c1, chl c2, chl c3 and chl c2 monoga-
lactosyldiacylglyceride esters) improves CHEMTAX and
allowed us to distinguish between four haptophytes
populations across the studied section. The inclusion of
four categories accommodated the natural split of the
haptophytes between the different ecosystems studied.

Fig. 8 Percentage of different phytoplankton groups contributing
to the total concentration of chl a across the transect (station 1–
19), estimated by interpretation of pigment HPLC data using the
CHEMTAX program, grouped in separate panels: a diatoms,
cryptophytes, dinoflagellates and chlorophyceans; b haptophytes
B, haptophytes C, haptophytes D and haptophytes E; c pelago-
phyceans, cyanobacteria and prasinophyceans
Calculated pigment ratios
Haptophytes B
The coastal water haptophytes (B) showed a pigment
profile type, with Fuco, chl c3 and chl c2 as the prin-
cipal components, and chl c2–MGDG Emiliania-type
and chl c2–MGDG Chrysochromulina-type as tracer-
associated pigments (Garrido et al. 2000; Zapata et al.
2001). Our calculated Fuco:chl a ratio (0.465) was
similar to that presented by one Chrysochromulina
species (0.487; C. kappa) with the same pigment profile
(Zapata et al. 2001).
Haptophytes C
The continental shelf (C) haptophytes contained Hex-
Fuco as the main carotenoid. It is remarkable that
1023
acyloxyfucoxanthin ratios obtained in this study were
very similar to those calculated by Wright et al. (1996)
for haptophyte populations from the Southern Ocean,
resembling E. huxleyi (haptophytes N). In addition, the
chl c2–MGDG Emiliania-type to chl a ratio (0.08) was
similar (0.11–0.13) to that observed for E. huxleyi cul-
tures (clone CCMP370) by Zapata et al. (2001). They
were also in the range estimated for low- (0.13) and
high-light (0.08)-exposed cultures of E. huxleyi
(clone CCMP370) (Galmozzi et al. 2001). Although,
strain-dependent differences and phenotypic variation in
pigment ratios were found in several strains of E. hux-
leyi, the geographic distribution of pigment patterns
indicated that Fuco was the major carotenoid in Indian
and Pacific Ocean strains, whereas Hex-Fuco is more
abundant in all the Atlantic strains (Stolte et al. 2000).
Haptophytes D
The calculated acyloxyfucoxanthin ratios for Phaeocys-
tis sp. populations in Malvinas Current waters were
similar to those obtained by Wright et al. (1996) for
Phaeocystis antarctica (haptophytes S) populations in
the Southern Ocean. Hex-Fuco was the main carotenoid
in both cases, but our data showed that the Phaeocystis
sp. population exhibited a higher Hex-Fuco:chl a ratio
(0.74) than that observed for P. antarctica (0.51), but
similar to that reported by van Leeuwe and Stefels
(1998) for iron-limited cultures of an Antarctic Phaeo-
cystis sp. strain . In addition to the acyloxyfucoxanthin
ratios, haptophytes D differ from haptophytes C in the
nature of the main non-polar chl c (unknown non-polar
chl c). Another unknown chl c–like derivative of inter-
mediate polarity was also present in this population.
Haptophytes E
Our calculated acyloxyfucoxanthin ratios for the ha-
ptophytes growing in subtropical waters (coccolitho-
phorids different to E. huxleyi) were similar to those
observed for Phaeocystis sp. populations, but they dif-
fered between Phaeocystis sp. and E. huxleyi in the
prevailing non-polar chl c. The ratio of chl c2–MGDG
Chrysochromulina-type to chl a (0.176) was in the range
estimated (0.00–0.269) for several Chrysochromulina
species studied by Zapata et al. (2001). As there is no
information on the distribution of this non-polar chl c2
among coccolithophorids, the calculated ratio should be
treated with caution.
Diatoms
The Fuco:chl a ratio (0.86) was higher than that calcu-
lated by Wright et al. (1996) for diatom populations
from the Australian sector of the Southern Ocean (0.60),
but much lower than that calculated for shade-adapted
diatoms (1.25) by Letelier et al. (1993). A similar ratio
variation range (0.479–1.218) was found by Schlüter
1024
et al. (2000) for coastal species. Also chl c1:chl a and
chl c2:chl a ratios (respectively 0.178 and 0.208) were
both in the range observed by Jeffrey (1972) for several
diatom species.
Pelagophyceans
Our initial pigment ratios were taken from Mackey et al.
(1998), obtained from surface pelagophyceans of the
western equatorial Pacific (1.33 for But:chl a and 0.166
for Fuco:chl a). The program left these ratios un-
changed. Our values obtained for both the chl c3:chl a
(0.24) and the chl c2:chl a (0.47) ratios were similar to
those calculated by Jeffrey and Wright (1997) from one
strain of Pelagoccus subviridis (respectively 0.25 and
0.44).
Cyanobacteria
The Zea:chl a ratio of cyanobacteria was highly depen-
dent on irradiance (Kana et al.1988; Bigidare et al. 1989;
Lutz et al. 2001), being consistent with its photopro-
tective function. Although large errors can be expected
in the calculated pigment ratios, our value (1.12) was
largely comparable with averaged values of literature
results, obtained at high and low light intensities
[Schlüter et al. 2000 (1.24); Mackey et al. 1998 (1.15);
Kana et al. 1988 (1.20)]. Our value was also similar to
the ratios obtained at medium light intensities [Moore
et al. 1995 (0.7 at 350 lE m)2 s)1); Morel et al. 1993
(0.71 at 100 lE m)2 s)1); Lutz et al. 2001 (1.00 at 100 lE
m)2 s)1)] for several strains of marine cyanobacteria
(Synechococcus spp.).
Cryptophytes
Whereas the Allo:chl a ratio, calculated from oceanic
phytoplankton, was in the range of 0.105–0.240 (Gieskes
and Kraay 1983; Wright et al. 1996; Mackey et al. 1998;
Peeken 1997), values up to 1.58 were found in phyto-
plankton communities of the turbid lower St. Lawrence
estuary (Roy et al. 1996). On the other hand, our cal-
culated ratio was similar to the one obtained for a
coastal strain of Plagioselmis prolonga (0.682), growing
at medium light intensity (Schlüter at al. 2000).
Prasinophyceans and chlorophyceans
A high inter-species variability was found in pigment
ratios (Mackey et al. 1996), attending the complex tax-
onomy of prasinophyceans. Some of them produce
common green algal carotenoids, whereas some others
synthesize unusual carotenoids for this algal class
(micromonadophytes), Pras being the most important
among them (Egeland et al. 1995). Additionally, some
chlorophyceans and prasinophyceans grown at low light
intensity do not always contain the accessory pigments
Lut, Zea and its epoxides (Egeland et al. 1995; Mackey
et al. 1996). Consequently, pigment analysis does not
permit a clear separation of prasinophyceans from
chlorophyceans. We included two groups of green algae
in our samples: chlorophyceans and prasinophyceans
lacking Pras (chlorophyceans), and Prasinophyceans
containing Pras (prasinophyceans). Despite regional
differences, species composition and variation owing to
the algae physiological state, our values calculated for
both chl b:chl a (1.156) and Pras:chl a (0.372) ratios in
prasinophyceans coincide with literature values (Mackey
et al. 1996; Peeken 1997).
Algal assemblages and environmental conditions
Both pigment HPLC patterns and microscopy-derived
information showed five different phytoplankton assem-
blages in the region during spring 1999. These assem-
blages displayed a clear spatial segregation imposed by the
extant environmental conditions. All of them presented a
complex community structure, comprising small-sized
cells from cyanobacteria, cryptophytes, chlorophyceans,
haptophytes and prasinophyceans, on which diatoms,
cryptophytes, or some haptophytes overlapped.
Several diatom species were dominant in the phyto-
plankton assemblage (Table 3) of the nutrient-rich and
high-silicate estuary water (Carreto et al. 1986), where a
high chl a concentration was measured. The dominance
of larger diatom cells was coincident with most of the
previous reports for this area (Negri et al. 1988; Elge
et al. 1990; Mesones 1991). The Prasinophyceae Pyra-
mimonas sp. and the Cryptophyta Cryptomonas sp. were
also very frequent in this eutrophic environment; the
latter codominating in some cases with diatom flora.
Some bloom-forming dinoflagellate species (e.g. Proro-
centrum minimum) were also present, although in rela-
tively low concentrations, indicating that factors other
than nutrients regulate the dinoflagellate blooms in this
fluctuating environment.
Total phytoplankton biomass was very low, and a
more diverse phytoplankton community was observed in
the coastal system. The slightly stratified coastal water
presented both the lowest absolute (<1 lM) and rela-
tive (N:P:Si ratio: of 0.1:1:1.4) nitrate surface concen-
tration (Carreto et al. 1986), as expected for late spring
after the diatom bloom (November), therefore explain-
ing the low chl a concentration. As in estuarine waters,
diatoms were dominant in the phytoplankton assem-
blage. Their dominance in the coastal zone was probably
related to transport in the buoyant plume from the
estuary, and then dilution by vertical mixing of the water
column. Although nitrate and silicate limitations induce
the replacement of diatoms by some haptophytes, the
chl a biomass of codominant haptophytes B was typi-
cally low within the coastal assemblage (B).
In contrast with the diatom dominance in the estu-
arial and coastal systems, a flagellate-dominated com-
munity was found over the continental shelf, almost

exclusively comprised by haptophytes resembling the
coccolithophorid E. huxleyi (haptophytes C). In spite of
E. huxleyi being dominant in assemblage C, the chl a
biomass estimated from microscopic cell counts
(assuming a cell content of 0.1 lg chl a per 106 cells;
Stolte et al. 2000) was three orders of magnitude lower
than that determined by HPLC. This difference could be
associated to the cell stage in the life cycle of E. huxleyi
(Paasche 2000). The E. huxleyi coccolith-forming cell is
the stage normally reported for the sea, and is the one
that completely dominates blooms. However, cells de-
void of coccoliths are practically indistinguishable from
other haptophytes and rarely, if ever, quantified in field
work (Campbell et al. 1994). The dominance of naked or
scaly E. huxleyi cells and/or the presence of other ha-
ptophytes, undetected by microscopy, could explain
such inconsistency between E. huxleyi cell biomass and
chl a. The only previously reported occurrence of an
E. huxleyi bloom in Argentine waters occurred in the
same area during austral spring (November) of 1989
(Gayoso 1995). However, patches of water with spectral
signatures identical to those of blooms of E. huxleyi
have been regularly observed on the shelf and shelf-
break off Argentina during late austral spring and
summer (Brown and Podestá 1997). In our data the most
remarkable hydrographic feature associated with the
E. huxleyi bloom was the existence of a two-layer water
column with a well-developed, shallow pycnocline near
20 m, resulting from the superposition of tow type of
waters. However, we can only speculate as to which
species contributed the most biomass in the deep pop-
ulation.
The phytoplankton community in the cold and
nutrient-rich water of the Malvinas Current, near the
shelf-break, was dominated by another haptophyte,
Phaeocystis sp. This species is also common on the
Argentine continental shelf, associated with coastal tidal
fronts as well as with the shelf-break front (Carreto et al.
1985, 1995; Carreto and Benavides 1989). Large differ-
ences in pigment profiles (authors’ unpublished results)
were observed after comparing a Phaeocystis sp. strain
isolated from a northern Argentine coastal area with our
field data (shelf-break), indicating some regional physi-
ological heterogeneity within the species. Phaeocystis
antarctica is a common microalgae in the Southern
Ocean; its spring blooms are yearly events in all seas
surrounding Antarctica (Palmisano et al. 1986; Wright
and Jeffrey 1987; Davidson and Marchant 1992).
Therefore, it is likely that the Phaeocystis sp. population
found in the sub-Antarctic waters of the Malvinas
Current is related to Southern Ocean populations. This
was supported by the observed pigment profiles (see
pigment ratios). The low diatom abundance observed in
this region was coincident with previous results that
showed the dominance of microflagellates in the Malv-
inas Current (Negri, personal communication).
The warm, nutrient-poor surface water of the Brazil
Current is generally characterized by a low new
production, except in the upper photic zone of the
1025
Brazil–Malvinas Confluence (Hubold 1980; Gayoso and
Podestá 1996; Brandini et al. 2000). It is well known that
nano- and picoplankton are the main components in
the phytoplankton communities of nutrient-depleted
oceanic waters. Coincidentally, our results showed that
cyanobacteria and haptophytes were the main contrib-
utors to chl a, each group providing approximately 30–
40% of total chl a. Nevertheless these groups’ biomass,
expressed as chl a absolute values, was low. For exam-
ple, the cyanobacterial contribution to chl a (up to
0.25 lg l)1) was on the same order of magnitude as that
observed in the estuary system. The haptophytes ob-
served in the samples comprised several non-identified
coccolithophorid species and low numbers of E. huxleyi.
In this area, pelagophycean, prasinophycean and cryp-
tophyte species appeared to be poor competitors, as each
group contributed about 5–10% of the total chl a. The
absence of diatoms and dinoflagellates in the subtropical
waters of the Brazil Current was noteworthy. This
observation contrasts with results from Gayoso and
Podestá (1996) on the phytoplankton species composi-
tion near the confluence of Brazil and Malvinas Cur-
rents. Gayoso and Podestá (1996) speculated that the
high diatom density (up to 5.5·106 cells l)1) observed in
the Malvinas retroflexion was due to the nutrient
enrichment of the subtropical assemblages from the
Malvinas Current.
Conclusions
Our results showed that the complementary use of pig-
ment analysis and microscopy is very useful tool in
characterizing phytoplankton communities of highly
diverse ecosystems, with a wide variability in species
distribution, similar to that represented in the studied
area.
The inclusion of the most abundant members of the
chl c pigment family (chl c1, chl c2, chl c3 and non-polar
types of chl c) in the pigment matrix improved the
CHEMTAX interpretation of field data. This improve-
ment allowed us to distinguish between four haptophyte
populations along the section, even though they had
similar pigment signatures. It is evident that the pre-
dominance of each subtype was spatially segregated and
that segregation was imposed by the environmental
conditions. The identification of the algae prevailing in
the estuary system (prasinophyceans without Pras) could
only be resolved by microscopy. Therefore, the optimal
use of pigment data is achieved if such data are com-
bined with microscopic examination of representative
samples.
Acknowledgements We are grateful to Dr. M.D. Mackey for pro-
viding the computer program CHEMTAX. We thank three
anonymous reviewers for providing helpful comments and sug-
gestions on the manuscript. This is contribution no. 1254 of
INIDEP.
1026
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