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- were subdominant (20–35%). The continental shelf
tion of phytoplankton communities were studied in a assemblage showed an almost exclusive dominance
section across the continental shelf between the Rı́o de (90%) of haptophytes resembling the coccolithophorid
La Plata and the oceanic waters of the Subtropical E. huxleyi. Another haptophyte (Phaeocystis sp.) was
Convergence, during late spring (November 1999). Al- dominant (75–85%) in the Malvinas Current assem-
gal communities were examined using light microscopy blage. The Brazil Current assemblage was characterized
and HPLC-derived (high-performance liquid chroma- by the codominance of cyanobacteria (45%) and ha-
tography) pigment concentrations. The CHEMTAX ptophytes (35%). These results are discussed in rela-
program was used to estimate the chlorophyll a (chl a) tionship to the complex hydrographic features of the
biomass of different algal classes. The inclusion of the area.
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 Introduction
novel approach four haptophyte populations were
distinguished across the studied section, even though The Rı́o de La Plata drains the second largest basin of
they had qualitatively similar pigment signatures, South America. It flows into the Atlantic Ocean with a
although one subtype lacked 19¢-hexanoyloxyfucoxan- total discharge of 20,000–25,000 m3 s)1 (Framiñan and
thin (Hex-Fuco). Five different phytoplankton assem- Brown 1996). The main characteristics of the Rı́o de La
blages, spatially segregated by the prevailing Plata estuary are its large spatial scale and the occur-
environmental conditions, were distinguished during rence of a quasi-permanent salt wedge regime gener-
the studied period. All of them showed a complex ating a border system (Carreto et al. 1986; Guerrero
community structure, formed by a background of et al. 1997; Mianzan et al. 2001). Tidal currents are
small-sized cells such as cyanobacteria, cryptophytes, typically below 45 cm s)1 (Framiñan et al. 1999), and
haptophytes and prasinophyceans, on which diatom, the estuary therefore becomes a typically two-layer
cryptophyte or some haptophyte blooms were over- system with strong vertical stratification that gradually
lapped. In the estuarine assemblage, where maximum weakens seaward (Mianzan et al. 2001). Although the
chl a concentrations where found, diatoms were always main flow of the Rı́o de La Plata is expected to have a
the dominant group (30–60% of total chl a), but NNE direction along the Uruguayan coastline, its tra-
cryptophytes (10–40%), prasinophyceans (2–20%) and jectory, forced by the dominant winds, fluctuated sea-
dinoflagellates (2–12%) were also relevant. In the sonally. Its influence may be detected, in spring and
coastal assemblage diatoms were also the dominant summer, in a southward direction along the coastal
group (35–45%), but haptophytes lacking Hex-Fuco region of Argentina (Guerrero et al. 1997) and/or along
the mild-shelf regime (Carreto et al. 1995). Occasionally
Communicated by O. Kinne, Oldendorf/Luhe its area of influence may extend to the continental
slope, where these estuarine waters contact the warm,
J. I. Carreto (&) Æ N. G. Montoya Æ H. R. Benavides subtropical Brazil Current water (Lusquiños and Val-
R. Guerrero Æ M. O. Carignan des 1971; Provost et al. 1995), and the nutrient-rich
Instituto Nacional de lnvestigación y Desarrollo Pesquero
(INIDEP), Paseo Victoria Ocampo no. 1, waters of the Malvinas flow (Negri et al. 1992), or it
7600 Mar del Plata, Argentina may be found between both currents. These waters can
E-mail: [email protected] be carried offshore by the return flux of the Malvinas
1014

Current (Provost et al. 1995) in an area referred to as papers, although the interpretation of HPLC field data is
the Subtropical Confluence. The time and intensity of not always a straightforward task (see Jeffrey et al. 1997).
the occurrence of these events may be dependent on the Recent developments in HPLC technology using C18
cyclical excursions of the main currents along the polymeric and C8 monomeric HPLC columns, and
continental slope, on the volume discharge of the river pyridine as modifier (Garrido and Zapata 1997; Jeffrey et
(Provost et al. 1995) and on the meteorological con- al. 1999; Zapata et al. 2000), have uncovered additional
ditions (Lusquiños and Valdes 1971; Carreto et al. chlorophyll c (chl c) pigments and acyloxyfucoxanthins
1986; Glorioso and Flather 1995; Guerrero et al. 1997). (Garrido and Zapata 1998; Egeland et al. 2000; Garrido
The strong contrast in various ocean properties et al. 2000; Zapata et al. 2001). The improved resolution
(e.g. salinity, sea surface temperature and nutrient of HPLC methodology and the potential utility of these
concentration), together with complex mesoscale pigments as biological markers (Zapata et al. 2001) could
patterns, make this area an important biogeographic facilitate the interpretation of field data.
boundary between phytoplankton communities of Several possibilities to estimate quantitatively the
brackish and oceanic waters of subtropical and sub- various phytoplankton groups from marked pigment
Antarctic origin. However, relatively few phytoplank- concentrations have been developed (Gieskes et al.
ton studies have been carried out in this area since 1988; Everitt et al. 1990; Letelier et al. 1993). A po-
Hentschel’s (1932) early data. Phytoplankton taxonomy tential source of errors in these approaches was that
and biogeography of dinoflagellates, diatoms and sili- pigment ratios might change as a consequence of long-
coflagellates have been previously studied by Balech term changes in light climate and nutrient availability
(1949, 1971, 1986), by Frenguelli and Orlando (1959), (Goericke and Montoya 1998; Mackey et al. 1998; van
Müller-Melchers (1959) and Lange (1985), respectively. Leeuwe and Stefels 1998). Mackey et al. (1996) intro-
Negri et al. (1988) and Bayssé et al. (1986) have studied duced the CHEMTAX program to obtain biomass
relative phytoplankton abundance and community estimations of algal groups that share common
structure of net samples and their relationship with pigments, taking into account the physiological and
environmental factors. Gayoso (1995) and Gayoso and genetic variation in pigment ratios. Nevertheless, inter-
Podestá (1996) reported on the phytoplankton abun- species (Jeffrey and Vesk 1997; Zapata et al. 2001) and
dance, although cells <5 lm in diameter were not in- inter-clones (Stolte et al. 2000) variation in pigment:chl
cluded in their analyses. The scenario resulting from a ratios, was also considerable. These findings indicated
these previous studies displayed a region where robust that the ratios chosen for the applications of the
diatoms, armored dinoflagellates, calcareous cocco- CHEMTAX calculations should, if at all possible,
lithoporids and Phaeocystis (Prymnesiophyceae) colo- reflect the dominant phytoplankton species present in
nies are the most frequent forms. The recognition of the sample (Schlüter et al. 2000).
the importance of delicate flagellates and minute coc- In the present work, the structure of surface phyto-
coid forms has come only recently, after more appro- plankton communities was studied during November
priate techniques for studying the picoplankton were 1999 in a section across the Argentine continental shelf
applied in this area (Silva and Negri 2000). between the Rı́o de la Plata and the oceanic waters of the
Picoplankton and many of the smaller nanoplankton Subtropical Convergence. Microscopic enumeration of
cells are difficult to study. Their small size and the ab- phytoplankton species, HPLC-derived pigment concen-
sence of clearly discernible features often preclude trations and the CHEMTAX software were used for the
identification by light microscopy, even to the class level, first time in the area to evaluate the distribution and
and many are too fragile to be adequately preserved abundance of the different phytoplankton groups. The
(Jeffrey and Vesk 1997). Fluorescence microscopy and spatial distribution of the different phytoplankton
flow cytometry have proven to be invaluable for the communities and their dominant species were analyzed
enumeration of cyanobacteria and prochlorophytes, but in relation to the complex environmental features of the
the identification of the less distinctive cells remains a region.
problem.
The characteristic pigment composition of phyto-
plankton groups and the development of modern of Materials and methods
high-performance liquid chromatography (HPLC)
methods has favored the present use of pigment analysis Sample collection
in phytoplankton community research (Gieskes and
Kraay 1983; Bidigare et al. 1990; Wright et al. 1996; Sampling procedures were performed from 6 to 11 November 1999,
on board the R.V. ‘‘Cap. E. Canepa’’ (Fig. 1a). Vertical profiles for
Schlüter et al. 2000). Pigment chemo-taxonomy has been temperature, salinity and fluorescence were obtained with a CTD-
especially useful in samples from oceanic areas (Veldhuis SBE 911 equipped with a Seapoint fluorometer. CTD data and in
and Kraay 1990; Letelier et al. 1993; Peeken 1997), and vivo fluorescence measurements were calibrated and reduced to
proved to be valuable for the characterization of small- 1-m bins. Light penetration was measured with a PUV 500 (Bio-
spherical Instruments) underwater radiometer. Surface water
sized phytoplankton in coastal marine areas (Schlüter samples (0 m depth) were collected with a clean plastic bucket, and
et al. 2000) and estuaries (Roy et al. 1996). The signature discrete sampling of the water column (5 m depth) for CTD cali-
role of the pigments has been summarized in various bration was performed with a Niskin bottle.
 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.
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 Data processing
November 1999 composite
The chl a of phytoplankton groups at each sampled station were
calculated using CHEMTAX, a matrix factorization program
Remote data on SST and ocean color
running under MATLAB (Mackey et al. 1996). All diagnostic
pigments detected, as well as those ambiguous markers indicative
Sea surface temperature (SST) field was derived from data collected
of only a few groups, were included in the program MATRIX.
by the Advanced Very High Resolution Radiometer (AVHRR)
Chl c1, chl c2, chl c3 and chl c2 monogalactosyldiacylglyceride es-
installed on the NOAA 11 polar satellite and processed by the
ters were also included, as the HPLC method used is very useful to
Argentine National Space Research Commission (CONAE) using a
separate the members of this pigment family. Prochlophytes were
multi-channel algorithm. Chlorophyll surface field was derived
not included in the data matrix, as divinyl chl a and b could not be
from data collected by SeaWiFS installed on the Orb View-2 polar
separated from monovinyl chl a and b with this method. The light-
satellite and processed to level 3 (standard mapped image) by
protecting carotenoids of the xanthophyll cycle, diadinoxanthin
CONAE.
(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).
Microscopic analysis A matrix using initial pigment:chl a ratios derived from the
literature was constructed (Wright et al. 1996; Jeffrey and Vesk
Surface water subsamples (0 m depth) for phytoplankton identifi- 1997; Peeken 1997; Mackey et al. 1998; Schlüter et al. 2000; Stolte
cation and cell counts were obtained from five selected stations and et al. 2000; Zapata et al. 2001), since data on pigment ratios for the
fixed with neutral Lugol’s iodine. Sample volumes of 50 and 100 ml dominant phytoplankton species in the area were not available.
were settled. Phytoplankton species were identified and enumerated Although the original data must be previously split into homoge-
with an Olympus IX-70 inverted microscope using bright-field neous groups, the samples were all analyzed together since the
optics and ·200, ·400, ·600 and ·1,000 (oil) magnifications. Algal small number of samples adversely affected the accuracy of the
cells were identified to species level when possible, except for 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)
 1017

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 Fig. 3 Chlorophyll a concentration (lg l)1) at the surface (0 m
depth) of occupied stations along the section, as determined by
(upper 15 m). Continental shelf waters are characterized HPLC
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). (Framiñan and Brown 1996). Chl a surface concentra-
Upper Malvinas waters, lying over the continental shelf tion was low in the coastal area near the estuary (sta-
break (stations 14–15), are characterized by a salinity of tions 7–11) (Fig. 3), but increased gradually with depth
33.8 psu and a temperature range from 6.3C to 10.3C. (Fig. 4b). Except for the estuarine area, the highest
Brazil waters have a salinity >34.5 psu and tempera- surface chlorophyll concentration (2.14 lg l)1) was
tures >16C (stations 16–19). At some stations of the measured in the over-shelf stations 12 and 13, where
Brazil waters and throughout the anticyclonic eddy de- 5.0 lg l)1 maximum was observed at 10–15 m depth,
scribed above, a variable layer of low-salinity water at the base of the mixed layer (Fig. 4c). The high chl a
coming from the continental shelf system capped the values in the photic zone of these stations appeared to be
surface waters. 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
Structure of phytoplankton communities 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-
Distribution of chl a geneous at these stations (Fig. 4d). In contrast, the
The SeaWiFS (Fig. 1b) imagery provided valuable minimum surface chl a values (Fig. 3) measured ‘‘in
information on the overall distribution of near-surface situ’’ in the oceanic region dominated by the subtropical
phytoplankton biomass in the region and its relation water of the Brazil Current (stations 16–19) were
with hydrography. The lowest chl a values were ob- accompanied by a maximum (1.8 lg l)1) at the base of
served in both the warm water of the Brazil Current, the photic zone (Fig. 4e).
where phytoplankton is nutrient limited, and in the
nutrient rich but highly turbulent waters of Malvinas Pigment profiles and microscopy data
Current. In the transition zone between these boundary
currents, the chlorophyll concentration was enhanced, Table 2 lists the microalgal pigments detected in
especially near the shelf-break, where filaments of high increasing elution order, spectral characteristics of pig-
chlorophyll concentrations were found (Gayoso and ment in the mobile phase are also included. The chro-
Podestá 1996; Brandini et al. 2000). These filaments matograms obtained (Fig. 5) from five selected stations
appear to be related to a band of high chlorophyll showed the elution patterns of a range of chlorophyll
concentrations present along the west side of the frontal and carotenoid pigments characteristic of the phyto-
region separating the Malvinas Current from shelf wa- plankton communities that prevail in the different
ters (Carreto et al. 1986, 1995). Except for this frontal studied ecosystems. The species composition of the
region, the over-shelf chlorophyll concentration was phytoplankton communities estimated by cell counts, in
slight and similar to that observed in the transition zone. these selected stations are presented in Table 3.
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 Phytoplankton communities
estuary stations (1–6) was much lower than that ob-
tained from SeaWiFS (Fig. 3), and did not vary signifi- Three main phytoplankton assemblages, closely related
cantly with depth, as observed in the vertical profiles of to the hydrography of the area, were differentiated on
station 2, representative of this sector (Fig. 4a). There- the basis of the dominant carotenoid: (1) estuarine and
fore, the optical signal produced by the dissolved or- coastal communities, (2) continental shelf and Malvinas
ganic matter and suspended sediments in the area was communities and (3) the Brazil Current community
likely to be stronger than the phytoplankton signal (Fig. 6a).
1018

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
Fig. 4 Vertical profiles of temperature (C), salinity (psu) and was observed in B; however, Hex-Fuco was rarely
chlorophyll a (lg l)1), from five selected representative stations of: observed in this assemblage (Fig. 6a). Moreover, as in
a estuarine (station 2), b coastal (station 7), c continental shelf
(station 12), d Malvinas Current (station 15) and e Brazil Current assemblage A, diatoms prevailed and no haptophytes
waters (station 16). Data derived from continuous CTD-fluoro- were found by light microscopy. Some clones of the
meter readings diatoms Thalassionema nitzschioides and Rhizosolenia
setigera (identified in our samples) are known to con-
tain chl c3 (Stauber and Jeffrey 1988). However, as
Estuarine and coastal communities diatom species composition was similar in both assem-
blages, the relative increase of chl c3 detected in
Fuco was the dominant carotenoid in the Rı́o de La assemblage B was not related to a change in the diatom
Plata estuarine and coastal area. Microscopic analysis flora. Another possibility was that some small or delicate
(Table 3) confirmed that several diatom species were haptophytes with Fuco, such as the main carotenoid,
dominant in this phytoplankton assemblage, explaining would have been present (Jeffrey and Vesk 1997). The
the observed high relative Fuco concentration. This 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, to that observed for the minor non-polar chl c (un-
Garrido et al. 2000) and chl c2–monogaloctosyldiacyl- known non-polar chl c) from E. huxleyi (clo-
glyceride from Chrysochromulina polylepis (chl c2- ne CCMP370). A compound with the same retention
MGDG Chrysochromulina-type, Zapata et al. 2001) as time and spectral properties (Table 2) was also present
associated trace pigments (Fig. 5b) supported this in assemblage C, although at a trace level. Also, trace
assumption. amounts of a pigment with a retention time
(rt=35.23 min) similar to that observed for chl c2–
Continental shelf and Malvinas communities MGDG Chrysochromulina-type (Zapata et al. 2001),
were present in both assemblages. Another unknown
Hex-Fuco was the dominant carotenoid in continental chl c–like derivative of intermediate polarity
shelf and Malvinas waters. Herein, the relative abundance (rt=23.29 min) was observed only in assemblage D
of the associated pigments also indicated at least two (Fig. 5f).
spatially segregated pigment associations (Fig. 5c, d). Microscopy showed (Table 3) that haptophytes were
The relative proportion of Fuco to Hex-Fuco in the the dominant group in both phytoplankton assem-
continental shelf assemblage (C) was lower than that blages. However, E. huxleyi was the dominant species
observed in the Malvinas assemblage (D) (Fig. 6a). In in assemblage C and Phaeocystis sp. was dominant in
addition, although the two assemblages presented sev- assemblage D. Some other unidentified Coccosphaerales
eral non-polar types of chl c (Fig. 5c, d), a different species were also present in assemblage D. Microscopy
proportion of these compounds was evident (Fig. 6e). showed that some diatom species were present in low
In assemblage C the HPLC retention time of the main abundances in both assemblages, although chl c1 was
compound (rt=33.36 min) was coincident with that of not detected in our chromatograms (Fig. 5c, d).
the principal non-polar chl c from E. huxleyi (chl c2– Microscopy also showed that the Prasinophyceae
MGDG Emiliania-type) (Garrido et al. 2000). The Micromonas sp. was present in both assemblages as a
retention time of the major non-polar chl c in assem- minor component, explaining the small amounts of
blage D was slightly slower (rt=32.41 min) and similar 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

the ratio of But-Fuco to total fucoxanthins was signifi-
Zea was the principal carotenoid in the community cantly higher than that observed for the other assem-
corresponding to the subtropical waters of the Brazil blages (Fig. 5e) pelagophyceans were expected to be
Current. Microscopy showed that picoplanktonic cy- present. Despite the error arising from the quantification
anobacteria (Synechococcus spp.) were the most abun- of low pigment amounts associated with green algae, a
dant cells in the phytoplankton assemblage. The relative significant relation between chl b and Pras was observed.
proportions of chl c3, Fuco, But-Fuco and Hex-Fuco On the other hand, Lut was only present in trace
were also high (Fig. 5e), indicating that haptophytes amounts and then was not quantified. The evidence
and/or pelagophyceans were abundant. Chl c2–MGDG indicated that, in this assemblage, chl b stemmed from
Chrysochromulina-type (Zapata et al. 2001) was the main prasinophyceans containing Pras. However, the Prasin-
non-polar chl c, although several non-polar types of ophyceae lacking Pras, Pyramimonas sp., was the only
chl c, with similar retention times to those observed in green alga observed by microscopy (Table 3). Another
assemblages C and D, were present in trace levels uncertainty is related to the HPLC method, as divinyl
(Fig. 5e). The pigment profile of the dominant hapto- chl a and b could not be separated from monovinyl chl a
phytes in assemblage E was different from that observed and b. Hence, the presence of picoplanktonic prochlo-
for E. huxleyi (C) and Phaeocystis sp. populations (D). rophytes could not be totally excluded. The unequivocal
Microscopy showed that, although E. huxleyi was also Cryptophyceae marker Allo was present in low amounts
present, some unidentified coccolithophorid species were in the samples from this assemblage (Fig. 6b), but this
the dominant haptophytes in this assemblage (Table 3). group was not detected by microscopy; cryptophytes
Pelagophyceans were not observed by light microscopy, probably could not withstand the fixation process
as these small cells can only be properly identified by (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 prevailed (35–45% of total chl a), as in the estuary com-
munity. In terms of chl a calculated by CHEMTAX,
The total and relative contribution of the various groups haptophyte B was the subdominant group (20–35% of
of phytoplankton to chl a, calculated by CHEMTAX total chl a). Cyanobacteria (5–15% of total chl a) and
are shown in Figs. 7 and 8, respectively. prasinophyceans (2–12% of total chl a) were also rel-
As expected from pigment results, diatoms were al- evant in decreasing order. The contribution of other
ways the dominant group (30–60% of total chl a) in the groups was small (chlorophyceans, prasinophyceans
phytoplankton community of the estuary assemblage lacking Pras, cryptophytes, and other haptophyte
(A), where a maximum chl a concentration was found. types).
In terms of chl a calculated by CHEMTAX, crypto- The algal group composition obtained by CHEM-
phytes were the subdominant group (10–40% of total TAX for the continental shelf area (C) showed an
chl a). In decreasing order, chlorophyceans (2–20% of almost total dominance (90% of total chl a) of ha-
total chl a) and dinoflagellates (2–12% of total chl a) ptophytes C, probably represented by the coccolitho-
were also relevant. The contribution of other groups phorid E. huxley. The contribution of other groups was
such as cyanobacteria (<10% of total chl a) and Pras- very low, with cyanobacteria the most important of
containing prasinophyceans (<5% of total chl a) was these groups (about 5% of total chl a).
small. A high dominance (75–85% of total chl a) of ha-
Total phytoplankton biomass was very low in the ptophytes D was detected by CHEMTAX in the
coastal assemblage (B), but the phytoplankton com- Malvinas Current assemblage (D), mainly represented
munity was more diverse. The pigment proportions by the flagellated cells of Phaeocystis sp. However,
detected with CHEMTAX showed that diatoms some differences between the two stations (14 and 15)
1022

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,
Fig. 6 Relative abundance of major pigments across the transect cyanobacteria and prasinophyceans
(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 Discussion
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-
As in most studies, the lack of local known pigment ratios
nia-type, chlorophyll c2–MGDG Chrysochromulina-type, unknown was a limitation for the application of CHEMTAX
non-polar chlorophyll c analysis. Nevertheless, our pigment ratio estimations are
consistent with those obtained by previous authors. The
CHEMTAX program assumes that each taxonomic cat-
can be expected after considering the subdominant egory has a consistent set of pigment ratios through all
groups. Haptophyte E was the subdominant group samples. However, some changes in both, the pigment
(15% of total chl a) at the oceanic station (15), whereas ratios within a given algae species and the different species
diatoms were subdominant (5% of total chl a) on the composition of an algae group, would be expected along
west side of the shelf-break (station 14). The contri- the section. Our results showed that the inclusion in the
bution of each of the other detected groups was <5%. pigment matrix of the most abundant member of the chl c
The algal group composition detected by CHEM- pigment family (chl c1, chl c2, chl c3 and chl c2 monoga-
TAX in the Brazil Current assemblage (E) showed the lactosyldiacylglyceride esters) improves CHEMTAX and
codominance of cyanobacteria (45%) and haptophytes allowed us to distinguish between four haptophytes
E (35%). The contribution of prasinophyceans, cryp- populations across the studied section. The inclusion of
tophytes and pelagophyceans was similar, each group four categories accommodated the natural split of the
representing about 10% of the total chl a. haptophytes between the different ecosystems studied.
 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-
Fig. 8 Percentage of different phytoplankton groups contributing
to the total concentration of chl a across the transect (station 1– mediate polarity was also present in this population.
19), estimated by interpretation of pigment HPLC data using the
CHEMTAX program, grouped in separate panels: a diatoms,
cryptophytes, dinoflagellates and chlorophyceans; b haptophytes Haptophytes E
B, haptophytes C, haptophytes D and haptophytes E; c pelago-
phyceans, cyanobacteria and prasinophyceans Our calculated acyloxyfucoxanthin ratios for the ha-
ptophytes growing in subtropical waters (coccolitho-
Calculated pigment ratios 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
Haptophytes B prevailing non-polar chl c. The ratio of chl c2–MGDG
Chrysochromulina-type to chl a (0.176) was in the range
The coastal water haptophytes (B) showed a pigment estimated (0.00–0.269) for several Chrysochromulina
profile type, with Fuco, chl c3 and chl c2 as the prin- species studied by Zapata et al. (2001). As there is no
cipal components, and chl c2–MGDG Emiliania-type information on the distribution of this non-polar chl c2
and chl c2–MGDG Chrysochromulina-type as tracer- among coccolithophorids, the calculated ratio should be
associated pigments (Garrido et al. 2000; Zapata et al. treated with caution.
2001). Our calculated Fuco:chl a ratio (0.465) was
similar to that presented by one Chrysochromulina
Diatoms
species (0.487; C. kappa) with the same pigment profile
(Zapata et al. 2001). The Fuco:chl a ratio (0.86) was higher than that calcu-
lated by Wright et al. (1996) for diatom populations
Haptophytes C from the Australian sector of the Southern Ocean (0.60),
but much lower than that calculated for shade-adapted
The continental shelf (C) haptophytes contained Hex- diatoms (1.25) by Letelier et al. (1993). A similar ratio
Fuco as the main carotenoid. It is remarkable that variation range (0.479–1.218) was found by Schlüter
1024

et al. (2000) for coastal species. Also chl c1:chl a and Lut, Zea and its epoxides (Egeland et al. 1995; Mackey
chl c2:chl a ratios (respectively 0.178 and 0.208) were et al. 1996). Consequently, pigment analysis does not
both in the range observed by Jeffrey (1972) for several permit a clear separation of prasinophyceans from
diatom species. chlorophyceans. We included two groups of green algae
in our samples: chlorophyceans and prasinophyceans
lacking Pras (chlorophyceans), and Prasinophyceans
Pelagophyceans containing Pras (prasinophyceans). Despite regional
differences, species composition and variation owing to
Our initial pigment ratios were taken from Mackey et al. the algae physiological state, our values calculated for
(1998), obtained from surface pelagophyceans of the both chl b:chl a (1.156) and Pras:chl a (0.372) ratios in
western equatorial Pacific (1.33 for But:chl a and 0.166 prasinophyceans coincide with literature values (Mackey
for Fuco:chl a). The program left these ratios un- et al. 1996; Peeken 1997).
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 Algal assemblages and environmental conditions
strain of Pelagoccus subviridis (respectively 0.25 and
0.44). Both pigment HPLC patterns and microscopy-derived
information showed five different phytoplankton assem-
Cyanobacteria blages in the region during spring 1999. These assem-
blages displayed a clear spatial segregation imposed by the
The Zea:chl a ratio of cyanobacteria was highly depen- extant environmental conditions. All of them presented a
dent on irradiance (Kana et al.1988; Bigidare et al. 1989; complex community structure, comprising small-sized
Lutz et al. 2001), being consistent with its photopro- cells from cyanobacteria, cryptophytes, chlorophyceans,
tective function. Although large errors can be expected haptophytes and prasinophyceans, on which diatoms,
in the calculated pigment ratios, our value (1.12) was cryptophytes, or some haptophytes overlapped.
largely comparable with averaged values of literature Several diatom species were dominant in the phyto-
results, obtained at high and low light intensities plankton assemblage (Table 3) of the nutrient-rich and
[Schlüter et al. 2000 (1.24); Mackey et al. 1998 (1.15); high-silicate estuary water (Carreto et al. 1986), where a
Kana et al. 1988 (1.20)]. Our value was also similar to high chl a concentration was measured. The dominance
the ratios obtained at medium light intensities [Moore of larger diatom cells was coincident with most of the
et al. 1995 (0.7 at 350 lE m)2 s)1); Morel et al. 1993 previous reports for this area (Negri et al. 1988; Elge
(0.71 at 100 lE m)2 s)1); Lutz et al. 2001 (1.00 at 100 lE et al. 1990; Mesones 1991). The Prasinophyceae Pyra-
m)2 s)1)] for several strains of marine cyanobacteria mimonas sp. and the Cryptophyta Cryptomonas sp. were
(Synechococcus spp.). also very frequent in this eutrophic environment; the
latter codominating in some cases with diatom flora.
Some bloom-forming dinoflagellate species (e.g. Proro-
Cryptophytes centrum minimum) were also present, although in rela-
tively low concentrations, indicating that factors other
Whereas the Allo:chl a ratio, calculated from oceanic than nutrients regulate the dinoflagellate blooms in this
phytoplankton, was in the range of 0.105–0.240 (Gieskes fluctuating environment.
and Kraay 1983; Wright et al. 1996; Mackey et al. 1998; Total phytoplankton biomass was very low, and a
Peeken 1997), values up to 1.58 were found in phyto- more diverse phytoplankton community was observed in
plankton communities of the turbid lower St. Lawrence the coastal system. The slightly stratified coastal water
estuary (Roy et al. 1996). On the other hand, our cal- presented both the lowest absolute (<1 lM) and rela-
culated ratio was similar to the one obtained for a tive (N:P:Si ratio: of 0.1:1:1.4) nitrate surface concen-
coastal strain of Plagioselmis prolonga (0.682), growing tration (Carreto et al. 1986), as expected for late spring
at medium light intensity (Schlüter at al. 2000). after the diatom bloom (November), therefore explain-
ing the low chl a concentration. As in estuarine waters,
Prasinophyceans and chlorophyceans diatoms were dominant in the phytoplankton assem-
blage. Their dominance in the coastal zone was probably
A high inter-species variability was found in pigment related to transport in the buoyant plume from the
ratios (Mackey et al. 1996), attending the complex tax- estuary, and then dilution by vertical mixing of the water
onomy of prasinophyceans. Some of them produce column. Although nitrate and silicate limitations induce
common green algal carotenoids, whereas some others the replacement of diatoms by some haptophytes, the
synthesize unusual carotenoids for this algal class chl a biomass of codominant haptophytes B was typi-
(micromonadophytes), Pras being the most important cally low within the coastal assemblage (B).
among them (Egeland et al. 1995). Additionally, some In contrast with the diatom dominance in the estu-
chlorophyceans and prasinophyceans grown at low light arial and coastal systems, a flagellate-dominated com-
intensity do not always contain the accessory pigments munity was found over the continental shelf, almost
 1025

exclusively comprised by haptophytes resembling the Brazil–Malvinas Confluence (Hubold 1980; Gayoso and
coccolithophorid E. huxleyi (haptophytes C). In spite of Podestá 1996; Brandini et al. 2000). It is well known that
E. huxleyi being dominant in assemblage C, the chl a nano- and picoplankton are the main components in
biomass estimated from microscopic cell counts the phytoplankton communities of nutrient-depleted
(assuming a cell content of 0.1 lg chl a per 106 cells; oceanic waters. Coincidentally, our results showed that
Stolte et al. 2000) was three orders of magnitude lower cyanobacteria and haptophytes were the main contrib-
than that determined by HPLC. This difference could be utors to chl a, each group providing approximately 30–
associated to the cell stage in the life cycle of E. huxleyi 40% of total chl a. Nevertheless these groups’ biomass,
(Paasche 2000). The E. huxleyi coccolith-forming cell is expressed as chl a absolute values, was low. For exam-
the stage normally reported for the sea, and is the one ple, the cyanobacterial contribution to chl a (up to
that completely dominates blooms. However, cells de- 0.25 lg l)1) was on the same order of magnitude as that
void of coccoliths are practically indistinguishable from observed in the estuary system. The haptophytes ob-
other haptophytes and rarely, if ever, quantified in field served in the samples comprised several non-identified
work (Campbell et al. 1994). The dominance of naked or coccolithophorid species and low numbers of E. huxleyi.
scaly E. huxleyi cells and/or the presence of other ha- In this area, pelagophycean, prasinophycean and cryp-
ptophytes, undetected by microscopy, could explain tophyte species appeared to be poor competitors, as each
such inconsistency between E. huxleyi cell biomass and group contributed about 5–10% of the total chl a. The
chl a. The only previously reported occurrence of an absence of diatoms and dinoflagellates in the subtropical
E. huxleyi bloom in Argentine waters occurred in the waters of the Brazil Current was noteworthy. This
same area during austral spring (November) of 1989 observation contrasts with results from Gayoso and
(Gayoso 1995). However, patches of water with spectral Podestá (1996) on the phytoplankton species composi-
signatures identical to those of blooms of E. huxleyi tion near the confluence of Brazil and Malvinas Cur-
have been regularly observed on the shelf and shelf- rents. Gayoso and Podestá (1996) speculated that the
break off Argentina during late austral spring and high diatom density (up to 5.5·106 cells l)1) observed in
summer (Brown and Podestá 1997). In our data the most the Malvinas retroflexion was due to the nutrient
remarkable hydrographic feature associated with the enrichment of the subtropical assemblages from the
E. huxleyi bloom was the existence of a two-layer water Malvinas Current.
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. Conclusions
The phytoplankton community in the cold and
nutrient-rich water of the Malvinas Current, near the Our results showed that the complementary use of pig-
shelf-break, was dominated by another haptophyte, ment analysis and microscopy is very useful tool in
Phaeocystis sp. This species is also common on the characterizing phytoplankton communities of highly
Argentine continental shelf, associated with coastal tidal diverse ecosystems, with a wide variability in species
fronts as well as with the shelf-break front (Carreto et al. distribution, similar to that represented in the studied
1985, 1995; Carreto and Benavides 1989). Large differ- area.
ences in pigment profiles (authors’ unpublished results) The inclusion of the most abundant members of the
were observed after comparing a Phaeocystis sp. strain chl c pigment family (chl c1, chl c2, chl c3 and non-polar
isolated from a northern Argentine coastal area with our types of chl c) in the pigment matrix improved the
field data (shelf-break), indicating some regional physi- CHEMTAX interpretation of field data. This improve-
ological heterogeneity within the species. Phaeocystis ment allowed us to distinguish between four haptophyte
antarctica is a common microalgae in the Southern populations along the section, even though they had
Ocean; its spring blooms are yearly events in all seas similar pigment signatures. It is evident that the pre-
surrounding Antarctica (Palmisano et al. 1986; Wright dominance of each subtype was spatially segregated and
and Jeffrey 1987; Davidson and Marchant 1992). that segregation was imposed by the environmental
Therefore, it is likely that the Phaeocystis sp. population conditions. The identification of the algae prevailing in
found in the sub-Antarctic waters of the Malvinas the estuary system (prasinophyceans without Pras) could
Current is related to Southern Ocean populations. This only be resolved by microscopy. Therefore, the optimal
was supported by the observed pigment profiles (see use of pigment data is achieved if such data are com-
pigment ratios). The low diatom abundance observed in bined with microscopic examination of representative
this region was coincident with previous results that samples.
showed the dominance of microflagellates in the Malv-
inas Current (Negri, personal communication). Acknowledgements We are grateful to Dr. M.D. Mackey for pro-
viding the computer program CHEMTAX. We thank three
The warm, nutrient-poor surface water of the Brazil anonymous reviewers for providing helpful comments and sug-
Current is generally characterized by a low new gestions on the manuscript. This is contribution no. 1254 of
production, except in the upper photic zone of the INIDEP.
1026

Framiñan M, Etala M, Acha EM, Guerrero RA, Lasta CA, Brown

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