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Biovolume Determination of Phytoplankton Guilds in Transitional Water Ecosystems of Mediterranean Ecoregion
Biovolume Determination of Phytoplankton Guilds in Transitional Water Ecosystems of Mediterranean Ecoregion
Biovolume Determination of Phytoplankton Guilds in Transitional Water Ecosystems of Mediterranean Ecoregion
RESEARCH ARTICLE
Corresponding author: Tel.: +39 0832 298604; Fax: +39 0832 298722; e-mail:
mariarosaria.vadrucci@unile.it
Abstract
1 - Conceptually, morphometric measurements of phytoplankton guilds seem to have major advantages as
descriptors of the ecological status of transitional water ecosystems (TW) with respect to classical
taxonomic descriptors. However, at present, standardized or common methodologies for the use of
morphometric descriptors do not exist.
2 - This paper aims to provide a starting point for the activation of standardized methods for the determination
of morphometric descriptors of phytoplankton as a quality element in TW in accordance with the new
directive of WFD 2000/60/EU.
3 - Phytoplankton biovolume is one of the most studied morphometric descriptors. It can be estimated by
associating the algae with similar geometric forms and determining the volume of these by measuring the
linear dimensions required for its calculation under the microscope. However, the lack of a standardized set
of geometric forms and equations for calculating biovolume causes difficulties and produces data that are
not comparable.
4 - A set of geometric models is suggested here for calculating the cell biovolumes of 201 phytoplankton
genera found in transitional water ecosystems of Mediterranean Ecoregion. The equations were designed to
minimize the effort of microscopic measurements. The main methodological problems, and the similarities
and differences between our own and previously published proposals are discussed.
Keywords: morphometric descriptors, algal biovolume, geometric model, phytoplankton.
Introduction
Phytoplankton are listed in the Water
Framework Directive (2000/60/EC) as a quality
element for determining the ecological status of
transitional water ecosystems. Nevertheless, at
this time, only taxonomic parameters (diversity
and abundance) are used as phytoplankton
descriptors in the monitoring plans for water
quality assessment. However, there are some
disadvantages, in using these descriptors as
indicators of the health status of transitional
waters, because taxonomic parameters shown an
TWB 2 (2007)
Vadrucci et al
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Vadrucci et al
Methods
Compilation of the unified floristic list.
We selected a set of geometric shapes for
determining biovolume in phytoplankton guilds
in transitional waters by consulting floristic lists
available for a number of different types of
transitional
water
ecosystems
in
the
Mediterranean eco-region, including salt-pans,
river deltas and coastal lagoons. These lists
came from our own projects and from the
literature (Vadrucci et al., 2004; Caroppo, 2000,
Facca et al., 2001). In the case of our projects,
the lists were drawn up by laboratories with
experience in the field of phytoplankton
analysis, such as the University of Lecce
Laboratory of Ecology, the Department of
Biological oceanography INOGS of Trieste, the
University of Tirana Laboratory of Botany
(Albania) and the Institute of Oceanology of the
Dinophyceae
18%
Euglenophyceae
3%
Xantophiceae
1%
Bacillariophyceae
32%
Cyanophyceae
8%
Cryptophyceae +
Coanoflagellati +
Kinetoplastidi
13%
Chrysophyceae+
Dictyochophyceae+
Haptophyceae
14%
Chlorophyceae+
Prasinophyceae
11%
Figure
1.
Relative
contribution of the main
phytoplankton
groups
in
transitional water ecosystems
in the Mediterranean ecoregion (data from a number of
credited
lists
of
phytoplankton
found
in
transitional ecosystems).
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87
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Vadrucci et al
88
TWB 2 (2007)
50
45
Number of genera
40
35
30
Simply shapes
Combined shapes
25
20
15
10
5
0
4
12
11
13
10
18
14
16
22
21
23
20
17
19
15
50
45
Number of genera
40
required of measurement of
cell thickness
35
30
25
20
15
10
5
0
4
18
14
11
13
16
21
17
19
15
12
10
20
22
23
Figure 2. Number of genera associated with simple geometric forms (light grey) and combined
geometric forms (dark grey). Number of genera that do not require (light grey) and that do require
(dark grey) measurement of cell thickness (B). The numbers on the x-axis are taken from Annex I.
Light halos, which affect the measurements of
the smallest cells, can be overcome by
increasing the magnification of the microscope.
In this protocol, we proposed to measure the
linear dimensions using image analysis systems.
Image Analysis systems include software that is
able to measure semi-automatically a series of
morphometric parameters of the phytoplankton
cells. The system acquires a digital image of the
microscopy field or part of the field, and the
boundary of the objects (cells) to be measured is
then traced on the digital image; the
measurement
module
then
provides
measurements of morphometric features such as
length, width, perimeter and circularity. In some
cases, they provide measurements of biovolume
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TWB 2 (2007)
Vadrucci et al
Conclusion
The use of morphometric parameters of
phytoplankton for defining the health of
transitional ecosystems status appears to be
validated by the quantity of experimental
evidence highlighting the sensitivity and
symptomatic responses of these parameters to
environmental forcing and pollution. However,
a good descriptor needs to be not only sensitive
and symptomatic of environmental risk but also
comparable and reliable; this is possible only if
there exists a standardize procedure for its
90
TWB 2 (2007)
References
Ansotegui A, Sarobe A., Trigueros J M,
Urrutxurtu I, Orive E. 2003. Size distribution
of algal pigments and phytoplankton
assemblages
in
a
coastalestuarine
environment: contribution of small eukaryotic
algae. J. Plankton Res., 25: 341-355.
Basset A, Sangiorgio F, Pinna M. 2004.
Monitoring with benthic macroinvertebrate:
advantage and disadvantages of body size
descriptors. Aquat. Conserv.: Mar. and
Freshwat. Ecosyst,. 14: 43-58.
Boyd CM, Johnson GW 1995. Precision of size
determination of resistive electronic particle
counters. J. Plankton Res., 17: 223-234.
Brown LM, Gargantini I, Brown DJ, Atkinnson
HJ, Govindarajan J, Vanlerberghe GC 1989.
Computer-based image analisys for the
automated counting and morphological
description of microalgae in culture. J. Appl.
Phycol., 1: 211-225.
91
TWB 2 (2007)
Vadrucci et al
Annexes
Annex I - Schedule for geometric shapes
showing: mathematical model, number and
types of linear dimensions to measure, genera to
which the shape is applied, notes on the
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Vadrucci et al
1) Sphere
simple
2) Prolate spheroid
Simple
V = /6 d 3
3) Ellipsoid
simple
h
d
V = /6 d 2 h
V = /6 a c h
1: d= diameter
2 : d= diameter; h= height
94
TWB 2 (2007)
Annex I-continued
Geometric shape
Type of geometric shape
4) Cylinder
Simple
5) Cone
Simple
d
z
V = (/6 a c h)-10%
3: a= length; c=width; h=height
V = /4 d 2 h
2: d= diameter; h=height
V = /12 d 2 h
2: d= diameter; z= height of cone
Group 1: Gyrosigma
Number of genera: 1
Percentage: 0.5%
This model was applied according
to Edlers (1979) suggestion.
Number of genera: 43
Percentage: 21.0%
This model is easy to apply and is generally calculated automatically by most
image analysis software. The shape is also used in other sets of geometric forms
for the above-mentioned genera, with the exception of Chaetoceros. Hillebrand
et al. (Hillenbrand et al., 1999) and Sun and Liu, (Sun and Liu 2003), proposed
the elliptic prism form, whereas Edler (Edler, 1979) proposed the ellipsoid form.
Here, we propose the cylindrical form, although it overestimates cell volume by
as much as 40 % depending on the TA/AA ratio. Nevertheless, it is adequate for
routine analysis (Montagnes and Franklin 2001).
Number of genera: 4
Percentage: 1.9%
This form was applied to just four genera, in accordance
with Hillebrand et al. (Hillebrand et al., 1999).
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Vadrucci et al
Annex I-continued
Geometric shape
Type of geometric shape
c
6) Truncated cone
simple
7) Parallelepiped
simple
d1
h
d2
Formula for volume calculus
V = /12 h (d 12 +d 1d 2+ d 2 2)
Number of genera: 1
Percentage: 0.5%
Number of genera: 10
Percentage: 4.7%
Number of genera: 19
Percentage: 8.61%
This form was introduced for the first time in Hillebrand et al.s paper
(Hillebrand et al., 1999) It provides a more accurate estimate of biovolume
for the above-mentioned genera, because it is more similar to the real shape
of the cell than the geometric form (parallelepiped) proposed by Edler
(Edler, 1979). In this set, we are agree with Hillebrand et al.s set of
geometric forms, including the exceptions for Navicula. This genus is quite
variable and therefore some species can require the use of a more
appropriate geometric form according to their shape (such as a box or prism
on a parallelogram base). The need to measure the third dimension can
render the application of this model difficult.
V=abc
V = /4 a b c
96
TWB 2 (2007)
Annex I-continued
Geometric shape
Type of geometric shape
11) Cube
Simple
c
b
m
h
l
V=abc
V= l m h
V = a3
Number of genera: 3
Percentage: 1.43%
Number of genera: 2
Percentage: 0.95%
Number of genera: 2
Percentage: 0.95%
Note
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TWB 2 (2007)
Vadrucci et al
Annex I-continued
Geometric shape
Type of geometric shape
b1
b
a
V = /4 a b c
V= [(b 1 +b b 1 b) a]/3
V = /6 d 2 h
Group 1: Epithemia,
Phaeodactylum
Number of genera: 3
Percentage: 1.43 %
For this form, we agree with the
proposal by Hillebrand et al.
(Hillebrand et al., 1999) Edler
(Edler, 1979) proposed ellipsoid
forms for the first two genera,
whereas Sun and Liu (Sun and
Liu, 2003) proposed the sickleshaped prism, but only for the
Eunotia genus (the other two
genera are not included in their
list). We consider the half elliptic
prism more suitable, because it has
the same number of linear
dimensions required but is more
similar to the real shape of the
cell.
Number of genera: 2
Percentage: 0.95 %
The model differs by those proposed by other authors, e.g.
Hillebrand et al. (Hillebrand et al., 1999), who proposed the
gomphonemoid form. This form requires four linear dimensions,
some of them very difficult to measure. For example, in his work,
the linear measurement f is the length of the transapically widest
part of the head pole. Sun and Liu (Sun and Liu, 2003) proposed
the same form but only for the Gomphonema genus; for the
Licmophora genus, they proposed the sickle-shaped cylinder form.
However, the volume of the sickle-shaped cylinder proposed by
Sun and Liu for Licmophora can overestimate the volume, because
it considers the two transapical views of the cell to be similar. We
propose the truncated pyramid shape for both genera, because: the
minor base, the major base and the height of a truncated pyramid
with a square base is easier to measure and the lower accuracy of
the shape is balanced by the greater replicability of the data.
Eunotia,
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TWB 2 (2007)
Annex I-continued
Geometric shape
Type of geometric shape
d1
z
d
d2
h
V = /6 h (d 1 +d 1 *d 2+ d 2 )
2
V = (/4 d h) + (/6 d z)
V = (/4 d h) + (/12 d 2 z)
Number of genera: 1
Percentage: 0.47 %
Number of genera: 2
Percentage: 0.95 %
Number of genera: 2
Percentage: 0.95%
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TWB 2 (2007)
Vadrucci et al
Annex I-continued
d2
d
h1
V = /12 d 2 (z+d)
h2
d1=d2
Z1
Z2
a c
z
d1
d4 hd5
h1
d1
Geometric shape
Type of geometric shape
Number of genera: 1
Percentage: 0.47 %
Number of genera: 1
Percentage: 0.47%
100
TWB 2 (2007)
Annex I-continued
Geometric shape
Type of geometric shape
a2
d
a
b2
h
Formula for volume calculus
V = (/4 d h) + (/6 d 3 )
V = (/12 a c) (h+z)
a1
c
V= c (a 1 b 1 + /4 a2 b 2)
2: d=diameter; h= height
Group 7: Euglena
Group 1: Climacosphenia
Number of genera: 2
Percentage: 0.95 %
Number of genera: 1
Percentage: 0.47 %
Number of genera: 1
Percentage: 0.47 %
Note
al.s
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TWB 2 (2007)
Vadrucci et al
Annex II
Genus
shape
C.U.
Group 1
Achnanthes
cell
Amphiprora
Amphora
Asterionella
Asterolampa
Asteromphalus
Bacillaria
Bacteriastrum
Bellerochea
Biddulphia
Campylodiscus
Cerataulina
Chaetoceros
Climacosphenia
Cocconeis
Corethron
Coscinodiscus
Coscinosira
Cyclotella
Cylindrotheca
Cymatopleura
Cymbella
Dactylosolen
Detonula
Diatoma
Dimerogramma
Diploneis
Ditylum
Ellerbeckia
Epithemia
Eucampia
Eunotia
Fragilaria
Fragilariopsis
Gomphonema
Grammatophora
Guinardia
Gyrosigma
Hemiaulus
Lauderia
Leptocylindrus
Licmophora
Lioloma
Lyrella
Mastogloia
Melosira
Navicula
Nitzschia
Paralia
Phaeodactylum
Pinnularia
Planktoniella
Pleurosigma
Porosira
Proboscia
Pseudonitzschia
Rhabdonema
Rhizosolenia
Skeletonema
Stauroneis
Stephanodiscus
Stictocyclus
8
3
7
4
4
7
4
10
8
8
4
4
23
3
21
4
4
4
19
7
3
4
4
8
8
8
4
4
12
8
12
8
8
13
8
4
3a
4
4
4
13
4
8
8
4
8
9
4
12
7
4
9
4
4
9
7
4
4
8
21
4
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
Genus
Striatella
Surrirella
Synedra
Tabellaria
Thalassionema
Thalassiosira
Thalassiothrix
Toxarium
Trachyneis
Triceratium
Group 2
Actinastrum
Ankistrodesmus
Carteria
Chlorogonium
Closterium
Crucigenia
Gonium
Halosphaera
Monoraphidium
Oocystis
Pachysphaera
Pavlova
Pediastrium
Pseudotetraedon
Pterosperma
Pyramimonas
Pyraminomonas
Scenedesmus
Schroderia
Staurastrum
Sticochoccus
Tetraedon
Tetraselmis
Trochiscia
Group 3
Acanthoica
Acanthosolenia
Apedinella
Braarudosphaera
Calciopappus
Calciosolenia
Calycomonas
Calyptrosphaera
Ceratolithus
Chrysochromulina
Coccolithos
Coronosphaera
Dichtyocha
Dinobryon
Ebria
Emiliana
Gephyrocapsa
Halopappus
Helladosphaera
Hermesinum
Mamiella
Monochrysis
Ochromonas
Ophiaster
Parapedinella
Phaeocystis
shape
3
4
C.U.
colony
cell
31
7
7
7
4
4
4
8
10
cell
cell
cell
cell
cell
cell
cell
cell
cell
16
16
1
14
8
11
2
1
14
2
2
2
2
7
1
5
5
2
14
15
4
7
2
1
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
colony
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
2
4
2
1
5
4
5
1
4
2
1
1
1
2
1
1
1
2
1
1
1
2
18
2
1
1
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
Genus
Pontosphaera
Prymnesium
Pseudopedinella
Rhabdosphaera
Syracolithus
Syracosphaera
Group 4
Bicosta
Chroomonas
Criptaulax
Criptoficee undet.
Cryptomonas
Hilea
Leucocryptos
Micromonas
Plagioselmis
Pseudobodo
Rhinomonas
Rhodomonas
Spumella
Group 5
Anabaena
Anabaenopsis
Aphanizomenon
Aphanizonium
Chroococcus
Coelosphaerium
Gloeocapsa
Merismopedia
Microcystis
Nodularia
Nostoc
Oscillatoria
Phormidium
Snowella
Spirulina
Synechoccocus
Tetrapedia
Woronichinia
Group 6
Alexandrium
Amphidinium
Amphisolenia
Blastodinium
Ceratium
Cochlodinium
Dinophysis
Diplopsalis
Exuviaella
Glenodinium
Goniaulax
Goniodoma
Gymnodinium
Gyrodinium
Heterocapsa
Katodinium
shape
1
U.C
cell
Genus
Lyngulodinium
shape
3
U.C.
cell
18
6
1
1
1
cell
cell
cell
cell
cell
4
18
18
4 or 1
2
2
18
18
18
1
18
18
18
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
Massartia
Minuscula
Nematodinium
Oblea
Oxyphysis
Oxyrrhis
Oxytoxum
4
4
4
4
1
2
1
11
10
1
4
4
4
4
2
4
1
1
1
filament
filament
filament
filament
cell
colony
single, colony
cell
part of colony
filament
cell
filament
filament
colony
filament
cell
cell
colony
3
18
3
1
14
2
2
8
14
18
14
3
3
5
3
1
18
3
9
18
1
14
14
10
14
11
14
12
3
13
1
2
3
3
2
2
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
3
22
17
17
14
8
cell
cell
cell
cell
cell
cell
cell
3
3
2
2
4
3
20
3
14
2
3
4
3
5
2
18
3
3
18
6
14
1
3
3
7
18
14
17
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
cell
Pachidinium
Peridinium
Phalacroma
Pheopolykrikos
Podolampas
Polykrikos
Porella
Pronoctiluca
Prorocentrum
Protoceratium
Protogonyaulax
Protoperidinium
Ptychodiscus
Pyrophacus
Scrippsiella
Torodinium
Warnowia
Group 7
Astasia
Euglena
Eutreptia
Eutreptiella
Lepocinclis
Phacus
Group 8
Meringosphaera
Exceptions:
1)
2)
3)
7)
8)
9)
10)
14)
102