Pandey 2017

Water Research 118 (2017) 39e58
Contents lists available at ScienceDirect
Water Research
journal homepage: www.elsevier.com/locate/watres
Review
The use of diatoms in ecotoxicology and bioassessment: Insights,

advances and challenges
Lalit K. Pandey a, b, Elizabeth A. Bergey c, Jie Lyu d, Jihae Park d, e, Soyeon Choi d,
Hojun Lee b, Stephen Depuydt e, Young-Tae Oh f, Sung-Mo Lee f, Taejun Han b, g, *
a
Institute of Green Environmental Research Center, 169, Gaetbeol-ro, Yeonsu-gu, Incheon 21999, South Korea
b
Department of Marine Sciences, Incheon National University, 119, Academy-ro, Yeonsu-gu, Incheon 22012, South Korea
c
Oklahoma Biological Survey and Department of Biology, University of Oklahoma, Norman, OK 73019, USA
d
Division of Life Sciences, Incheon National University, 119, Academy-ro, Yeonsu-gu, Incheon 22012, South Korea
e
Department of Plant Biotechnology and Bioinformatics, Ghent University Global Campus, 119, Songdomunwha-ro, Yeonsu-gu, Incheon 21985, South Korea
f
Institute of Public Health and Environment, 471, Seohae-daero, Jung-gu, Incheon 22320, Republic of Korea
g
Ghent University Global Campus, 119, Songdomunwha-ro, Yeonsu-gu, Incheon 21985, South Korea
a r t i c l e i n f o a b s t r a c t
Article history: Diatoms are regularly used for bioassessment and ecotoxicological studies in relation to environmental
Received 25 September 2016 and anthropogenic disturbances. Traditional taxonomical diatom parameters (cell counts, biovolume
Received in revised form estimates, species richness, diversity indices and metrics using sensitive and tolerant diatom species) are
28 January 2017
regularly used for these studies. In the same context, very less focus was given on new endpoints of
Accepted 30 January 2017
Available online 2 February 2017
diatoms (life-forms, nuclear anomalies, alteration in photosynthetic apparatus shape, motility, lipid
bodies, size reduction and deformities), in spite of their numerous merits, such as, their easiness,
quickness, cheapness, global acceptation and no especial training in diatom taxonomy. In this review we
Keywords:
Biomonitoring
analyzed 202 articles (from lab and field studies), with the aim to investigate the bioassessment and
Ecotoxicology ecotoxicological advancement taken place in diatom research especially in terms of exploring new
Nuclear anomalies endpoints along with the traditional taxonomical parameters in a perspective which can greatly enhance
Lipid bodies the evaluation of fluvial ecosystem quality for biomonitoring practices.
Size reduction © 2017 Elsevier Ltd. All rights reserved.
Deformities
Contents
1. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 40
2. Traditional endpoints . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 41
2.1. Diatom diversity and abundances . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 41
2.2. Cell densities and biovolume of the community . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 42
2.3. Sensitive and tolerant species . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 43
2.4. Life-forms and ecological guilds . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 44
3. New endpoints . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 44
3.1. Alterations in cell integrity . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 44
3.1.1. Nuclear anomalies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 44
3.1.2. Alteration in the cell membrane and cytoplasmic content . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 45
3.1.3. Alteration in chloroplasts (shape, size, color and number) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 46
3.2. Lipid bodies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 47
3.3. Alteration in frustule size . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 49
3.4. Morphological deformities . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 51
4. Conclusion and future prospective . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 54
* Corresponding author. Department of Marine Science, Incheon National Uni-

versity, 119, Academy-ro, Yeonsu-gu, Incheon 22012, Republic of Korea.
E-mail address: [email protected] (T. Han).
http://dx.doi.org/10.1016/j.watres.2017.01.062
0043-1354/© 2017 Elsevier Ltd. All rights reserved.
40 L.K. Pandey et al. / Water Research 118 (2017) 39e58
Acknowledgements . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 54
Supplementary data . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 54
Conflict of interest . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 54
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 54
1. Introduction adverse effects of chemicals at the population and community

level, thus they are directly linked to biodiversity. According Knauer
Globally, diatoms are used to assess the ecological status of and Hommen (2012), two common metrics - species richness and
aquatic systems because diatoms have a cosmopolitan nature, short Shannon-Wiener index (H’) - showed low bioassessment value for
life span and quick response to environmental and anthropogenic community structure, whereas total abundance and the abundance
disturbances (Stevenson et al., 2010). Such use of diatoms is of the dominant species demonstrated high sensitivity.
meaningful, given that diatoms are the chief primary producers in High diatom community variability is an inherent characteristic
waterbodies, contributing 40% of the primary productivity of the of complex test systems, such as in experimental micro- and
oceans and contributing approximately 20% of global carbon fixa- mesocosm studies, and consequently, this variability complicates
tion (Hildebrand, 2008). Various organisms (macroinvertebrates, interpretation of studies (Campbell et al., 1999). Therefore, evalu-
fishes, macrophytes and algae, including diatoms) have been used ation of test systems is preferably not based on biodiversity metrics
for biomonitoring around the world, but among these organisms, alone, but is complemented by other parameters (e.g., functional
diatoms are the most suitable for assessing the chemical status of metrics) that are quick, easy and can be used globally. Functional
waterbodies (McCormick and Cairns, 1994; Stevenson et al., 2010). metrics are pertinent because they are closely linked to some reg-
For example, Hering et al. (2006) found that diatoms were sensitive ulatory and supporting ecosystem services, and may be less vari-
to nutrient and organic matter contamination, whereas fishes, able among replicates. Furthermore, even if the structural integrity
macroinvertebrates and macrophytes were more sensitive to hy- of the community is altered, some functional parameters (such as
drological changes in aquatic ecosystems. Indeed, the high sensi- primary productivity and nutrient cycling) may be unaffected and
tivity of diatoms to organic toxicants (atrazine, metolachlor, vice versa. Combining information from both structural and func-
simazine, phenols and PAHs) (Blanco and Be cares, 2010), organic tional metrics enhances the sensitivity and predictive power of
matter (especially nutrients, nitrate and phosphate) (Stevenson studies aimed at bioassessment and risk assessment. Indeed, the
et al., 2010; Stevenson, 2014; Morin et al., 2016) and inorganic use of both structural and functional parameters provides sensitive
contamination (heavy metals) (Hirst et al., 2002; De Jonge et al., and powerful early warning tools for evaluating sub-lethal effects
2008; Morin et al., 2012) has been reported from different parts of exposure to toxins (Renzi et al., 2014). However, structural
of the world. A major advantage of using diatoms in environmental metrics (cell counts and biovolume, species composition and
studies, including ecotoxicology, is that diatom assemblages are abundance) had high variation due to variation among replicates,
specious and can be used to investigate the effects of toxicants at which resulted in statistically weak relationships with the studied
different levels of ecological organization (community, population, stressors (Kraufvelin, 1998; Knauer et al., 2005). Varying division
and individual levels) (McCormick and Cairns, 1994; Debenest et al., rates of individual diatom species (which is often related to their
2013; Stevenson, 2014). Because of these characteristics, a variety of size; Lavoie et al., 2006) is a prime reason for high variation among
national and international agencies have recommended diatoms as test replicates, in part because accurate quantification of diatom
a biomonitoring tool for assessing the ecological status of rivers and frustules is still a major constrain for ecotoxicological studies. Thus,
streams (Kelly, 1998; Stevenson et al., 2010). there is an urgent need to develop or incorporate more sensitive
Diatoms are characterized by their robust, ornamented, species- metrics for assessing early exposure to stress, specifically metrics
specific siliceous frustules, which are preserved and dependably that measure physiological or morphological changes in diatom
replicated in successive generations (Falasco et al., 2009). Species species, making functional metrics a priority for further research
identification is based on these siliceous frustules. Even in fossilized (Renzi et al., 2014).
form, diatom frustules are a useful tool to assess the paleoclimatic For bioassessment and ecotoxicological practices several new
conditions (Mackay, 2007). Although used for biomonitoring diatom parameters have been reported in the last decade; param-
worldwide, few studies have related diatom diversity in Polar Re- eters that are very promising but sporadically used or reported.
gions to global climate change (Anderson, 2000; Bopp et al., 2005; These parameters are nuclear anomalies (Debenest et al., 2008;
Alvain et al., 2013). Licursi and Go mez, 2013), alteration in the cell membrane, cyto-
In spite of their diversity, beauty, ecological importance and plasmic content and photosynthetic apparatus (Chang et al., 2011;
biomonitoring potential, diatoms are globally underutilized as a Armbrecht et al., 2014; Wood et al., 2014), changes in lipid body
tool for risk assessment and for evaluating management options for formation (Pandey et al., 2015) and the classification of diatoms
fluvial ecosystems (Stevenson et al., 2010). The main reason behind using various life-forms and ecological guilds (Passy, 2007; Rimet
this underutilization is the limited types of metrics traditionally and Bouchez, 2011). Relationships among these unconventional
used in bioassessment. Traditional metrics for diatoms include parameters have been found in some studies. For example, diatom
biovolume, cell density and relative abundance (with special motility was associated with the size/number of lipid bodies (Wang
reference to indicator species), whereas the newer behavioral, et al., 2013), size reduction can be associated with frustule defor-
physiological and functional metrics are rarely used. These newer mity (Hasle and Syversten, 1996), lipid bodies metrics and frustule
metrics have advantages in understanding the dynamics of bio- deformities may vary with cytoplasmic anomalies (Renzi et al.,
logical communities (Giddings et al., 2002), especially because 2014) and frustule deformity is associated with lipid body charac-
these metrics demonstrate sublethal effects that are not apparent teristics (Pandey et al., 2015). This correlation among unconven-
in diatom counts (the basis of traditional metrics). tional metrics demonstrates shared sensitivity and indicates their
Community structural metrics are useful for predicting possible efficiency in measuring responses to toxicants, especially early
L.K. Pandey et al. / Water Research 118 (2017) 39e58 41
effects and cellular responses to high toxicant doses. These metrics analysis of how loss of diversity may be linked to chemical
may allow early detection of stress after exposure to doses below contamination would be useful in refining risk-assessment pro-
those needed to cause cell death, which is the endpoint detected by cedures. Effects of contaminants on diatom communities have been
traditional methods (Renzi et al., 2014). In addition, these uncon- frequently evaluated using various diversity indices, such as species
ventional parameters are also reported to be effective for bio- richness, Shannon index, evenness, and Jaccard similarity index. For
assessment practices. For example, Gillett et al. (2009, 2011) example, Gold et al. (2002) used a translocation experiment and
reported percent live diatoms in the biofilm to be a cost-effective reported lower cell density, species richness and Shannon index of
bioassessment tool. Similarly, Morin et al. (2016) advocated the the periphytic diatom community at the severely polluted sites.
use of impaired cytoplasmic content in live diatoms for the bio- Similarly, Verb and Vis (2005) reported lower diversity (in terms of
assessment of fresh waterbodies. Furthermore, various life-forms species richness and Shannon index) of periphytic diatoms at the
and ecological guilds of diatoms are regularly investigated for acid mine drainage (AMD) impacted sites in the USA. Recently,
assessing the ecological health of waterbodies (Medley and Morin et al. (2015) found significantly higher values for various
Clements, 1998; Passy, 2007; B-Be res et al., 2014). These newer diversity indices for periphytic diatoms (species richness and
metrics are quick, easy, require less human expertise, have good Shannon index) from the control sites (Firmi and Moulin) than from
reproducibility, have standard protocols and, most importantly, can contaminated sites (Joanis and Usine for metals and Decazeville for
be adopted world-wide. However, the use of these metrics for high nutrient loads) of the Riou-Mort River, South West France.
ecotoxicological and bioassessment testing needs further study and Similarly, Luís et al. (2011) reported lower Shannon index values of
demonstration in order to better determine concentration- periphytic diatom communities examined at acid mine drainage
dependent and/or time-dependent responses and to gain wider sites in Portugal. Moreover, reproduction of diatom species may be
acceptance (Dickman, 1998; Renzi et al., 2014). inhibited under stress, resulting in decreases in species richness
The use of molecular approaches (next-generation sequencing, and diversity, as has often been reported at metal polluted sites
DNA barcode, DNA fingerprinting) in diatom-based biomonitoring (Deniseger et al., 1986; Genter and Lehman, 2000).
practices has recently received significant attention (Kermarrec These biodiversity parameters are regularly used but sometimes
et al., 2013, 2014; Manoylov, 2014; Visco et al., 2015). De- do not provide consistent information about the known impair-
velopments of next-generation sequencing technologies offer the ment of waterbodies (Blanco et al., 2012). For example, Hirst et al.
possibility to use molecular barcoding for fast and reliable biodi- (2002) showed that species richness, evenness and diversity were
versity surveys based on environmental samples. Next-generation not significantly related to the chemical characteristics of stream
sequencing approaches have tremendous potential for diatom- water polluted by mining activities in United Kingdom. In the same
based monitoring, such as ease in species identification, compari- context, De Jonge et al. (2008) reported statistically insignificant
son of species inventories, real-time assessment of living commu- correlations between periphytic diatom community parameters
nities and reduction in sample processing time (Visco et al., 2015). (diversity, evenness and different indices) and the heavy metal load
Similarly, DNA fingerprinting may be used to measure the genetic of water and sediment in a lowland river in Flanders, Belgium.
diversity in populations of diatoms (e.g., Ditylum brightwellii) and Similarly, Duong et al. (2008) found no difference in the diversity
may also help understand the relationship between diatom blooms indices of periphytic diatom communities examined at the refer-
and environmental conditions (Rynearson and Armbrust, 2000). ence (Firmi) sites and contaminated (Joanis) sites of the Riou-Mort
Molecular bioassessment of diatoms is a promising field but at this River, South West France. Duong et al. (2010) also found no sig-
point, we cannot yet adopt them for many reasons. The identity of nificant difference in terms of species richness (S) and diversity (H0 )
most taxa in genetic reference libraries has not been rigorously between the control periphytic diatom community and that
evaluated, so reference library based taxonomy may not be accurate contaminated with 100 mg L1 Cd after one week of colonization.
(Manoylov, 2014). Lastly, the high cost of reagents per megabase However, significantly higher metric values were recorded in the
sequencing output and the reading of homopolymer regions are control community than in the contaminated community at the
additional issues with molecular bioassessment that need to be fourth and sixth week of the experiment.
addressed (Claesson et al., 2010). As a consequence, the number of Other studies showed mixed responses of diatoms against
field-collected samples analyzed for bioassessment with molecular different types of stresses. For example, Medley and Clements
tools has been limited (Kermarrec et al., 2013). (1998) reported significantly lower species diversity (p < 0.05)
Incorporating the new metrics along with the traditionally used but the relation was insignificant in terms of cell density, species
metrics will make diatom ecotoxicological and bioassessment richness and abundance of dominant taxa at the Zn polluted sites in
practices easier and more rapid, reproducible, cheap and globally the Colorado Rocky Mountain streams (USA), whereas Hill et al.
accepted. Thus, in this review, we explore the present status of (2000) were unable to relate species richness with metal concen-
various structural and functional metrics of diatoms used in eco- tration in a Rocky Mountain stream polluted with heavy metals.
toxicology and bioassessment studies and discuss their merits and Gold et al. (2003a) reported higher Shannon index (H’), the same
demerits in order to raise the profile of diatoms as an effective tool species richness and a low cell density at the metal polluted (Cd and
for biomonitoring practices. Zn) sites in comparison to reference sites. In contrast, Sabater
(2000) found that heavy metals in water and sediments were
2. Traditional endpoints significantly and negatively affecting various diatom metrics
(evenness, diversity and diatom indices). In addition, they also re-
2.1. Diatom diversity and abundances ported marked and long-lasting effects of heavy metals on the
periphytic diatom communities from impacted versus reference
Analysis of available literature does not support a direct cause- sites of the Guadiamar River, South-West Spain. This inconsistency
effect relationship between chemical pollution and diatom di- of diatom diversity indices and lack of significant correlations be-
versity. In general, the relationship between diversity and chemical tween diversity, evenness and environmental variables is in
contamination is not always a simple linear positive/negative as- accordance with the “Intermediate Disturbance Hypothesis”, which
sociation and results are not always strong enough to indicate a assumes a parabolic relation between diversity and water quality,
management approach that would most effectively conserve the with the highest diatom diversity at intermediate pollution levels
health of the ecosystem (Ricciardi et al., 2009). Thus, a careful (Van de Vijver and Beyens, 1998; Hirst et al., 2002).
Ricciardi et al. (2009) reviewed the status of biological com- combinations of environmental variables, along which the relative
munities in rivers in relation to chemical contamination and re- distributions of species assemblages. Ordination plots help in
ported two major constrains in diversity and pollution studies. visualizing the pattern of community variation, distributions of
First, they found that the use of a single diversity index or small species and assemblages among environmental variations, and can
number of diversity indices was inadequate. The different biodi- be used for indicating species-environment relationships. (ter
versity indices assess different types of information and using only Braak, 1986; ter Braak, 1987; ter Braak and Smilauer, 2002).
one of these indices (types of information) is not enough to Furthermore, ordination and clustering can be used to show as-
adequately define biological diversity. Multivariate-analysis studies semblages that differ from other assemblages, which may be
that combine both conventional and taxonomy-based indices and caused by anthropogenic impacts (e.g. Chessman, 1986; Stevenson
possibly different trophic levels provide a more comprehensive and White, 1995). Thus, incorporation of these analyses (any one)
view of the diversity status of an ecosystem and its potential provides a simple, easy-to-understand description of the percent
response to pollutants. Second, they advocated the use of coupled change in species relative abundances between assemblages at
chemical and biological analyses to evaluate water-quality status assessed and reference sites.
and the effect of chemicals on fluvial biological diversity, thus Diatom diversity indices and their relative abundance in the
promoting a multidisciplinary approach. Further, they recom- diatom communities are widely used to indicate the presence and
mended incorporating a variety of types of biodiversity indices, extent of organic and inorganic contaminants in fluvial ecosystems.
especially when using algae (diatoms) for assessing the chemical In most studies, these routinely used indices provide a statistically
status of aquatic systems. According to Stevenson et al. (2010), weak relationship between diatom diversity and pollutants. Thus,
composite indices that incorporate both the richness and evenness in the light of this result, there is an urgent need to incorporate new
elements of diversity are needed to characterize the kind and types of diatom-based indices (e.g., a taxonomic composition based
severity of pollution in fluvial ecosystems. Similarly, Blanco and index), which can provide reliable information using statistical
Becares (2010) found that the simultaneous use of several diatom tools. Improvement in taxonomic resolution of diatom species, use
indices is required to detect and evaluate the huge spectrum of of composite indices (richness, evenness, diversity and dominance),
potential pollutants in river basins. increased cell counting (3000e8000 valves) and simultaneous use
Changes in species composition tend to be the most sensitive of several indices to detect and evaluate the huge spectrum of
response when comparing impacted and reference diatom as- potential pollutants have been proposed to improve the efficacy of
semblages (Stevenson, 1984; Jüttner et al., 1996). Species compo- diatom-based ecotoxicological assessments (Blanco and Be cares,
sition is often examined as relative abundance of diatom species in 2010; Stevenson et al., 2010; Rimet and Bouchez, 2011).
the community; a metric frequently used for deciphering the
ecological health of waterbodies (Medley and Clements, 1998; Gold 2.2. Cell densities and biovolume of the community
et al., 2002, 2003a; Morin et al., 2008a; De Jonge et al., 2008;
Stevenson et al., 2010; Duong et al., 2012; Morin et al., 2012; Changes in community cell densities and biovolume represent
Arini et al., 2012a,b,c; Pandey et al., 2014, 2015, 2016; Gautam an important tool for diatom-based community bioassessment
et al., 2017). Lavoie et al. (2006) reported relative abundance as a (Pandey et al., 2014). Diatom densities are estimated using various
more informative tool than other community parameters (espe- methods, such as counting cells with a Spencer's haemocytometer
cially biovolume of the community) for biomonitoring. In this at 450 magnification, and densities are converted to biovolume by
study, analyses conducted on diatoms grouped by size showed that applying geometrical volume formulas for generalized diatom
small and large taxa have similar responses to the water chemistry shapes (Hillebrand et al., 1999). Alteration in the cell density and
variables (total phosphorus, soluble phosphorus, total dissolved biovolume of the diatom in biofilms has been variously attributed
nitrogen, nitrates, ammonia, temperature, dissolved oxygen, con- to current velocity (Ghosh and Gaur, 1998), diatom emigration and
ductivity and dissolved organic carbon). Furthermore, they found immigration (Stevenson and Peterson, 1991), grazers (Colletti et al.,
that biovolume estimation analysis provided no extra information 1987; Arini et al., 2012b), pesticides (Pere
s et al., 1996; Debenest
but required considerably more time, money and human expertise. et al., 2009; Debenest et al., 2010; Morin et al., 2010) and heavy
Similarly, Litchman et al. (2009) concluded that data on the relative metals (Gold et al., 2002; Pandey et al., 2014). Cell density and
abundance of different diatom sizes in diverse aquatic ecosystems biovolume changes (either single or together) following the
may indicate high variation in distribution, reflecting a variety of translocation of diatom assemblages between non-impacted and
immediate, system-specific selective pressures. Stevenson et al. impacted locations has been used to assess the level of disturbance
(2010) also found relative abundance of diatom genera and spe- in the waterbodies (Gold et al., 2002, 2003a,b; Duong et al., 2012;
cies as the most valuable attribute of diatom assemblages because Arini et al., 2012a,b,c). Under heavy metal stress, translocation-
several multimetric indices of biotic and ecological condition have based studies indicated differential response for the direction of
been developed for bioassessment using relative abundance. translocation. Specifically, in moving from polluted to unpolluted
Furthermore, Schindler (1990) reported that in most field sampling conditions, total diatom density showed rapid changes (in 2
situations, species composition should be more sensitive to weeks), but this change was much slower in the opposite direction,
changes in environmental conditions than changes in biomass or in that the worsening water quality imposed on the very dense
metabolic rates, especially when stresses have existed long enough biofilm matrix from non-polluted conditions took more than 4
for immigration of new species and accrual of rare taxa that are weeks to show clear modifications, despite the presence of con-
stress-tolerant. taminants, the poor conditions of oxygenation, and high levels of
Relative abundance data can become informative through the nutrients (Gold et al., 2002; Duong et al., 2012).
use of ordination, clustering and similarity indices (Stevenson et al., Although diatom densities in biofilms have been widely re-
2010). More specifically, ordination-based multivariate analysis ported as a relevant bioindicator of heavy metal contamination
(e.g., correspondence analysis, detrended correspondence analysis, (Gold et al., 2003a,b; Morin et al., 2008a, 2010; Duong et al., 2010),
non-metric multidimensional scaling, principal component anal- the usefulness as a bioassessment tool has not been consistent and
ysis and distance-based redundancy analysis) was developed to has been criticized. Major reasons for this criticism is not including
relate community composition to measured variation in the envi- biotic variable (e.g., immigration, emigration, grazers etc.)
ronment. Ordination plots obtained in the analyses are linear (Stevenson and Peterson, 1991; Sabater et al., 2002; Arini et al.,
2012b) and abiotic variables (substrates, pH, conductivity, alka- (Hillebrand et al., 1999; Takabayashi et al., 2006). Accurate frustule
linity) (Medley and Clements, 1998; Potapova and Charles, 2005) counts require taxonomic expertise of diatoms, a skill that is often
that can also strongly influence diatom densities. For example, hampered by inadequate or not readily assessable taxonomic re-
heavy metal contamination can indirectly affect diatom cell density sources. Furthermore, small size of diatoms, even of the most
by reducing the density of grazers in contaminated sites, which can common taxa, can be difficult to identify using light microscopy
lead to the unexpected finding in some field studies that higher and hence making the investigation more complicated. Some
diatom cell densities occurred at contaminated sites than at refer- diatom species or individuals may be in valve, girdle, or tilted ori-
ence sites (Guasch et al., 2016). Sabater et al. (2002) suggested the entations when viewed and diatoms in different orientation look
need for closer attention to grazing effects on biofilm function, markedly different from each other, making diatom identification
since grazing can affect the performance of biofilms in the more complicated. Additionally, counts require a lot of time (and,
amelioration of river water quality. Diatom cell density response to hence, expense). More specifically, investigation of diatom com-
pesticide exposure may not be consistent (Debenest et al., 2010). munity biovolume is regularly criticized as it requires time-
For instance, atrazine exposure may decrease cell density in some consuming measurements of diatom frustule dimensions (length,
centric diatom species, and increase cell density for some pennate width and height) followed by estimation of biovolume, using
diatom species (Be rard and Benninghoff, 2001; Be rard et al., 2004). formulae based on generalized diatom shape, which will again
In contrast, another herbicide, isoproturon, consistently impacts increase the time and cost of research.
diatom cell densities (Pe re
s et al., 1996; Schmitt-Jansen and
Altenburger, 2005; Debenest et al., 2009). 2.3. Sensitive and tolerant species
Biovolume estimation of diatom communities has been used for
bioassessment, as an alternative to cell counts for representing the Diatoms inhabit all types of waterbodies and contain troves of
level of stress in diatom biofilms (Ricart et al., 2009; Pandey et al., diversity in terms of taxonomy, morphology and ecology (Larras
2015). Cell biovolume has the advantage of measuring the contri- et al., 2014a). Due their rapid turnover time, diatoms respond
bution of each algal group or taxa relative to primary production quickly and also recovery much faster than other algae (Cattaneo
(Hillebrand et al., 1999; Lavoie et al., 2004; Cloern and Dufford, et al., 2004; Morin et al., 2008a; Stevenson et al., 2010). Different
2005; Larras et al., 2014b). Cell biovolume may have another species of diatoms respond differently and characteristically to
advantage. Under stress, diatom density may increase or be un- environmental (e.g., pH, salinity, ionic content, dissolve organic
changed because of the disproportionate growth of small and carbon; Sabater et al., 2007; Potapova and Charles, 2007) and
adnate diatoms (Medley and Clements, 1998; Gold et al., 2002, anthropogenic (e.g., nutrients, heavy metals, pesticides, herbicides;
2003a,b; Duong et al., 2012; Arini et al., 2012b,c; Morin et al., Kelly and Wilson, 2004; Falasco et al., 2009; Larras et al., 2013)
2008a; Morin et al., 2012; Leguay et al., 2016; Pandey and Bergey, stressors (Fig. S1). Because of these characteristics, diatoms are
2016), which may lead to erroneous findings based on cell used globally for assessing the ecological health of waterbodies.
counts; that is, that the investigated system may not be under any Although individual diatom species may be good indicators of
type of stress. In this case, use of community biovolume would be certain conditions, the genus level is often also useful. For example,
more appropriate for bioassessment than cell counts. As an at the genus level, certain diatoms (Eunotia, Fragilaria, Navicula,
example, Arini et al. (2012b) reported a gradual increase in cell Nitzschia and Pinnularia) can be excellent bioindicators for acid
density with gradual decrease in diatom biovolume of the com- mine drainage (AMD) (DeNicola, 2000; Morin et al., 2012; Pandey
munity at sites heavily contaminated with heavy metals in a lab- and Bergey, 2016). Similarly, Larras et al. (2014a) found diatom
oratory experiment. Similarly, Barral-Fraga et al. (2016) reported sensitivity to herbicides and reported that centric and araphid di-
appreciable reduction in mean (approximately halved) and total atoms (Thalassiosirales and Fragilariales, respectively) included the
diatom biovolume in a diatom dominated biofilm exposed to most sensitive species whereas most resistant species were mainly
arsenic versus the control, which had no apparent difference in pennates (Cymbellales, Naviculales and Bacillariales). Meanwhile,
biofilm cell density. Alternatively, no pattern in density and bio- Van dam et al. (1994) found that the genera Nitzschia and Ach-
volume of the diatom community have been reported under stress nanthidium were characteristic of polluted waters receiving either
(Arini et al., 2012b,c). Chemical exposure can produce similar re- nutrients or organic biodegradable materials, Pinnularia and Cym-
sults. Ricart et al. (2009) reported a concentration dependent bella were found proportionally more numerous in pristine condi-
decrease in the diatom biovolume under exposure to the herbicide tions than polluted ones and the abundant genus Navicula was
diuron, which was associated with the predominance of smaller ambiguous in terms of the water quality.
growth forms in the biofilm (Sabater and Admiraal, 2005). In In contrast, many authors suggest that species-level identifica-
addition, Ricart et al. (2009) found that the EC50 values for the tion of diatoms is adequate for routine biomonitoring (Pe re
s et al.,
biovolume were extremely low, indicating that this was a highly 1996; Guasch et al., 1998; Dorigo et al., 2004; Morin et al., 2009).
sensitive parameter to diuron exposure, and that effects might be Rimet and Bouchez (2012) reviewed the status of diatoms as in-
significant in nature. Furthermore, NEC and EC50 values obtained dicators of river pollution and found that species-level studies were
with the community biovolumes were generally lower than the more common than genus-level studies; indeed, community,
ones obtained using short-term single-species tests, as registered in assemblage, group and taxa level studies were more frequent than
the US EPA database (i.e. Gatidou and Thomaidis 2007 genus level-studies.
(EC50 ¼ 27 mg L1, Navicula forcipata, growth); Ma et al., 2002 Ecological assessments at finer taxonomic levels are very useful.
(EC50 ¼ 4.3 mg L1, Chlorella vulgaris, growth); Podola and For example, the species Eunotia exigua and Achnanthidium minu-
Melkonian 2005 (EC50 ¼ 6.4 mg L1, Cryptomonas sp., chlorophyll- tissium are tolerant of acidic and heavy metal contaminated waters,
a fluorescence). This comparison emphasizes the relevance of us- i.e., occurring at AMD sites (DeNicola, 2000). Similarly, several
ing natural communities to test environmental effects. diatom species are regularly reported in lotic environments
Traditionally used community parameters (e.g., cell density, exposed to toxic events, and these species are considered as in-
biovolume) provide useful information about the nature of com- dicators of organic matter (Palmer, 1969), pesticides (Debenest
munity but there is a major drawback. These parameters are et al., 2010; Rimet and Bouchez, 2011; Roubeix et al., 2011a,b;
investigated by counting 500e1000 diatom frustules in each sam- 2012) and metal pollution. Such tolerant taxa include Achnanthi-
ple. The quality of this well-accepted practice has been criticised dium minutissimum, Fragilaria spp., Nitzschia palea and Ulnaria ulna
(previously Synedra ulna) (Palmer, 1969; Ruggiu et al., 1998; Gold environmental concentrations in lotic mesocosm experiments.
et al., 2002; Passy, 2007; Morin et al., 2008a,b; da Silva et al., Similarly, Roubeix et al. (2012) reported the dominance of pioneer
2009; Debenest et al., 2010; Rimet and Bouchez, 2011; Luís et al., or low profile guilds (with Achnanthidium minutissimum, Cocconeis
2011; Roubeix et al., 2011b, 2012; Morin et al., 2012; Pandey and placentula and Planothidium rostratum) and motile forms or guilds
Bergey, 2016). (Nitzschia palea) in the periphytic diatom communities exposed to
This short literature review illustrates that diatoms show varied metolachlor and acetochlor. In contrast, pesticides, including
response to different types of contaminants. These responses may chloroacetanilides, modify the 3-D structure of more complex
vary depending on the biogeographical region of study (Smol and (multispecies) biofilms by reducing biofilm thickness and the dis-
Glew, 1992), in part because of taxonomic ambiguity of some tribution of the life forms; i.e., replacement of sensitive diatom
widely distributed species (e.g., Achnanthidium minutissimum, species like Achnanthidium minutissimum or Eolimna minima
Nitzschia fonticola, Sellaphora pupula, Caloneis sp., Nitzschia sp. and (pioneer forms or low profile guild) with more tolerant species like
Gomphonema sp.) (Mann and Droop, 1996; Potapova et al., 2004; Nitzschia lanceolata (motile forms) under metolachlor exposure
Trobajo et al., 2006, 2013; Potapova and Hamilton, 2007; Pandey (Roubeix et al., 2011a). Similarly, Ricart et al. (2009) showed that
et al., 2016). Although diatom checklists and taxonomic ratings diuron modified the composition of periphytic diatom commu-
are regionally available for assessing the water quality (especially nities. After 1 month at a concentration of 7 ppm, F. capucina (high
nutrient enrichment) in waterbodies (Palmer, 1969; Van Dam et al., profile) was eliminated, but remained in control conditions (though
1994), no such well-developed resources exist for assessing heavy at very low abundance), the abundance of A. minutissimum (low
metal stress and organic contaminations, which is an urgent need. profile) was reduced and Nitzschia palea (motile guild) appeared
indifferent to the presence of diuron. Recently, Larras et al. (2014b)
2.4. Life-forms and ecological guilds reported that natural seasonal variation of diatom diversity (and so
of sensitivity) significantly affect the community response to
Diatoms have different strategies that resist or tolerate envi- mixture of herbicides, as protective threshold for the herbicides
ronmental stresses, such as grazing, flow disturbance, and nutrient mixture obtained in the winter community was not found same for
resource limitation, which may differentially affect different life- the summer community based on their structural parameters.
forms: benthic, planktonic, mobility, colonial, tube-dwelling, Thus, a significant difference was examined in terms of diatom
stalked and adnate. Additionally, some species can be considered community shift under heavy metal stress and organic contami-
pioneers (early colonists), whereas others may be late successional nation (herbicides or pesticides) namely, heavy metal stress
species (Rimet and Bouchez, 2011). In contrast to these life forms, resulted in more pioneer life forms or low profile guilds whereas
ecological guilds comprises group of species that live in same organic contamination promoted motile forms.
environment and exploit the same resources but are adapted in Recognizing the roles of both life-forms and ecological guilds
different ways to abiotic factors (Passy, 2007; Rimet and Bouchez, within biofilms enables better understanding of species-
2011; B-Be res et al., 2014). These biological traits provide addi- environment relationships and interactions (Passy, 2007; Passy
tional information about the structure and architecture of biofilms. and Larson, 2011; Berthon et al., 2011; Rimet and Bouchez, 2011;
Sporadic attempts have been made to explore life history char- B-Beres et al., 2014). The major advantage of incorporating one or
acteristics and morphological growth forms of diatoms against both biological metrics into ecotoxicological studies is their ease of
natural (Passy and Larson, 2011; Gottschalk and Kahlert, 2012; use because there are so few categories in each metric. With few
Stenger-Kova cs et al., 2013; B-Be
res et al., 2014; Svensson et al., categories and application to all biofilms, these biological metrics
2014) and anthropogenic (Passy, 2007; Berthon et al., 2011; Larras often display robust and predictable responses to ecological gra-
et al., 2014b) disturbances in fluvial ecosystems. The need to dients. Biological metrics are often characteristic of the genus level
incorporate these parameters in the ecotoxicological studies has for diatoms and therefore are easy to use for routine monitoring
been regularly advocated (Medley and Clements, 1998; Rimet and purposes. These diatom metrics transform complex ecological in-
Bouchez, 2011; Elias et al., 2015). Physico-chemical changes formation at species level into a clearer dataset with more robust
within fluvial biofilms produce different successional patterns. trends and more easily testable hypotheses (Potapova and Charles,
According to Passy and Larson (2011) algal colonization or succes- 2007; Rimet and Bouchez, 2011; Rimet and Bouchez, 2012). Thus,
sion is result of stresses; driven primarily by nutrient supply and biological metrics should be incorporated into the suite of diatom
secondarily by current velocity of laboratory stream. On the other metrics used in effective water quality bioassessment in rivers.
hand, anthropogenic disturbances also have significant effects in Indeed, Medley and Clements (1998) suggested incorporating
terms of changing the successional patterns of diatom species in periphyton community responses to pollutants in terms of life-
the community, especially under heavy metal contamination. For history traits, ecological strategies, and morphological forms as a
example, Medley and Clements (1998) found that small and adnate tool for water quality assessment.
(pioneer) diatom species, such as Achnanthidium minutissimum and
Fragilaria vaucheriae, dominate in the streams polluted with Zn. 3. New endpoints
Field studies of periphytic diatom communities collected from acid
mine drainage areas regularly show the dominance of pioneer 3.1. Alterations in cell integrity
diatom taxa, especially Achnanthidium minutissimum (Cattaneo
et al., 2004; Lavoie et al., 2012; Cantonati et al., 2014; Pandey 3.1.1. Nuclear anomalies
et al., 2016; Luís et al., 2011; Morin et al., 2012). Pandey et al. According to Bidle and Falkowski (2004), planktonic photosyn-
(2015) reported dominance of pioneer forms in lab cultured thetic microorganisms undergo programmed cell death (PCD) in
diatom communities under Cu and Zn stress (100 ppm). In com- response to environmental stress. Chromatin condensation,
parison to the heavy metal stress, diatom biological metrices were migration to the edge of the nuclear membrane and fragmentation
more studied under organic contamination (such as, diuron, are the chief morphological markers of PCD (Bidle and Falkowski,
metolachlor, acetochlor, azoxystrobin and tebuconazole). For 2004). Very few reports indicate how intracellular organelles,
example, Rimet and Bouchez (2011) reported dominance of motile, more specifically the nucleus of live diatoms, react to different
low-profile and mucous tube forms of diatoms under herbicides environmental and anthropogenic disturbances (Coombs et al.,
(diuron) and fungicides (azoxystrobin, tebuconazole) exposure at 1968; Casotti et al., 2005). To the best of our knowledge, Desai
et al. (2006) is the first report of genotoxic effects of metals in di- Nitzschia palea after Cr (IV) treatment (Fig. 1). Investigation of nu-
atoms. Using comet assays on Chaetoceros tenuissimus, the authors clear abnormalities in diatoms reveals the effects of pollutants
found that an elevated cadmium level damaged the nuclear ma- (metals and herbicides) on the microproducers forming the basal
terial in cells and that the degree of injury increased with pro- trophic level of in fluvial ecosystems, which helps in understanding
gressive exposure. Debenest et al. (2008) studied the effect of the mechanisms of change in biofilm composition, especially
herbicide maleic hydrazide on a freshwater benthic diatom com- increasing dominance of the most tolerant diatom species, as well
munity in the laboratory by exposing the community to three factors that might lead to morphological anomalies in diatom
herbicide concentrations, using an exposure time of 6 h, followed frustules (Debenest et al., 2008; Licursi and Go mez, 2013).
by a 24 h recovery period. After the maleic hydrazide treatment, a
dose-dependent increase in nuclear alterations was observed
(abnormal nucleus location, micronucleus, multinuclear cell or 3.1.2. Alteration in the cell membrane and cytoplasmic content
disruption of the nuclear membrane) and the difference between Diatoms are well known for their transparent silica frustules
the control and highest dose of herbicide-treated samples was and, as a result, it is very easy to observe the intracellular changes
mez (2013)
statistically significant (Fig. 1). Similarly, Licursi and Go (in chloroplast, cytoplasm, lipid bodies and vacuoles) under light
reported alteration in nuclear region (abnormal nucleus location microscopy. Alterations of cell membrane of diatoms are used to
and nuclear membrane damage) in the epipsammic diatoms Fal- determine whether the examined cell is alive or dead (Radchenko
lacia pygmaea and Navicula novaesiberica after 7 days of Cr (IV) and Il'yash, 2006; Manoylov, 2014). Morin et al. (2012) reported
(chromium) treatment, but the difference was not statistically that metal toxicity on diatoms is linked to different steps in the
significant after 3 days of Cr treatment. Effects of chromium circulation of the toxicant across the membrane (especially uptake
exposure differ among taxa, as no nuclear alterations were found in mechanisms) and inside the cell, inducing perturbations in the
normal functioning of structural/functional intracellular
Fig. 1. Alterations in cell integrity as nuclear anomalies. The effect of maleic hydrazide (MH) on the diatom genera (Nitzschia (a), Gomphonema (b) and Navicula (c)) with normal (N),
abnormal nuclear location (ANL), fragmented nucleus (FN), micronucleus (MN) and broken nucleus membrane (BNM) (nucleus stained in blue with Hoescht 33342, chloroplasts
appear in red). Scale bar- 10 mm. Source- Debenest et al. (2008). The effect of hexavalent chromium on the cells of Fallacia pymaea (d) and Navicula novaesiberica (e) with normal (N),
abnormal nuclear location (ANL), fragmented nucleus (FN), micronucleus (MN) and broken nucleus membrane (BNM) (nucleus stained in blue with Hoescht 33342, chloroplasts
appear in red). Scale bar- 10 mm. Source-Licursi and Go mez (2013). (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of
this article.)
components. Percent alteration in cell membranes was calculated permeability, photosynthetic activity, pigmentation and growth. In
by measuring the changes in the perimeter of the cell membrane this study, fully disintegrated cell membranes were considered
with respect to the control specimens by using Motic (Motic, BML non-viable. Wei et al. (2014) found that a large blank space
series, Hong Kong) or Image J software (Fig. 2). Alterations was appeared between the plasma membrane and the cell wall in Cu
rated on the scale: 0% denoted intact cell membrane state, 25, 50, treated cells (Phaeodactylum tricornutum), indicating that the
75, and 100% denoted altered membrane state in less than one- plasma membrane was damaged and separated from the cell wall.
quarter, a quarter to a half, a half to three-quarters, and more Compared to the control, the mitochondria and nuclei in the Cu-
than three-quarters of the live diatom cells, respectively. Franklin treated cells were swollen and their membranes appeared
et al. (2006) reviewed the status of programmed cell death in smeared. More recently, Morin et al. (2016) found that in the lower
phytoplankton ecology and advocated the use of cell viability as a Matanza-Riachuelo basin (Argentina), strong symptoms of eutro-
health indicator. Membrane integrity indicates the viability of live phication and high amounts of organic matter and heavy metals
diatom cells and, as such, can be used to indicate health in studies can be diagnosed through high percentages of cytological abnor-
of species succession and food-web structure (Veldhuis et al., malities and thus advocated the need for incorporating cytological
2001). Drum and Hopkin (1966) studied the degree to which abnormalities (cytoplasmic content impaired) in diatoms for the
plasmolysis in the anterior and posterior ends of the protoplast detection of changes of the water quality associated with toxic
affect cell adhesion and movement by the diatom Pleurosigma pollution.
angulatum. Bidle and Falkowski (2004) reviewed the status of cell This testing methodology of using cell membrane and other
death in planktonic and photosynthetic microorganisms and visible cytological features to relate diatom health and, possibly, the
concluded that under adverse environmental conditions (nutrient ecological status of fluvial ecosystems with the effects of toxins is
deprivation, intense light, excessive salt concentrations or oxidative relatively quick and takes little expertise in diatom taxonomy
stress), a wide variety of phytoplankton undergo apoptosis (pro- (Pandey et al., 2014). Cytoplasmic alterations or autolysis of cyto-
grammed cell death), with early symptoms including cell plasmic content (a viability assay) (comparing images of controls
shrinkage. Chang et al. (2011) quantified cell integrity of the diatom and treated samples; treated samples showed contracted or irreg-
Nitzschia palea and the cyanobacterium Microcystis aeruginosa ular shapes compared to the control) adds a new and important
before and after b-cyclocitral addition and they found that cells of approach in addressing the physiological condition within a single
N. palea ruptured at a much lower concentration (5e10 ppm) of b- population and thereby provides information on the health status
cyclocitral than in M. aeruginosa (200e1000 ppm). Armbrecht et al. of diatom communities, which can be a tool for both ecotoxico-
(2014) studied cell death and aggregate formation in a giant diatom logical and bioassessment studies relating to diatom biology.
Coscinodiscus wailesii and reported autolysis in the cytoplasm of
C. wailesii under Si and N deficiency. In a laboratory experiment 3.1.3. Alteration in chloroplasts (shape, size, color and number)
with Ditylum brightwelli and Chaetoceros calcitrans, Veldhuis et al. In diatoms, chloroplasts intactness is rarely considered as a tool
(2001) found correlations among changes in membrane for ecological bioassessment or for ecotoxicological assessments
Fig. 2. Alteration in the cell membrane and cytoplamic content. (a) Percent decrease in the photosynthetic apparatus (PDPA) of live Navicula, (b) Percent decrease in the photo-
synthetic apparatus (PDPA) of live Amphora under Cu stress under laboratory conditions. Scale bar- 8 mm (The results presented were obtained by the authors of the present ms).
(Arini et al., 2012b,c). Sporadic attempts have been made to same methodology (intactness of chloroplast),Wood et al. (2016)
establish live (with intact chloroplast) and dead (with empty reported that natural benthic diatom communities respond to
frustules) diatoms (Fig. 2) as a tool for bioassessment and the eight common herbicides (atrazine, simazine, hexazinone, tebu-
ecological health of waterbodies (Gillett et al., 2009, 2011). Gener- thiuron, diuron, MCPA, 2,4-D and glyphosate) in which the most
ally, smaller chloroplast volume and less thylakoid surface density sensitive taxa were Gomphonema spp., Encyonema gracilis and
were observed in diatoms under light and nutrient stress (Rosen Navicula cryptotenella was the most tolerant to herbicide exposure.
and Lowe, 1984; Janssen et al., 2001). Lommer et al. (2012) re- There was no significant effect of the different herbicide modes of
ported shrinkage of the chloroplast system in iron-limited Tha- action at the community level. In several studies, alterations in the
lassiosira oceanica cells. They found that Fe limited cells had a diatom chloroplast morphology was clearly observed under
reduced number of two chloroplasts, instead of the normal four. mez, 2013;
different stresses, such as, heavy metals (Licursi and Go
Renzi et al. (2014) also reported alteration in the photosynthetic Pandey and Bergey, 2016) and herbicides (Debenest et al., 2008).
complex (by using confocal and optical microscopy along with cell Alteration in diatoms chloroplast morphology (shape, size and
fluorescence) as a sensitive and powerful early warning tool for number) is a rapid and simple toxicity testing method to obtain
evaluating sub-lethal effects of Cu, Zn and methylene blue active sensitivity data for multiple taxa within a natural benthic diatom
substance (MBAS). Arini et al. (2012b,c) reported that under Cd and community in a relatively short period of time, warranting further
Zn toxicity showed no significant effect on the health of diatom development of chloroplast metrics as an assessment tool. Note,
cells assessed in terms of presence and absence of chloroplasts. however, that the number and shape of chloroplasts may vary be-
However, under organic contamination, the effect on chloroplast tween species.
was more apparent. For example, Wood et al. (2014) developed an
excellent toxicity test for diatom cells by using the intactness of
stained chloroplasts for assessing the toxicity of atrazine over a 3.2. Lipid bodies
period of 48 h. Cells were classified depending on the condition of
the stained chloroplast i.e., if it appeared more than 50% intact then Different classes of algae are characterized by the type of food
it was classed as a healthy cell, and if the chloroplast was <50% reserve stored inside their cells (Barsanti et al., 2008). Diatoms
intact or absent then it was classed as unhealthy. In this study they (Bacillariophyceae) are unique in storing lipids, which occur in
found that exposure to their highest treatment of atrazine intracellular lipid bodies (LBs), as a reserve food material (Figs. 3
(500 mg L1) produced significant declines in healthy cells of the and 4).
most sensitive genera: Gomphonema declined by 74%, Amphora by In diatoms, LBs become more prominent (in terms of number as
62%, Cymbella by 54% and Ulnaria by 34%, compared to control well as in size) under various types of environmental and anthro-
levels. In contrast, the genera, Eunotia, Achnanthidium and Navicula, pogenic perturbations (Ramachandra et al., 2009; Hildebrand et al.,
had no statistically significant decline in cell health. By using the 2012) (Tables 1 and 2). Increased oil reserves may aid cell survival
during unfavorable conditions. In an early observation, Evans
Fig. 3. Lipid bodies visualization in the periphytic community dominated with diatoms and stained with BODIPY fluorescent dye. Green fluorescence is BODIPY staining of neutral
lipids, red is chlorophyll fluorescence. Scale bar- 10 mm. (The results presented were obtained by the authors of the present ms). (For interpretation of the references to colour in this
figure legend, the reader is referred to the web version of this article.)
Fig. 4. Light micrographs of lipid bodies in 29 diatom species. (1) Navicula veneta, (2) N. gregaria, (3) N. decussis, (4) N. schoenfeldii, (5) N. capitatoradiata, (6) N. cincta, (7) N.
accomoda, (8) N. gregaria, (9) N. phyllepta, (10) Craticula cuspidata, (11) Gyrosigma exilis, (12) G. nodiferum, (13) Navicula incertata, (14) Achnanthidium exiguum, (15) Pinnularia
subcapitata, (16) Tryblionella debilis, (17) Cocconeis placentula, (18) Nitzschia palea, (19) N. linearis, (20) N. amphibia, (21) N. sigmoidea, (22) N. sigmoidea, (23) Bacillaria paradoxa, (24)
Nitzschia intermedia, (25) N. frustulum, (26) N. filiformis, (27) N. inconspicua, (28) Tryblionella apiculata, (29) Plagiotropis lepidotropis, (30) Haslea ostrearia, (31) Bacillaria paradoxa and
(32) Gyrosigma spencerii. Scale bar-5mm. (The results presented were obtained by the authors of the present ms).
(1960) reported that Pinnularia biceps f. minutissima cells along et al., 2012; Frada et al., 2013). In the same context, Julius and
pond edges that had large oil globules survived desiccation better Theriot (2007) reported that under environmental stress, LB in-
than cells with fewer lipids. Cells lacking oil globules sometimes duction in diatom cells is associated with modification in chloro-
plasmolysed and were unable to recover when put back into a plast number. They also proposed that the chlorophyll to lipid ratio
liquid medium. could be used as a tool to measure the health of diatom cells.
LB inductions are apparent under nitrogen starvation (Jiang In contrast to the effects of nutrients, the effects of heavy metal
et al., 2012; Guerra et al., 2013), which may also lead to a lower stress on LBs is relatively unexplored (Pandey et al., 2015). Joux-
growth rate, an association meaning that reducing nitrogen may Arab et al. (2000) reported higher lipid content in the diatom
not be a good mechanism for increasing lipid yield (Hildebrand Haslea ostrearia under Cu stress in comparison to the control
Table 1
Biovolume and number of lipid bodies in five diatom species exposed to metals (Cu and Zn; 100 mg L1) under laboratory conditions. See Fig. 7 (14, 1, 6, 20 and 19). Source:
Pandey et al., 2015.
Diatom species *Biovolume (mm3 Cell1) of lipid bodies (number of lipid bodies) % contribution of lipid bodies to total cell volume
under metal stress
Control Cu Zn Control Cu Zn
Achnanthidium exiguum 2* (1) 6* (2)e23(2) 4* (1)e16(2) 7 20e77 13e53

Navicula gregaria 4 (2) 20(4)e90(4) 13(3)e68(4) 2 9e41 6e31
Navicula recens 3 (2) 12(2)e67(2) 12(2)e61(2) 1.5 6e34 6e31
Nitzschia amphibia 4 (2) 52(6)e165(6) 45(4)e145(4) 1.2 16e50 14e44
Nitzschia linearis 4 (0) 16(6)e126(5) 16(5)e126(5) 1.3 5e40 5e40
Table 2
Number of lipid bodies and % biovolume contribution of lipid bodies per cell in three common diatom species examined at the metalliferous sites (Khetri and Zawar) of
Rajasthan, India. See Fig. 7 (15, 19, 21 and 22). Source: Pandey and Bergey, 2016.
Diatom species No. of lipid bodies (diameter in mm) % contribution of lipid bodies to total cell volume
under metal stress
Control Metal stress (Cu, Zn) Control Metal stress (Cu, Zn)
Pinnularia subcapitata 2 (0.1e0.5) 4 (1e3) 2e5 15e60

Nitzschia linearis 4 (0.1e0.32) 4 (1e2.8) 4e8 14e43
Nitzschia sigmoidea 5 (0.1e0.2) 10 (1e2) 2e5 12e25
treatment in a laboratory study, but the difference was not statis- only an improved energy capturing capacity of diatoms, but can
tically significant. Similarly, Lelong et al. (2013) described increased also act as a buoyancy regulator, as a reserve food material
lipid content in Cu-starved cells of Pseudo-nitzschia, but no effect (Smetacek, 2001), an aid in diatom movement (Wang et al., 2013),
was observed in severe Fe-limited Pseudo-nitzschia cells. In reservoir for toxicants and as an energy storing body that can be
contrast, mild Fe limitation produced a 100% increase in lipid utilized in recovering from different types of environmental and
content but under Cu and Fe co-limitation, lipid content (per cell anthropogenic stresses. Lipid bodies and their induction in living
and per unit biovolume) decreased slightly. More recently, Pandey diatoms form a very promising basis for easy and rapid ecological
et al. (2015) reported induction of LBs (increased number and size) assessments as well as for biomonitoring of fluvial ecosystem.
in a lab cultured phytoplanktonic community under Cu and Zn
stress, which was also associated with deformities in the silica 3.3. Alteration in frustule size
frustules of live diatoms. Similarly, Pandey and Bergey (2016) re-
ported higher LBs (number and biovolume) at mining sites severely Diatoms are well known for their robust, species-specific,
polluted with Cu and Zn. The number and size of LBs have also been ornamented silica frustules, which are dependably replicated
explored as a stress indicator in other algal classes, such as cya- from generation to generation (Falasco et al., 2009). However, these
nobacteria (Peramuna and Summers, 2014), green algae (Andrade silica frustules are occasionally susceptible to alteration under
et al., 2004; Liu et al., 2008; Wang et al., 2009) and di- various types of environmental (Winder et al., 2009; Svensson
noflagellates (Weng et al., 2014). et al., 2014) and anthropogenic stress. These alterations include
The mechanism causing induction of lipid bodies in diatoms is the production of various morphological forms (phenotypic plas-
not yet known. LB formation is associated with the degradation or ticity), size reduction and even frustule deformation (Kociolek and
alteration of the photosynthetic apparatus of diatoms (Julius and Stoermer, 2010; Cox et al., 2012; Morin et al., 2012). According to
Theriot, 2007; Lommer et al., 2012; d’Ippolito et al., 2015). Lipid Kociolek and Stoermer (2010), variation in diatom cell wall
bodies may also be a site for accumulating heavy metals (Pb and Cu) morphology can be either size-dependent or size-free. In the same
by forming lipid complexes with the toxicants (Lombardi and context, Snoeijs et al. (2002) and Busse and Snoeijs (2003) found
Vieira, 1999, 2000). Lipids are used as both a reserve food mate- that small and large diatom species residing in the same benthic
rial and as a buoyancy organelle; buoyancy that counters the heavy community respond differently to environmental variation (Si,
weight of the silica frustules and aids in the maintenance of a depth nutrients, phosphate, salinity and wave action). Size reduction is a
in the water column appropriate for accessing light and nutrients. normal feature of diatoms due to their vegetative mode of repro-
Buoyancy due to lipid bodies has also been reported in the plank- duction (Laney et al., 2012), in which one cell of the original size and
tonic green alga Botryococcus floating on the surface of a small lake a slightly smaller cell are produced during each cell division.
(Belcher, 1968). Variation in size may be adaptive or circumstantial (Kociolek and
Induction of lipid bodies in diatoms apparently has multiple Stoermer, 2010). An example of an environmental effect was re-
functions, including use in combating stress. Walsby and Reynolds ported by Trobajo et al. (2011), who found that saline concentration
(1980) concluded that the reduction in density due to intracellular significantly affected diatom width (and stria density) in Nitzschia
lipid accumulation contributes to a reduced rate of sinking in di- frustulum and N. pusilla, but had no effect on valve length in any of
atoms, but they are also of the view that buoyancy-regulation is not the five studied species.
the primary function of lipid bodies. Significant induction of lipids Heavy metal enrichment is a major factor intensifying the
(40% of the cell volume) inside live diatom cells can indicate its poor naturally occurring phenomenon of size reduction in diatom frus-
health i.e., increase in LB volume with decrease in chloroplast tules (Fig. 5; Table 3) and can result in significant reduction in
volume (Liang et al., 2015; Gautam et al., 2017). Alternatively, Wang diatom size (Morin et al., 2012; Cantonati et al., 2014). Cattaneo
et al. (2013) reported that induction of LBs in live benthic diatoms et al. (2004) reported significant size reduction in four diatom
assisted their movement. Thus, lipid body induction indicates not species (Achnanthidim minutissimum, Asterionella formosa,
Fig. 5. Photographic documentation of alteration in frustule size in 14 commonly occurring diatom species under different environmental and anthropogenic disturbances. (1)
Navicula salinarium, (2) Navicula recens, (3) Diploneis interrupta, (4) Surirella stalagama, (5) Tryblionella coarctata, (6) Diploneis elliptica, (7) Surirella robusta, (8) Surirella gemma, (9)
Meloneis mimallis var. mimallis, (10) Encyonema sp., (11) Amphora sp., (12) Cocconeis placentula, (13) Meloneis akytos and (14) M. mimallis var. zephyria. Scale bar- 5 mm (The results
presented were obtained by the authors of the present ms).
Table 3
Investigation of % change in cell length in 14 different diatom species under environmental and anthropogenic disturbances. The diatom samples were collected from
various contaminated (nutrient and heavy metals) waterbodies of Korea during 2014e2016. Percent change in length is percent loss of length relative to the longest cell
measured.
Diatom species No. of frustules examined % change in length
1. Cocconeis placentula 50 0e25

2. Diploneis elliptica 50 0e85
3. Diploneis interrupta 50 5e58
4. Encyonema minutum 50 0e55
5. Meloneis akytos 50 5e80
6. Meloneis gorgis 50 0e90
7. Meloneis mimallis var. mimallis 50 10e80
8. Meloneis mimallis var. zephyria 50 0e10
9. Navicula recens 50 15e55
10. Navicula salinarium 50 10e60
11. Surirella gemma 50 5e65
12. Surirella robusta 50 0e40
13. Surirella stalagma 50 5e65
14. Tryblionella coarctata 50 0e60
Brachysira vitrea and Tabellaria flocculosa) collected from the heavy of diatom cells to become longer. These changes could be explained
metal (Cd, Cu, Fe, Pb and Zn) polluted sediments of a watershed of by an absence of cytokinesis that causes individual T. pseudonana
Lac Dufault (Que bec, Canada) and Morin and Coste (2006) reported cells to elongate, accumulate more plastids and become polyploid.
significant reduction in two abundant diatom species, Gompho- Frustule morphology may differ strikingly between the largest
nema parvulum and Nitzschia palea, collected from metal (Cd and and smallest cells in a size series of a single species, or even a single
Zn) polluted sites (Joanis and Usine) of Riou Mort and Riou Viou clone (Cox et al., 2012), which is a challenge to community analysis
streams of South West France. Similarly, Luís et al. (2011) re- based on diatom size. Furthermore, size ambiguity in diatoms il-
ported > 60% size reduction in Brachysira vitrea at the abandoned lustrates that molecular bioassessment simply based on a DNA code
mining area in Portugal, which is highly contaminated with heavy may be inadequate to describe diatoms because the expression of
metals (in both sediments and surface water). Barral-Fraga et al. proteins depends on the unique set of environmental conditions in
(2016) reported significant size reduction in different diatom spe- which algae live and affects diatom shapes, adaptations, and sur-
cies (Amphipleura pellucida, Nitzschia dissipata, Nitzschia fonticola vival (Will and Rubinoff, 2004; Sluys, 2013). In addition, the identity
and Nitzschia palea) treated with arsenic in a laboratory of most diatom taxa in the reference libraries of genetic sequences
experiment. has not been rigorously evaluated, so the reference library taxon-
The exact mechanism of intensified size reduction in diatom omy may not be accurate for molecular bioassessment (Manoylov,
frustules is uncertain, but according to Morin et al. (2012), size 2014).
reduction can be the result of a higher cell division rate that is
inherent to organisms inhabiting stressed ecosystems. Climate 3.4. Morphological deformities
warming in conjunction with increased organic loads could also
facilitate increased phosphate availability (Wilhelm and Adrian, Morphological abnormalities in diatom frustules are globally
2008), that could result in a decrease in dissolved Si:P and assessed under various environmental (aging, crowding, tides,
decrease in light availability, potential favouring smaller diatoms, mechanical pressure, low current velocity, drought, light intensity,
especially in late spring and summer (Kilham et al., 1996; Finkel temperature, moisture, salinity, pH, long-term culture, Si and nu-
et al., 2009). The phenomenon of significant reduction in diatom trients) and anthropogenic (heavy metals, herbicides and pesti-
frustule size has potential use in ecotoxicological studies, but cides) disturbances (Falasco et al., 2009) (Figs. 6 and 7). The
further work is needed. Size reduction occurs more frequently than proportion of valve abnormalities was inferred directly from the
cell deformity (see next section), but it is difficult to segregate the taxonomical counts, by determining the percentage of individuals
effects of stress-related reduction from the natural tendency of that had unusual shape and/or ornamentation of the frustule. De-
diatom populations to get smaller and smaller with successive cell formations in diatoms were first correlated with physical variables
divisions. Deformity and size reduction in diatoms often occur such as temperature, current velocity, flow and rainfall, rather than
together and if we able to establish that significant size reduction is with chemical variables (Go mez and Licursi, 2003). Recent studies
due to stress, as has been done previously (Cattaneo et al., 2004), reported significant relationships between anthropogenic stresses
then size reduction might prove to be a valuable biomonitoring (herbicides; Roubeix et al., 2011b; heavy metals; Pandey et al.,
tool. 2014) and abnormalities in diatom frustules. For example, under
Larger cell size in diatoms has been reported under different natural conditions, the percent of deformed frustules was signifi-
types of stress, such as light (Olson et al., 1986), temperature cantly lower (0.35%) than under heavy metal stress (1.0e4.0%)
(Montagnes and Franklin, 2001), long-term culturing (Rose and (Falasco et al., 2009; Morin et al., 2012; Pandey et al., 2014),
Cox, 2013) and heavy metals (Stauber and Florence, 1987; Levy whereas organic contamination was associated with an interme-
et al., 2008), although some studies do not indicate a clear size diate deformity percent of between 0.32 and 1.5% (Morin et al.,
trend (Montagnes and Franklin, 2001; Levy et al., 2007). Greater cell 2009; Debenest et al., 2008; Roubeix et al., 2011a).
size has been reported mainly in marine centric diatoms (Aulaco- Deformities in diatoms are often not associated with species
seira sp. Biddulphia aurita, Cosinodiscus sp., Lauderia borealis, P. tri- that dominant under various anthropogenic stresses. For example,
cornutum, Skeletonema costatum, Stephanopyxis turris, Thalassiosira Roubeix et al. (2011a) reported a higher percent of deformity in a
weissflogii, and T. psuedonana). In converse, pennate diatoms in any sub-dominant diatom species, i.e., Surirella angusta under diuron
aquatic ecosystem are rarely reported as having larger than ex- exposure. Similarly, Duong et al. (2008) reported deformities in the
pected cell size (Reavie and Barbiero, 2013). diatom species Fragilaria capucina, Gomphonema parvulum and
Diatom size and changes in size (and surface area) might be Ulnaria ulna, which was numerically average or sub-dominant in
associated to nutrient absorption, especially in nutrient-limited abundance. Dziengo-Czaja et al. (2008) also reported deformities in
(oligotrophic) environments. Increasing frustules length or diam- diatom species that were not prevalent in the nutrient (phosphorus
eter is also considered an adaptation to reduce sinking rates and nitrite) enriched (organic matters) habitat. In contrast, de-
(Miklasz and Denny, 2010), which may be enhanced by a longer ice- formities in dominant diatom species (Achnanthes minutissima and
free season and stronger late-summer stratification. According to Brachysira vitrea) occurred in waterbodies contaminated with
Kerrigan et al. (2015), climate warming and increases in the dura- heavy metals (AMD sites) (Cattaneo et al., 1998, 2004; Luís et al.,
tion of the growing season and stratified period may allow greater 2011).
accumulation of nutrients during transient mixing events in the Araphid and monoraphid forms are more susceptible to defor-
spring and fall overturns (Kilham et al., 1996), which may perhaps mation than other morphological forms. For example, in the diatom
favor larger-sized diatom species in the community. The relation- genera Achnanthidium (Achnanthidium minutissimum), Fragilaria
ship between cell size and genome size is of significant importance (Fragilaria capucina and F. rumpens) and Ulnaria (Ulnaria ulna),
in diatoms. For example, Koester et al. (2010) examined two iso- deformed frustules have been associated with various types of
lated population of Ditylum brightwellii from New Zealand and anthropogenic perturbations (Gold et al., 2002, 2003b; Nunes et al.,
found that their increased sizes are directly related with amount of 2003; Duong et al., 2008; Morin et al., 2008a; Roubeix et al., 2011b;
DNA content in the cells. Similarly, Van Tol et al. (2016) recently Lavoie et al., 2012; Cantonati et al., 2014; Pandey et al., 2014, 2015;
reported a symbiotic association between bacteria (Croceibacter Pandey and Bergey, 2016). In contrast, Morin et al. (2008b) reported
atlanticus) and the marine diatom Thalassiosira pseudonana, which more deformities in raphid forms (66%) than araphid ones (33%) in
resulted in an inhibition of cell division and induces the mean size a freshwater biofilm exposed to Cd. However, under herbicide
Fig. 6. Morphological deformities observed in living diatom frustules examined under different types of environmental and anthropogenic perturbations. (1) Navicula veneta, (2)
Nitzschia linearis, (3) Nitzschia amphibia, (4) Fragilaria capucina, (5) Diatoma vulgare, (6) Diatoma vulgare, (7) Nitzschia filiformis, (8) Gomphonema pseudoaugur, (9) Fragilaria capucina
and (10) Ulnaria ulna. First frustules in each species is normal while rest ones are deformed. Scale bar- 8 mm. Source- Pandey et al., 2015; Pandey and Bergey, 2016; Gautam et al.,
2017.
contamination, deformities were equally shared mainly between addition, permanent slide preparation results in an inability to
raphid and araphid forms (Debenest et al., 2008; Roubeix et al., examine all sides of individual diatoms for deformities. Examining
2011b). all four views (two valve and two girdle views) of diatom frustules
The relationship of diatom size to deformity has been debated. would almost certainly increase the percent of deformed cells
Morphological abnormalities are often missed in small diatoms counted in samples with deformities.
when diatoms are viewed with light microscopy (Morin et al., In diatoms, deformities are quantified as percentages by
2008c; Manoylov, 2014). Studies using scanning electron micro- counting 500 or 1000 frustules in the community. In order to make
scopy indicate that small diatoms are as susceptible to deformities deformities more informative, Falasco et al. (2009) broadly classi-
as larger species (Morin et al., 2008c). However, according to Lavoie fied deformities into 8 types, a classification refined by other re-
et al. (2012), the lower percentage of deformed frustules in the searchers (Arini et al., 2012b,c; Pandey et al., 2014, 2015),
small diatom Achnanthidium minutissimum may signify that small culminating into four types i.e., deformities in valve (type I), striae
species are less susceptible to morphological deformations, (type II), raphe (type III) and mixed types (type IV) (Table S1). All
although Cantonati et al. (2014) extensively studied (and found) four types of deformity were examined under heavy metal
deformities in this species from 8 different sites in Europe and contamination by Cu and Zn in the field and laboratory conditions
Canada. Furthermore, the length/width ratio of diatom species also (Pandey et al., 2014, 2015) (Table 4). Reports of deformities in di-
plays a role, especially in araphid forms, as species with higher atoms exposed to heavy metal pollution indicate primarily of Type
length/width ratios are generally more prone to deformation under mez and Licursi
1 (valve outline) and Type 2 (striae) deformities. Go
stress than other shapes of diatom frustules. (2003) reported the occurrence of all four types of deformities
Another issue to consider is the low percentage of deformed (valve, striations, raphe and mixed) in the diatom species Pinnularia
frustules even under stress conditions, which is the major concern gibba in a periphytic biofilm in an area with Cu and Zn contami-
in assessing the use of deformities as a biomonitoring tool. Incor- nation. Arini et al. (2012a,b) similarly reported prevalence of
rect identification of diatom species and the lack of proper training deformed valves (type 1), deformed striations (type 2) and mixed
to discriminate deformed frustules from normal ones are main deformities (type 4) in the periphytic diatom community contam-
reasons for lower counts of deformity in diatom assemblages. In inated with Cd and Zn, and that type 3 (deformed raphe)
Fig. 7. Normal (first frustules) and Deformed frustules in 29 diatom species examined under different types of environmental and anthropogenic purturbations. (1) Achnanthidium
breviceps, (2) Mastogloia smithii, (3) Luticola muticopsis, (4) Nitzschia compressa, (5) Planothidium frequentissimum, (6) Diatoma vulgare, (7) Diatoma vulgare, (8) Diatoma sp., (9)
Tryblionella apiculata, (10) Cymbella tumida, (11) Cocconeis placentula, (12) Eolimna subminuscula, (13) Achnanthidium exiguum, (14) Fragilaria sp., (15) Tabularia fasciculata, (16)
Fragilaria capucina, (17) Nitzschia clausii, (18) Ulnaria ulna, (19) Fragilariforma bicapitata, (20) Brachysira microcephala, (21) Caloneis bacillum, (22) Caloneis bacillum (Girdle view), (23)
Nitzschia palea, (24) Ulnaria ulna, (25) Cymbella turgida, (26) Gomphonema pseudoaugur, (27) Ulnaria ulna, (28) Brachysira brebissonii and (29) Gomphonema parvulum. First frustules
in each species is normal while rest ones are defromed. Scale bar- 7 mm (The results presented were obtained by the authors of the present ms).
Table 4 deformities were present in low percentages. In field studies,

Relative proportions (%) of various deformities examined under in situ Pandey et al. (2014) and Pandey and Bergey (2016) found that
(45e165 mg cm2 d1 for Cu; 42e150 mg cm2 d1 for Zn in 14 days) and laboratory
deformed raphes (type 3) were more prevalent under Cu stress
(100 mg L1 after 7 days) condition under Cu and Zn stress (separately) in the
periphytic diatoms (Pandey et al., 2014, 2015; Pandey and Bergey, 2016). than other deformities, but that deformed striations (type 2) and
mixed deformities (type 4) were more prevalent under Zn stress.
In situ Lab
Laboratory experiments using planktonic communities produced
Control Stressed Control Stressed similar results [i.e., Cu stress lead to higher prevalence of deformed
Total deformity (%) 0.1 3.0 2 10 raphes (type 3), whereas Zn exposure resulted in a higher preva-
Type 1 (Valve) 70 76 80 40 lence of deformed striae (type 2)]. Arini et al. (2013) ran an
Type 2 (Striation) 30 5 20 20 experiment that described the development and persistence of
Type 3 (Raphe) 0 17 0 20
Type 4 (Mixed) 0 2 0 20
deformities in a population of the diatom Planothidium fre-
quentissimum within a Cd-impacted population, in terms of the
viability of deformed frustules, their reproduction capabilities, and References

the time for the population to return to normal forms after the
Alvain, S., Que re, C.L., Bopp, L., Racault, M.F., Beaugrand, G., Dessailly, D.,
cessation of pollution. During the 21 days of Cd treatment, de-
Buitenhuis, E.T., 2013. Rapid climatic driven shifts of diatoms at high latitudes.
formities of the striae and mixed anomalies appeared first, followed Remote Sens. Environ. 132, 195e201. http://dx.doi.org/10.1016/j.rse.2013.01.014.
by alterations in central region and valve outlines. After an addi- Anderson, N.J., 2000. Diatoms, temperature and climatic change. Eur. J. Phycol. 35,
tional 28 days with no Cd exposure, a reduction in deformed 307e314. http://dx.doi.org/10.1080/09670260010001735911.
Andrade, L.R., Marcos Farina, M., Filho, G.M.A., 2004. Effects of copper on Enter-
frustules was observed but deformities were still present. De- omorpha flexuosa (Chlorophyta) in vitro. Ecotoxicol. Environ. Saf. 58, 117e125.
formations of the striae appeared to be the most sustainable http://dx.doi.org/10.1016/S0147-6513(03)00106-4.
phenotype, since they were still significantly higher than in refer- Arini, A., Feurtet-Mazel, A., Morin, S., Maury-Brachet, R., Coste, M., Delmas, F., 2012a.
Remediation of a watershed contaminated by heavy metals: a 2-year field
ence cultures at the end of the decontamination phase for the biomonitoring of periphytic biofilms. Sci. Total Environ. 425, 242e253. http://
previously exposed the Cd cultures. Thus, deformities remain after dx.doi.org/10.1016/j.scitotenv.2012.02.067.
numerous cell divisions. Arini, A., Feurtet-Mazel, A., Maury-Brachet, R., Pokrovsky, O.S., Coste, M., Delmas, F.,
2012b. Recovery potential of periphytic biofilms translocated in artificial
Another issue to consider is the low percentage of deformed
streams after industrial contamination (Cd and Zn). Ecotoxicology 21,
frustules even under stress conditions, which is the major concern 1403e1414. http://dx.doi.org/10.1007/s10646-012-0894-3.
in the use of deformities as a biomonitoring tool. Furthermore, Arini, A., Feurtet-Mazel, A., Maury-Brachet, R., Coste, M., Delmas, F., 2012c. Field
translocation of diatom biofilms impacted by Cd and Zn to assess decontami-
reports of higher incidences of deformed frustules are often from
nation and community restructuring capacities. Ecol. Indic. 18, 520e531. http://
field sites that are concomitantly contaminated with different dx.doi.org/10.1016/j.ecolind.2012.01.005.
toxicants. Thus, it is not always possible to attribute deformities to Arini, A., Durant, F., Coste, M., Delmas, F., Feurtet-Mazel, A., 2013. Cadmium
any particular contaminant or group of contaminants (Falasco et al., decontamination and reversal potential of teratological forms of the diatom
Planothidium frequentissimum (Bacillariophyceae) after experimental contami-
2009), which ultimately leads to weak statistical relationships be- nation. J. Phycol. 49, 361e370. http://dx.doi.org/10.1111/jpy.12044.
tween deformities and the contaminants (Cattaneo et al., 2004). In Armbrecht, L.H., Smetacek, V., Assmy, P., Klaas, C., 2014. Cell death and aggregate
situ studies using diffusing substrates, such as nutrient diffusing formation in the giant diatom Coscinodiscus wailesii (Gran &Angst, 1931). J. Exp.
Mar. Biol. Ecol. 452, 31e39. http://dx.doi.org/10.1016/j.jembe.2013.12.004.
substrates (NDSs) (Pringle, 1990) or metal diffusing substrates Barsanti, L., Coltelli, P., Evangelista, V., Frassanito, A.M., Passarelli, V., Vesentini, N.,
(MDSs) (Pandey et al., 2014), are an excellent tool to assess the Gualtieri, P., 2008. The world of algae. In: Evangelista, V., Barsanti, L.,
effect of single toxicants or combinations of toxicants on frustule Frassanito, A.M., Passarelli, V., Gualtieri, P. (Eds.), Algal Toxins: Nature, Occur-
rence, Effect and Detection. Springer ScienceþBusiness Media B.V, Netherlands,
abnormalities in benthic diatoms. pp. 1e15.
res, V., To
B-Be €ro€k, P., Ko kai, Z., Krasznai, E.T., To
thme resz, B., Bacsi, I., 2014.
Ecological diatom guilds are useful but not sensitive enough as indicators of
4. Conclusion and future prospective extremely changing water regimes. Hydrobiologia 738, 191e204. http://
dx.doi.org/10.1007/s10750-014-1929-y.
Barral-Fraga, L., Morin, S., Rovira, M.D.M., Urrea, G., Magellan, G., Guasch, H., 2016.
Traditional community structural metrics using diatoms (for
Short-term arsenic exposure reduces diatom cell size in biofilm communities.
bioassessment and ecotoxicological studies) can provide effective Environ. Sci. Pollut. R. 23, 4257e4270.
diagnostic information about fluvial ecosystems. However, the Belcher, J.H., 1968. Notes on the physiology of Botryococcus braunii Kützing. Arch.
extensive time (and financial) requirements, necessity of expertise Mikrobiol. 61, 335e346. http://dx.doi.org/10.1007/BF00409670.
rard, A., Benninghoff, C., 2001. Pollution-induced community tolerance (PICT) and
Be
in diatom taxonomy and the need for statistical validation means seasonal variations in the sensitivity of phytoplankton to atrazine in nanocosms.
that the use of structural metrics are often not feasible at the local Chemosphere 45, 427e437. http://dx.doi.org/10.1016/S0045-6535(01)00063-7.
rard, A., Rimet, F., Capowiez, Y., Leboulanger, C., 2004. Procedures for determining
Be
level, and can sometimes make the use of diatom metrics unpop-
the pesticide sensitivity of indigenous soil algae: a possible bioindicator of soil
ular. Conversely, recently developed non-taxonomical metrics contamination? Arch. Environ. Contam. Toxicol. 46, 24e31. http://dx.doi.org/
effectively deal with these shortcomings, as these metrics are 10.1007/s00244-003-2147-1.
relatively quick, easy, cost-effective, reproducible and are based on Berthon, V., Bouchez, A., Rimet, F., 2011. Using diatom life-forms and ecological
guilds to assess organic pollution and trophic level in rivers: a case study of
globally accepted protocols. Furthermore, non-taxonomic metrics rivers in south-eastern France. Hydrobiologia 673, 259e271. http://dx.doi.org/
allow for comparisons between different sites and even 10.1007/s10750-011-0786-1.
geographical locations (countries, continents etc.), as they are in- Bidle, K.D., Falkowski, P.G., 2004. Cell death in planktonic, photosynthetic microor-
ganisms. Nat. Rev. Microbiol. 2, 643e655. http://dx.doi.org/10.1038/nrmicro956.
dependent of the taxonomic similarity or differences. Thus, Blanco, S., Becares, E., 2010. Are biotic indices sensitive to river toxicants? A com-
refinement of current protocols, especially through incorporation parison of metrics based on diatoms and macro-invertebrates. Chemosphere
of new metrics, is needed to improve bioassessment and ecotoxi- 79, 18e25. http://dx.doi.org/10.1016/j.chemosphere.2010.01.059.
Blanco, S., Cejudo-Figueiras, C., Tudesque, L., Be cares, E., Hoffmann, L., Ector, L.,
cological assessment studies of diatoms and for developing efficient
2012. Are diatom diversity indices reliable monitoring metrics? Hydrobiologia
and effective biomonitoring strategies. 695, 199e206. http://dx.doi.org/10.1007/s10750-012-1113-1.
Bopp, L., Aumont, O., Cadule, P., Alvain, S., Gehlen, M., 2005. Response of diatoms
distribution to global warming and potential implications: a global model
Acknowledgements study. Geophys. Res. Lett. 32, L19606. http://dx.doi.org/10.1029/2005GL023653.
Busse, S., Snoeijs, P., 2003. Gradient responses of diatom communities in the
Bothnian Sea (northern Baltic Sea), with emphasis on responses to water
This work was supported by Post-Doctoral Research Program movement. Phycologia 42, 451e464. http://dx.doi.org/10.2216/i0031-8884-42-
through Incheon National University (2015). We are grateful to Dr. 5-451.1.
J.C. Taylor (North-West University, South Africa) for generous gift of Campbell, P.J., Arnold, D.J.S., Brock, T.C.M., Grandy, N.J., Heger, W., Heimbach, F.,
Maund, S.J., Streloke, M. (Eds.), 1999. Guidance Document on Higher-tier
Pleurax. Aquatic Risk Assessment for Pesticides (HARAP). SETAC Press, Brussels.
Cantonati, M., Angeli, N., Virtanen, L., Wojtal, A.Z., Gabriell, J., Falasco, E., Lavoie, I.,
Morin, S., Marchetoo, A., Fortin, C., Smirnova, S., 2014. Achnanthidium minu-
Appendix A. Supplementary data tissimum (Bacillariophyta) valve deformities as indicators of metal enrichment
in diverse widely-distributed freshwater habitats. Sci. Total Environ. 75,
201e215. http://dx.doi.org/10.1016/j.scitotenv.2013.10.018.
Supplementary data related to this article can be found at http:// Casotti, R., Mazza, S., Brunet, C., Vantrepotte, V., Ianora, A., Miralto, A., 2005. Growth
dx.doi.org/10.1016/j.watres.2017.01.062. inhibition and toxicity of the diatomaldehyde 2-trans,4-trans-decadienal on the
Thalassiosira weissflogii (Baccilariophyceae). J. Phycol. 41, 7e20. http://
dx.doi.org/10.1111/j.1529-8817.2005.04052.x.
Conflict of interest Cattaneo, A., Asioli, A., Comoli, P., Manca, M., 1998. Organisms' response in a
chronically polluted lake supports hypothesized link between stress and size.
Limnol. Oceanogr. 43, 1938e1943. http://dx.doi.org/10.4319/lo.1998.43.8.1938.
The authors declare that they have no conflict of interest.
Cattaneo, A., Couillard, Y., Wunsam, S., Courcelles, M., 2004. Diatom taxonomic and indicators of water pollution in the western part of Puck Bay southern Baltic
morphological changes as indicators of metal pollution and recovery in Lac Sea. Oceanol. Hydrobiol. St. 37, 119e132. http://dx.doi.org/10.2478/v10009-007-
Dufault (Que bec, Canada). J. Paleolimnol. 32, 163e175. http://dx.doi.org/ 0042-1.
10.1023/B: JOPL.0000029430.78278.a5. Elias, C.M., Calapez, A.R., Almeida, S.F.P., Feio, M.J., 2015. From perennial to temporal
Chang, D.-W., Hsieh, M.-L., Chen, Y.-M., Lin, T.-F., Chang, J.-S., 2011. Kinetics of cell streams: an extreme drought as a driving force of freshwater communities'
lysis for Microcystis aeruginosa and Nitzschia palea in the exposure to b- traits. Mar. Freshw. Res. 66, 469e480. http://dx.doi.org/10.1071/MF13312.
cyclocitral. J. Hazard. Mater. 185, 1214e1220. http://dx.doi.org/10.1016/ Evans, J.H., 1960. Further investigations of the algae of pond margins. Hydrobiologia
j.jhazmat.2010.10.033. 15, 384e394. http://dx.doi.org/10.1007/BF00046420.
Chessman, B.C., 1986. Diatom flora of an Australian river system: spatial patterns Falasco, E., Bona, F., Badino, G., Hoffmann, L., Ector, L., 2009. Diatom teratological
and environmental relationships. Freshw. Biol. 16, 805e819. http://dx.doi.org/ forms and environmental alterations: a review. Hydrobiologia 623, 1e35.
10.1111/j.1365-2427.1986.tb01018.x. http://dx.doi.org/10.1007/s10750-008-9687-3.
Claesson, M.J., Wang, Q., O'Sullivan, O., Greene-Diniz, R., Cole, J.R., Ross, R.P., Finkel, Z.V., Vaillancourt, C.-J., Irwin, A.J., Reavie, E.D., Smol, J.P., 2009. Environ-
O'Toole, P.W., 2010. Comparison of two next-generation sequencing technolo- mental control of diatom community size structure varies across aquatic eco-
gies for resolving highly complex microbiota composition using tandem vari- systems. Proc. R. Soc. Lond. B 276, 1627e1634. http://dx.doi.org/10.1098/
able 16S rRNA gene regions. Nucl. Acids Res. 38, 1e13. http://dx.doi.org/10.1093/ rspb.2008.1610.
nar/gkq873. Frada, M.J., Burrows, E.H., Wyman, K.D., Falkowski, P.G., 2013. Quantum re-
Cloern, J.E., Dufford, R., 2005. Phytoplankton community ecology: principles applied quirements for growth and fatty acid biosynthesis in the marine diatom
in San Francisco Bay. Mar. Ecol. Prog. Ser. 285, 11e28. http://dx.doi.org/10.3354/ Phaeodactylum tricornutum (Bacillariophyceae) in nitrogen replete and limited
meps285011. conditions. J. Phycol. 49, 381e388. http://dx.doi.org/10.1111/jpy.12046.
Colletti, P., Blinn, D.W., Pickart, A., Wagner, V.T., 1987. Influence of different densities Franklin, D.J., Brussaard, C.P.D., Berges, J.A., 2006. What is the role and nature of
of the mayfly grazer Heptagenia criddlei on lotic diatom communities. J.N.Am. programmed cell death in phytoplankton ecology? Eur. J. Phycol 41, 1e14.
Benthol. Soc. 6, 270e280. http://dx.doi.org/10.2307/1467314. http://dx.doi.org/10.1080/09670260500505433.
Coombs, J., Lauritis, J.A., Darley, W.M., Volcani, B.E., 1968. Studies on the biochem- Gatidou, G., Thomaidis, N.S., 2007. Evaluation of single and joint toxic effects of two
istry and fine structure of silica shell formation in diatoms. Z. für Pflanzen- antifouling biocides, their main metabolites and copper using phytoplankton bio-
physiol. Bd./Int. J. Plant Physiol. 59, 274e284. assays. Aquat. Toxicol. 85,184e191. http://dx.doi.org/10.1016/j.aquatox.2007.09.002.
Cox, E.J., Willis, E., Bentley, K., 2012. Integrated stimulation with experimentation is Gautam, S., Pandey, L.K., Vinayak, V., Arya, A., 2017. Morphological and physiological
a powerful tool for understanding diatom valve morphologenesis. BioSystems alterations in the diatom Gomphonema pseudoaugur due to heavy metal stress.
109, 450e459. http://dx.doi.org/10.1016/j.biosystems.2012.05.012. Ecol. Indic. 72, 67e76. http://dx.doi.org/10.1016/j.ecolind.2016.08.002.
d'Ippolito, G., Sardo, A., Paris, D., Vella, F.M., Adelfi, M.G., Botte, P., Gallo, C., Genter, R.B., Lehman, R.M., 2000. Metal toxicity inferred from algal population
Fontana, A., 2015. Potential of lipid metabolism in marine diatoms for biofuel density, heterotrophic substrate use, and fatty acid profile in a small stream.
production. Biotechnol. Biofuels 8, 1e10. http://dx.doi.org/10.1186/s13068-015- Environ. Toxicol. Chem. 19, 869e878. http://dx.doi.org/10.1002/etc.5620190413.
0212-4. Ghosh, M., Gaur, J.P., 1998. Current velocity and the establishment of stream algal
da Silva, E.F., Almeida, S.F.P., Nunes, M.L., Luís, A.T., Borg, F., Hedlund, M., de S
a, C.M., periphyton communities. Aquat. Bot. 60, 1e10. http://dx.doi.org/10.1016/S0304-
Patinha, C., Teixeira, P., 2009. Heavy metal pollution downstream the aban- 3770(97)00073-9.
doned Coval da Mo mine (Portugal) and associated effects on epilithic diatom Giddings, J.M., Brock, T.C.M., Heger, W., Heimbach, F., Maund, S.J., Norman, S.M.,
communities. Sci. Total Environ. 407, 5620e5636. http://dx.doi.org/10.1016/ Ratte, H.T., Scha €fers, C., Streloke, M. (Eds.), 2002. Community-level Aquatic
j.scitotenv.2009.06.047. System Studiesdinterpretation Criteria (CLASSIC). SETAC Press, Pensacola, FL.
De Jonge, M., Van de Vijver, B., Blusta, R., Lieven Bervoets, L., 2008. Responses of Gillett, N., Pan, Y., Parker, C., 2009. Should only live diatoms be used in the bio-
aquatic organisms to metal pollution in a lowland river in Flanders: a com- assessment of small mountain streams? Hydrobiologia 620, 135e147. http://
parison of diatoms and macroinvertebrates. Sci. Total Environ. 407, 615e629. dx.doi.org/10.1007/s10750-008-9624-5.
http://dx.doi.org/10.1016/j.scitotenv.2008.07.020. Gillett, N.D., Pan, Y., Manoylov, K.M., Stevenson, R.J., 2011. The role of live diatoms in
Debenest, T., Silvestrea, J., Coste, M., Delmas, F., Pinelli, E., 2008. Herbicide effects on bioassessment: a large-scale study of Western US streams. Hydrobiologia 665,
freshwater benthic diatoms: induction of nucleus alterations and silica cell wall 79e92. http://dx.doi.org/10.1007/s10750-011-0606-7.
abnormalities. Aquat. Toxicol. 88, 88e94. http://dx.doi.org/10.1016/j.aquatox. Gold, C., Feurtet-Mazel, A., Coste, M., Boudou, A., 2002. Field transfer of periphytic
2008.03.011. diatom communities to assess short-term structural effects of metals (Cd, Zn) in
Debenest, T., Pinelli, E., Coste, M., Silvestre, J., Mazzella, N., Madigouc, C., Delmas, F., rivers. Water Res. 36, 3654e3664. http://dx.doi.org/10.1016/S0043-1354(02)
2009. Sensitivity of freshwater periphytic diatoms to agricultural herbicides. 00051-9.
Aquat. Toxicol. 93, 11e17. http://dx.doi.org/10.1016/j.aquatox.2009.02.014. Gold, C., Feurtet-Mazel, A., Coste, M., Boudou, A., 2003a. Effects of cadmium stress
Debenest, T., Silvestre, J., Coste, M., Pinelli, E., 2010. Effects of pesticides on fresh- on periphytic diatom communities in indoor artificial streams. Freshw. Biol. 48,
water diatoms. In: Whitacre, D.M. (Ed.), Reviews of Environmental Contami- 316e328. http://dx.doi.org/10.1046/j.1365-2427.2003.00980.x.
nation and Toxicology. Springer, Berlin, pp. 87e103. http://dx.doi.org/10.1007/ Gold, C., Feurtet-Mazel, A., Coste, M., Boudou, A., 2003b. Impacts of Cd and Zn on
978-1-4419-1352-4_2. the Development of periphytic diatom communities in artificial streams located
Debenest, T., Silvestrea, J., Pinelli, E., 2013. Diatoms in ecotoxicology. In: Fe rard, J.-F., along a river pollution gradient. Arch. Environ. Contam. Toxicol 44, 189e197.
Blaise, C. (Eds.), Encylopedia of Aquatic Ecotoxicology. Springer, Netherlands, http://dx.doi.org/10.1007/s00244-002-2024-3.
pp. 295e304. Gomez, N., Licursi, M., 2003. Abnormal forms in Pinnularia gibba (Bacillariophyceae)
DeNicola, D.M., 2000. A review of diatoms found in highly acidic environments. in a polluted lowland stream from Argentina. Nova Hedwig. 77, 389e398.
Hydrobiologia 433, 111e122. http://dx.doi.org/10.1023/A:1004066620172. http://dx.doi.org/10.1127/0029-5035/2003/0077-0389.
Deniseger, J., Austin, A., Lucey, W.P., 1986. Periphyton communities in a pristine Gottschalk, S., Kahlert, M., 2012. Shifts in taxonomical and guild composition of
mountain stream above and below heavy metal mining operations. Freshw. littoral diatom assemblages along environmental gradients. Hydrobiologia 694,
Biol. 16, 209e218. http://dx.doi.org/10.1111/j.1365-2427.1986.tb00965.x. 41e56. http://dx.doi.org/10.1007/s10750-012-1128-7.
Desai, S.R., Verlecar, X.N., Goswami, N.U., 2006. Genotoxicity of cadmium in marine Guasch, H., Ivorra, N., Lehmann, V., Paulsson, M., Real, M., Sabater, S., 1998. Com-
diatom Chaetoceros tenuissimus using the alkaline Comet assay. Ecotoxicology munity composition and sensitivity of periphyton to atrazine in flowing waters:
15, 359e363. http://dx.doi.org/10.1007/s10646-006-0076-2. the role of environmental factors. J. Appl. Phycol. 10, 203e213. http://dx.doi.org/
Dickman, M.D., 1998. Benthic marine diatom deformities associated with contam- 10.1023/A:1008035212208.
inated sediments in Hong Kong. Environ. Int. 24, 749e759. http://dx.doi.org/ Guasch, H., Ricart, M., Lo pez-Doval, J., Bonnineau, C., Proia, L., Morin, S., Mun
~ oz, I.,
10.1016/S0160-4120(98)00060-9. Romaní, A.M., Sabater, S., 2016. Influence of grazing on triclosan toxicity to
Dorigo, U., Bourrain, X., Be rard, A., Leboulanger, C., 2004. Seasonal changes in the stream periphyton. Freshw. Biol. 16, 2002e2012. http://dx.doi.org/10.1111/
sensitivity of river microalgae to atrazine and isoproturon along a contamina- fwb.12797.
tion gradient. Sci. Total Environ. 318, 101e114. http://dx.doi.org/10.1016/S0048- Guerra, L.T., Levitan, O., Frada, M.J., Sun, J.S., Falkowski, P.G., Dismukes, G.C., 2013.
9697(03)00398-X. Regulatory branch points affecting protein and lipid biosynthesis in the diatom
Drum, R.W., Hopkin, J.T., 1966. Diatom locomotion: an explanation. Protoplasma 62, Phaeodactylum tricornutum. Biomass Bioenerg. 59, 306e315. http://dx.doi.org/
1e33. http://dx.doi.org/10.1007/BF01254629. 10.1016/j.biombioe.2013.1010.1007.
Duong, T.T., Morin, S., Herlory, O., Feurtet-Mazel, A., Coste, M., Boudou, A., 2008. Hasle, G.R., Syversten, E.E., 1996. Marine diatoms. In: Tomas, C.R. (Ed.), Identifying
Seasonal effects of cadmium accumulation in periphytic diatom communities of Marine Diatoms and Dinoflagellates. Academic Press, California, pp. 5e385.
freshwater biofilms. Aquat. Toxicol. 90, 19e28. http://dx.doi.org/10.1016/ Hering, D., Johnson, R.K., Kramm, S., 2006. Assessment of European streams with
j.aquatox.2008.07.012. diatoms, macrophytes, macroinvertebrates and fish: a comparative metric-
Duong, T.T., Morin, S., Coste, M., Olivier Herlory, O., Feurtet-Mazel, A., Boudou, A., based analysis of organism response due to stress. Freshw. Biol. 51,
2010. Experimental toxicity and bioaccumulation of cadmium in freshwater 1757e1785. http://dx.doi.org/10.1111/j.1365-2427.2006.01610.x.
periphytic diatoms in relation with biofilm maturity. Sci. Total Environ. 408, Hildebrand, M., 2008. Diatoms, biomineralization processes, and genomics. Chem.
552e562. http://dx.doi.org/10.1016/j.scitotenv.2009.10.015. Rev. 108, 4855e4874. http://dx.doi.org/10.1021/cr078253z.
Duong, T.T., Coste, M., Feurtet-Mazel, A., Dang, D.K., Ho, C.T., Le, T.P.Q., 2012. Re- Hildebrand, M., Davis, A.K., Smith, S.R., Traller, J.C., Abbriano, R., 2012. The place of
sponses and structural recovery of periphytic diatom communities after short- diatoms in the biofuels industry. Biofuels 3, 221e240. http://dx.doi.org/10.4155/
term disturbance in some rivers (Hanoi, Vietnam). J. Appl. Phycol. 24, BFS.11.157.
1053e1065. http://dx.doi.org/10.1007/s10811-011-9733-9. Hill, B.H., Williamgham, W.T., Parrish, L.P., McFarland, B.H., 2000. Periphyton com-
Dziengo-Czaja, M., Koss, J., Matuszak, A., 2008. Teratological forms of diatoms as munity responses to elevated metal concentrations in a Rocky Mountain stream.
Hydrobiologia 428, 161e169. http://dx.doi.org/10.1023/A:1004028318542. metal bioaccumulation. Environ. Toxicol. Chem. 35, 1489e1501. http://
Hillebrand, H., Durselen, C., Kirschtel, D., Pollingher, U., Zohary, T., 1999. Biovolume dx.doi.org/10.1002/etc.3292.
calculation for pelagic and benthic microalgae. J. Phycol. 35, 403e424. http:// Lelong, A., Bucciarelli, E., He garet, H., Soudant, P., 2013. Iron and copper limitations
dx.doi.org/10.1046/j.1529-8817.1999.3520403.x. differently affect growth rates and photosynthetic and physiological parameters
Hirst, H., Ingrid, J.U., Jüttner, I., Ormerod, S.J., 2002. Comparing the responses of of the marine diatom Pseudo-nitzschia delicatissima. Limnol. Oceanogr. 58,
diatoms and macroinvertebrates to metals in upland streams of Wales and 613e623. http://dx.doi.org/10.4319/lo.2013.58.2.0613.
Cornwall. Freshw. Biol. 47, 1752e1765. http://dx.doi.org/10.1046/j.1365- Levy, J., Stauber, J.L., Jolley, D.F., 2007. Sensitivity of marine microalgae to copper:
2427.2002.00904.x. the effect of biotic factors on copper adsorption and toxicity. Sci. Total Environ.
Janssen, M., Bathke, L., Marquardt, J., Krumbein, W.E., Rhiel, E., 2001. Changes in the 387, 141e154. http://dx.doi.org/10.1016/j.scitotenv.2007.07.016.
photosynthetic apparatus of diatoms in response to low and high light in- Levy, J., Angel, B., Stauber, J.L., Poon, W.L., Simpson, S.L., Cheng, S., Jolley, D.F., 2008.
tensities. Int. Microbiol. 4, 27e33. http://dx.doi.org/10.1007/s101230100005. Uptake and internalisation of copper by three marine microalgae: comparison
Jiang, Y., Yoshida, T., Quigg, A., 2012. Photosynthetic performance, lipid production and of copper-sensitive and copper-tolerant species. Aquat. Toxicol. 89, 82e93.
biomass composition in response to nitrogen limitation in marine microalgae. http://dx.doi.org/10.1016/j.aquatox.2008.06.003.
Plant Physiol. Biochem. 54, 70e77. http://dx.doi.org/10.1016/j.plaphy.2012.02.012. Liang, Y., Osada, K., Sunaga, Y., Yoshino, T., Bowler, C., Tanaka, T., 2015. Dynamic oil
Joux-Arab, L., Berthet, B., Robert, J.M., 2000. Do toxicity and accumulation of copper body generation in the marine oleaginous diatom Fistulifera solaris in response
change during size reduction in the marine pennate diatom Haslea ostrearia? to nutrient limitation as revealed by morphological and lipidomic analysis.
Mar. Biol. 136, 323e330. http://dx.doi.org/10.1007/s002270050690. Algal Res. 12, 359e367. http://dx.doi.org/10.1016/j.algal.2015.09.017.
Julius, M.L., Theriot, E.C., 2007. The diatoms: a primer. In: Smol, J.P., Stoermer, E.F. Licursi, M., Go mez, N., 2013. Short-term toxicity of hexavalent-chromium to epi-
(Eds.), The Diatoms: Applications for the Environmental and Earth Sciences, psammic diatoms of a microtidal estuary (Río de la Plata): responses from the
second ed.s. Cambridge University Press, London, pp. 8e22. http://dx.doi.org/ individual cell to the community structure. Aquat. Toxicol. 134e135, 82e91.
10.1017/CBO9780511763175.003. http://dx.doi.org/10.1016/j.aquatox.2013.03.007.
Jüttner, I., Rothfritz, H., Omerod, S.J., 1996. Diatoms as indicators of river water Litchman, E., Klausmeier, C.A., Yoshiyama, K., 2009. Contrasting size evolution in
quality in the Nepalese Middle Hills with consideration of the effects of habitat- marine and freshwater diatoms. Proc. Natl. Acad. Sci. 106, 2665e2670. http://
specific sampling. Freshw. Biol. 36, 475e486. http://dx.doi.org/10.1046/j.1365- dx.doi.org/10.1073/pnas.0810891106.
2427.1996.00101.x. Liu, Z.Y., Wang, G.C., Zhou, B.C., 2008. Effect of iron on growth and lipid accumu-
Kelly, M.G., 1998. Use of the trophic diatom index to eutrophication in rivers. Water lation in Chlorella vulgaris. Bioresour. Technol. 99, 4717e4722. http://dx.doi.org/
Res. 32, 236e242. http://dx.doi.org/10.1016/S0043-1354(97)00157-7. 10.1016/j.biortech.2007.09.073.
Kelly, M.G., Wilson, S., 2004. Effect of phosphorus stripping on water chemistry and Lombardi, A.T., Vieira, A.A.H., 1999. Lead- and copper-complexing extracellular li-
diatom ecology in an eastern lowland river. Water Res. 38, 1559e1567. http:// gands released by Kirchneriella aperta (Chlorococcales, Chlorophyta). Phycologia
dx.doi.org/10.1016/j.watres.2003.12.022. 38, 283e288. http://dx.doi.org/10.2216/i0031-8884-38-4-283.1.
Kermarrec, L., Franc, A., Rimet, F., Chaumeil, P., Humbert, J.F., Bouchez, A., 2013. Lombardi, A.T., Vieira, A.A.H., 2000. Copper complexation by cyanophyta and
Next-generation sequencing to inventory taxonomic diversity in eukaryotic chlorophyta exudates. Phycologia 39, 118e125. http://dx.doi.org/10.2216/i0031-
communities: a test for freshwater diatoms. Mol. Ecol. Resour. 13, 607e619. 8884-39-2-118.1.
http://dx.doi.org/10.1111/1755-0998.12105. Lommer, M., Specht, M., Roy, A.-S., Kraemer, L., Andreson, R., Gutowska, M.A.,
Kermarrec, L., Franc, A., Rimet, F., Chaumeil, P., Frigerio, J.M., Humbert, J.F., Wolf, J., Bergner, S.V., Schilhabel, M.B., Klostermeier, U.C., Beiko, R.G.,
Bouchez, A., 2014. A next-generation sequencing approach to river bio- Rosenstiel, P., Hippler, M., LaRoche, J., 2012. Genome and low-iron response of
monitoring using benthic diatoms. Freshw. Sci. 33, 349e363. http://dx.doi.org/ an oceanic diatom adapted to chronic iron limitation. Genome Biol. 13, 1e20.
10.1086/675079. http://dx.doi.org/10.1186/gb-2012-13-7-r66.
Kerrigan, E.A., Irwin, A.J., Finkel, Z.V., 2015. Community- and population-level Luís, A.T., Teixeira, P., Almeida, S.F.P., Ector, L., Matos, J.X., Ferreira da Silva, E.A., 2011.
changes in diatom size structure in a subarctic lake over the last two cen- Environmental impact of mining activities in the Lousal area (Portugal):
turies. PeerJ 3, e1074. http://dx.doi.org/10.7717/peerj.1074. chemical and diatom characterization of metal-contaminated stream sediments
Kilham, S.S., Theriot, E.C., Fritz, S.C., 1996. Linking planktonic diatoms and climate and surface water of Corona stream. Sci. Total Environ. 409, 4312e4325. http://
change in the large lakes of the Yellowstone ecosystem using resource theory. dx.doi.org/10.1016/j.scitotenv.2011.06.052.
Limnol. Oceanogr. 41, 1052e1062. http://dx.doi.org/10.4319/lo.1996.41.5.1052. Ma, J., Xu, L., Wang, S., Zheng, R., Jin, S., Huang, S., Huang, Y., 2002. Toxicity of 40
Knauer, K., Maise, S., Thoma, G., Hommen, U., Gonzalez-Valero, J., 2005. Long-term herbicides to the green algae Chlorella vulgaris. Ecotoxicol. Environ. Saf. 51,
variability of zooplankton populations in aquatic mesocosms. Environ. Toxicol. 128e132. http://dx.doi.org/10.1006/eesa.2001.2113.
Chem. 24, 1182e1189. http://dx.doi.org/10.1897/04-010R.1. Mackay, A.W., 2007. The paleoclimatology of Lake Baikal: a diatom synthesis and
Knauer, K., Hommen, U., 2012. Sensitivity, variability, and recovery of functional and prospectus. Earth-Sci. Rev. 82, 181e215. http://dx.doi.org/10.1016/j.earscirev.
structural endpoints of an aquatic community exposed to herbicides. Ecotox- 2007.03.002.
icol. Environ. Saf. 78, 178e183. http://dx.doi.org/10.1016/j.ecoenv.2011.11.019. Mann, D.G., Droop, S.J.M., 1996. Biodiversity, biogeography and conservation of
Kociolek, J.P., Stoermer, E.F., 2010. Variation and polymorphism in diatoms: the diatoms. Hydrobiologia 336, 19e32. http://dx.doi.org/10.1007/BF00010816.
triple helix of development, genetics and environment. A review of the litera- Manoylov, K.M., 2014. Taxonomic identification of algae (morphological and mo-
ture. Vie Milieu Life Environ. 60, 75e87. lecular): species concepts, methodologies, and their implications for ecological
Koester, J.A., Swalwell, J.E., Dassow, P.V., Armbrust, E.V., 2010. Genome size differ- bioassessment. J. Phycol. 50, 409e424. http://dx.doi.org/10.1111/jpy.12183.
entiates co-occurring populations of the planktonic diatom Ditylum brightwellii McCormick, P.V., Cairns Jr., J., 1994. Algae as indicators of environmental change.
(Bacillariophyceae). BMC Evol. Biol. 10 (1) http://dx.doi.org/10.1186/1471-2148- J. Appl. Phycol. 6, 509e526. http://dx.doi.org/10.1007/BF02182405.
10-1. Medley, C.L., Clements, W.H., 1998. Responses of diatom communities to heavy
Kraufvelin, P., 1998. Model ecosystem replicability challenged by the ‘‘soft’’ reality of metals in streams: the influence of longitudinal variation. Ecol. Appl. 8, 631e644.
a hard bottom mesocosm. J. Exp. Mar. Biol. Ecol. 222, 247e267. http:// http://dx.doi.org/10.1890/1051-0761(1998)008[0631:RODCTH]2.0.CO;2.
dx.doi.org/10.1016/S0022-0981(97)00143-3. Miklasz, K.A., Denny, M.W., 2010. Diatom sinking speeds: improved predictions and
Laney, S.R., Olson, R.J., Sosik, H.M., 2012. Diatoms favor their younger daughters. insight from a modified Stokes' law. Limnol. Oceanogr. 55, 2513e2525. http://
Limnol. Ocean. 57, 1572e1578. http://dx.doi.org/10.4319/lo.2012.57.5.1572. dx.doi.org/10.4319/lo.2010.55.6.2513.
Larras, F., Montuelle, B., Bouchez, A., 2013. Assessment of toxicity thresholds in Montagnes, D.J.S., Franklin, D.J., 2001. Effect of temperature on diatom volume,
aquatic environments: does benthic growth of diatoms affect their exposure growth rate, and carbon and nitrogen content: reconsidering some paradigms.
and sensitivity to herbicides? Sci. Total Environ. 463e464, 469e477. http:// Limnol. Oceanogr. 46, 2008e2018.
dx.doi.org/10.1016/j.scitotenv.2013.06.063. Morin, S., Coste, M., 2006. Metal-induced shifts in the morphology of diatoms from
Larras, F., Keck, F., Montuelle, B., Rimet, F., Bouchez, A., 2014a. Linking diatom the Riou Mort and Riou Viou streams (South west France). In: Acs, E., Kiss, K.T.,
sensitivity to herbicides to phylogeny: a step forward for biomonitoring? En- Padiska, J., Szabo, K. (Eds.), Use of Algae for Monitoring Rivers VI. Hungarian
viron. Sci. Technol. 48, 1921e1930. http://dx.doi.org/10.1021/es4045105. Algological Society, God, Hungary, Balatonfured, pp. 91e106.
Larras, F., Montuelle, B., Rimet, F., Che vre, N., Bouchez, A., 2014b. Seasonal shift in Morin, S., Coste, M., Hamilton, P.B., 2008c. Scanning electron microscopy observa-
the sensitivity of a natural benthic microalgal community to a herbicide tions of small pinnate diatoms exposed to high cadmium concentrations.
mixture: impact on the protective level of thresholds derived from species J. Phycol. 44, 1512e1518. http://dx.doi.org/10.1111/j.1529-8817.2008.00587.x.
sensitivity distributions. Ecotoxicology 23, 1109e1123. http://dx.doi.org/ Morin, S., Duong, T.T., Dabrin, A., Coynel, A., Herlory, O., Baudrimont, M., Delmas, F.,
10.1007/s10646-014-1254-2. Durrieu, G., Scha €fer, J., Winterton, P., Blanc, G., Coste, M., 2008a. Long-term
Lavoie, I., Vincent, W.F., Pienitz, R., Painchaud, J., 2004. Benthic algae as bioindicators survey of heavy-metal pollution, biofilm contamination and diatom community
of agricultural pollution in the streams and rivers of southern Que bec (Canada). structure in the Riou Mort watershed, South-West France. Environ. Pollut. 151,
Aquat. Ecosyst. Health 7, 43e58. http://dx.doi.org/10.1080/14634980490281236. 532e542. http://dx.doi.org/10.1016/j.envpol.2007.04.023.
Lavoie, I., Campeau, S., Fallu, M., Dillon, P.J., 2006. Diatoms and biomonitoring: Morin, S., Duong, T.T., Herlory, O., Feurtet-Mazel, A., Coste, M., 2008b. Cadmium
should cell size be accounted for? Hydrobiologia 573, 1e16. http://dx.doi.org/ toxicity and bioaccumulation in freshwater biofilms. Arch. Environ. Contam.
10.1007/s10750-006-0223-z. Toxicol. 54, 173e186. http://dx.doi.org/10.1007/s00244-007-9022-4.
Lavoie, I., Lavoie, M., Fortin, C., 2012. A mine of information: benthic algal com- Morin, S., Bottin, M., Mazzella, N., Macary, F., Delmas, F., Winterton, P., Coste, M.,
munities as biomonitors of metal contamination from abandoned tailings. Sci. 2009. Linking diatom community structure to pesticide input as evaluated
Total Environ. 425, 231e241. http://dx.doi.org/10.1016/j.scitotenv.2012.02.057. through a spatial contamination potential (Phytopixal): a case study in the
Leguay, S., Lavoie, I., Levy, J.L., Fortin, C., 2016. Using biofilms for monitoring metal Neste river system (South-West France). Aquat. Toxicol. 94, 28e39. http://
contamination in lotic ecosystems: the protective effects of hardness and pH on dx.doi.org/10.1016/j.aquatox.2009.05.012.
Morin, S., Pesce, S., Tili, A., Coste, M., Montuelle, B., 2010. Recovery potential of Ricciardi, F., Bonnineau, C., Faggiano, L., Geiszinger, A., Guasch, H., Lopez-Doval, J.,
periphytic communities in a river impacted by a vineyard watershed. Ecol. Mun ~ oz, I., Proia, L., Ricart, M., Romaní, A., Sabater, S., 2009. Is chemical
Indic. 10, 419e426. http://dx.doi.org/10.1016/j.ecolind.2009.07.008. contamination linked to the diversity of biological communities in rivers?
Morin, S., Berta Bonet, B., Corcoll, N., Guasch, H., Bottin, M., Coste, M., 2015. Cu- Trends Anal. Chem. 28, 592e602. http://dx.doi.org/10.1016/j.trac.2009.02.007.
mulative stressors trigger increased vulnerability of diatom communities to Rimet, F., Bouchez, A., 2011. Use of diatom life-forms and ecological guilds to assess
additional disturbances. Microb. Ecol. 70, 585e595. http://dx.doi.org/10.1007/ pesticide contamination in rivers: lotic mesocosm approaches. Ecol. Indic. 11,
s00248-015-0602-y. 489e499. http://dx.doi.org/10.1016/j.ecolind.2010.07.004.
Morin, S., Cordonier, A., Lavoie, I., Arini, A., Blanco, S., Duong, T.T., Torne s, E., Rimet, F., Bouchez, A., 2012. Biomonitoring river diatoms: implications of taxonomic
Bonet, B., Corcoll, N., Faggiano, L., Laviale, M., Pe re
s, F., Becares, E., Coste, M., resolution. Ecol. Indic. 15, 92e99. http://dx.doi.org/10.1016/j.ecolind.2011.09.014.
Feurtet-Mazel, A., Fortin, C., Guasch, H., Sabater, S., 2012. Consistency in diatom Rose, D.T., Cox, E.J., 2013. Some diatom species do not show a gradual decrease in
response to metal-contaminated environments. In: Guasch, H., Ginebreda, A., cell size as they reproduce. Fundam. Appl. Limnol. 182/2, 117e122. http://
Geiszinger, A. (Eds.), Emerging and Priority Pollutants in Rivers. Springer- dx.doi.org/10.1127/1863-9135/2013/0406.
Verlag, Berlin, pp. 117e146. Rosen, B.H., Lowe, R.L., 1984. Physiological and ultrastructural responses of Cyclo-
Morin, S., Go mez, N., Torne s, E., Licursi, M., Rosebery, J., 2016. Benthic diatom tella meneghiniana (Bacillariophyta) to light intensity and nutrient stress.
monitoring and assessment of freshwater environments: standard methods J. Phycol. 20, 173e183. http://dx.doi.org/10.1111/j.0022-3646.1984.00173.x.
and future challenges. In: Romaní, A.M., Guasch, H., Balaguer, M.D. (Eds.), Roubeix, V., Mazzella, N., Schouler, L., Fauvelle, V., Morin, S., Coste, M., Delmas, F.,
Aquatic Biofilms: Ecology, Water Quality and Water Treatment. Caister Aca- Margoum, C., 2011a. Variations of periphytic diatom sensitivity to the herbicide
demic Press, U.K, pp. 111e124. http://dx.doi.org/10.21775/9781910190173.06. diuron and relation to species distribution in a contamination gradient: im-
Nunes, M.L., da Silva, E.F., Almeida, S.F.P., 2003. Assessment of water quality in the plications for biomonitoring. J. Environ. Monit. 13, 1768e1774. http://dx.doi.org/
Caima and Mau River basins (Portugal) using geochemical and biological Indices. 10.1039/c0em00783h.
Water Air Soil Pollut. 149, 227e250. http://dx.doi.org/10.1023/A:1025636106890. Roubeix, V., Mazzella, N., Me chin, B., Coste, M., Delmas, F., 2011b. Impact of the
Olson, R.J., Watras, C., Chisholm, S.W., 1986. Patterns of individual cell growth in herbicide metolachlor on river periphytic diatoms: experimental comparison of
marine centric diatoms. J. Gen. Microbiol. 132, 1197e1204. descriptors at different biological organization levels. Ann. Limnol. Int. J. Lim. 47,
Palmer, C.M., 1969. A composite rating of algae tolerating organic pollution. 1e11. http://dx.doi.org/10.1051/limn/20110080.
J. Phycol. 5, 78e82. http://dx.doi.org/10.1111/j.1529-8817.1969.tb02581.x. Roubeix, V., Fauvelle, V., Tison-Rosebery, J., Mazzella, N., Coste, M., Delmas, F., 2012.
Pandey, L.K., Kumar, D., Yadav, A., Rai, J., Gaur, J.P., 2014. Morphological abnormalities Assessing the impact of chloroacetanilide herbicides and their metabolites on
in periphytic diatoms as a tool for biomonitoring of heavy metal pollution in a periphyton in the Leyre River (SW France) via short term growth inhibition
river. Ecol. Indic. 36, 272e279. http://dx.doi.org/10.1016/j.ecolind.2013.08.002. tests on autochthonous diatoms. J. Environ. Monit. 14, 1655e1663. http://
Pandey, L.K., Han, T., Gaur, J.P., 2015. Response of a phytoplanktonic assemblage to dx.doi.org/10.1039/c2em10887a.
copper and zinc enrichment in microcosm. Ecotoxicology 24, 573e582. http:// Ruggiu, D., Luglie , A., Cattaneo, A., Panzani, P., 1998. Paleoecological evidence for
dx.doi.org/10.1007/s10646-014-1405-5. diatom response to metal pollution in Lake Orta (N. Italy). J. Paleolimnol. 20,
Pandey, L.K., Bergey, E.A., 2016. Exploring the status of motility, lipid bodies, de- 333e345. http://dx.doi.org/10.1023/A:1007929926526.
formities and size reduction in periphytic diatom community from chronically Rynearson, T.A., Armbrust, E.V., 2000. DNA fingerprinting reveals extensive genetic
metal (Cu, Zn) polluted waterbodies as a biomonitoring tool. Sci. Total Environ. diversity in a field population of the centric diatom Ditylum brightwellii.
550, 372e381. http://dx.doi.org/10.1016/j.scitotenv.2015.11.151. Limnol. Oceanogr. 45, 1329e1340. http://dx.doi.org/10.4319/lo.2000.45.6.1329.
Pandey, L.K., Ojha, K.K., Singh, P.K., Singh, C.S., Dwivedi, S., Bergey, E.A., 2016. Di- Sabater, S., 2000. Diatom communities as indicators of environmental stress in the
atoms image database of India (DIDI): a research tool. Environ. Technol. Inn. 5, Guadiamar River, S-W. Spain, following a major mine tailings spill. J. Appl.
148e160. http://dx.doi.org/10.1016/j.eti.2016.02.001. Phycol. 12, 113e124. http://dx.doi.org/10.1023/A:1008197411815.
Passy, S.I., 2007. Diatom ecological guilds display distinct and predictable behavior Sabater, S., Guasch, H., Romaní, A., Isabel Mun ~ oz, I., 2002. The effect of biological
along nutrient and disturbance gradients in running waters. Aquat. Bot. 86, factors on the efficiency of river biofilms in improving water quality. Hydro-
171e178. http://dx.doi.org/10.1016/j.aquabot.2006.09.018. biologia 469, 149e156. http://dx.doi.org/10.1023/A:1015549404082.
Passy, S.I., Larson, C.A., 2011. Succession in stream biofilms is an environmentally Sabater, S., Admiraal, W., 2005. Periphyton as biological indicators in managed
driven gradient of stress tolerance. Microb. Ecol. 62, 414e424. aquatic ecosystems. In: Azim, M.E., Verdegem, M.C.J., van Dam, A.A.,
Peramuna, A., Summers, M.L., 2014. Composition and occurrence of lipid droplets in Beveridge, M.C.M. (Eds.), Periphyton: Ecology Exploitation and Management.
the cyanobacterium Nostoc punctiforme. Arch. Microbiol. 196, 881e890. http:// CABI Publishing, Cambridge, pp. 159e177.
dx.doi.org/10.1007/s00203-014-1027-6. Sabater, S., Guasch, H., Ricart, M., Romaní, A., Vidal, G., Klünder, C., Schmitt-
re
Pe s, F., Florin, D., Grollier, T., Feurtet-Mazel, A., Coste, M., Ribeyre, F., Ricard, M., Jansen, M., 2007. Monitoring the effect of chemicals on biological communi-
Boudou, A., 1996. Effects of the phenylurea herbicide isoproturon on periphytic ties.The biofilm as an interface. Anal. Bioanal. Chem. 387, 1425e1434. http://
diatom communities in freshwater indoor microcosms. Environ. Pollut. 94, dx.doi.org/10.1007/s00216-006-1051-8.
141e152. http://dx.doi.org/10.1016/S0269-7491(96)00080-2. Schindler, D.W., 1990. Experimental perturbations of whole lakes as tests of hy-
Podola, B., Melkonian, M., 2005. Selective real-time herbicide monitoring by an potheses concerning ecosystem structure and function. Oikos 57, 25e41. http://
array chip biosensor employing diverse microalgae. J. Appl. Phycol. 17, 261e271. dx.doi.org/10.2307/3565733.
http://dx.doi.org/10.1007/s10811-005-4945-5. Schmitt-Jansen, M., Altenburger, R., 2005. Toxic effects of isoproturon on periphyton
Potapova, M.G., Charles, D.F., Ponder, K.C., Winter, D.M., 2004. Quantifying species communities e a microcosm study. Estuar. Coast. Shelf Sci. 62, 539e545. http://
indicator values for trophic diatom indices: a comparison of approaches. Hydro- dx.doi.org/10.1016/j.ecss.2004.09.016.
biologia 517, 25e41. http://dx.doi.org/10.1023/B: HYDR.0000027335.73651.ea. Sluys, R., 2013. The unappreciated, fundamentally analytical nature of taxonomy
Potapova, M., Charles, D.F., 2005. Choice of substrate in algae-based water-quality and the implications for the inventory of biodiversity. Biodivers. Conserv. 22,
assessment. J. N. Am. Benthol. Soc. 24, 415e427. http://dx.doi.org/10.1899/03- 1095e1105. http://dx.doi.org/10.1007/s10531-013-0472-x.
111.1. Smetacek, V., 2001. A watery arms race. Nature 411 (745). http://dx.doi.org/10.1038/
Potapova, M., Charles, D.F., 2007. Diatom metrics for monitoring eutrophication in 35081210.
rivers of the United States. Ecological Indicators 7, 48e70. http://dx.doi.org/ Smol, J.P., Glew, J.R., 1992. Paleolimnology. In: Nierenberg, W.A. (Ed.), Encylopedia of
10.1016/j.ecolind.2005.10.001. Earth System Science 3. Academic press, San Diego, pp. 551e564.
Potapova, M., Hamilton, P.B., 2007. Morphological and ecological variation within Snoeijs, P., Busse, S., Potapova, M., 2002. The importance of diatom cell size in
the Achnanthidium minutissimum (Bacillariophyceae) species complex. J. Phycol. community analysis. J. Phycol. 38, 265e272. http://dx.doi.org/10.1046/j.1529-
43, 561e575. http://dx.doi.org/10.1111/j.1529-8817.2007.00332.x. 8817.2002.01105.x.
Pringle, C.M., 1990. Nutrient Spatial Heterogeneity: effects on community structure, Stauber, J.L., Florence, T.M., 1987. Mechanism of toxicity of ionic copper and copper
physiognomy, and diversity of stream algae. Ecology 71, 905e920. http://www. complexes to algae. Mar. Biol. 94, 511e519. http://dx.doi.org/10.1007/
jstor.org/stable/1937362. BF00431397.
Radchenko, I.G., Il’yash, L.V., 2006. Growth and photosynthetic activity of diatom Stenger-Kov acs, C., Lengyel, E., Crossetti, L.O., Uveges, V., Padisak, J., 2013. Diatom
Thalassiosira weissflogii at decreasing salinity. Biol. Bull. 33, 242e247. http:// ecological guilds as indicators of temporally changing stressors and distur-
dx.doi.org/10.1134/S106235900603006X. bances in the small Torna-stream. Hung. Ecol. Indic. 24, 138e147. http://
Ramachandra, T.V., Mahapatra, D.M., Karthick, B., Gordon, R., 2009. Milking diatoms dx.doi.org/10.1016/j.ecolind.2012.06.003.
for sustainable energy: biochemical engineering versus gasoline-secreting Stevenson, R.J., 1984. Epilithic and epipelic diatoms in the Sandusky River, with
diatom solar panels. Ind. Eng. Chem. Res. 48, 8769e8788. http://dx.doi.org/ emphasis on species diversity and water quality. Hydrobiologia 114, 161e175.
10.1021/ie900044j. Stevenson, R.J., Peterson, C.G., 1991. Emigration and immigration can be important
Reavie, E.D., Barbiero, R.P., 2013. Recent changes in abundance and cell size of determinants of benthic diatom assemblages in streams. Freshw. Biol. 26,
pelagic diatoms in the North American Great Lakes. Phytotaxa 127, 150e162. 279e294. http://dx.doi.org/10.1111/j.1365-2427.1991.tb01735.x.
http://dx.doi.org/10.11646/phytotaxa.127.1.15. Stevenson, R.J., White, K.D., 1995. A comparison of natural and human determinants
Renzi, M., Roselli, L., Giovani, A., Focardi, S.E., Basset, A., 2014. Early warning tools of phytoplankton communities in the Kentucky River basin, USA. Hydrobiologia
for ecotoxicity assessment based on Phaeodactylum tricornutum. Ecotoxicology 297, 201e216. http://dx.doi.org/10.1007/BF00019285.
23, 1055e1072. http://dx.doi.org/10.1007/s10646-014-1249-z. Stevenson, R.J., Pan, Y., Vandam, H., 2010. Assessing environmental conditions in
Ricart, M., Barcelo , D., Geiszinger, A., Guasch, H., de Alda, M.L., Romaní, A.M., rivers and streams with diatoms. In: Smol, J.P., Stoermer, E.F. (Eds.), The Di-
Vidal, G., Villagrasa, M., Sabater, S., 2009. Effects of low concentrations of the atoms: Applications for the Environmental and Earth Sciences, second ed.s.
phenylurea herbicide diuron on biofilm algae and bacteria. Chemosphere 76, Cambridge University Press, London, pp. 57e85. http://dx.doi.org/10.1017/
1392e1401. http://dx.doi.org/10.1016/j.chemosphere.2009.06.017. CBO9780511763175.005.
Stevenson, J., 2014. Ecological assessments with algae: a review and synthesis. Soil Pollut. 161, 227e265. http://dx.doi.org/10.1007/s11270-005-4285-8.
J. Phycol. 50, 437e461. http://dx.doi.org/10.1111/jpy.12189. Visco, J.A., Gentil, L.A.P., Cordonier, A., Esling, P., Pillet, L., Pawlowski, J., 2015. Envi-
Svensson, F., Norberg, J., Snoeijs, P., 2014. Diatom cell size, coloniality and motility: ronmental monitoring: inferring the diatom index from next-generation
trade-offs between temperature, salinity and nutrient supply with climate sequencing data. Environ. Sci. Technol. 49, 7597e7605. http://dx.doi.org/10.1021/
change. PLoS One 9, e109993. http://dx.doi.org/10.1371/journal.pone.0109993. es506158m.
Takabayashi, M., Lew, K., Johnson, A., Marchi, A.L., Dugdale, R., Wilkerson, F.P., 2006. Walsby, A.E., Reynolds, C.S., 1980. Sinking and floating. In: Morris, I. (Ed.), The
The effect of the nutrient availability and temperature on chain length of the Physiological Ecology of Phytoplankton. Blackwell Scientific Publications, New
diatom, Skeletonema costatum. J. Plankton Res. 28, 831e840. http://dx.doi.org/ York, pp. 371e412.
10.1093/plankt/fbl018. Wang, J., Cao, S., Du, C., Chen, D., 2013. Underwater locomotion strategy by a benthic
ter Braak, C.J.F., 1986. Canonical correspondence analysis: a new eigenvector tech- pinnate diatom Navicula sp. Protoplasma 250, 1203e1212. http://dx.doi.org/
nique for multivariate direct gradient analysis. Ecology 67, 1167e1179. http:// 10.1007/s00709-013-0502-2.
dx.doi.org/10.2307/1938672. Wang, Z.T., Ullrich, N., Joo, S., Waffenschmidt, S., Goodenough, U., 2009. Algal lipid
ter Braak, C.J.F., 1987. The analysis of vegetation-environment relationships by ca- bodies: stress induction, purification and biochemical characterization in wild-
nonical correspondence analysis. Vegetatio 69, 69e77. http://dx.doi.org/ type and starchless Chlamydomonas reinhardtii. Eukaryot. Cell 8, 1856e1868.
10.1007/978-94-009-4061-1_7. http://dx.doi.org/10.1128/EC.00272-09.

ter Braak, C.J.F., Smilauer, P., 2002. CANOCO Reference Manual and CanDraw for Wei, Y., Zhu, N., Lavoie, M., Wang, J., Qian, H., Fu, Z., 2014. Copper toxicity to
Windows User's Guide. Software for Canonical Community Ordination (version Phaeodactylum tricornutum: a survey of the sensitivity of various toxicity end-
4.5). Biometris, Wageningen and Cesk Bude
e jovice. points at the physiological, biochemical, molecular and structural levels. Bio-
Trobajo, R., Mann, D.G., Chepurnov, V.A., Clavero, E., Cox, E.J., 2006. Taxonomy, life metals 27, 527e537. http://dx.doi.org/10.1007/s10534-014-9727-6.
cycle, and auxosporulation of Nitzschia Fonticola (Bacillariophyta)1. J. Phycol. 42, Weng, L.C., Pasaribu, B., Lin, I.P., Tsai, C.H., Chen, C.S., Jiang, P.L., 2014. Nitrogen
1353e1372. http://dx.doi.org/10.1111/j.1529-8817.2006.00291.x. deprivation induces lipid droplet accumulation and alters fatty acid met-
Trobajo, R., Rovira, L., Mann, D.G., Cox, E.J., 2011. Effects of salinity on growth and on albolism in symbiotic dinoflagellates isolated from Aiptasia pulchella. Sci. Rep. 4,
valve morphology of five estuarine diatoms. Phycol. Res. 59, 83e90. http:// 1e8. http://dx.doi.org/10.1038/srep05777.
dx.doi.org/10.1111/j.1440-1835.2010.00603.x. Wilhelm, S.W., Adrian, R., 2008. Impact of summer warming on the thermal char-
Trobajo, R., Rovira, L., Euctor, L., Wetzel, C.E., Kelly, M., Mann, D.G., 2013. acteristics of a polymictic lake and consequences for oxygen, nutrients and
Morphology and identity of some ecologically important small Nitzschia spe- phytoplankton. Freshw. Biol. 53, 226e237. http://dx.doi.org/10.1111/j.1365-
cies. Diatom Res. 28, 37e59. http://dx.doi.org/10.1080/0269249X.2012.734531. 2427.2008.01980.x.
Van dam, H., Mertens, A., Sinkeldam, J., 1994. A coded checklist and ecological in- Will, K.W., Rubinoff, D., 2004. Myth of the molecule: DNA barcodes for species
dicator values of freshwater diatoms from The Netherlands. Neth. J. Aquat. Ecol. cannot replace morphology for identification and classification. Cladistics 20,
28, 117e133. http://dx.doi.org/10.1007/BF02334251. 47e55. http://dx.doi.org/10.1111/j.1096-0031.2003.00008.x.
Van de Vijver, B., Beyens, L., 1998. Diatoms and water quality in the Kleine Nete, a Winder, M., Reuter, J.E., Schladow, S.G., 2009. Lake warming favours small-sized
Belgian lowland stream. Limnologica 28, 145e152. planktonic diatom species. Proc. R. Soc. B 276, 427e435. http://dx.doi.org/
Van Tol, H.M., Amin, S.A., Armbrust, E.V., 2016. Ubiquitous marine bacterium in- 10.1098/rspb.2008.1200.
hibits diatom cell division. ISME J. http://dx.doi.org/10.1038/ismej.2016.112. Wood, R.J., Mitrovic, S.M., Kefford, B.J., 2014. Determining the relative sensitivity of
Veldhuis, M., Kraay, G., Timmermans, K., 2001. Cell death in phytoplankton: cor- benthic diatoms to atrazine using rapid toxicity testing: a novel method. Sci. Total
relation between changes in membrane permeability, photosynthetic activity, Environ. 485e486, 421e427. http://dx.doi.org/10.1016/j.scitotenv.2014.03.115.
pigmentation and growth. Eur. J. Phycol. 36, 167e177. http://dx.doi.org/10.1080/ Wood, R.J., Mitrovic, S.M., Lim, R.P., Kefford, B.J., 2016. How benthic diatoms within
09670260110001735318. natural communities respond to eight common herbicides with different
Verb, R.G., Vis, M.L., 2005. Periphyton assemblages as bioindicators of mine- modes of action. Sci. Total Environ. 557e558, 636e643. http://dx.doi.org/
drainage inunglaciated Western Allegheny Plateau lotic systems. Water Air 10.1016/j.scitotenv.2016.03.142.

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Contents lists available at ScienceDirect

The use of diatoms in ecotoxicology and bioassessment: Insights,

* Corresponding author. Department of Marine Science, Incheon National Uni-

1. Introduction adverse effects of chemicals at the population and community

Achnanthidium exiguum 2* (1) 6* (2)e23(2) 4* (1)e16(2) 7 20e77 13e53

Pinnularia subcapitata 2 (0.1e0.5) 4 (1e3) 2e5 15e60

Diatom species No. of frustules examined % change in length

1. Cocconeis placentula 50 0e25

Table 4 deformities were present in low percentages. In ﬁeld studies,

viability of deformed frustules, their reproduction capabilities, and References

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