Gut passage and insect grazer selectivity of lotic diatoms

SUMMARY 1. Grazing experiments were conducted in a small Kentucky stream to determine if epilithic diatom taxa differed in susceptibility to grazing by the larval caddisfly, Neophylax autumnus , and whether ingested taxa varied in digestibility. Live/dead ratios of diatom cells from N. autumnus faecal material were compared with live/dead ratios of cells taken from adjacent epilithic habitats to determine diatom digestibility. Selectivity was studied by comparing relative abundances of diatom taxa on substrata with the relative abundances of these taxa in grazer faecal material.
2. 73% of the diatoms collected from epilithic habitats contained intact chloroplasts and were characterized as live cells. Only 42% of diatoms eliminated in caddisfly faeces were living. The five dominant diatom taxa did not differ in digestibility. Elimination of viable diatoms in grazer faecal material may contribute to diatom drift in streams, and thus supply individuals for downstream recolonization. Failure to document live/dead ratios of diatoms from grazed substrata can lead to overestimation of the digestibility of taxa and misinterpretation of results.
3. Grazers in this investigation were more successful at ingesting large, high-profile diatom taxa (i.e. Cymbella, Meridion and Gomphonema ) and less able to remove small, adnate forms ( Achnanthes minutissima ). Such 'selectivity'may be one mechanism by which A. minutissima maintains dominance in this system.     

Stream epilithic, epipelic and epiphytic diatoms: habitat fidelity and use in biomonitoring

We compared the use of epilithic, epiphytic, and epipelic diatom communities in stream biomonitoring by investigating species composition and relationships with measured water quality variables in two tributaries of the Grand River, Ontario, Canada. Although ordination showed some separation of the epilithon from other habitats, clear habitat preferences and seasonality were not identified. Canonical correspondence analysis showed that measured water quality variables explained the major gradients in the diatom data for all three habitats. The diatoms from each habitat were strongly related to alkalinity, suspended solids, biological oxygen demand and conductivity, and in addition, epipelic and epiphytic diatoms showed significant relationships with total phosphorus. It was concluded that although some species were more abundant in one habitat than the others, the community structure of the epilithic, epiphytic and epipelic habitats from these streams was not consistently different. The results indicate no apparent benefit to sampling discrete habitats for water quality monitoring using diatoms, however, the best relationship are obtained using the sum of the diatom data from all three habitats.     

Benthic diatoms as indicators of eutrophication in tropical streams

Diatoms are frequently used as indicators of eutrophication in temperate systems, but little is known about their application to impacted African tropical systems. Five streams located within Gombe Stream National Park and five streams supporting human settlements draining into Lake Tanganyika, East Africa, were investigated for species composition, richness and diversity of epilithic algae. In addition, a trophic diatom index (TDI) developed for monitoring European rivers was applied to these tropical systems. 54 specific and infraspecific diatom taxa representing 20 genera were identified for all sites with Achnanthes s.l., Gomphonema and Navicula s.l. being the most common genera. Species richness varied between 10 and 21 in disturbed streams and 13 and 19 in undisturbed streams. Nutrients were significantly enriched in streams draining the deforested watersheds but indices of diversity and evenness (Shannon H, J and Simpson–Yule D, E) did not show any significant differences between streams in forested and deforested watersheds. Significant differences were observed between pooled data for the TDI between forested and deforested watersheds. Analysis of percent pollution tolerant diatom taxa indicates that organic pollution of streams in deforested watersheds may be contributing to eutrophication. This study shows that African diatoms, cosmopolitan or resembling well-known North American and European taxa, allows for trophic indices tailored to the autecological preferences of species to be applied to new regions, although intensive studies on these African taxa will lead to more accurate results. Measures of species-richness and diversity, historically used to describe the state of an ecosystem, may not be suitable to evaluate streams which are not grossly polluted. Electronic supplementary material Electronic supplementary material is available for this article at and accessible for authorised users.     

THE IMPORTANCE OF DIATOM CELL SIZE IN COMMUNITY ANALYSIS1

The large variation in size and shape in diatoms is shown by morphometric measurements of 515 benthic and pelagic diatom species from the Baltic Sea area. The largest mean cell dimension (mostly the apical axis) varied between 4.2 and 653 μm, cell surface area between 55 and 344,000 μm², and cell volume between 21 and 14.2 × 10⁶μm³. The shape‐related index, length to width ratio, was between 1.0 and 63.3 and the shape‐ and size‐related index, surface area to volume ratio, was between 0.02 and 3.13. Diatom community analysis by multivariate statistics is usually based on counts of a fixed number of diatom valves with species scores irrespective of cell size. This procedure underestimates the large species for two reasons. First, the importance of a species with higher cell volume is usually larger in a community. Second, larger species usually have lower abundances and their occurrence in the diatom counts is stochastic. This article shows that co‐occurring small and large diatom species can respond very differently to environmental constraints. Large epiphytic diatoms responded most to macroalgal host species and small epiphytic diatoms most to environmental conditions at the sampling site. Large epilithic diatoms responded strongly to salinity, whereas small epilithic diatoms did so less clearly. The conclusion is that different scale‐dependent responses are possible within one data set. The results from the test data also show that important ecological information from diatom data can be missed when the large species are neglected or underestimated.     

Harmonization is more important than experience—results of the first Nordic–Baltic diatom intercalibration exercise 2007 (stream monitoring)

The goal of this study was a harmonization of diatom identification and counting among diatomists from the Scandinavian and Baltic countries to improve the comparison of diatom studies in this geographical area. An analysis of the results of 25 diatomists following the European standard EN 14407 during an intercalibration exercise showed that a high similarity was achieved by harmonization and not because of a long experience with diatoms. Sources of error were wrong calibration scales, overlooking of small taxa, especially small Navicula s.l., misidentifications (Eunotia rhomboidea was mistaken for Eunotia incisa) and unclear separation between certain taxa in the identification literature. The latter was discussed during a workshop with focus on the Achnanthes minutissima group, the separation of Fragilaria capucina var. gracilis from F. capucina var. rumpens, and Nitzschia palea var. palea from N. palea var. debilis. The exercise showed also that the Swedish standard diatom method tested here worked fine with acceptable error for the indices IPS (Indice de Polluo-sensibilité Spécifique) and ACID (ACidity Index for Diatoms) when diatomists with a low similarity (Bray–Curtis <60%) with the auditor in at least one of the samples are excluded.     

Alpine freshwater ecosystems in a protected area: a source of diatom diversity

The “All Taxa Biodiversity Inventories” (ATBIs) project coordinated by the European Distributed Institute of Taxonomy (EDIT) aims to achieve a baseline biodiversity assessment of flora and fauna in those regions characterized by a lack of knowledge and a high potential for biodiversity. Within the framework of the ATBIs, the aim of this study was to analyse the diatom flora and ecology of a complex of freshwater ecosystems in the Maritime Alps Natural Park (Italy), designated as a Special area of Conservation under the European Habitat Directive. We sampled epilithic and epiphytic diatoms in different habitats in 24 sites: shallow lake, springs, and streams. Our analysis resulted in a list of 138 diatom taxa, highlighting the great biodiversity and the complex structure of the investigated diatom communities. The taxa list included a wide range of uncommon species, including some recorded for the first time in North-Western Italy. Among the different habitats the highest level of diversity was found in the more lentic waters, in particular in limnocrene springs. These results show that the diatom communities of pristine and undisturbed high-mountain environments are rich and complex, despite the severe environmental conditions.     

Seasonality of macroalgae and epilithic diatoms in spring-fed streams in Texas, USA

A seasonal study of two spring-fed stream systems in south-central Texas was undertaken over a 15-month period from June 1996 to September 1997. Relative abundance of the epilithic diatom flora, percent cover of macroalgae, and several physical and chemical conditions were monitored in one 20-m stretch in each of the streams at approximately two-month intervals. Six additional spring-fed stream segments were sampled in May 1996 for macroalgae and measured for the same conditions, in order to determine how representative the macroalgal floras of the study sites were of south-central and western Texas. Conditions in the two stream segments were relatively constant over the 15 months, except for maximum depth; this finding is similar those reported for other spring-fed systems worldwide. Sixty-eight diatom taxa and nine macroalgae taxa were identified from the San Marcos River, which compares to 46 diatom taxa and six macroalgae taxa from the Comal River. Several diatom and macroalgae species were significantly correlated to stream conditions (e.g. Dichotomosiphon tuberosus and Hildenbrandia angolensis with daylength, and Amphora pediculus with maximum depth). Twenty-one macroalgal taxa were identified from the six additional spring-fed steam segments. Conditions in these streams were very similar to those measured in the San Marcos River and Comal River, except that they had a higher average pH _x= 8.1 versus _x= 7.6). The epilithic diatom flora of the San Marcos River and the Comal River contains many of the diatom and some macroalgal taxa commonly reported from springs globally. This revised version was published online in July 2006 with corrections to the Cover Date.     

Taxonomy‐free molecular diatom index for high‐throughput eDNA biomonitoring

Current biodiversity assessment and biomonitoring are largely based on the morphological identification of selected bioindicator taxa. Recently, several attempts have been made to use eDNA metabarcoding as an alternative tool. However, until now, most applied metabarcoding studies have been based on the taxonomic assignment of sequences that provides reference to morphospecies ecology. Usually, only a small portion of metabarcoding data can be used due to a limited reference database and a lack of phylogenetic resolution. Here, we investigate the possibility to overcome these limitations using a taxonomy‐free approach that allows the computing of a molecular index directly from eDNA data without any reference to morphotaxonomy. As a case study, we use the benthic diatoms index, commonly used for monitoring the biological quality of rivers and streams. We analysed 87 epilithic samples from Swiss rivers, the ecological status of which was established based on the microscopic identification of diatom species. We compared the diatom index derived from eDNA data obtained with or without taxonomic assignment. Our taxonomy‐free approach yields promising results by providing a correct assessment for 77% of examined sites. The main advantage of this method is that almost 95% of OTUs could be used for index calculation, compared to 35% in the case of the taxonomic assignment approach. Its main limitations are under‐sampling and the need to calibrate the index based on the microscopic assessment of diatoms communities. However, once calibrated, the taxonomy‐free molecular index can be easily standardized and applied in routine biomonitoring, as a complementary tool allowing fast and cost‐effective assessment of the biological quality of watercourses.     

Uncertainty of using habitat fidelity in biomonitoring based on benthic diatoms - the Raška River case study

Epilithic diatoms are frequently recommended for river biomonitoring, while much less emphasis is placed on epiphytic communities. This study considers use of epiphytic and epilithic diatom communities from the Ra?ka River in biomonitoring. A total of 212 diatom taxa were recorded in both communities. Dominant diatoms were Achnanthidium minutissimum (Kützing) Czarnecki, A. affine (Grunow) Czarnecki, A. subatomus (Hustedt) Lange-Bertalot, Amphora pediculus (Kützing) Grunow, Diatoma vulgaris Bory, Gomphonema tergestinum (Grunow) Fricke, Cocconeis placentula var. lineata (Ehrenberg) Van Heurck, Melosira varians Agardh and Navicula tripunctata (Müller) Bory. Redundancy analysis (RDA) was used to detect the relationships between diatoms, some environmental factors and sampling sites. Diatoms of the epiphytic community showed a clearly grouping in relation to the sampling sites as compared with diatoms of the epilithic community. Species common to both communities showed a similar correlation with some environmental variables (BOD, pH, NH₄⁺, TP, NO₃^? and conductivity), with three exceptions (Diatoma vulgaris, Cymbella compacta, and Encyonema silesiacum). Pearson’s correlation coefficient showed correlation between selected environmental variables and diatom indices calculated using OMNIDIA 6.2 software (TDI, IPS, CEE, GENRE, TID, SID, SHE and IDSE/5). Diatoms of the epiphytic community are more clearly clustered in relation to the sampling sites as compared with diatoms of the epilithic community. According to our results there is still “place” in biomonitoring for epiphytic community. The present study highlights the necessity of sampling both epiphytic and epilithic communities because substrate specificity could play important role in biomonitoring.     

Is the biological classification of benthic diatom communities concordant with ecotypes?

In this study a biological classification based on diatom communities (a posteriori classification) was used to search for the most appropriate a priori classification in reference conditions, on the basis of 31 reference sites encompassing eight watersheds in the NE Iberian Peninsula. We considered three different a priori systems for comparison: (i) a typological classification (ecotypes and subecotypes) based on geomorphology and water flow; (ii) the watershed as a unit, irrespective of its size; and (iii) a classification based on the geographical distance between sites under the assumption that biological characteristics are increasingly similar in geographically closer sites. Classification in ecotypes and subecotypes was the most robust with minor differences between them. The watershed (hydrological units) classification was significant though weaker than the typological classification and there was a larger environmental variation within watersheds than within ecotypes. Finally, results showed that geographically closer sites were not more biologically similar in terms of diatom community composition. We can conclude that the ecotypological environmental-based classifications developed by the local water agency did adequately describe the biological classification based on epilithic diatoms.     

Relationships Between Environmental Variables and Benthic Diatom Assemblages in California Central Valley Streams (USA)

This study examines distributional patterns of benthic diatom assemblages in relation to environmental characteristics in streams and rivers in the California Central Valley ecoregion. Benthic diatoms, water quality, and physical habitat conditions were characterized from 53 randomly selected sites. The stream sites were characterized by low mid-channel canopy cover and high channel substrate embeddedness. The waters at these sites were enriched with minerals and turbidity varied from 1.3 to 185.0 NTU with an average of 13.5 NTU. A total of 249 diatom taxa were identified. Average taxa richness was 41 with a range of 7–76. The assemblages were dominated by Staurosira construens (11%), Epithemia sorex (8%), Cocconeis placentula (7%), and Nitzschia amphibia (6%). Multivariate analyses (cluster analysis, classification tree analysis, and canonical correspondence analysis) all showed that benthic diatom assemblages were mainly affected by channel morphology, in-stream habitat, and riparian conditions. The 1st CCA axis negatively correlated with mean wetted channel width (r = −0.66) and thalweg depth (r = −0.65) (Table 4). The 2nd axis correlated with % coarse substrates (r=0.60). Our results suggest that benthic diatoms can be used for assessing physical habitat alterations in streams.     

Ecological background of diatom functional groups: Comparability of classification systems

Functional classification based on species traits brought a revolution in community ecology, and also boosted phytoplankton and in phytobenton (diatom) research. Several studies stressed the usefulness of phytoplankton functional groups in ecological status assessment, and there is also a strong emphasis to use combined traits in ecological assessments of diatom assemblages. The Combined Eco-Morphological Functional Groups (CEMFGs) help to reveal species-environmental correlations, which can be hidden, controlling traits separately. Nowadays, there are three types of functional guild classifications simultaneously used in the literature: (i) the original classification proposed by Passy (O); (ii) Passy’s classification complemented with a separated planktic guild (P); (iii) the refined guild classification by Rimet and Bouchez, also containing the planktic guild (RB). One of the most important criteria of the combined functional groups is the well-defined ecological frame of these combined groups; thus it is vital to harmonise the classification of the taxa into guild based combined eco-morphological functional groups (CEMFGs). In this study we tested the similarities and dissimilarities of the correlations between environmental factors and CEMFGs created in the (i)-(iii) classifications. Samples were collected in 138 sampling sites on lowland rivers and streams in the Hungarian Lowland Region. Strong correlations were anticipated between the functional groups and environmental factors due to the presence of common and/or abundant planktic taxa. Our results validated the necessity of a separated planktic guild in ecological assessments of diatom assemblages. Further relevant differences between the correlations of CEMFGs-P or CEMFGs-RB and abiotic factors were hypothesised in the case of those functional groups which contained reassigned taxa with high frequency and/or abundance. Our results confirmed this hypothesis. Furthermore, the present study also highlighted the relevance of well-defined trait classification. Abundant and/or frequent taxa, which are able to change their life forms, can modify significantly the relationship between the functional group containing them and environmental factors. In the future, both field and laboratory studies should focus on revealing the circumstances, which cause the mentioned changes in traits of diatoms.     

The revised edition of a computerized plankton counter for plankton,periphyton and sediment diatom analyses

A revised computer counting program is presented with faster processing and more efficient features to enhance counting precision. The program is now capable of counting epilithic communities and sediment samples for diatom analysis. This version (2.3) is available for APPLE, IBM, TANDY-RADIO SHACK, and HYPERION systems using standard Basic commands. New improvements include, density and biovolume calculations, a Chi square transect test for randomness, D'Agostino's test for a normal taxa distribution and a three file output with raw and processed data.     

Regional distribution of diatom assemblages in the headwater streams of Luxembourg

The aim of this study was to determine the influence of environmental variables on the structure of benthic diatom assemblages, and to propose type assemblages of diatoms characterizing unpolluted headwater streams of Luxembourg. A total of 289 diatom samples were collected in the headwater streams of Luxembourg. At each sampling site, physical and chemical variables were also measured. The relationships between environmental variables and the distribution of the taxa were estimated using univariate and multivariate statistical analyses. On the basis of geology and stream water chemistry, Luxembourg could be separated in two regions. The streams in the schistose northern part of Luxembourg presented a low carbonate hardness (median 3.5° F); the streams in the southern part of Luxembourg presented a high carbonate hardness (median 21.8° F) notably connected to the presence of sandstone and limestone substrata. A Twinspan classification carried out on diatom assemblages defined two groups of samples corresponding to these two regions. The carbonate hardness that is related to the nature of the geological substratum appeared to be the major structuring variable for the assemblage composition. Anthropogenic pollution was a secondary structuring variable for diatom assemblages since each group could be subdivided in subgroups presenting statistically different nutrient and organic matter concentrations. The diatom assemblages of these subgroups were characterised by differences of saprobic and trophic preferences. On the basis of these results, two type assemblages of diatoms are proposed for the unpolluted headwater streams in the two regions of Luxembourg. The presence of these different type assemblages in Luxembourg shows the necessity to adapt diatom bioindication to the different regions of the country.     

Epilithic diatoms as indicators in tropical African rivers (Lake Victoria catchment)

We investigated epilithic diatoms in rivers draining to the Nyanza Bay in Lake Victoria (Kenya) with the aim of determining environmental gradients in the assemblages and testing the usefulness of diatom metrics from temperate regions. Spatial and temporal variations of assemblages were studied in 12 sites of three rivers. Kibos, Nyando, and Kisat rivers contained 224 diatom taxa collected on seven sampling occasions over 4 years. Species richness showed a marginal decrease downstream and was the lowest at sites with high conductivity and ammonia–nitrogen levels. Two-Way Indicator Species Analysis and Canonical Correspondence Analysis revealed two major groups of river sites. Conductivity, alkalinity, turbidity, dissolved oxygen, and silicate were the most important variables influencing species distribution. Ecological diatom metrics of temperate regions and the Specific Pollution sensitivity Index showed good relationships with environmental variables. Both diatom assemblages and averaged indicator values were strong in predicting sites of ecological deterioration and in differentiating an upstream site of better quality (drinking water supply of Kisumu), thereby confirming epilithic diatoms as suitable water quality indicators in equatorial rivers. The use of metrics initially designed for temperate rivers, however, appears less valuable in good quality tropical rivers because potential indicators are not considered.     

Seasonality and distribution of epilithic diatoms,macroalgae and macrophytes in a spring-fed stream system in Ontario,Canada

A study of the epilithic diatom, macroalgal and macrophyte communities from a spring-fed stream in Ontario, Canada was undertaken from September 1996 to July 1997. The relative abundance of the epilithic diatom flora, percent cover of macroalgal and macrophyte taxa, and several physical and chemical stream conditions were monitored along a 20-m stretch at each of four sites, approximately every 2 months. Several stream conditions were relatively constant over the sampling period (pH, maximum width and maximum depth), while others exhibited a distinct seasonal pattern (water temperature, specific conductance and daylength) and some fluctuated strongly with no discernable seasonal pattern (turbidity, current velocity). A total of 124 taxa were identified from the four sites, including 79 epilithic diatoms, three macroalgal diatom species (large gelatinous masses), one cyanobacterium, two red algae, eight green algae, one chrysophyte alga, one tribophyte alga, three mosses, three horsetails and 23 angiosperm taxa. Species richness was positively correlated to stream channel maximum width and depth, indicating that the total number of species tends to increase in a downstream direction. Distribution of several diatom and macroalgal species was significantly correlated to stream conditions (e.g. Gomphonema parvulum and Phormidium subfuscum with current velocity); however, the vast majority of species did not display seasonal variation in abundance that could be explained by changes in stream conditions. Many of the taxa identified from Blue Springs Creek are common elsewhere in North America.     

Using algal metrics and biomass to evaluate multiple ways of defining concentration-based nutrient criteria in streams and their ecological relevance

We examined the utility of nutrient criteria derived solely from total phosphorus (TP) concentrations in streams (regression models and percentile distributions) and evaluated their ecological relevance to diatom and algal biomass responses. We used a variety of statistics to characterize ecological responses and to develop concentration-based nutrient criteria (derived from ecological effects) for streams in Connecticut, USA, where urbanization is the primary cause of watershed alteration. Mean background TP concentration in the absence of anthropogenic land cover was predicted to be 0.017 mg/l, which was similar to the 25th percentile of all study sites. Increased TP concentrations were significantly correlated with altered diatom community structure, decreased percent low P diatoms and diatoms sensitive to impervious cover, and increased percent high P diatoms, diatoms that increase with greater impervious cover, and chlorophyll a (P < 0.01). Variance partitioning models showed that shared effects of anthropogenic land cover and chemistry (i.e., chemistry affected by land cover) represented the majority of explained variation in diatom metrics and chlorophyll a. Bootstrapped regression trees, threshold indicator taxa analysis, and boosted regression trees identified TP concentrations at which strong responses of diatom metrics and communities occurred, but these values varied among analyses. When considering ecological responses, scientifically defensible and ecologically relevant TP criteria were identified at (1) 0.020 mg/l for designating highest quality streams and restoration targets, above which sensitive taxa steeply declined, tolerant taxa increased, and community structure changed, (2) 0.040 mg/l, at which community level change points began to occur and sensitive diatoms were greatly reduced, (3) 0.065 mg/l, above which most sensitive diatoms were lost and tolerant diatoms steeply increased to their maxima, and (4) 0.082 mg/l, which appeared to be a saturated threshold, beyond which substantially altered community structure was sustained. These criteria can inform anti-degradation policies for high quality streams, discharge permit decisions, and future strategies for watershed development and managment. Our results indicated that management practices and decisions at the watershed scale will likely be important for improving degraded streams and conserving high quality streams. Results also emphasized the importance of incorporating ecological responses and considering the body of evidence from multiple conceptual approaches and statistical analyses for developing nutrient criteria, because solely relying on one approach could lead to misdirected decisions and resources.     

Use of benthic diatom communities to evaluate water quality in rivers of southern Poland

Biological and chemical data were processed to estimate trophic stage and degree of pollution in several streams and rivers in southern Poland. The majority were eutrophic and some of them heavily polluted; only a few were oligo-mesotrophic. The differences in the water quality of the rivers were reflected by different types of diatom community and also by the values for some diatom indices, which were calculated using the latest version of the 'Omnidia' database software. Except for the Sládeček's index, all diatom indices correlated significantly with organic load (COD), oxygen concentration, conductivity and most of the measured ions. Some indices showed a significant negative correlation with trophic level (expressed by NH₄-N and PO₄-P). In general, IPS (Specific Pollution Sensitivity Index) and GDI (Generic Diatom Index) indices gave the best results. Among the investigated diatom communities, only a few taxa indicated oligo-mesotrophy and oligo-β-mesosaprobity. Most of the sites were characterised by a greater relative contribution of eutraphent and tolerant ones as well as α-mesosaprobic and polysaprobic diatoms. This study suggests that the structure of benthic diatom communities and diatom indices, especially GDI, can be applied for monitoring rivers in Poland. This revised version was published online in June 2006 with corrections to the Cover Date.     

PERIPHYTIC DIATOM ASSEMBLAGES FROM HIGH ARCTIC PONDS1

Epiphytic, epilithic, and surface sediment diatom assemblages were identified and enumerated from 35 study ponds on CapeHerschel (78°37″N, 74°42″W), east-central Ellesmere Island, Canada. All the sites are shallow (maximum depth <2 m), clear, oligotrophic, and freshwater. The ponds freeze completely for 10 months of the year. Major ion concentrations are relatively similar among the 35 sites, although environmental gradients exist. Over 130 diatom taxa from 28 genera were identified in the periphyton samples. Marked differences in species composition were evident among the ponds. Moreover, many of the diatoms exhibited varying degrees of microhabitat specificity. Variance partitioning by canonical correspondence analysis showed that 26% of the total variance exhibited by diatom species composition could be accounted for by the measured environmental variables (i.e. 10.2% by habitat and 15.8% by water chemistry). Pondwater alkalinity best explained the distribution of taxa, and weighted averaging regression and calibration were used to develop a transfer function to infer pondwater alkalinity from the diatom assemblages. Other important environmental variables included [Na⁺] for the epilithic and [SiO₂] for the epiphytic assemblages.