Things We Like: Human Preferences among Similar Organisms and Implications for Conservation

Human preferences will increasingly determine many species’ prospects for survival. However, aside from a small number of survey-based studies of preference among disparate taxa, human species preferences have received little attention. I determined human aesthetic preferences among a relatively homogenous group, the penguins, from representation in all recently published, comprehensive, popular books of photographs of penguins (n = 4 books; 304 photographs). Representation of visually distinguishable types of penguins, measured by total photograph area, was highly skewed and rankings were highly concordant across books, suggesting large and commonly held differences in aesthetic appeal. Multiple regression analysis indicated that amount of warm color was the only significant determinant of representation, and warm color was highly correlated (r ² = 0.96) with mean representation of the penguin types. Body size and neotenic form, traits found to influence human preferences among other animals, were not significant, suggesting that the bases of human species preferences differ by species type. The results of this study indicate that human aesthetic preferences discriminate finely among species and may be based on minor features. Conservationists must be vigilant to the potential for aesthetic responses to influence conservation efforts.     

Denitrification in Aquatic Environments: A Cross-system Analysis

A meta-analysis was conducted on 136 data sets of denitrification rates (DR) recorded both during the period of highest water temperature and monthly in five types of aquatic ecosystems: oceans, coastal environments, estuaries, lakes and rivers. There was a gradual increase of DR from the ocean to rivers and lakes at both scales, with the rivers showing the highest DR variability. Denitrification peaked during summertime and showed highest seasonal variability in lakes and rivers. High concentrations of nitrate and interstitially-dissolved organic carbon as well as low oxygen concentration in the overlying water enhanced DR both during summer and at a seasonal scale whereas total phosphorus did at the seasonal scale only. There was a positive linear relationship between overlying nitrate and DR over the range of 1–970 μmol NO₃ (r ² = 0.86, P = 0.001). DR in lakes and rivers might reach values doubling those in the more denitrifying terrestrial ecosystems (e.g. agrosystems). Discrepancies in DR and its controlling factors between site-specific studies and this meta-analysis may arise from environmental variability at two, often confounded, scales of observation: the habitat and the ecosystem level. Future studies on denitrification in aquatic environments should address the topic of spatial heterogeneity more thoroughly.     

Simplified Method for Dissolved DNA Determination in Aquatic Environments

A method was developed for the determination of dissolved DNA in aquatic environments. The method is based upon the concentration of dissolved DNA by ethanol precipitation of 0.2-μm-pore-size filtered water. The DNA in concentrated extracts was quantified by the fluorescence of Hoechst 33258-DNA complexes. Fluorescence not attributable to DNA was corrected for by DNase I digestion of the extracts and averaged 25% of the total fluorescence for all samples. The effectiveness of the procedure for concentrating dissolved DNA was demonstrated by the efficient (>90%) recovery of internal standards. Concentrations of dissolved DNA from a variety of marine and freshwater environments ranged from 0.2 to 44 μg/liter, with the highest values being obtained for estuarine and river environments. The method is simple, specific for DNA, and more sensitive than previously described methods for the determination of extracellular DNA.     

Ecology of Vibrio mimicus in Aquatic Environments

[This corrects the article on p. 2077 in vol. 55.].     

Diatoms: A Novel Source for the Neurotoxin BMAA in Aquatic Environments

Amyotrophic lateral sclerosis (ALS) or Lou Gehrig’s disease is a neurological disorder linked to environmental exposure to a non-protein amino acid, β-N-methylamino-L-alanine (BMAA). The only organisms reported to be BMAA-producing, are cyanobacteria – prokaryotic organisms. In this study, we demonstrate that diatoms – eukaryotic organisms – also produce BMAA. Ultra-high-performance liquid chromatography coupled with tandem mass spectrometry revealed the occurrence of BMAA in six investigated axenic diatom cultures. BMAA was also detected in planktonic field samples collected on the Swedish west coast that display an overrepresentation of diatoms relative to cyanobacteria. Given the ubiquity of diatoms in aquatic environments and their central role as primary producers and the main food items of zooplankton, the use of filter and suspension feeders as livestock fodder dramatically increases the risk of human exposure to BMAA-contaminated food.     

Rate of Bacterial Mortality in Aquatic Environments

A method is proposed which provides a minimum estimate of the rate of bacterial mortality in growing natural populations of planktonic bacteria. This estimate is given by the rate of decrease of radioactivity from the DNA of a [³H]thymidine-labeled natural assemblage of bacteria after all added thymidine has been exhausted from the medium. Results obtained from river water, estuarine water, and seawater show overall bacterial mortality rates in the range 0.010 to 0.030 h⁻¹, in good agreement with the range of growth rates measured in the same environments. Use of selective filtration through Nuclepore filters (pore size, 2 μm) allowed us to determine the contribution of microzooplankton grazing to overall bacterial mortality. Grazing rates estimated by this method ranged from 0 to 0.02 h⁻¹.     

Methods for Detection of Conjugative Plasmid Transfer in Aquatic Environments

Donor and recipient counter selection was evaluated by selecting bacteria that received plasmid RP4 by conjugation on filters and in lake water microcosms. Three counter selection systems were compared; (i) Use of antibiotic-resistant recipients, (ii) use of an auxotrophic donor, and (iii) use of a donor with chromosomal suicide genes. Transfer efficiencies of transconjugants per recipient obtained with the three different counter selection systems in filter-matings were not significantly different. Some nalidixic acid-resistant recipients became partly sensitive to nalidixic acid after receiving the plasmid. Use of an auxotrophic donor was a feasible and easy way to recover indigenous transconjugants. A strain with two copies of the suicide gene gef was successfully eliminated in filter-matings, but elimination of the donor in microcosms by induction of the suicide genes did not succeed. Thus, this counter selection system was not usable in microcosm experiments. Received: 3 March 1998 / Accepted: 15 May 1998     

Molecular Barcoding of Aquatic Oligochaetes: Implications for Biomonitoring

Aquatic oligochaetes are well recognized bioindicators of quality of sediments and water in watercourses and lakes. However, the difficult taxonomic determination based on morphological features compromises their more common use in eco-diagnostic analyses. To overcome this limitation, we investigated molecular barcodes as identification tool for broad range of taxa of aquatic oligochaetes. We report 185 COI and 52 ITS2 rDNA sequences for specimens collected in Switzerland and belonging to the families Naididae, Lumbriculidae, Enchytraeidae and Lumbricidae. Phylogenetic analyses allowed distinguishing 41 lineages separated by more than 10 % divergence in COI sequences. The lineage distinction was confirmed by Automatic Barcode Gap Discovery (ABGD) method and by ITS2 data. Our results showed that morphological identification underestimates the oligochaete diversity. Only 26 of the lineages could be assigned to morphospecies, of which seven were sequenced for the first time. Several cryptic species were detected within common morphospecies. Many juvenile specimens that could not be assigned morphologically have found their home after genetic analysis. Our study showed that COI barcodes performed very well as species identifiers in aquatic oligochaetes. Their easy amplification and good taxonomic resolution might help promoting aquatic oligochaetes as bioindicators for next generation environmental DNA biomonitoring of aquatic ecosystems.     

Protein Method for Investigating Mercuric Reductase Gene Expression in Aquatic Environments

A colorimetric assay for NADPH-dependent, mercuric ion-specific oxidoreductase activity was developed to facilitate the investigation of mercuric reductase gene expression in polluted aquatic ecosystems. Protein molecules extracted directly from unseeded freshwater and samples seeded with Pseudomonas aeruginosa PU21(Rip64) were quantitatively assayed for mercuric reductase activity in microtiter plates by stoichiometric coupling of mercuric ion reduction to a colorimetric redox chain through NADPH oxidation. Residual NADPH was determined by titration with phenazine methosulfate-catalyzed reduction of methyl thiazolyl tetrazolium to produce visible formazan. Spectrophotometric determination of formazan concentration showed a positive correlation with the amount of NADPH remaining in the reaction mixture (r² = 0.99). Mercuric reductase activity in the protein extracts was inversely related to the amount of NADPH remaining and to the amount of formazan produced. A qualitative nitrocellulose membrane-based version of the method was also developed, where regions of mercuric reductase activity remained colorless against a stained-membrane background. The assay detected induced mercuric reductase activity from 10² CFU, and up to threefold signal intensity was detected in seeded freshwater samples amended with mercury compared to that in mercury-free samples. The efficiency of extraction of bacterial proteins from the freshwater samples was (97 ± 2)% over the range of population densities investigated (10² to 10⁸ CFU/ml). The method was validated by detection of enzyme activity in protein extracts of water samples from a polluted site harboring naturally occurring mercury-resistant bacteria. The new method is proposed as a supplement to the repertoire of molecular techniques available for assessing specific gene expression in heterogeneous microbial communities impacted by mercury pollution.     

Introduction to the Symposium: Ontogenetic Strategies of Invertebrates in Aquatic Environments

This symposium presents different ecological and physiologicalstrategies used by invertebrates to successfully adapt to aquaticenvironments. Adaptation has been studied mainly in adult animals,but the papers comprising the symposium emphasize ontogeneticstrategies, starting from the principle that natural selectionacts on all stages of development. Adaptive strategies may thusdiffer strikingly between developmental stages of the same organism.Invertebrates offer a wide array of ecophysiological modelsfor study, and these are exemplified by the contributions tothe symposium, which are briefly summarized. Future researchin the field will 1, expand the number of models for comparativepurposes; 2, examine the strategies, not only of larvae andjuveniles, but also of embryos, eggs and reproductive cells;and 3, investigate the genetic basis of ontogenetic strategies.     

Detection of the Antiviral Drug Oseltamivir in Aquatic Environments

Oseltamivir (Tamiflu®) is the most important antiviral drug available and a cornerstone in the defence against a future influenza pandemic. Recent publications have shown that the active metabolite, oseltamivir carboxylate (OC), is not degraded in sewage treatment plants and is also persistent in aquatic environments. This implies that OC will be present in aquatic environments in areas where oseltamivir is prescribed to patients for therapeutic use. The country where oseltamivir is used most is Japan, where it is used to treat seasonal flu. We measured the levels of OC in water samples from the Yodo River system in the Kyoto and Osaka prefectures, Japan, taken before and during the flu-season 2007/8. No OC was detected before the flu-season but 2–58 ng L⁻¹ was detected in the samples taken during the flu season. This study shows, for the first time, that low levels of oseltamivir can be found in the aquatic environment. Therefore the natural reservoir of influenza virus, dabbling ducks, is exposed to oseltamivir, which could promote the evolution of viral resistance.     

Intra- and Interannual Vegetation Change: Implications for Long-Term Research

To draw reliable conclusions from forest restoration experiments, it is important that long‐term measurements be repeatable or year‐to‐year variability may interfere with the correct interpretation of treatment effects. We used permanent plots in a long‐term restoration study in southwestern Colorado to measure herbaceous and shrub vegetation at three dates within a single year (June, July, and August), and between years (2003 and 2005), on untreated control plots in a warm, dry mixed conifer forest. Growing season precipitation patterns were similar between 2003 and 2005, so differences in vegetation should be related primarily to differences in the sampling month. Significant indicator species for each sampling month were present within a single year (2005), primarily reflecting early‐season annuals. We found no significant differences for total species abundance (2005). Species richness, abundance, and indicator species were significantly different between years for different sampling months indicating that sampling should be conducted within a similar time frame to avoid detecting differences that are not due to treatment effects or variations in year‐to‐year climate. These findings have implications for long‐term research studies where the objectives are to detect changes over time in response to treatments, climate variation, and natural processes. Long‐term sampling should occur within a similar phenological time frame each year over a short amount of time and should be based on the following criteria: (1) the sampling period is congruent with research objectives such as detecting rare species or peak understory abundance and (2) the sampling period is feasible in regard to personnel and financial constraints.     

Detection of Mercury in Aquatic Environments Using EPRE Reporter Zebrafish

It has been proposed that transgenic zebrafish could be designed to detect low levels of chemical contaminants that cause oxidative stress in aquatic environments, such as heavy metals or pesticides. In this paper, we describe such a transgenic zebrafish that produces a luciferase–green fluorescent protein (LUC–GFP) fusion protein under conditions of oxidative stress. The reporter gene expression is under the regulation of the electrophile responsive element (EPRE), which activates gene expression in response to oxidative stressors. The GFP component of this fusion protein allows us to visually detect reporter gene activity in live animals to determine if activity is localized to a particular tissue. The luciferase component is capable of returning a quantitative assessment of reporter gene activity that allows us to determine if reporter gene activity is directly correlated to the concentration of the chemical inducer. We have tested this reporter construct in both transient and stable transgenic fish after exposure to a range of HgCl₂ concentrations. GFP expression from the EPRE–LUC–GFP construct was inducible in transient assays but was below the limit of detection in stable lines. In contrast, we observed inducible luciferase activity in both transient assays and stable lines treated with HgCl₂. We conclude that the EPRE is capable of driving reporter gene expression in a whole animal assay under conditions of oxidative stress. Furthermore, expression was induced at HgCl₂ concentrations that do not result in obvious morphological defects, making this approach useful for the detection of low levels of oxidative contaminants in aquatic environments.     

The Challenges of Living in Hypoxic and Hypercapnic Aquatic Environments

Organisms living in coastal waters, and especially estuaries,have long been known to have behavioral or physiological mechanismsthat enable them toexist in water containing low amounts ofoxygen. However, the respiratory consumption of oxygen thatgenerates hypoxia is also responsible for producing significantamounts of carbon dioxide. An elevation of carbon dioxide pressurein water will cause a significant acidosis in most aquatic organisms.Thus, the combination of low oxygen and elevated carbon dioxidethat occurs in estuaries represents a significant environmentalchallenge to organisms living in this habitat. Organisms maymaintain oxygen uptake in declining oxygen conditions by usinga respiratory pigment and/or by making adjustments in the convectiveflow of water and blood past respiratory surfaces (i.e., increasecardiac output and ventilation rate). Severe hypoxia may resultin an organism switching partially or completely to anaerobicbiochemical pathways to sustain metabolic rate. There is alsoevidence to suggest that organisms lower their metabolism duringhypoxic stress. Elevated water CO₂ (hypercapnia) produces anacidosis in the tissues of organisms that breathe it. This acidosismay be wholly or partially compensated (i.e., mechanisms returnpH to pre-exposure levels), or may be uncompensated. Some studieshave examined the effects on organisms of exposure simultaneouslyto hypoxia and hypercapnia. This article reviews some of thespecific adaptations and responses of organisms to low oxygen,to high carbon dioxide, and to the cooccurrence of low oxygenand high carbon dioxide     

Combined Approach for Characterization of Uncultivated Magnetotactic Bacteria from Various Aquatic Environments

Both magnetic collection and “race track” purification techniques were highly effective for selective enrichment of magnetotactic bacteria (MTB) from complex communities, as suggested by amplified ribosomal DNA restriction analysis and denaturing gradient gel electrophoresis combined with sequence analysis of 16S rRNA genes. Using these purification methods, the occurrence and diversity of MTB in microcosms from various marine and freshwater environments were assayed by using a combined microscopic, molecular, and cultivation approach. Most microcosms were dominated by magnetotactic cocci. Consistently, the majority of retrieved 16S RNA sequences were affiliated with a distinct cluster in the Alphaproteobacteria. Within this lineage the levels of sequence divergence were <1 to 11%, indicating genus-level diversity between magnetotactic cocci from various microcosms, as well as between MTB from different stages of succession of the same microcosms. The community composition in microscosms underwent drastic succession during incubation, and significant heterogeneities were observed between microcosms from the same environmental sources. A novel magnetotactic rod (MHB-1) was detected in a sediment sample from a lake in northern Germany by fluorescence in situ hybridization. MHB-1 falls into the Nitrospira phylum, displaying 91% 16S rRNA sequence similarity to “Magnetobacterium bavaricum.” In extensive cultivation attempts, we failed to isolate MHB-1, as well as most other MTB present in our samples. However, although magnetotactic spirilla were not frequently observed in the enrichments, 10 novel isolates of the genus Magnetospirillum which had not routinely been isolated in pure culture before were obtained.     

Woody Debris Volume Depletion Through Decay: Implications for Biomass and Carbon Accounting

Woody debris decay rates have recently received much attention because of the need to quantify temporal changes in forest carbon stocks. Published decay rates, available for many species, are commonly used to characterize deadwood biomass and carbon depletion. However, decay rates are often derived from reductions in wood density through time, which when used to model biomass and carbon depletion are known to underestimate rate loss because they fail to account for volume reduction (changes in log shape) as decay progresses. We present a method for estimating changes in log volume through time and illustrate the method using a chronosequence approach. The method is based on the observation, confirmed herein, that decaying logs have a collapse ratio (cross-sectional height/width) that can serve as a surrogate for the volume remaining. Combining the resulting volume loss with concurrent changes in wood density from the same logs then allowed us to quantify biomass and carbon depletion for three study species. Results show that volume, density, and biomass follow distinct depletion curves during decomposition. Volume showed an initial lag period (log dimensions remained unchanged), even while wood density was being reduced. However, once volume depletion began, biomass loss (the product of density and volume depletion) occurred much more rapidly than density alone. At the temporal limit of our data, the proportion of the biomass remaining was roughly half that of the density remaining. Accounting for log volume depletion, as demonstrated in this study, provides a comprehensive characterization of deadwood decomposition, thereby improving biomass-loss and carbon-accounting models.     

On the Long-Term Fitness of Cells in Periodically Switching Environments

Because all the cell populations are capable of making switches between different genetic expression states in response to the environmental change, Thattai and van Oudenaarden (Genetics 167, 523–530, 2004) have raised a very interesting question: In a constantly fluctuating environment, which type of cell population (heterogeneous or homogeneous) is fitter in the long term? This problem is very important to development and evolution biology. We thus take an extensive analysis about how the cell population evolves in a periodically switching environment either with symmetrical time-span or asymmetrical time-span. A complete picture of the phase diagrams for both cases is obtained. Furthermore, we find that the systems with time-dependent cellular transitions all collapse to the same set of dynamical equations with the modified parameters. Furthermore, we also explain in detail how the fitness problem bears much resemblance to the phenomenon, stochastic resonance, in physical sciences. Our results could be helpful for the biologists to design artificial evolution experiments and unveil the mystery of development and evolution.     

Measurements of Diel Rates of Bacterial Secondary Production in Aquatic Environments

Measurements of bacterial secondary production were carried out during 13 diel studies at one coastal marine station and in five lakes differing with respect to nutrient concentration and primary production. Bacterial secondary production was measured in situ every 3 to 5 h by [³H]thymidine incorporation into DNA. In some of the diel studies, these results were compared with results obtained from dark ¹⁴CO₂ uptake and frequency of dividing cells. Only minor diel changes were observed. The rate of [³H]thymidine incorporation into DNA and the frequency of dividing cells varied from 23 to 194% of the diel mean. The dark CO₂ uptake rate varied from 12 to 259% of the diel mean. An analysis of variance demonstrated that no specific time periods during 24 h showed significantly different production rates, supporting the idea that bacterial activities in natural assemblages are controlled by a variety of events. The best correction (r² = 0.74) was obtained between the [³H]thymidine incorporation and frequency of dividing cells procedures from the lake water samples. The actual production rates calculated by [³H]thymidine incorporation into DNA were appreciably lower than those obtained by the frequency of dividing cells and the dark CO₂ uptake techniques. Diel rates of bacterial production are discussed in relation to sampling frequency, statistical errors, and choice of method.