Survival,Growth and Condition of Freshwater Mussels: Effects of Municipal Wastewater Effluent

Freshwater mussels (Family Unionidae) are among the most imperiled group of organisms in the world, with nearly 65% of North American species considered endangered. Anthropogenic disturbances, including altered flow regimes, habitat alteration, and pollution, are the major driver of this group''s decline. We investigated the effects of tertiary treated municipal wastewater effluent on survivorship, growth, and condition of freshwater mussels in experimental cages in a small Central Texas stream. We tested the effluent effects by measuring basic physical parameters of native three ridge mussels (Amblema plicata) and of non-native Asian clams (Corbicula fluminea), before and after 72-day exposure at four sites above and below a municipal wastewater treatment plant outfall. Survivorship and growth of the non-native Asian clams and growth and condition indices of the native three ridge mussels were significantly higher at the reference site above the outfall than in downstream sites. We attribute this reduction in fitness below the outfall to elevated nutrient and heavy metal concentrations, and the potential presence of other untested-for compounds commonly found in municipal effluent. These results, along with an absence of native mussels below the discharge, indicate a significant negative impact of wastewater effluent on both native and non-native mussels in the stream.     

Using Bioassays and Species Sensitivity Distributions to Assess Herbicide Toxicity towards Benthic Diatoms

Although benthic diatoms are widely used in ecological studies of aquatic systems, there is still a dearth of data concerning species sensitivities towards several contaminants. Within the same community, different species may respond differently depending on their physiological and ecological characteristics. This lack of knowledge makes specific appropriate risk assessment impossible. To find out whether species sensitivity distribution (SSD) could be used to estimate the risk of herbicide toxicity for diatoms, we need to know whether their sensitivity depends on their physiological and ecological characteristics. We carried out single-species bioassays on 11 diatom species exposed to 8 herbicides. Dose-responses relationships were used to extrapolate the Effective Concentration 5 (EC(5)) and the Effective Concentration 50 (EC(50)) for each exposure. These data were used to fit a SSD curve for each herbicide, and to determine the Hazardous concentration 5 (HC(5)) and 50 (HC(50)). Our results revealed a high level of variability of the sensitivity in the set of species tested. For photosystem-II inhibitor (PSII) herbicides, diatoms species displayed a typical grouping of sensitivity levels consistent with their trophic mode and their ecological guild. N-heterotroph and "motile" guild species were more tolerant of PSII inhibitors, while N-autotroph and "low profile" guild species were more sensitive. Comprehensive SSD curves were obtained for 5 herbicides, but not for sulfonylurea herbicides or for dimetachlor, which had toxicity levels that were below the range of concentration tested. The SSD curves provided the following ranking of toxicity: diuron> terbutryn> isoproturon> atrazine> metolachlor. The HC that affected 5% of the species revealed that, even at the usual environmental concentrations of herbicides, diatom assemblages could be affected, especially by isoproturon, terbutryn, and diuron.     

Food-Web Structure in Relation to Environmental Gradients and Predator-Prey Ratios in Tank-Bromeliad Ecosystems

Little is known of how linkage patterns between species change along environmental gradients. The small, spatially discrete food webs inhabiting tank-bromeliads provide an excellent opportunity to analyse patterns of community diversity and food-web topology (connectance, linkage density, nestedness) in relation to key environmental variables (habitat size, detrital resource, incident radiation) and predators:prey ratios. We sampled 365 bromeliads in a wide range of understorey environments in French Guiana and used gut contents of invertebrates to draw the corresponding 365 connectance webs. At the bromeliad scale, habitat size (water volume) determined the number of species that constitute food-web nodes, the proportion of predators, and food-web topology. The number of species as well as the proportion of predators within bromeliads declined from open to forested habitats, where the volume of water collected by bromeliads was generally lower because of rainfall interception by the canopy. A core group of microorganisms and generalist detritivores remained relatively constant across environments. This suggests that (i) a highly-connected core ensures food-web stability and key ecosystem functions across environments, and (ii) larger deviations in food-web structures can be expected following disturbance if detritivores share traits that determine responses to environmental changes. While linkage density and nestedness were lower in bromeliads in the forest than in open areas, experiments are needed to confirm a trend for lower food-web stability in the understorey of primary forests.     

Subsampling Herbarium Collections to Assess Geographic Diversity Gradients: A Case Study with Endemic Orchidaceae and Rubiaceae in Cameroon

We compiled herbarium specimen data to provide an improved characterization of geographic patterns of diversity using indices of species diversity and floristic similarity based on rarefaction principles. A dataset of 3650 georeferenced plant specimens belonging to Orchidaceae and Rubiaceae endemic to Atlantic Central Africa was assembled to assess species composition per half‐degree or one‐degree grid cells. Local diversity was measured by the expected number of species (S_k) per grid cell found in subsamples of increasing size and compared with raw species richness (S_R). A nearly unbiased estimator of the effective number of species per grid cell was also used, allowing quantification of ratios of ‘true diversity’ between grid cells. Species turnover was measured using a presence/absence‐based similarity index (Sørensen) and an abundance‐based index that corrects for sampling bias (NNESS). Our results confirm that the coastal region of Cameroon is more diverse in endemic species than those more inland. The southern part of this coastal forest is, however, as diverse as the more intensively inventoried northern part, and should also be recognized as an important center of endemism. A strong congruence between Sørensen and NNESS similarity matrices lead to similar delimitations of floristic units. Hence, heterogeneous sampling seems to confer more bias when measuring patterns of local diversity using raw species richness than species turnover using Sørensen index. Overall, we argue that subsampling methods represent a useful way to assess diversity gradients using herbarium specimens while correcting for heterogeneous sampling effort. Abstract in French is available in the online version of this article.     

A Whole Cell Bioreporter Approach to Assess Transport and Bioavailability of Organic Contaminants in Water Unsaturated Systems

Bioavailability of contaminants is a prerequisite for their effective biodegradation in soil. The average bulk concentration of a contaminant, however, is not an appropriate measure for its availability; bioavailability rather depends on the dynamic interplay of potential mass transfer (flux) of a compound to a microbial cell and the capacity of the latter to degrade the compound. In water-unsaturated parts of the soil, mycelia have been shown to overcome bioavailability limitations by actively transporting and mobilizing organic compounds over the range of centimeters. Whereas the extent of mycelia-based transport can be quantified easily by chemical means, verification of the contaminant-bioavailability to bacterial cells requires a biological method. Addressing this constraint, we chose the PAH fluorene (FLU) as a model compound and developed a water unsaturated model microcosm linking a spatially separated FLU point source and the FLU degrading bioreporter bacterium Burkholderia sartisoli RP037-mChe by a mycelial network of Pythium ultimum. Since the bioreporter expresses eGFP in response of the PAH flux to the cell, bacterial FLU exposure and degradation could be monitored directly in the microcosms via confocal laser scanning microscopy (CLSM). CLSM and image analyses revealed a significant increase of the eGFP expression in the presence of P. ultimum compared to controls without mycelia or FLU thus indicating FLU bioavailability to bacteria after mycelia-mediated transport. CLSM results were supported by chemical analyses in identical microcosms. The developed microcosm proved suitable to investigate contaminant bioavailability and to concomitantly visualize the involved bacteria-mycelial interactions.     

重要环境因子对小球藻去除污水中氮磷的影响

对小球藻去除污水中氮磷的性能进行了研究,考察了初始氮磷浓度、氮磷比、光照条件和pH等因素对其去除效率的影响。结果表明,在初始氮磷浓度分别在35 mg/L和7 mg/L以下时去除率接近100%;氮磷比为5∶1和10∶1,小球藻第4 d基本完全去除了污水中的氨态氮,而氮磷比对小球藻的除磷能力没有显著影响;在初始氨态氮或总磷浓度相同的条件下,光照条件(L/D为24 h∶0 h和12 h∶12 h)对氮磷去除效果的无明显差异性（P＞0.05）,而随着氮磷浓度的增加,连续光照条件逐渐展现出去除氮磷的优势;小球藻在pH 7～8范围内对氨态氮的去除率最佳,pH 5～7范围内,对总磷的去除率最佳。     

Phosphoprotein from Dentin. New Approaches to Achieve and Assess Purity

Phosphoprotein extracted from rat incisors was purified by passage through a sulfonated polystyrene column. The phosphoprotein that emerged in the void volume contained 54% phosphoserine + serine and 36% aspartic acid and, in contrast to that obtained by DEAE-cellulose chromatography, was devoid of proline, valine, isoleucine, leucine, tyrosine, phenylalanine and arginine. Gel electrophoresis of the material purified on sulfonated polystyrene columns gave one major phosphate-containing band which would not stain with Coomassie Blue. EDTA or acetic acid demineralization yielded phosphoprotein preparations with identical compositions and electrophoretic properties. These data show that purification procedures reported earlier are insufficient.     

A Modified RNA-Seq Approach for Whole Genome Sequencing of RNA Viruses from Faecal and Blood Samples

To date, very large scale sequencing of many clinically important RNA viruses has been complicated by their high population molecular variation, which creates challenges for polymerase chain reaction and sequencing primer design. Many RNA viruses are also difficult or currently not possible to culture, severely limiting the amount and purity of available starting material. Here, we describe a simple, novel, high-throughput approach to Norovirus and Hepatitis C virus whole genome sequence determination based on RNA shotgun sequencing (also known as RNA-Seq). We demonstrate the effectiveness of this method by sequencing three Norovirus samples from faeces and two Hepatitis C virus samples from blood, on an Illumina MiSeq benchtop sequencer. More than 97% of reference genomes were recovered. Compared with Sanger sequencing, our method had no nucleotide differences in 14,019 nucleotides (nt) for Noroviruses (from a total of 2 Norovirus genomes obtained with Sanger sequencing), and 8 variants in 9,542 nt for Hepatitis C virus (1 variant per 1,193 nt). The three Norovirus samples had 2, 3, and 2 distinct positions called as heterozygous, while the two Hepatitis C virus samples had 117 and 131 positions called as heterozygous. To confirm that our sample and library preparation could be scaled to true high-throughput, we prepared and sequenced an additional 77 Norovirus samples in a single batch on an Illumina HiSeq 2000 sequencer, recovering >90% of the reference genome in all but one sample. No discrepancies were observed across 118,757 nt compared between Sanger and our custom RNA-Seq method in 16 samples. By generating viral genomic sequences that are not biased by primer-specific amplification or enrichment, this method offers the prospect of large-scale, affordable studies of RNA viruses which could be adapted to routine diagnostic laboratory workflows in the near future, with the potential to directly characterize within-host viral diversity.     

Toxicity and Sublethal Effects of Phthalides Analogs to Rhyzopertha dominica

Phthalides and their precursors have demonstrated a large variety of biological activities. Eighteen phthalides were synthesized and tested on the stored grain pest Rhyzopertha dominica. In the screening bioassay, compounds rac‐(2R,2aS,4R,4aS,6aR,6bS,7R)‐7‐bromohexahydro‐2,4‐methano‐1,6‐dioxacyclopenta[cd]pentalen‐5(2H)‐one ( 15 ) and rac‐(3R,3aR,4R,7S,7aS)‐3‐(propan‐2‐yloxy)hexahydro‐4,7‐methano‐2‐benzofuran‐1(3H)‐one ( 17 ) showed mortality similar to the commercial insecticide, Bifenthrin® (≥90 %). The time (LT₅₀) and dose (LD₅₀) necessary to kill 50 % of the R. dominica population were determined for the most efficacious phthalides 15 and 17 . Compound 15 presented the lowest LD₅₀ (1.97 μg g^?1), being four times more toxic than Bifenthrin® (LD₅₀=9.11 μg g^?1). Both compounds presented an LT₅₀ value equal to 24 h. When applied at a sublethal dose, both phthalides (especially compound 15 ), reduced the emergence of the first progeny of R. dominica. These findings highlight the potential of phthalides 15 and 17 as precursors for the development of insecticides for R. dominica control.     

A Proposed Structure of Modified Nucleosides Expected to Have High Antiviral Activity and Low Toxicity

This article describes modified nucleosides with deactivated 3′-OH and modified at two or more positions. These are expected to have high antiviral activity as well as have low toxicity.     

Sediment Toxicity Bioassays Using Ruditapes decussatus Embryos and Larvae to Assess the Contamination in Four Sites on the Coast of Tunisia

This paper aims to assess the marine sediment quality along the Tunisian coast using clam Ruditapes decussatus embryos and larval bioassays tests. Elutriate samples prepared with sediment from four sites (Monastir Lagoon, Chebba, Mahres, and Zarat) were used for chemical analysis and clam bioassays tests (embryogenesis, larval growth. and metamorphosis). For embryogenesis test, five elutriates concentrations (12%, 25%, 50%, 75%, and 100%) were prepared. Metal concentrations were highest at the Mahres and Monastir Lagoon sites. Clam embryogenesis success was significantly (p < 0.05) reduced in all elutriates preparations at concentrations up to 50% except Chebba, where inhibition was evident at only the highest concentration (100%). Compared to the control (9.4 μm day^?1), larval growth rate was significantly (p < 0.05) reduced only in two sites, Monastir Lagoon (6.8 μm day^?1) and Zarat (5.6 μm day^?1). However, larval survival was lower (42.9%) in elutriates prepared from Mahres sediment compared to other studied sites and control treatment. We did not note any significant (p > 0.05) effect of elutriate preparations on clam metamorphosis; conversely, survival in this stage was significantly affected in three sites (Monastir: 59.0 ± 6.7%; Zarat: 52.1 ± 6.3%, and Mahres: 64.1 ± 5.4%) compared to the control (83.2 ± 4.7%). This work shows that both clam embryos and larvae are sensitive to contaminants and can be used to evaluate sediment contamination and monitor pollution.     

Utilizing Longitudinal Measures of Fetal Growth to Create a Standard Method to Assess the Impacts of Maternal Disease and Environmental Exposure

Impaired or suboptimal fetal growth is associated with an increased risk of perinatal morbidity and mortality. By utilizing readily available clinical data on the relative size of the fetus at multiple points in pregnancy, including delivery, future epidemiological research can improve our understanding of the impacts of maternal, fetal, and environmental factors on fetal growth at different windows during pregnancy. This study presents mean and standard deviation ultrasound measurements from a clinically representative US population that can be utilized for creating Z-scores to this end. Between 2006 and 2012, 18, 904 non-anomalous pregnancies that received prenatal care, first and second trimester ultrasound evaluations, and ultimately delivered singleton newborns at Brigham and Women’s hospital in Boston were used to create the standard population. To illustrate the utility of this standard, we created Z-scores for ultrasound and delivery measurements for a cohort study population and examined associations with factors known to be associated with fetal growth. In addition to cross-sectional regression models, we created linear mixed models and generalized additive mixed models to illustrate how these scores can be utilized longitudinally and for the identification of windows of susceptibility. After adjustment for a priori confounders, maternal BMI was positively associated with increased fetal size beginning in the second trimester in cross-sectional models. Female infants and maternal smoking were associated with consistently reduced fetal size in the longitudinal models. Maternal age had a non-significant association with increased size in the first trimester that was attenuated as gestation progressed. As the growth measurements examined here are widely available in contemporary obstetrical practice, these data may be abstracted from medical records by investigators and standardized with the population means presented here. This will enable easy extension of clinical data to epidemiologic studies investigating novel maternal, fetal, and environmental factors that may impact fetal growth.     

Abundance and Biomass Responses of Microbial Food Web Components to Hydrology and Environmental Gradients within a Floodplain of the River Danube

This study investigated the relationships of time-dependent hydrological variability and selected microbial food web components. Samples were collected monthly from the Kopa?ki Rit floodplain in Croatia, over a period of 19 months, for analysis of bacterioplankton abundance, cell size and biomass; abundance of heterotrophic nanoflagellates and nanophytoplankton; and concentration of chlorophyll a. Similar hydrological variability at different times of the year enabled partition of seasonal effects from hydrological changes on microbial community properties. The results suggested that, unlike some other studies investigating sites with different connectivity, bacterioplankton abundance, and phytoplankton abundance and biomass increased during lentic conditions. At increasing water level, nanophytoplankton showed lower sensitivity to disturbance in comparison with total phytoplankton biomass: this could prolong autotrophic conditions within the floodplain. Bacterioplankton biomass, unlike phytoplankton, was not impacted by hydrology. The bacterial biomass less affected by hydrological changes can be an important additional food component for the floodplain food web. The results also suggested a mechanism controlling bacterial cell size independent of hydrology, as bacterial cell size was significantly decreased as nanoflagellate abundance increased. Hydrology, regardless of seasonal sucession, has the potential to structure microbial food webs, supporting microbial development during lentic conditions. Conversely, other components appear unaffected by hydrology or may be more strongly controlled by biotic interactions. This research, therefore, adds to understanding on microbial food web interactions in the context of flood and flow pulses in river-floodplain ecosystems.     

A Comprehensive Behavioral Test Battery to Assess Learning and Memory in 129S6/Tg2576 Mice

Transgenic Tg2576 mice overexpressing human amyloid precursor protein (hAPP) are a widely used Alzheimer’s disease (AD) mouse model to evaluate treatment effects on amyloid beta (Aβ) pathology and cognition. Tg2576 mice on a B6;SJL background strain carry a recessive rd1 mutation that leads to early retinal degeneration and visual impairment in homozygous carriers. This can impair performance in behavioral tests that rely on visual cues, and thus, affect study results. Therefore, B6;SJL/Tg2576 mice were systematically backcrossed with 129S6/SvEvTac mice resulting in 129S6/Tg2576 mice that lack the rd1 mutation. 129S6/Tg2576 mice do not develop retinal degeneration but still show Aβ accumulation in the brain that is comparable to the original B6;SJL/Tg2576 mouse. However, comprehensive studies on cognitive decline in 129S6/Tg2576 mice are limited. In this study, we used two dementia mouse models on a 129S6 background—scopolamine-treated 129S6/SvEvTac mice (3–5 month-old) and transgenic 129S6/Tg2576 mice (11–13 month-old)–to establish a behavioral test battery for assessing learning and memory. The test battery consisted of five tests to evaluate different aspects of cognitive impairment: a Y-Maze forced alternation task, a novel object recognition test, the Morris water maze, the radial arm water maze, and a Y-maze spontaneous alternation task. We first established this behavioral test battery with the scopolamine-induced dementia model using 129S6/SvEvTac mice and then evaluated 129S6/Tg2576 mice using the same testing protocol. Both models showed distinctive patterns of cognitive impairment. Together, the non-invasive behavioral test battery presented here allows detecting cognitive impairment in scopolamine-treated 129S6/SvEvTac mice and in transgenic 129S6/Tg2576 mice. Due to the modular nature of this test battery, more behavioral tests, e.g. invasive assays to gain additional cognitive information, can easily be added.     

Environmental Life Cycle Costing and Sustainability: Insights from Pollution Abatement and Resource Recovery in Wastewater Treatment

The relationship between environmental life cycle costing (ELCC) and sustainability was explored using two detailed wastewater case studies. The case studies were selected to increase the tension between existing market values and values for sustainability; the first case study considered incremental change to an existing plant and the second considered a paradigm shift in wastewater treatment. Pollution control provided the greatest cost savings for the first case study and provided a “win‐win” result—meeting existing standards and saving money. However, benefits for pollution control beyond current standards were not captured, which emphasized the role of standards to internalize as well as limit the values considered in ELCC. In the second case study, the value of water had the potential to change the focus of wastewater design from pollution abatement to resource recovery. However, social acceptance of recovered water and market access for resources created large risk for investment. The ELCC was also sensitive to the discount rate which limited longer‐term considerations. Other sustainability values such as scarcity and ecological thresholds were not captured. The ELCC code of practice suggests including such costs if likely in the foreseeable future; defining these values may also clarify the role of ELCC to evaluate sustainability over the life cycle.     

Archaeal and Bacterial Communities Respond Differently to Environmental Gradients in Anoxic Sediments of a California Hypersaline Lake,the Salton Sea

Sulfidic, anoxic sediments of the moderately hypersaline Salton Sea contain gradients in salinity and carbon that potentially structure the sedimentary microbial community. We investigated the abundance, community structure, and diversity of Bacteria and Archaea along these gradients to further distinguish the ecologies of these domains outside their established physiological range. Quantitative PCR was used to enumerate 16S rRNA gene abundances of Bacteria, Archaea, and Crenarchaeota. Community structure and diversity were evaluated by terminal restriction fragment length polymorphism (T-RFLP), quantitative analysis of gene (16S rRNA) frequencies of dominant microorganisms, and cloning and sequencing of 16S rRNA. Archaea were numerically dominant at all depths and exhibited a lesser response to environmental gradients than that of Bacteria. The relative abundance of Crenarchaeota was low (0.4 to 22%) at all depths but increased with decreased carbon content and increased salinity. Salinity structured the bacterial community but exerted no significant control on archaeal community structure, which was weakly correlated with total carbon. Partial sequencing of archaeal 16S rRNA genes retrieved from three sediment depths revealed diverse communities of Euryarchaeota and Crenarchaeota, many of which were affiliated with groups previously described from marine sediments. The abundance of these groups across all depths suggests that many putative marine archaeal groups can tolerate elevated salinity (5.0 to 11.8% [wt/vol]) and persist under the anaerobic conditions present in Salton Sea sediments. The differential response of archaeal and bacterial communities to salinity and carbon patterns is consistent with the hypothesis that adaptations to energy stress and availability distinguish the ecologies of these domains.The vast majority of cultured Archaea isolates are characterized as extremophiles, which thrive under environmental extremes of temperature, pH, salinity, and oxygen availability. Unlike Bacteria, these organisms are well defined by select physiologies or catabolic activities. Cultivated halophilic archaea are obligate aerobes, and with a few exceptions (58), most 16S rRNA gene sequences affiliated with this physiological group have been recovered primarily from environments with oxygen present. Thermophilic archaea, many of which utilize hydrogen-based metabolisms, have temperature requirements that preclude their survival and growth in more moderate environments. Other archaeal physiological groups include acidophiles, which thrive in acidic and mostly high-temperature environments, the obligate anaerobic methanogens, which are capable of competing with Bacteria when more energetically favorable electron acceptors are not available (i.e., sulfate), and methane-oxidizing archaea, which require methane for energy production. Recent work on several Crenarchaeota isolates points to nitrification as their primary energy metabolism, but these organisms have been detected in cold, predominantly aerobic environments, such as open ocean waters and soil (47), and in hyperthermophilic environments (24).Several archaeal groups identified using only 16S rRNA genes, for which no current isolates exist, have been detected in anaerobic sediments of the marine subsurface (6), estuaries (42), freshwater (46), and salt lakes (29). While their physiology and catabolism remain a source of speculation, the environmental distribution patterns of these mesophilic, presumably anaerobic, groups seemingly exclude the physiological and catabolic types outlined above. That is, the persistence of diverse archaeal populations in anoxic sediments at moderate temperature and salinity and at circumneutral pH with only trace levels of methane strongly suggests that alternative metabolic or physiological activities must characterize these populations.Saline lakes are ubiquitous and can be found on all continents. Although many saline lakes are labeled “extreme” environments, microbial diversity within their sediments is often equivalent to that reported for studies of freshwater and marine systems (28). Most studies of the microbial ecology within saline lakes have focused on gradients within the water column, with very few studies on patterns within the sediments. Specifically, these studies have examined how changes in water column salinity lead to shifts in microbial productivity and diversity (8). However, particle-associated microbial communities are known to differ fundamentally from water column or free-living populations (1, 18). These observed differences could be explained by the type and strength of environmental gradients that microbial communities in sediments experience, as opposed to those encountered by pelagic communities.Sediments contain strong environmental gradients, such as time (e.g., sediment age at depth), nutrient and carbon availability, and the dominant terminal electron-accepting process (TEAP) resulting from the sequential use of available oxidants by the microbial community (41). These gradients can lead to changes in the dominant microbial groups (i.e., a shift from sulfate reducers to methanogens with depth and age). Many saline lakes are highly productive and shallow and experience large fluctuations in water level due to climatic changes or to changes in inflows due to urban and agricultural activities. Changes in lake level can lead to dramatic shifts in mixing regimens, nutrient cycling, and water chemistry. Historic fluctuations in water column salinity are often recorded within the sediments in the form of evaporite deposits, which may act as additional sources of ionic loading of the water column (62). These sedimentary salinity gradients may modulate the metabolic activity of some microbial groups. For example, Oren (44) proposed bioenergetic constraints as a possible explanation for the reduced activity or absence of some microbial groups within high-salinity environments. Thus, saline lake sediments are excellent natural laboratories in which to study changes and adaptations of microbial communities due to large-scale changes in environmental gradients.The Salton Sea is a large (980 km²), eutrophic, moderately hypersaline (48 to 50 g liter⁻¹), terminal lake located 69 m below sea level in the Salton Basin, CA. Several large lakes have formed in the Salton Basin over geologic history, the most recent of which was Lake Cahuilla ca. 300 years ago (7). The current lake was unintentionally created in 1905-1907, when the Colorado River flooded the Salton Basin for a period of 16 months. Profundal sediments are highly sulfidic, and sulfate reduction is suspected to be the dominant TEAP within these sediments (54). Based on elemental analysis (51) and ¹³⁷Cs activity (37) of sediment layers, a depth of ∼22 cm marks the point when flooding of the Salton Basin occurred. Sediment above this depth represents the ca. 102 years of historical change within the Salton Sea, including a shift from a water column salinity of 35 g liter⁻¹ to the hypersaline conditions that currently exist. Sediments below this depth consist of low-carbon, gypsum-rich evaporite deposits that were present on the older dry lake bed prior to the formation of the current lake. A previous study reported several strong geochemical gradients within pore water across this relatively small depth range (62).In this work, a suite of cultivation-independent techniques and geochemical analyses was utilized to correlate shifts in abundance, community structure, and diversity of Archaea and Bacteria in Salton Sea sediments with changes in environmental gradients. Large differences in abundance and community structure patterns of Archaea and Bacteria were found along the gradients. In addition, the majority of archaeal sequences retrieved were affiliated with previously described but as yet uncultivated groups identified from various marine sedimentary environments. This indicates that these groups are able to tolerate the higher salinity and anaerobic conditions characteristic of Salton Sea sediments. Fundamental differences between the metabolic capacities and ecologies of Archaea and Bacteria are discussed to explain these patterns.     

Upscaling from Rhizosphere to Whole Root System: Modelling the Effects of Phospholipid Surfactants on Water and Nutrient Uptake

While the rhizosphere presents a different chemical, physical and biological environment to bulk soil, most experimental and modelling investigations of plant growth and productivity are based on bulk soil parameters. In this study, water and nutrient acquisition by wheat (Triticum aestivum L.) roots was investigated using rhizosphere- and root-system-scale modelling. The physical and chemical properties of rhizosphere soil could be influenced by phospholipid surfactants in the root mucilage. Two models were compared: a 2-dimensional (2D) Finite Element Method rhizosphere model, and a 3-dimensional (3D) root architecture model, ROOTMAP. ROOTMAP was parameterised to reproduce the results of the detailed 2D model, and was modified to include a rhizosphere soil volume. Lecithin (a phospholipid surfactant) could be exuded into the rhizosphere soil volume, decreasing soil water content and hydraulic conductivity at any given soil water potential, and decreasing phosphate adsorption to soil particles. The rhizosphere-scale modelling (5 × 5 mm² soil area, 10 mm root length, uptake over 12 h) predicted a reduction in water uptake (up to 16% at 30 kPa) and an increase in phosphate uptake (up to 4%) with lecithin exudation into the rhizosphere, but little effect on nitrate uptake, with only a small reduction in dry soil (1.6% at 200 kPa). The 3D root model reproduced the water (y = 1.013x, R² = 0.996), nitrate (y = 1x, R² = 1) and phosphate (y = 0.978x, R² = 0.998) uptake predictions of the rhizosphere model, providing confidence that a whole root system model could reproduce the dynamics simulated by a Finite Element Method rhizosphere model. The 3D root architecture model was then used to scale-up the rhizosphere dynamics, simulating the effect of lecithin exudation on water, nitrate and phosphate acquisition by a wheat root system, growing over 41 d. When applied to growing and responsive roots, lecithin exudation increased P acquisition by up to 13% in nutrient-rich, and 49% in relatively nutrient-poor soil. A comparison of wheat (Triticum aestivum L.) and lupin (Lupinus angustifolius L.) root architectures, suggested an interaction between the P acquisition benefit of rhizosphere lecithin and root architecture, with the more highly-branched wheat root structure acquiring relatively more P in the presence of lecithin than the sparsely-branched lupin root system.     

Antibiotic Resistance Characteristics of Environmental Bacteria from an Oxytetracycline Production Wastewater Treatment Plant and the Receiving River

We characterized the bacterial populations in surface water receiving effluent from an oxytetracycline (OTC) production plant. Additional sampling sites included the receiving river water 5 km upstream and 20 km downstream from the discharge point. High levels of OTC were found in the wastewater (WW), and the antibiotic was still detectable in river water downstream (RWD), with undetectable levels in river water upstream (RWU). A total of 341 bacterial strains were isolated using nonselective media, with the majority being identified as Gammaproteobacteria. The MICs were determined for 10 antibiotics representing seven different classes of antibiotics, and the corresponding values were significantly higher for the WW and RWD isolates than for the RWU isolates. Almost all bacteria (97%) from the WW and RWD samples demonstrated multidrug-resistant (MDR) phenotypes, while in RWU samples, these were less frequent (28%). The WW and RWD isolates were analyzed for the presence of 23 tetracycline (tet) resistance genes. The majority of isolates (94.2% and 95.4% in WW and RWD, respectively) harbored the corresponding genes, with tet(A) being the most common (67.0%), followed by tet(W), tet(C), tet(J), tet(L), tet(D), tet(Y), and tet(K) (in the range between 21.0% and 40.6%). Class I integrons were detected in the majority of WW and RWD isolates (97.4% and 86.2%, respectively) but were not associated with the tet genes. We hypothesize that the strong selective pressure imposed by a high concentration of OTC contributes to the wide dissemination of tetracycline resistance genes and other antibiotic resistance genes, possibly through mobile genetic elements.The widespread emergence of antibiotic resistance, particularly multidrug resistance (MDR), among bacterial pathogens has become one of the most serious challenges in clinical therapy (22, 46). Some pathogens, such as MDR Klebsiella pneumoniae and Acinetobacter baumannii, are now virtually untreatable with current antibiotics (14, 30). Acquisition of resistance genes through horizontal transfer has been found to be ubiquitous in clinical pathogens (22). Environmental bacteria have been shown to be a reservoir of antibiotic resistance genes and a potential source of novel resistance genes in clinical pathogens (10, 12). Horizontal transfer of genes between bacterial strains could be facilitated by mobile genetic elements, such as plasmids, transposons, bacteriophages, integrons, insertion elements (IS), and genomic islands (13). Some elements, including class I integrons, conjugative plasmids, and transposons, are frequently linked to antibiotic resistance as they harbor rather diverse resistance genes and possibly promote the distribution of these genes in phylogenetically diverse bacteria (29). In light of the potential health risk, many studies have focused on antibiotic-resistant bacteria recovered from various ecosystems (1, 18, 45). Environments that contain antibiotic residues are particularly worrisome because antibiotics could exert selective pressure and might contribute to the appearance of resistant bacteria. Hospital sewage was once considered the major source of antibiotics in aquatic environments, followed by municipal, agricultural, and aquacultural wastewater (WW), which have also been shown to be important sources of these compounds and resistant bacteria (40). It has also been reported that treated antibiotic production wastewater contains much higher concentrations of antibiotic residues than other aquatic environments (20, 26, 27) and can serve as an important reservoir of resistant bacteria and genes (25).In the current study, we investigated resistance profiles of bacterial isolates from a unique wastewater treatment plant (WWTP) that is used solely for treating oxytetracycline (OTC), avermectin, and ivermectin production wastewater from the facility of the North China Pharmaceutical Group Corporation in Hebei Province, China. Avermectin and ivermectin are both broad-spectrum antiparasitic agents without any antibacterial or antifungal activities. The effluent of the WWTP is discharged directly into the receiving river without disinfection, and residual levels of OTC in wastewater and downstream sampling points nearly approach those in human blood after drug administration (15, 26). Thus, the characteristics of bacterial strains in heavily OTC-contaminated wastewater and surface water could be unique. In a 1-year (2002-2003) survey of antibiotic resistance in 140 clinical enterococcal isolates from a hospital in this region, the prevalence of tetracycline resistance ranged from 14.9% to 25.0% (47). This rate increased to 47.9 to 75.9% for 302 clinical enterococcal isolates from the same hospital over the following 3 years (2004-2006) (28), indicating that tetracycline resistance among human pathogens in this region is becoming more prevalent. Elucidation of the resistance characteristics of bacterial isolates from OTC wastewater and surface water might help explain the prevalence of increased resistance among human pathogens. To gain a comprehensive understanding of this relationship, we isolated bacterial strains from wastewater (WW) and river water downstream (RWD) and river water upstream (RWU) samples using nonselective culture media. Previously, it was shown that long-term administration of a single antibiotic can lead to MDR, a phenomenon that has been observed in bacteria obtained from the urinary tract, intestine, and other niches in both clinical therapy and livestock raising (11, 23, 34). Thus, the combination of long-term treatment and discharge of antibiotic production wastewater might contribute to MDR in environmental bacteria in this specific ecosystem. To test this hypothesis, we determined the resistance profiles of bacterial isolates for 10 antibiotics representing seven classes.OTC belongs to the tetracycline class of antibiotics, which includes tetracycline, chlortetracycline, doxycycline, minocycline, and glycylcyclines (6). This class of antibiotics represents broad-spectrum agents that act against a range of Gram-positive and Gram-negative bacteria by inhibiting protein synthesis. The tetracyclines have played an important role in human and veterinary medicine, and some have been used as growth promoters for livestock and aquaculture. However, resistance to tetracyclines in many commensal and pathogenic bacteria emerged soon after the widespread application of this class of antibiotic. More than 40 types of tetracycline resistance determinants have been described to date and grouped into three main classes: energy-dependent membrane-associated efflux proteins, which export tetracycline out of the cell; ribosomal protection proteins, which interact with the ribosome and disrupt the tetracycline binding site; and tetracycline inactivation enzymes (6, 37). Many studies of tetracycline resistance (tet) genes in various environmental niches have been reported. Resistance genes encoding efflux proteins and ribosomal protection proteins have been identified in phylogenetically diverse bacterial genera. Extensive reviews with detailed information about tet genes and their distribution are available (4, 6, 24, 37). Notably lacking, however, is information about tet genes in aquatic environments with high levels of antibiotic residue. Studies have shown that several tet genes are associated with mobile genetic elements (6, 37) and that the SOS response induced by antibiotics and other factors can promote horizontal dissemination of mobile genetic elements like genomic islands, as well as integron recombination, among bacterial populations (5, 17, 41). In the current study, the presence and distribution of 23 tet genes in bacterial isolates from wastewater and river water were determined by PCR. Class I integrons were also examined in the bacterial isolates to evaluate the possibility of horizontal transfer of these resistance genes and/or the promoted dissemination of mobile genetic elements in such heavily antibiotic-polluted environments. We also compared the characteristics of the recovered bacteria with those of isolates from samples from water upstream of the discharge site in the river. Our results reveal the potential environmental influence of wastewater discharge on acquired antibiotic resistance.     

Life Cycle Environmental and Cost Impacts of Using an Algal Turf Scrubber to Treat Dairy Wastewater

Using algae to simultaneously treat wastewater and produce energy products has potential environmental and economic benefits. This study evaluates the life cycle energy, greenhouse gas (GHG) emissions, eutrophication potential, and cost impacts of incorporating an algal turf scrubber (ATS) into a treatment process for dairy wastewater. A life cycle inventory and cost model was developed to simulate an ATS treatment system where harvested algae would be used to generate biogas for process heat and electricity generation. Modeling results show that using an ATS significantly reduces eutrophication impacts by reducing chemical oxygen demand, nitrogen, and phosphorus in the wastewater. With low water recirculation rates through the ATS and high algae productivity, inclusion of the ATS results in net energy displacement and a reduction of GHG emissions compared to a system with no ATS. However, if high water recirculation rates are used or if algae biosolids from the digester are dried, the system results in a net increase in energy consumption and GHG emissions. The life cycle treatment cost was estimated to be $1.42 USD per cubic meter of treated wastewater. At this cost, using an ATS would only be cost effective for dairies if they received monetary credits for improved water quality on the order of $3.83 per kilogram of nitrogen and $9.57 per kilogram of phosphorus through, for example, nutrient trading programs.