Leached Compounds from Smoked Cigarettes and Their Potential for Bioaccumulation in Rainbow Trout (Oncorhynchus mykiss).

Cigarette butts are one of the most prevalent forms of litter worldwide and may leach toxic compounds when deposited in aquatic environments. Previous studies demonstrated that smoked cigarette leachate is toxic toward aquatic organisms. However, the specific bioavailable chemicals from the leachate and the potential for human and wildlife exposure through the food chain were unknown. Using a nontargeted analytical approach based on GC×GC/TOF-MS, 43 compounds were confirmed to leach from smoked cigarettes when exposed to a water source. Additionally, the bioaccumulation potential of organic contaminants in an edible fish, rainbow trout (Oncorhynchus mykiss), was assessed through direct exposure to the leachate of smoked cigarettes at 0.5 CB/L for 28 days. There was a significant reduction in fish mass among the exposed rainbow trout vs the control group (χ2 (1) = 5.3, p = 0.021). Both nontargeted and targeted chemical analysis of representative fish tissue identified four tobacco alkaloids, nicotine, nicotyrine, myosmine, and 2,2'-bipyridine. Their average tissue concentrations were 466, 55.4, 94.1, and 70.8 ng/g, respectively. This study identifies leached compounds from smoked cigarettes and demonstrates the uptake of specific chemicals in rainbow trout, thus suggesting a potential for accumulation in food webs, resulting in human and wildlife exposure.

Physiological and transcriptomic effects of hexafluoropropylene oxide dimer acid in Caenorhabditis elegans during development.

Per- and polyfluoroalkyl substances (PFAS) are chemicals resistant to degradation. While such a feature is desirable in consumer and industrial products, some PFAS, including perfluorooctanoic acid (PFOA), are toxic and bioaccumulate. Hexafluoropropylene oxide dimer acid (HFPO-DA), an emerging PFAS developed to replace PFOA, has not been extensively studied. To evaluate the potential toxicity of HFPO-DA with a cost- and time-efficient approach, we exposed C. elegans larvae for 48 h to 4 × 10-9-4 g/L HFPO-DA in liquid media and measured developmental, behavioral, locomotor, and transcriptional effects at various exposure levels. Worms exposed to 1.5-4 g/L HFPO-DA were developmentally delayed, and progeny production was significantly delayed (p < 0.05) in worms exposed to 2-4 g/L HFPO-DA. Statistically significant differential gene expression was identified in all fourteen HFPO-DA exposure groups ranging from 1.25 × 10-5 to 4 g/L, except for 6.25 × 10-5 g/L. Among 10298 analyzed genes, 2624 differentially expressed genes (DEGs) were identified in the developmentally delayed 4 g/L group only, and 78 genes were differentially expressed in at least one of the thirteen groups testing 1.25 × 10-5-2 g/L HFPO-DA exposures. Genes encoding for detoxification enzymes including cytochrome P450 and UDP glucuronosyltransferases were upregulated in 0.25-4 g/L acute exposure groups. DEGs were also identified in lower exposure level groups, though they did not share biological functions except for six ribosomal protein-coding genes. While our transcriptional data is inconclusive to infer mechanisms of toxicity, the significant gene expression differences at 1.25 × 10-5 g/L, the lowest concentration tested for transcriptional changes, calls for further targeted analyses of low-dose HFPO-DA exposure effects.

Assessing Toxicity and in Vitro Bioactivity of Smoked Cigarette Leachate Using Cell-Based Assays and Chemical Analysis.

Smoked cigarettes are the most prevalent form of litter worldwide, often finding their way into oceans and inland waterways. Cigarette smoke contains more than 4000 individual chemicals, some of them carcinogenic or otherwise toxic. We examined the cytotoxicity, genotoxicity, aryl hydrocarbon receptor (AhR), estrogen receptor (ER), and p53 response pathways of smoked cigarette leachate in vitro. Both seawater and freshwater leachates of smoked cigarettes were tested. Cytotoxicity and genotoxicity were negligible at 100 smoked cigarettes/L, while statistically significant AhR, ER, and p53 responses were observed in the extracts of both leachates, suggesting a potential risk to human health through exposure to cigarette litter in the environment. To identify responsible chemicals for the AhR response, an effect directed analysis approach was coupled with nontargeted chemical analysis based on comprehensive two-dimensional gas chromatography coupled to time-of-flight mass spectrometry (GC × GC/TOF-MS). Eleven compounds potentially responsible for the AhR response were identified. Among them, 2-methylindole was partially responsible for the AhR response.

Application of constructed wetlands for wastewater treatment in tropical and subtropical regions (2000-2013).

Constructed wetlands (CWs) have been successfully used for treating various wastewaters for decades and have been identified as a sustainable wastewater management option for developing countries. With the goal of promoting sustainable engineered systems that support human well-being but are also compatible with sustaining natural (environmental) systems, the application of CWs has become more relevant. Such application is especially significant for developing countries with tropical climates, which are very conducive to higher biological activity and productivity, resulting in higher treatment efficiencies compared to those in temperate climates. This paper therefore highlights the practice, applications, and research of treatment wetlands under tropical and subtropical conditions since 2000. In the present review, removal of biochemical oxygen demand (BOD) and total suspended solid (TSS) was shown to be very efficient and consistent across all types of treatment wetlands. Hybrid systems appeared more efficient in the removal of total suspended solid (TSS) (91.3%), chemical oxygen demand (COD) (84.3%), and nitrogen (i.e., 80.7% for ammonium (NH)4-N, 80.8% for nitrate (NO)3-N, and 75.4% for total nitrogen (TN)) as compared to other wetland systems. Vertical subsurface flow (VSSF) CWs removed TSS (84.9%), BOD (87.6%), and nitrogen (i.e., 66.2% for NH4-N, 73.3% for NO3-N, and 53.3% for TN) more efficiently than horizontal subsurface flow (HSSF) CWs, while HSSF CWs (69.8%) showed better total phosphorus (TP) removal compared to VSSF CWs (60.1%). Floating treatment wetlands (FTWs) showed comparable removal efficiencies for BOD (70.7%), NH4-N (63.6%), and TP (44.8%) to free water surface (FWS) CW systems.

Application of constructed wetlands for wastewater treatment in developing countries--a review of recent developments (2000-2013).

Inadequate access to clean water and sanitation has become one of the most pervasive problems afflicting people throughout the developing world. Replication of centralized water-, energy- and cost-intensive technologies has proved ineffective in resolving the complex water-related problems resulting from rapid urbanization in the developing countries. Instead constructed wetlands (CWs) have emerged and become a viable option for wastewater treatment, and are currently being recognized as attractive alternatives to conventional wastewater treatment methods. The primary objective of this review is to present a comprehensive overview of the diverse range of practice, applications and researches of CW systems for removing various contaminants from wastewater in developing countries, placing them in the overall context of the need for low-cost and sustainable wastewater treatment systems. Emphasis of this review is placed on the treatment performance of various types of CWs including: (i) free water surface flow CW; (ii) subsurface flow CW; (iii) hybrid systems; and, (iv) floating treatment wetland. The impacts of different wetland design and pertinent operational variables (e.g., hydraulic loading rate, vegetation species, physical configurations, and seasonal variation) on contaminant removal in CW systems are also summarized and highlighted. Finally, the cost and land requirements for CW systems are critically evaluated.

Toxicity assessment of hexafluoropropylene oxide-dimer acid on morphology, heart physiology, and gene expression during zebrafish (Danio rerio) development.

Hexafluoropropylene oxide-dimer acid (HFPO-DA) is one of the emerging replacements for the "forever" carcinogenic and toxic long-chain PFAS. HFPO-DA is a polymerization aid used for manufacturing fluoropolymers, whose global distribution and undetermined toxic properties are a concern regarding human and ecological health. To assess embryotoxic potential, zebrafish embryos were exposed to HFPO-DA at concentrations of 0.5-20,000 mg/L at 24-, 48-, and 72-h post-fertilization (hpf). Heart rate increased significantly in embryos exposed to 2 mg/L and 10 mg/L HFPO-DA across all time points. Spinal deformities and edema phenotypes were evident among embryos exposed to 1000-16,000 mg/L HFPO-DA at 72 hpf. A median lethal concentration (LC50) was derived as 7651 mg/L at 72 hpf. Shallow RNA sequencing analysis of 9465 transcripts identified 38 consistently differentially expressed genes at 0.5 mg/L, 1 mg/L, 2 mg/L, and 10 mg/L HFPO-DA exposures. Notably, seven downregulated genes were associated with visual response, and seven upregulated genes were expressed in or regulated the cardiovascular system. This study identifies biological targets and molecular pathways affected during animal development by an emerging, potentially problematic, and ubiquitous industrial chemical.

Locomotion and brain gene expression exhibit sex-specific non-monotonic dose-response to HFPO-DA during Drosophila melanogaster lifespan.

BACKGROUND: Legacy per- and polyfluoroalkyl substances (PFAS), known for their environmental persistence and bio-accumulative properties, have been phased out in the U.S. due to public health concerns. A newer polymerization aid used in the manufacture of some fluoropolymers, hexafluoropropylene oxide-dimer acid (HFPO-DA), has lower reported bioaccumulation and toxicity, but is a potential neurotoxicant implicated in dopaminergic neurodegeneration. OBJECTIVE: We investigated HFPO-DA's bio-accumulative potential and sex-specific effects on lifespan, locomotion, and brain gene expression in fruit flies. METHODS: We quantified bioaccumulation of HFPO-DA in fruit flies exposed to 8.7 × 104 µg/L of HFPO-DA in the fly media for 14 days via UHPLC-MS. Long-term effect on lifespan was determined by exposing both sexes to 8.7 × 102 - 8.7 × 105 µg/L of HFPO-DA in media. Locomotion was measured following 3, 7, and 14 days of exposures at 8.7 × 101 - 8.7 × 105 µg/L of HFPO-DA in media, and high-throughput 3'-end RNA-sequencing was used to quantify gene expression in fly brains across the same time points. RESULTS: Bioaccumulation of HFPO-DA in fruit flies was not detected. HFPO-DA-induced effects on lifespan, locomotion, and brain gene expression, and lowest adverse effect level (LOAEL) showed sexually dimorphic patterns. Locomotion scores significantly decreased in at least one dose at all time points for females and only at 3-day exposure for males, while brain gene expression exhibited non-monotonic dose-response. Differentially expressed genes correlated to locomotion scores revealed sex-specific numbers of positively and negatively correlated genes per functional category. CONCLUSION: Although HFPO-DA effects on locomotion and survival were significant at doses higher than the US EPA reference dose, the brain transcriptomic profiling reveals sex-specific changes and neurological molecular targets; gene enrichments highlight disproportionately affected categories, including immune response: female-specific co-upregulation suggests potential neuroinflammation. Consistent sex-specific exposure effects necessitate blocking for sex in experimental design during HFPO-DA risk assessment.

Effect of feeding strategies on pharmaceutical removal by subsurface flow constructed wetlands.

This study presents findings on an assessment of the effect of continuous and batch feeding strategies on the removal of selected pharmaceuticals from synthetic wastewater. Six mesocosm-scale constructed wetlands, including three horizontal subsurface flow constructed wetlands and three sand filters, were set up at the campus of Nanyang Technological University, Singapore. The findings showed that ibuprofen and diclofenac removal in the wetlands was significantly ( < 0.05) enhanced in the batch versus continuous mode. In contrast, naproxen and carbamazepine showed no significant differences ( > 0.05) in elimination under either feeding strategy. Our results also clearly showed that the presence of plants exerts a stimulatory effect on pharmaceutical removal for ibuprofen, diclofenac, and naproxen in batch and continuous mode. Estimation of the quantitative role of this stimulatory effect on pharmaceutical elimination of batch operation as compared with the effect of the presence of the higher plant alone showed that batch operation may account for 40 to 87% of the contribution conferred by the aquatic plant. The findings of this study imply that where maximal removal of pharmaceutical compounds is desired, periodic draining and filling might be the preferred operational strategy for full-scale, subsurface flow constructed wetlands.

Nutrient removal in tropical subsurface flow constructed wetlands under batch and continuous flow conditions.

The aim of this investigation was to evaluate the influence of batch versus continuous flow on the removal efficiencies of chemical oxygen demand (COD), nitrogen (N) and total phosphorus (TP) in tropical subsurface flow constructed wetlands (SSF CW). The quantitative role of the higher aquatic plants in nutrient removal in these two operational modes was also investigated. Results indicated no significant difference (p > 0.05) in COD removal between batch and continuous flow modes for either the planted or unplanted treatments. Furthermore, the batch-loaded planted wetlands showed significantly (p < 0.05) higher ammonium removal efficiencies (95.2%) compared with the continuously fed systems (80.4%), most probably because the drain and fill batch mode presented systematically more oxidized environmental conditions. With respect to TP removal, for both planted and unplanted beds, there was significant enhancement (p < 0.05) in batch flow operation (69.6% for planted beds; 39.1% for unplanted beds) as compared to continuous flow operation (46.8% for planted beds; 25.5% for unplanted beds). In addition, at a 4-day hydraulic retention time (HRT), the presence of plants significantly enhanced both ammonia oxidation and TP removal in both batch and continuous modes of operation as compared to that for unplanted beds. An estimation of the quantitative role of aeration from drain and fill operation at a 4-day HRT, as compared to rhizosphere aeration by the higher aquatic plant, indicated that drain and fill operation might account for only less than half of the higher aquatic plant's quantitative contribution of oxygen (1.55 g O2 per m2 per day for batch flow versus 1.13 g O2 per m2 per day for continuous flow).

Effect of secondary treatment at the South Bay Ocean Outfall (SBOO) on microbial ocean water quality near the US-Mexico border.

In this study, density plume visualizations and statistical comparisons were made of enterococci bacteria (the main marine recreational microbial water quality indicator) densities, both before and after the upgrade of the discharge from the South Bay Ocean Outfall (SBOO) to secondary treatment level, so that the effect of this upgrade on ocean microbial water quality could be assessed. During the dry weather (bathing) season, reduction in enterococci densities was rather limited with only 2 shore stations and one kelp station showing significant reductions, and none showing increased compliance frequency. During the wet weather season, although the signature of land-based sources of bacterial pollution were evident, a majority of both shore (7 of the 11 stations) and kelp (4 of the 7 stations) stations showed statistically significant (p ≤ 0.05) reductions enterococci densities pointing to the role of the upgrade to secondary treatment in improving microbial water quality.

Contaminant concentrations and risks associated with the Pacific oyster in the highly urbanized San Diego Bay.

Contaminant concentrations in filter-feeding shellfish may indicate the health of coastal waters and consumption risks. Widespread expansion of the Pacific oyster (Crassostrea gigas) and its popularity as food make it a useful sentinel. We surveyed intertidal Pacific oysters in San Diego Bay, California for contaminants during summer 2018 and winter 2019. We compared contaminants in Pacific oyster to California mussel from California's State Mussel Watch Program (1993-2003) and human consumption thresholds. Contaminants such as neonicotinoid and chlorinated pesticides, selenium, and several metals were higher in Pacific oysters in summer, while PBDEs, benzylbutyl phthalate, and plastics were higher in winter. Contaminant levels were generally lower in Pacific oyster than mussel except for copper and zinc. Bay-wide PCB concentrations in oysters exceeded thresholds but individual samples (locations) also met or surpassed chlordane, PCB and PAH thresholds. Monitoring and risk assessments that consider species' biology, season, location, effects of multiple contaminants, and human consumption patterns will contribute to more effective consumption guidelines.

Toxicity of cigarette butts, and their chemical components, to marine and freshwater fish.

BACKGROUND: Cigarette butts are the most common form of litter, as an estimated 4.5 trillion cigarette butts are thrown away every year worldwide. Many chemical products are used during the course of growing tobacco and manufacturing cigarettes, the residues of which may be found in cigarettes prepared for consumption. Additionally, over 4000 chemicals may also be introduced to the environment via cigarette particulate matter (tar) and mainstream smoke. METHODS: Using US Environmental Protection Agency standard acute fish bioassays, cigarette butt-derived leachate was analysed for aquatic toxicity. Survival was the single endpoint and data were analysed using Comprehensive Environmental Toxicity Information System to identify the LC50 of cigarette butt leachate to fish. RESULTS: The LC50 for leachate from smoked cigarette butts (smoked filter + tobacco) was approximately one cigarette butt/l for both the marine topsmelt (Atherinops affinis) and the freshwater fathead minnow (Pimephales promelas). Leachate from smoked cigarette filters (no tobacco), was less toxic, with LC50 values of 1.8 and 4.3 cigarette butts/l, respectively for both fish species. Unsmoked cigarette filters (no tobacco) were also found to be toxic, with LC50 values of 5.1 and 13.5 cigarette butts/l, respectively, for both fish species. CONCLUSION: Toxicity of cigarette butt leachate was found to increase from unsmoked cigarette filters (no tobacco) to smoked cigarette filters (no tobacco) to smoked cigarette butts (smoked filter + tobacco). This study represents the first in the literature to investigate and affirm the toxicity of cigarette butts to fish, and will assist in assessing the potential ecological risks of cigarette butts to the aquatic environment.

Municipal solid waste management in China: status, problems and challenges.

This paper presents an examination of MSW generation and composition in China, providing an overview of the current state of MSW management, an analysis of existing problems in MSW collection, separation, recycling and disposal, and some suggestions for improving MSW systems in the future. In China, along with urbanization, population growth and industrialization, the quantity of municipal solid waste (MSW) generation has been increasing rapidly. The total MSW amount increased from 31.3 million tonnes in 1980 to 212 million tonnes in 2006, and the waste generation rate increased from 0.50 kg/capita/day in 1980 to 0.98 kg/capita/year in 2006. Currently, waste composition in China is dominated by a high organic and moisture content, since the concentration of kitchen waste in urban solid waste makes up the highest proportion (at approximately 60%) of the waste stream. The total amount of MSW collected and transported was 148 million tonnes in 2006, of which 91.4% was landfilled, 6.4% was incinerated and 2.2% was composted. The overall MSW treatment rate in China was approximately 62% in 2007. In 2007, there were 460 facilities, including 366 landfill sites, 17 composing plants, and 66 incineration plants. This paper also considers the challenges faced and opportunities for MSW management in China, and a number of recommendations are made aimed at improving the MSW management system.

Fluorescence-based monitoring of anthropogenic pollutant inputs to an urban stream in Southern California, USA.

Fluorescence spectroscopy has been increasingly used to detect sewage and other anthropogenic contaminants in surface waters. Despite progress in successfully detecting bacterial and sewage inputs to rivers over diverse spatial scales, the use of fluorescence-based in-situ sensors to track contaminant inputs during storm events and to discern bacterial contamination from background natural organic matter (NOM) fluorescence have received less attention. A portable, submersible fluorometer equipped with tryptophan (TRP)-like and humic-like fluorescence sensors was used to track inputs of untreated wastewater added to natural creek water in a laboratory sewage spill simulation. Significant, positive correlations were observed between TRP fluorescence, the TRP:humic ratio, percent wastewater, and Escherichia coli concentrations, indicating that both the TRP sensor and the TRP:humic ratio tracked wastewater inputs against the background creek water DOM fluorescence. The portable fluorometer was subsequently deployed in an urban creek during a storm in 2018. The peak in TRP fluorescence was found to increase with the rising limb of the hydrograph and followed similar temporal dynamics to that of caffeine and fecal indicator bacteria, which are chemical and biological markers of potential fecal pollution. Results from this study demonstrate that tracking of TRP fluorescence intensity and TRP:humic ratios, with turbidity correction of sensor outputs, may be an appropriate warning tool for rapid monitoring of sewage or other bacterial inputs to aquatic environments.

Temporal pattern in levels of the neonicotinoid insecticide, imidacloprid, in an urban stream.

Imidacloprid is a widely used insecticide with high runoff potential posing a significant threat to aquatic ecosystems. In order to determine the spatial and temporal concentrations of imidacloprid in Forester Creek, a tributary to the San Diego River, surface water samples were collected from two sites under wet-weather and dry-weather conditions. Solid phase extraction and liquid chromatography tandem mass spectrometry were utilized to quantify imidacloprid levels in all samples. Imidacloprid was detected with 100% frequency in surface water samples from Forester Creek with a median concentration of 16.9 ng/L (range: 3.8-96.8 ng/L). Over 60% of samples exceeded U.S. EPA's chronic exposure benchmark (10 ng/L). Temporal analysis displayed significantly higher levels in wet-weather than dry-weather (median 45.6 ng/L vs. 8.2 ng/L (p < 0.05)), demonstrating the influence of wet-weather runoff on stream quality. Imidacloprid generally followed a first flush pattern with the highest levels observed on the rising portion of the hydrograph as compared to the remainder of the storm, further indicating that the build-up and wash off from land surfaces during storms is a major source of imidacloprid into urban surface waters. To our knowledge, the present study is the first to document this first flush pattern for imidacloprid in an urban stream in southern California. Such data on the occurrence and levels of imidacloprid in this urban stream contribute to the limited knowledge on imidacloprid in urban environments and will promote a better understanding of sources and effects of the neonicotinoid pesticide within the southern California region.

Nationwide satellite training for public health professionals: Web-based follow-up.

INTRODUCTION: Few studies have rigorously evaluated the effectiveness of health-related continuing education using satellite distribution. This study assessed participants' professional characteristics and their changes in knowledge, attitudes, and actions taken after viewing a public health preparedness training course on mass vaccination broadcast nationally by satellite. METHODS: A three-part survey design was used to evaluate training effectiveness. Paper-based questionnaires were distributed at a stratified sample of downlink facilities to collect knowledge and attitude responses before and after the broadcast. Six weeks later, willing participants were invited by e-mail to complete a Web-based follow-up questionnaire to assess actions taken as a result of viewing the broadcast. Paired t-tests and McNemar's chi-square were used to compare changes in attitude from before to after the broadcast, after the broadcast to follow-up, and before the broadcast to follow-up. RESULTS: A total of 544 paper-based questionnaires were received from 59 of 70 sampled sites. The pre- and post assessments administered the day of the broadcast demonstrated statistically significant knowledge gain (p < .001) and an increased belief immediately following the broadcast that mass vaccination clinics are important to their organizations' public health activities (p < .001). Of the 291 respondents who completed the follow-up online questionnaire, 44% (n = 128) reported they implemented or planned some form of action after the broadcast. Reported actions were higher among public health workers most likely to be directly involved in preparedness and immunization activities. DISCUSSION: This evaluation assessed (1) participants' professional characteristics, (2) knowledge gain, (3) self-reported actions taken following the broadcast, (4) program satisfaction, and (5) suggested improvements for future satellite broadcast programs. The study's methodology of using a Web-based survey for follow-up is a relatively economical tool for assessing longer-term continuing education program objectives.

Quantitation of hepatitis A virus and enterovirus levels in the lagoon canals and Lido beach of Venice, Italy, using real-time RT-PCR.

In order to assess the microbial water quality of the lagoon canals of Venice, Italy and nearby beach on Lido island, a study was conducted using real-time RT-PCR to enumerate levels of hepatitis A virus (HAV) and enteroviruses in these marine waters over a 3-year period from 2003 to 2005. A total of 17 sites (9 lagoon canal and 8 beach sites) were assayed. For the canals of the Venice Lagoon, 78% were positive for both HAV and enteroviruses, with levels ranging from 75 to 730 and 3 to 1,614 genome copies/L, respectively. At Lido beach, HAV was never detected, but enteroviruses were detected in all Lido beach samples at levels ranging from 2 to 71 genome copies/L. There was a statistically significant correlation between thermotolerant coliform densities and HAV levels (p=0.0002), but the relationship between thermotolerant coliform densities and enterovirus levels was not significant (p>0.05). Despite the fact that enteroviruses were detected at low levels in the surfzone at Lido beach, the risk for enteroviral infection (calculated using the beta-Poisson model) for recreational exposure from swimming, was in the range of 1.9 x 10(-3) - 6.1 x 10(-2), yielding a disease risk at or below the level (5% for gastroenteritis) deemed acceptable by European Guide standards.

Characterization of microbial communities in wetland mesocosms receiving caffeine-enriched wastewater.

A 454 high-throughput pyrosequencing approach was used to characterize the structures of microbial communities in wetland mesocosms receiving caffeine-enriched wastewater at a concentration of 250 µg L(-1). The removal efficiencies of caffeine in the planted beds (93.0 %) were significantly (p < 0.05) higher than those in the unplanted beds (81.4 %). Bacterial diversity was decreased by 25 and 22.4 %, respectively, in both planted and unplanted mesocosms after 210-day operation. The results of taxonomic analyses suggested that chronic exposure of wetland ecosystems to caffeine could lead to moderate shifts in microbial community composition. In total, 2156 operational taxonomic units (OTUs) were generated and 20 phyla comprising 260 genera were identified. The major phylogenetic groups at phylum level included Firmicutes (39 %), Actinobacteria (25.1 %), Proteobacteria (17.1 %), Synergistetes (5.6 %), and Chloroflexi (5.5 %). Bacilli and Synergistia increased in abundance in the planted mesocosms, while for the unplanted mesocosms, Actinobacterial, Clostridia and Betaproteobacteria exhibited increased proportion under the exposure of caffeine. At genus level, Propionibacterium, Staphylococcus, Bacillus, and Streptococcus were found to be increased in abundance after caffeine treatment. As for the response of fungal community to caffeine enrichment, genus like Cladosporium, Emericellopsis, Aspergillus, and Phoma were found to be resistant to caffeine disturbance. When compared to the microbial community between planted and unplanted mesocosms, a distinct community profile for both bacteria and fungi community was observed. The presence of plants had a remarkable effect on the structure of microbial community, helping buffer against the stress associated with caffeine exposure.

Ibuprofen removal in horizontal subsurface flow constructed wetlands: treatment performance and fungal community dynamics.

The treatment performance of ibuprofen (IBP)-enriched wastewater by horizontal subsurface flow constructed wetlands planted with cattail (Typha angustifolia) and unplanted control mesocosms was investigated. Removal efficiencies of IBP were significantly (p < .05) enhanced in the planted mesocosms (78.5%) compared to those in the unplanted beds (57.9%). An 18S rRNA gene high-throughput pyrosequencing approach was used to investigate the effects of IBP on the structure of the fungal community in these wetland systems. The overall diversity of the fungal community was reduced under the IBP exposure. Taxonomic analysis revealed that 62.2% of the fungal sequences were affiliated with Basidiomycota, followed by Ascomycota (37.4%) at the phylum level. Uncultured fungus (48.2%), Chaetomium sp. (14.2%), Aspergillus sp. (12.4%), Trichoderma sp. (5.7%), Cladosporium sp. (5.4%), and Emericellopsis sp. (5.2%) were identified as dominant genera. At the genus level, a distinct profile of the fungal community in the IBP-enriched mesocosms was observed as compared to the control beds, and as well specific fungal genera were enhanced in the planted beds, regardless of IBP enrichment. However, despite these differences, the composition of the fungal community (as measured by Bray-Curtis similarity) was mostly unaffected by the significant IBP enrichment. On the other hand, a consistent similarity pattern of fungal community structure in the planted mesocosms suggests that the presence of higher macrophytes in the wetland systems may well help shape the fungal community structure.

Detection and quantification of hepatitis A virus in seawater via real-time RT-PCR.

A real-time RT-PCR method utilizing SYBR Green chemistry was developed to detect and enumerate hepatitis A virus (HAV) in ocean water. Ocean water samples were taken at the Tijuana River mouth (Tijuana, Mexico) and Imperial Beach pier (1.4 km north of the Tijuana River mouth in San Diego, California) following four separate rain events. A total of eight samples were collected, one from each location, each consisting of 4 l of ocean water. Using conventional RT-PCR and primers based on the conserved sequence at the VP3-VP1 genes of HAV, a 247 bp cDNA was amplified from six out of eight rain event water samples. HAV cDNA (confirmed by sequence analysis) was cloned into a TOPO vector (Invitrogen, Carlsbad, CA), and four primer sets were designed for application in SYBR Green real-time RT-PCR. The water samples were shown to contain inhibitors that affected real-time RT-PCR amplifications, however diluting the cDNA solution enabled successful amplification. Using real-time RT-PCR, HAV could be detected in all eight samples. Depending on the rain event, the viral load in these samples varied from 90 to 3523 copies of HAV/L of ocean water near the mouth of the Tijuana River, and 347 to 2656 copies/l near the Imperial Beach pier. The sensitivity, quantitative ability and the high throughput nature of SYBR Green real-time RT-PCR will be useful in monitoring HAV contamination in seawater.