Enhanced remediation of BDE-209 in contaminated mangrove sediment by planting and aquaculture effluent.

Toxic and persistent flame retardant (BDE-209) and aquaculture effluent (AE) are ubiquitous in coastal environments, but how their co-existence influences their fate is not yet investigated. This study investigated AE effects on remediation and uptake of BDE-209 by Kandelia obovata (Ko) and Avicennia marina (Am), true and dominant mangrove species. After 12-months, a significant removal of BDE-209 was achieved in planted mangrove sediment and the removal was significantly enhanced by AE addition, possibly due to the enhancement of nitrogen (N) and phosphorous (P) content in sediment. Residual percentages of parent BDE-209 in Ko and Am planted sediments without AE were 61.4% and 70.9%, respectively, but decreased to 46.9% and 48.0% with AE addition after 12-months. A similar trend was found in unplanted sediment, with 86.5% and 65.3% of BDE-209 retained in sediments without and with AE addition, respectively. The results demonstrated that AE addition not only increased the debromination of BDE-209 in all treated sediments with the production of debrominated congeners (de-PBDEs) like di- to nona-BDEs in unplanted and planted sediments, but also enhanced the take up of BDE-209 in Ko root, and de-PBDEs in both Ko and Am, thus enhancing the phytoremediation of BDE-209 in contaminated sediments.

Enhancement effect of nanoscale zero-valent iron addition on microbial degradation of BDE-209 in contaminated mangrove sediment.

Chemical and biological methods have been employed to remedy polybrominated diphenyl ether contamination, but the removal of decabromodiphenyl ether (BDE-209) by either method still has limitations. The present study aims to evaluate the combined effect of nanoscale zero-valent iron (nZVI) (from 0.1 to 10%) reduction and microbial debromination on BDE-209 removal in mangrove sediments under an anaerobic condition. During the 12-months incubation, nZVI significantly enhanced BDE-209 removal, with 17.03% to 41.99% reduction in sterilized sediments. The reduction was even higher in non-sterilized sediments with living indigenous microorganisms, achieving 15.80%, 33.50%, 55.83% and 66.95% removal of BDE-209 at 0 (control without nZVI), 0.1%, 1% and 10% nZVI, respectively. In control sterilized sediments, no debromination was found, and debromination occurred according to spiked levels of nZVI, with BDE-153 being the dominant congener. The concentrations of debrominated congeners in non-sterilized sediments also increased with nZVI levels, but were significantly higher than the respective sterilized sediment. The relative proportions of different debrominated congeners in non-sterilized sediments depended on nZVI levels, with BDE-99 being the dominant congener in low nZVI amended sediments but shifted to BDE-153 under high nZVI. Higher concentrations of ferrous iron (Fe2+) were detected in both sterilized and non-sterilized sediments spiked with more nZVI, and their concentrations significantly correlated with BDE-209 removal. Growth of total bacteria in sediments with 1% and 10% nZVI was inhibited within first two months, but their numbers resumed to that in the control at the end of 12 months. The present study demonstrates the synergy between chemical and microbiological methods, and a combination of nZVI and indigenous microorganisms could be an efficient and feasible mean to remedy BDE-209 in contaminated sediments.

In situ hybridization to detect spatial gene expression in medaka.

A whole-animal tissue section in situ hybridization (ISH) system with radio-labeled probes was developed to detect differential gene expression among tissues of the small, oviparous teleost fish, Japanese medaka (Oryzias latipes). Because of its tissue- and gender-specific expression, gonadal aromatase (CYP19a) was selected as a model gene to demonstrate the potential of the system. The ISH system was validated with a 7d exposure to the model aromatase inhibitor, fadrozole. Fadrozole did not affect the magnitude of gene expression in testes, but significantly up-regulated CYP19a gene expression in ovaries. These results were confirmed with quantitative real-time-polymerase chain reaction (RT-PCR). Histological evaluation revealed that females exposed to 100microg/L fadrozole lacked mature oocytes. Male gonadal morphology was normal in all treatments. The ISH method developed in this study allowed tissue-specific resolution of gene expression in a whole animal model, as well as the ability to analyze cellular morphological detail in the same organism.

Development of a marine fish model for studying in vivo molecular responses in ecotoxicology.

A protocol for fixation and processing of whole adult marine medaka (Oryzias melastigma) was developed in parallel with in situ hybridization (ISH) and immunohistochemistry (IHC) for molecular analysis of in vivo gene and protein responses in fish. Over 200 serial sagittal sections (5microm) can be produced from a single adult medaka to facilitate simultaneous localization and quantification of gene-specific mRNAs and proteins in different tissues and subcellular compartments of a single fish. Stereological analysis (as measured by volume density, V(v)) was used to quantify ISH and IHC signals on tissue sections. Using the telomerase reverse transcriptase (omTERT) gene, omTERT and proliferating cell nuclear antigen (PCNA) proteins as examples, we demonstrated that it is possible to localize, quantify and correlate their tissue expression profiles in a whole fish system. Using chronic hypoxia (1.8+/-0.2 mgO(2)L(-1) for 3 months) as an environmental stressor, we were able to identify significant alterations in levels of omTERT mRNA, omTERT protein, PCNA (cell proliferation marker) and TUNEL (apoptosis) in livers of hypoxic O. melastigma (p<0.05). Overall, the results suggest that O. melastigma can serve as a model marine fish for assessing multiple in vivo molecular responses to stresses in the marine environment.

Molecular cloning and characterization of a hypoxia-responsive CITED3 cDNA from grass carp.

We have isolated a 1586-bp full-length CITED3 cDNA from grass carp which specifies for a cAMP-responsive element-binding protein/p300-interacting transactivator with glutamic acid (E)/aspartic acid (D)-rich C-terminal domain protein. The cDNA, designated as gcCITED3, has an open reading frame of 762 bp and encodes a protein of 253 amino acids with a predicted molecular mass of 28.3 kDa and pI of 6.4. Pairwise comparison showed that gcCITED3 shares high sequence identity with the CITED3 of zebrafish (94%), chicken (72%) and Xenopus (59%). Northern blot analysis indicated that gcCITED3 is most highly expressed and responsive to hypoxia in the carp kidney. Hypoxic induction was also observed in heart, albeit at a lower level. This is the first report on the isolation of a hypoxia-responsive CITED3 gene from fish.

Rapid detection of six types of bacterial pathogens in marine waters by multiplex PCR.

A rapid multiplex PCR (m-PCR) method that allows the simultaneous detection, in a single tube, of six commonly encountered waterborne pathogens is developed. The target genes used were: the aerolysin (aero) gene of Aeromonas hydrophila, the invasion plasmid antigen H (ipaH) gene of Shigella flexneri, the attachment invasion locus (ail) gene of Yersinia enterocolitca, the invasion plasmid antigen B (ipaB) gene of Salmonella typhimurium, the enterotoxin extracellular secretion protein (epsM) gene of Vibrio cholerae and a species-specific region of the 16S-23S rDNA (Vpara) gene of Vibrio parahaemolyticus were used as the gene targets. Multiplex PCR using the six pairs of primers produced specific amplicons of the expected sizes from mixed populations of reference bacterial strains in seawater and from pure cultures. The m-PCR assay was specific and rapid, with a turnaround time of < 12 h. The detection limit of the assay for the bacterial targets was estimated at 10(0)-10(2) cfu. Multiplex PCR analysis was performed on 19 seawater samples collected around Hong Kong and the results indicated significant levels of four bacterial pathogens at several sites where primary sewage wastes are discharged, and the levels of which showed no correlation with E. coli counts. Overall, both laboratory and field validation results demonstrated that the m-PCR assay developed in this study could provide a cost-effective and informative supplement to conventional microbiological methods for routine monitoring and risk assessment of water quality.

Minimizing the exposure of airborne pathogens by upper-room ultraviolet germicidal irradiation: an experimental and numerical study.

There has been increasing interest in the use of upper-room ultraviolet germicidal irradiation (UVGI) because of its proven effectiveness in disinfecting airborne pathogens. An improved drift flux mathematical model is developed for optimizing the design of indoor upper-room UVGI systems by predicting the distribution and inactivation of bioaerosols in a ventilation room equipped with a UVGI system. The model takes into account several bacteria removal mechanisms such as convection, turbulent diffusion, deposition and UV inactivation. Before applying the model, the natural die-off rate and susceptibility constants of bioaerosols were measured experimentally. Two bacteria aerosols, Escherichia coli and Serratia marcescens, were tested for this purpose. It was found out that the general decay trend of the bioaerosol concentration predicted by the numerical model agrees well with the experimental measurements. The modelling results agree better with experimental observations for the case when the UVGI inactivation mechanism dominates at the upper-room region than for the case without UVGI. The numerical results also illustrate that the spatial distribution of airborne bacteria was influenced by both air-flow pattern and irradiance distribution. In addition to predicting the local variation of concentration, the model assesses the overall performance of an upper-room UVGI system. This model has great potential for optimizing the design of indoor an upper-room UVGI systems.