环境DNA(eDNA)宏条形码技术对枝角类浮游动物物种鉴定及其生物量监测研究

环境DNA(eDNA)宏条形码(Metabarcoding)技术越来越多地被应用于环境中物种定性识别,但如何定量监测物种在环境中的丰度尚未得到解决。本研究以太湖流域常见的5种浮游动物拟同形溞、大型溞、蚤状溞、多刺裸腹溞、老年低额溞为研究对象,建立了一种基于eDNA宏条形码技术的物种定量方法,并与实时荧光定量PCR(qPCR)相比较,研究了eDNA宏条形码技术多物种定量的准确性。结果表明,PCR引物对eDNA宏条形码的物种检测和定量影响显著。313 bp COI313引物对浮游动物物种覆盖度高,但是物种间DNA扩增的偏好性大,不适用于eDNA宏条形码定量检测。基于COI序列重新设计的短COI116引物能够同时检测出所有5个物种。荧光定量PCR(qPCR)物种拷贝数与物种相对占比呈正相关。eDNA宏条形码所检出每个物种的序列数与qPCR定量拷贝数高度一致。综上,eDNA宏条形码技术可实现对浮游动物物种的半定量检测,在生物多样性监测和生物完整性评价有显著的应用价值。     

Characteristics of major volatile organic hazardous air pollutants in the urban air of Kaohsiung city

The concentrations and characteristics of volatile organic hazardous air pollutants (HAPs) in the urban city of Kaohsiung from motor vehicles and dense pollutant sources has become a national concern. To continuously monitor volatile organic HAPs, sampling sites were selected near the four air-quality monitoring stations established by Ethe nvironmental Protection Administration of Taiwan ROC, namely Nan-tz, Tso-ying, San-min and Hsiao-kang, from north to south. An on-site automated online monitor of volatile organic compounds (VOCs) was used for continuous monitoring. This study performed two consecutive days of 24-h monitoring of five volatile organic HAPs form August to October 2005 at the four monitoring sites, which cover the northern, central, and southern areas of Kaohsiung city. The average monitored concentration was 2.78–4.84 ppb for benzene, 5.90–9.66 ppb for toluene, 3.62–5.90 ppb for ethylbenzene, 3.73–5.34 ppb for m,p-xylene, 3.38–4.22 ppb for o-xylene, and 4.48–7.00 ppb for styrene. The average monitored concentrations of the major volatile organic HAPs tended to follow the pattern San-min > Nan-tz > Hsiao-kang > Tso-ying. Among all the species monitored in this study, toluene had the highest ambient concentration, followed by styrene, m,p-xylene, ethylbenzene, o-xylene, and benzene. The results showed that the concentration at night was higher than that in the day for toluene at Nan-tz, San-min, Hsiao-kang, and for benzene at Nan-tz and Hsiao-kang.     

Carbon paste electrode modified with chromium thiopental for the potentiometric flow injection analysis of chromium (III)

A chemically modified carbon paste electrode for chromium (III) based on the formation of ion-association complex of chromium with 5-ethyl-6-oxo-5-pentan-2-yl-sulfanyl-pyrimidin-4-olate (thiopental, THP) as electroactive ion-exchanger (Cr-THP) was prepared and investigated. The electrode has a linear dynamic range of 1.7?×?10^?6–1.3?×?10^?2?mol?L^?1, with a Nernstian slope of 19.6?±?0.5?mV per decade and a detection limit of 9.0?×?10^?7?mol?L^?1. It has a fast response time of <15?s and can be used for at least 7 weeks without any considerable divergence in potential. The proposed sensor revealed good selectivity for Cr(III) over a wide variety of mono-, di-, and trivalent cations and could be used in the pH range of 2.3–6.5. The proposed electrode was used in batch measurements and in flow injection analysis. The Cr-CMCPE was successfully applied for the determination of Cr(III) in real samples (wastewater of hot dip galvanizing unit and alloys). The present electrode was also used as an indicator electrode in potentiometric titration of Cr(III). The results were highly satisfactory.     

Degradation of azo dyes in water by Electro-Fenton process

The degradation of the azo dyes azobenzene, p-methyl red and methyl orange in aqueous solution at room temperature has been studied by an advanced electrochemical oxidation process (AEOPs) under potential-controlled electrolysis conditions, using a Pt anode and a carbon felt cathode. The electrochemical production of Fenton's reagent (H₂O₂, Fe²⁺) allows a controlled in situ generation of hydroxyl radicals (^·OH) by simultaneous reduction of dioxygen and ferrous ions on the carbon felt electrode. In turn, hydroxyl radicals react with azo dyes, thus leading to their mineralization into CO₂ and H₂O. The chemical composition of the azo dyes and their degradation products during electrolysis were monitored by high performance liquid chromatography (HPLC). The following degradation products were identified: hydroquinone, 1,4-benzoquinone, pyrocatechol, 4-nitrocatechol, 1,3,5-trihydroxynitrobenzene and p-nitrophenol. Degradation of the initial azo dyes was assessed by the measurement of the chemical oxygen demand (COD). Kinetic analysis of these data showed a pseudo-first order degradation reaction for all azo dyes. A pathway of degradation of azo dyes is proposed. Specifically, the degradation of dyes and intermediates proceeds by oxidation of azo bonds and aromatic ring by hydroxyl radicals. The results display the efficiency of the Electro-Fenton process to degrade organic matter. Electronic Publication     

城镇污水处理厂进出水对日本青鱂早期阶段亚慢性毒性效应

采用日本青鳉胚胎及幼鱼暴露评估污水处理厂进出水的综合毒性效应。水样采自广州市3个污水处理厂的进水口和出水口。96 h青鳉幼鱼急性毒性试验表明:6个水样均没有对青鳉幼鱼产生急性毒性效应。14 d胚胎暴露试验结果:3个污水处理厂的进水及1号和3号污水处理厂的出水均引起胚胎死亡,胚胎孵化率均低于70%且均与对照组产生了显著性差异(p0.05)。3个厂的进水和1号、2号厂出水的鱼苗畸形率与对照组也有显著差异(p0.05),其中3号厂进水鱼苗畸形率高达(33.73±8.94)%。而6个水样的胚胎孵化时间均与对照组无显著差异(p0.05),均在9 d左右。7 d鱼苗暴露试验结果:1、2、3号3个水厂进水的鱼苗死亡率分别是(30.16±2.75)%,(24.07±1.60)%,(37.90±4.77)%,与对照组产生明显差异(p0.05)。出水对鱼苗的致死效应明显低于进水,但出水对鱼苗的致死效应相对于对照组仍有显著差异(p0.05)。     

New methodological strategies for detecting bacterial indicators

The development of monitoring strategies for the early warning of seawater pollution, with particular reference to faecal and hydrocarbon contamination, has been the specific goal of research carried out within the Cluster 10-SAM (Advanced Systems for Coastal Marine Monitoring) Project, funded by the Italian Ministry for University and Scientific Research. Advanced analytical approaches have been designed and applied to detect bacterial species that have been selected as potential indicators of pollution in seawater samples. We report the results obtained using the fluorescent antibody and enzymatic assays for the detection of Escherichia coli, and a real-time PCR protocol for monitoring marine areas for hydrocarbon pollution. Immunofluorescence and enzymatic methods revealed the occurrence of different faecal pollution levels, reaching 10⁵ E. coli cells 100 ml^?1 in the Straits of Messina. Real-time PCR results corresponded to the different degree of oil pollution of the analysed samples. The specificity and speed make these methods promising for the detection and evaluation of marine pollution.     

Evaluating pollution-related damage and restoration success in urban forests with participatory monitoring and digital tools

Peri-urban forest monitoring requires indicators of vegetation damage. An example is the sacred fir (Abies religiosa) forests surrounding Mexico City, which have been heavily exposed to tropospheric ozone, a harmful pollutant, for over 4 decades. We developed a participatory monitoring system with which local community members and scientists generated data on ozone tree damage. Santa Rosa Xochiac rangers (13) used the digital tool KoboToolBox to record ozone damage to trees, tree height, tree ages, tree condition, tree position, and whether the tree had been planted. Thirty-five percent of the trees (n = 1765) had ozone damage. Younger trees had a lower percentage of foliage damaged by ozone than older trees (p < 0.0001), and asymptomatic trees tended to be younger (p < 0.0001). Symptomatic trees were taller than asymptomatic trees of the same age (R²_c = 0.43, R²_m = 0.27). Involving local communities facilitated forest monitoring and using digital technology improved data quality. This participatory system can be used to monitor forest condition change over time and thus aids restoration efforts driven by government or local communities’ interests, facilitating local decision-making.     

Development of a complex-based flow injection spectrophotometric method for determination of the herbicide pinoxaden in environmental samples

A flow injection (FI) spectrophotometric method for the determination of the herbicide pinoxaden (PXD) has been proposed. PXD was converted in alkaline media with hydroxylamine hydrochloride to the hydroxamate salt. The salt was reacted with iron(III) chloride, and the absorbance of the red colored tris iron hydroxamate complex was measured at 500?nm using a FI system. The method was found to be linear between 0.5 and 40?mg?L^?1 with a molar absorptivity of 1.53?×?10^4?L?mol^?1?cm^?1. The limit of detection and limit of quantification were found to be 0.1?±?0.01?mg?L^?1 and 0.6?±?0.05?mg?L^?1, respectively. Any interference of fenoxaprop-p-ethyl (FE) was avoided by the separation of PXD by liquid chromatography with a mixture of dichloromethane and n-hexane (1?:?1) as eluent. The method was applied to the determination of PXD in soil, water, and wheat grains with percent recoveries of 98?±?2, 100?±?2, and 98?±?5, respectively. Sample throughput of 60 samples per hour was achieved under optimized conditions.     

Eyes and extraocular photoreceptors in midwater cephalopods and fishes: Their roles in detecting downwelling light for counterillumination

The means of detecting downwelling light for counterillumination in several midwater animals has been examined. Eyes and extraocular photoreceptors (drosal photosensitive vesicles in the enoploteuthid squid Abraliopsis sp. B and pineal organs in the myctophid fish Myctophum spinosum) were alternately exposed to overhead light or covered by a small opaque shield above the animal and the bioluminescent response of the animal was monitored. Covering either the eyes or the extraocular photoreceptors resulted in a reduction in the intensity of counterillumination. Preliminary experiments examining the bioluminescent feedback mechanism for monitoring intensity of bioluminescence during counterillumination in the midwater squid Abralia trigonura indicated that the ventral photosensitive vesicles are responsible for bioluminescent feedback.     

MBR工艺处理螺旋霉素制药废水过程中抗生素耐药菌与抗性基因的研究

抗生素废水含有大量的抗生素耐药菌(antibiotic resistant bacteria,ARB)与抗性基因(antibiotic resistance genes,ARGs),处理排放后可能增强受纳环境的微生物抗性,因此有必要深入研究抗生素废水处理过程中ARB与ARGs的削减效果及其影响因素。本研究采用膜生物反应器(membrane bioreactor,MBR)工艺处理螺旋霉素制药废水,考察了不同水力停留时间(hydraulic retention time,HRT)对ARB与ARGs削减效果的影响。结果表明,虽然在HRT=30 h时MBR对COD与氨氮的去除率略高于HRT=40 h,但HRT=40 h时,不仅总异养菌与肠球菌的去除效果更佳,出水肠球菌耐药率及携带的抗性基因检出率也更低,而且MBR对废水中erm B、erm F、erm X、mef A、ere A、mph B和转移元件ISCR 1、Tn 916/1545相对丰度的削减效果更好。这表明长HRT更有利于MBR工艺削减螺旋霉素废水的耐药菌与抗性基因。     

Abundance and distribution of antibiotic resistance genes in a full-scale anaerobic–aerobic system alternately treating ribostamycin,spiramycin and paromomycin production wastewater

The occurrence of antibiotic-resistant bacteria and antibiotic resistance genes (ARGs) has been intensively investigated for wastewater treatment systems treating single class of antibiotic in recent years. However, the impacts of alternately occurring antibiotics in antibiotic production wastewater on the behavior of ARGs in biological treatment systems were not well understood yet. Herein, techniques including high-capacity quantitative PCR and quantitative PCR (qPCR) were used to investigate the behavior of ARGs in an anaerobic–aerobic full-scale system. The system alternately treated three kinds of antibiotic production wastewater including ribostamycin, spiramycin and paromomycin, which referred to stages 1, 2 and 3. The aminoglycoside ARGs (52.1–79.3%) determined using high-capacity quantitative PCR were the most abundant species in all sludge samples of the three stages. The total relative abundances of macrolide–lincosamide–streptogramin (MLS) resistance genes and aminoglycoside resistance genes measured using qPCR were significantly higher (P < 0.05) in aerobic sludge than in sewage sludge. However, the comparison of ARGs acquired from three alternate stages revealed that MLS genes and the aminoglycoside ARGs did not vary significantly (P > 0.05) in both aerobic and anaerobic sludge samples. In aerobic sludge, one acetyltransferase gene (aacA4) and the other three nucleotidyltransferase genes (aadB, aadA and aadE) exhibited positive correlations with intI1 (r ² = 0.83–0.94; P < 0.05), implying the significance of horizontal transfer in their proliferation. These results and facts will be helpful to understand the abundance and distribution of ARGs from antibiotic production wastewater treatment systems.

     

Community dynamics of ammonia oxidizing bacteria in a full-scale wastewater treatment system with nitrification stability

To determine whether the functional stability of nitrification was correlated to a stable community structure of ammonia oxidizing bacteria (AOB) in a full-scale wastewater treatment plant, the AOB community dynamics in a wastewater treatment system was monitored over one year. The community dynamics were investigated using specific PCR followed by terminal restriction fragment length polymorphism (T-RFLP) analysis of the amoA gene. The T-RFLP results indicated that during the period of nitrification stability, the AOB community structure in the full-scale wastewater treatment system was relatively stable, and the average change rate every 15 d of the system was 6.6%±5.8%. The phylogenetic analysis of the cloned amoA gene showed clearly that the dominant AOB in the system was Nitrosomonas spp. The results of this study indicated that throughout the study period, the AOB community structure was relatively stable in the full-scale wastewater treatment system with functional stability of nitrification.     

Application of electrochemical reactor divided by cellulosic membrane for optimized simultaneous removal of phenols,chromium, and ammonia from tannery effluents

The post treatment of simulated tannery wastewater was evaluated in an electrochemical oxidation process under galvanostatic conditions. A continuous flow reactor divided by a cellulosic membrane consisted of Ti/SnO₂–Sb anodes and iron cathodes was used. Central composite design and response surface methodology (RSM) were applied to investigate the effects of six operational parameters, namely initial concentration of total phenols (TPh), total chromium (TCr), total ammonia nitrogen (TAN), flow rate (Q), current intensity (I), and electrode surface area (A). Effectiveness of the innovative cellulosic membrane was proven by considerable pH variations in the anolyte and catholyte chambers. A faster removal rate was observed for TPh and TAN, followed by TCr. The treatment level was very sensitive to Q and I in the studied ranges. RSM showed the removal efficiencies of 78.14%, 63.42%, and 86.09% for TPh, TCr, and TAN, respectively, are achieved under optimal conditions with consumption of only 9.03 kWh m^?3 electrical energy. Chlorinated compounds such as chloroform, 2,4-dichlorophenol, and chlorobenzene were detected as the degradation intermediates. According to the obtained results, electrolysis in the divided cell with cellulosic membrane is a practical, cost-effective method for advanced treatment of tannery effluents.     

Detection of polychlorinated biphenyls employing chemical dechlorination followed by biphenyl whole cell sensing system

Polychlorinated biphenyls (PCBs) are a mixture of 209 individual chlorinated compounds commonly known as PCB congeners. These compounds are hydrophobic and are persistent in the environment. Their use was banned in the US a few decades ago because of harmful health effects. Therefore, detection of PCBs in environmental samples is increasingly important. To that end, we have developed a two-step simple and sensitive method for the detection of total PCBs. Specifically, our method involves dechlorination of PCBs to biphenyl followed by detection of biphenyl using a whole cell sensing system as the detection system. The whole cell sensing system consists of cells of the strain Pseudomonas pseudoalcaligenes KF707 harboring plasmid pSD7000. Plasmid pSD7000 contains the gene of lacZ, a reporter protein under the control of the bph operon. The detection is achieved through the emission of light afforded by the expression of reporter protein triggered by the presence of biphenyl. Due to the fact that this operon is activated only by few PCB congeners, a chemical dechlorination method was employed to convert all PCBs to biphenyl, and thus all the PCBs present in a given sample are able to be detected. The results showed that PCB congeners were rapidly (30?min) and efficiently (>98.5%) dechlorinated to biphenyl using a Mg/K₂PdCl₆ catalyst, and the biphenyl could be subsequently quantified using the whole cell sensing system. This hybrid analytical method that combines classical dechlorination with novel biosensing methods may find applications in the on-site monitoring of PCBs contamination levels.     

印染废水处理过程及排放水对草履虫遗传毒性评价

草履虫作为指示生物评估印染废水处理过程及排放水的毒性效应。急性毒性结果显示,原水、厌氧工段印染废水、好氧工段印染废水对草履虫的半数致死浓度(24h-LC₅₀)分别为54%、41%、98%;遗传毒性结果显示,印染废水原水、经处理的排放水在6.25%、12.25%稀释梯度下对草履虫造成的DNA损伤较对照组呈显著升高(p<0.01),而排放水对草履虫的DNA损伤较原水无显著性差异;草履虫增殖速率结果显示,印染废水排放水50%稀释梯度下对草履虫增殖速率有抑制作用。暴露于印染废水排放水中10%的草履虫其游动行为发生改变。以上结果表明,印染废水在经过处理后,理化指标均达到国家印染废水行业排放标准,但排放水仍对草履虫产生急性毒性,且遗传毒性较原水没有显著降低。水生生物毒性测试能有效补充理化指标评价,为印染废水行业全面达标排放提供依据。     

Analysis of air pollution at Shuaiba industrial area in Kuwait

A study has been conducted over a period of one year on measurements of air pollution in the Shuaiba Industrial Area (SIA) of Kuwait. The study included analysis of pollutant behaviour relative to the wind speed and direction. SIA comprises several large scale industries including three petroleum refineries, two power plants, two fertilizer plants, a cement plant, a chlorine and soda plant, a commercial harbour and two large oil loading terminals. Measurements of 15 parameters have been carried out every 5 minutes using a mobile laboratory fitted with an automatic calibrator and a data storage system. The pollutants studied include methane, non‐methane hydrocarbons (NMHC), carbon monoxide, carbon dioxide, nitrogen oxides (NO, NO₂, and NO _x ), sulphur dioxide, ozone and suspended dust. Meteorological parameters monitored simultaneously include wind speed and direction, air temperature, relative humidity, solar radiation, and barometric pressure. The air quality data collected using the mobile laboratory have been used to calculate the diurnal and monthly variations in the major primary and secondary pollutants. Distribution levels of these pollutants relative to wind direction and speed have also been used in the analysis. The results show large diurnal variations in some pollutant concentrations. Generally, two types of concentration variations have been found, depending on whether the species is a primary or a secondary pollutant. Diurnal variations with two maxima were observed in the concentrations of primary pollutants including NO, SO₂, NMHC, CO and suspended dust, whereas a single maximum was observed for secondary pollutants such as O₃and NO₂. The monthly variations of SO₂and NO _x showed maximum values during the warm months. However, ozone showed a quite marked seasonal variation with maxima during spring and late summer and a minimum during the early summer. The results also indicated a common source for NO _x , SO₂, NMHC, CO and suspended dust to the North‐West (NW) of the monitoring station. Moreover for NO _x and SO₂, another less significant source is to the South‐South‐West (SSW) and South‐West (SW) of the monitoring station.     

Coupled electrocoagulation–electro-Fenton for efficient domestic wastewater treatment

This article reports the first use of coupled electrocoagulation and electro-Fenton (EF-EC) to clean domestic wastewater. Domestic wastewater contains high amounts of organic, inorganic and microbial pollutants that cannot be usually treated in a single step. Here, to produce an effluent suitable for discharge in a single process step, a hybrid process combining electrocoagulation and electro-Fenton was simultaneously used to decrease chemical oxygen demand (COD), turbidity and total suspended solids (TSS) from domestic wastewater. The electrocoagulation–electro-Fenton process was firstly tested for the production of H₂O₂ using Ti–IrO₂ and vitreous carbon- or graphite electrodes arranged at the anode and the cathode, respectively. The concentration of H₂O₂ recorded at 1.5 A of current intensity during 60 min of electrolysis using vitreous carbon- and graphite electrodes at the cathode was 4.18 and 1.62 mg L^?1, respectively. By comparison, when the iron electrode was used at the anode, 2.05 and 1.06 mg L^?1 of H₂O₂ were recorded using vitreous carbon and graphite, respectively. The H₂O₂ concentration decrease was attributed to hydroxyl radical formation generated by the Fenton reaction. Electro-Fenton using iron electrode at the anode and vitreous carbon at the cathode with a current density imposed of 0.34 A dm^?2 ensures the removal efficiency of 50.1 % COD_T, 70.8 % TSS and 90.4 % turbidity. The electrocoagulation–electro-Fenton technique is therefore a promising secondary treatment to simultaneously remove organic, inorganic and microbial pollutants from domestic, municipal and industrial wastewaters.     

Duckweed: an effective tool for phyto-remediation

Effective wastewater treatment through conventional methods that rely on heavy aeration are expensive to install and operate. Duckweed is capable of recovering or extracting nutrients or pollutants and is an excellent candidate for bio-remediation of wastewaters. Such plants grow very fast, utilizing wastewater nutrients and also yield cost effective protein-rich biomass as a by-product. Duckweeds being tiny surface-floating plants are easy to harvest and have an appreciable amount of protein (15%–45% dry mass basis) and a lower fiber (7%–14% dry mass basis) content. Besides nutrient extraction, duckweeds has been found to reduce total suspended solids, biochemical oxygen demand (BOD), and chemical oxygen demand in wastewater significantly. Depending on the initial concentrations of nutrients, duckweed-covered systems can remove nitrate (NO₃^?) at daily rates of 120–590 mg NO₃^? m^?2 (73%–97% of initial concentration) and phosphate (PO₄^?) at 14–74 mg PO₄^? m^?2 (63%–99% of initial concentration). Removal efficiencies within 3 days of 96% and 99% have been reported for BOD and ammonia (NH₃). Besides several genera of duckweeds (Spirodela, Lemna, Wolffia), other surface-floating aquatic plants like water hyacinth (Eichhornia) are well known for their phyto-remediation qualities.     

Estimation of the density of a clustered point pattern using a distance method

Distance-based methods use point-to-point distances or random-location-to-point distances in a cloud of points to estimate characteristics of the point pattern. One such characteristics is the density of points. The difficulty with distance-based density estimators is that their distribution depends on the spatial pattern of points. In particular, the distribution of distances is untractable for usual clustered patterns, that are often observed in natural systems. Here, we propose a density estimator for clustered patterns, based on the random-location-to-pth-point distance X _p . An approximate expression for the distribution function, F _p , of X _p was obtained by identifying the first two moments of the count of individuals in disks for a given point process with the first two moments of a negative binomial distribution. The approximate expression of F _p was then used to derive a maximum-likelihood estimator of the intensity of the point process. The quality of the approximation of F _p was assessed for homogeneous Poisson processes (for which the expression of F _p is exact) and for Matérn processes. The intensity estimator based on Matérn processes was then used to estimate tree density in a tree savanna in Mali, and it compared favorably with six robust estimators found in the literature.