Bacterial biosensors for rapid and effective monitoring of biodegradation of organic pollutants in wastewater effluents

Significant amounts of toxic substances which are hazardous to animals, plants, microorganisms, and other living organisms including humans are released annually into aquatic and terrestrial environments, mostly from improper wastewater discharges. Early detection of such pollutants in wastewater effluents and proper monitoring before their final release into the environment is therefore necessary. In this study, two whole-cell bacterial biosensors were constructed by transforming competent cells of Shigella flexneri and Shigella sonnei with pLUX plasmids and evaluated for their potential to monitor wastewater samples undergoing degradation by measuring bioluminescence response using a microplate luminometer. Both bacterial biosensors were found to be extremely sensitive to the wastewater samples, with different patterns, concomitant with those of the COD removals demonstrated at the different days of the degradation. Generally higher bioluminescence values were obtained at the later days of the degradation period compared to the initial values, with up to 571.76% increase in bioluminescence value obtained at day 5 for 0.1% (v/v) effluent concentration. Also, a steady decrease in bioluminescence was observed for the bacterial biosensors with increasing time of exposure to the wastewater effluent for all the sampling days. These biosensor constructs could therefore be applicable to indicate the bioavailability of pollutants in a way that chemical analysis cannot, and for in situ monitoring of biodegradation. This has great potential to offer a risk assessment strategy in predicting the level of bioremediation required during municipal wastewater treatment before their final discharge into the aquatic milieu.     

Metabolic and bacterial diversity in soils historically contaminated by heavy metals and hydrocarbons

The aim of this study was to characterize soils contaminated by different levels of heavy metals and hydrocarbons (Madonna Dell'Acqua, Pisa, Italy). The soils were chemically and biochemically analysed by measuring the standard chemical properties and some enzyme activities related to microbial activity (dehydrogenase activity) and the soil carbon cycle (total and extracellular beta-glucosidase activities). The metabolic capacities of soil microorganisms to degrade hydrocarbons through catechol 2,3-dioxygenase were also described. The microbial diversity of contaminated and uncontaminated soils was estimated by denaturing gradient gel electrophoresis (DGGE) of amplified 16S rDNA sequences. The PCR/single-strand conformation polymorphism (PCR/SSCP) method was used to estimate the genetic diversity of PAH-degrading genes in both contaminated and uncontaminated soils. A greater bacterial diversity and lower catechol 2,3-dioxygenase activity was detected in unpolluted soils. The complexity of the microbial community (Shannon and Simpson indices) as well as the dehydrogenase soil activity negatively correlated with contamination levels. The greatest PAH-degrading gene diversity and the most intense catechol 2,3-dioxygenase activity were found in the soils with the highest levels of hydrocarbons. Heavy metals and hydrocarbon pollution has caused a genetic and metabolic alteration in microbial communities, corresponding to a reduction in microbial activity. A multi-technique approach combining traditional biochemical methods with molecular-based techniques, along with some methodological improvements, may represent an important tool to expand our knowledge of the role of microbial diversity in contaminated soil.     

Effect of wastewater irrigation on vegetables in relation to bioaccumulation of heavy metals and biochemical changes

The present study was conducted to determine the heavy metal contamination in soil with accumulation in edible parts of plants and their subsequent changes in biochemical constituents due to wastewater irrigation. Though the wastewater contains low levels of the heavy metals (Fe, Mn, Pb, Cd, and Cr), the soil and plant samples show higher values due to accumulation. The trend of metal accumulation in wastewater-irrigated soil is in the order: Fe > Pb > Mn > Cr > Cd. Of the three species Colocasia esculentum, Brassica nigra, and Raphanus sativus that are grown, the order of total heavy metal accumulation in roots is Raphanus sativus > Colocasia esculentum, while in shoots the order is Brassica nigra > Colocasia esculentum > Raphanus sativus. The enrichment factor (EF) of the heavy metals in contaminated soil is in the sequence of Cd (3) > Mn (2.7) > Cr (1.62) > Pb (1.46) > Fe (1.44), while in plants EF varies depending upon the species and plant part. C. esculentum and R. sativus show a higher EF for Cr and Cd. All plants show a high transfer factor (TF > 1) for Cd signifying a high mobility of Cd from soil to plant whereas the TF values for Pb are very low as it is not bioavailable. Results of the biochemical parameters show decrease in total chlorophyll and total amino acid levels in plants and an increase in amounts of soluble sugars, total protein, ascorbic acid, and phenol except B. nigra for protein in plants grown in soil irrigated with wastewater as compared to control site.     

Comparison of the suitability of two lichen species and one higher plant for monitoring airborne heavy metals

We compared the capacity to accumulate airborne heavy metals of two lichens (Flavoparmelia caperata and Parmotrema chinense) and one higher plant (Nerium oleander) at a very densely populated urban site near Naples. After 15, 45, 75, and 120 days of exposure at four sites with different levels of air pollution, equal portions of thalli and 20 leaves were collected, and four environmentally significant elements, Fe, Cu, Zn, and Pb, were measured by inductively coupled plasma analysis. To compare the accumulation rates of lichens and the vascular plant, we determined an index of relative accumulation rate of pollutants during time and the ratio between the concentrations of each element in exposed samples to that of control samples (exposed-to-control ratio). Our data indicate F. caperata as being the most suitable bioaccumulator, followed by P. chinense. N. oleander was also found to be a useful heavy metal biomonitor though not suitable as a bioaccumulator.     

Inventory compilation and distribution of heavy metals in wastewater from small-scale industrial areas of Delhi, India

Delhi has the highest cluster of small-scale industries (SSI) in India. There are generally less stringent rules for the treatment of waste in SSI due to less waste generation within each individual industry. This results in SSI disposing of their wastewater untreated into drains and subsequently into the river Yamuna, which is a major source of potable water in Delhi, thus posing a potential health and environmental risk to the people living in Delhi and downstream. To study the quantity, quality and distribution of heavy metals in liquid waste from industrial areas, wastewater, suspended materials and bed sediments were collected from industrial areas and from the river Yamuna in Delhi. This study has also focused on the efficiency of production processes in small-scale industries in India. Heavy metals such as Fe, Mn, Cu, Zn, Ni, Cr, Cd, Co and Pb were detected using a GBC 902 atomic absorption spectrometer. The concentration of heavy metals observed was as follows: Fe 2-212, Mn 0.3-39, Cu 0.2-20, Zn 0.2-5, Ni 0.6-6, Cr 0.2-53, Cd 0.08-0.2, Co 0.013-0.55, Pb 0.3-0.7 mg L(-1) in wastewater; Fe 5842-78 000, Mn 585-10 889, Cu 206-7201, Zn 406-9000, Ni 22-3621, Cr 178-10 533, Co 17-114, Cd 13-141, Pb 67-50 171 mg kg(-1) in suspended material; and Fe 3000-84000, Mn 479-1230, Cu 378-8127, Zn 647-4010, Ni 164-1582, Cr 139-3281, Co 20-54, Cd 37-65, Pb 228-293 mg kg(-1) in bed residues. This indicates that SSI could be one of the point sources of metals pollution in the river system.     

Speciation of heavy metals in biosolids of wastewater treatment plants at Mysore,Karnataka, India

Urban wastewater treatment leads to the generation of large quantities of biosolids. Accumulation of biosolids is a problem of environmental relevance due to the existence of heavy metals in the biosolids. Determination of total metal in biosolid provides information relating pollution levels. Determination of their mobilization capacity and behaviour in the environment is an important task. An experimental approach commonly used for studying the mobility, transport and bioavailability of metal in biosolids is the use of selective sequential extraction procedure. In the present study an attempt has been made to study the heavy metal properties in biosolid samples collected from urban wastewater treatment plants located at Mysore, Karnataka. Few heavy metals selected for the present study are cadmium, chromium, copper, iron, nickel and zinc. The concentration of these metals in biosolids and their partition in different fractions are studied. The speciation of metals based on the sequential extraction scheme was carried out. The concentration of heavy metals is lower than that established by European legislation. The residual fraction has the maximum percentage of heavy metals whereas, only a small fraction of heavy metals (Fe, Zn and Cd) are extracted in the most soluble fractions, exchangeable and carbonate fractions.     

Profiling and monitoring of DOM in brewery wastewater and treated wastewater

Dissolved organic matter (DOM) in raw and treated wastewater from two breweries in Thailand was profiled and monitored for the purpose of water reclamation. The wastewater and the effluent from the use of an upflow anaerobic sludge blanket (UASB) and activated sludge (AS) were collected and analyzed through a resin fractionation method using the fluorescent excitation?Cemission matrix (FEEM) technique. The results revealed that the major organic fractions in the brewery wastewater were hydrophobic acid (HPOA) and hydrophilic base (HPIB), accounting for 65% of total dissolved organic carbon (DOC) mass for brewery A and 56% of total DOC mass for brewery B. The FEEM results indicated that the organic matter in the wastewaters of both breweries were mainly composed of tryptophan-like substances, represented by peaks C (230 nm_Ex/340?C365 nm_Em) and D (265?C295 nm_Ex/315?C390 nm_Em), and humic-like substances, represented by peaks E (290 nm_Ex/400 nm_Em), F (330?C335 nm_Ex/395?C410 nm_Em), and G (255?C265 nm_Ex/435?C455 nm_Em). The analysis revealed that the reduction of DOM occurred mostly during the UASB treatment where most of the DOM reduction resulted from the removal of the HPOA and HPIB fractions. The HPOA fraction, a group of humic-like substances, is of particular concern when reclaiming treated brewery wastewater, and although it was reduced by more than 80% of its initial amount, it was still a dominant DOM fraction in the effluents.     

Seasonal monitoring of heavy metals and physicochemical characteristics in a lentic ecosystem of subtropical industrial region, India

Assessment of seasonal changes in surface water quality is an important aspect for evaluating temporal variations of lentic ecosystem (lakes and reservoirs) pollution due to industrial effluent discharge. In this study, nine metals and 15 physicochemical parameters, collected from four sampling sites in a tropical lake receiving the discharge from thermal power plant, coal mine, and chloralkali industry, during the years from 2004 to 2005, were analyzed. For greater efficacy in monitoring of heavy metals, particle-induced X-ray emission has been used during present investigation. Different statistical techniques like analysis of variance, Pearson correlation, principal component analysis, and factor analysis were employed to evaluate the seasonal correlations of physicochemical parameters. Most of the metals and physicochemical parameters monitored in the present study exhibited high spatial and temporal variability. Pertaining to metal pollution, the most polluted site was Belwadah, i.e., waters and sediments had the highest concentration of all the relevant metals. The reference site was characterized by the presence of low concentrations of metals in waters and in sediments. Based on the high metal concentration recorded in lake ambient, drinking, bathing, and irrigation water should not be used by the local people at the effluent discharge points.     

Assessment of heavy metals in aquatic sediments

The concentration of heavy metals in the bottom sediment and interstitial water collected from two reservoirs in Singapore was found to be enriched. A distribution coefficient,K _d , was used to assess the chemical stability of heavy metals in the sediments. Numerical models were used to assess (1) the redistribution of heavy metals in a changing environment, and (2) long-term self clean-up capabilities of a reservoir.     

Environmental monitoring of heavy metals and arsenic from Ag-Pb-Zn mining: a case study over two millennia

2000 years of mining activity at Wiesloch, Germanyleft behind a legacy of mining wastes, some of which haveextremely high contents of toxic elements like As, Cd, Tl,Sb, Pb and Zn. To evaluate their long-term impact ondifferent environmental compartments, the detailedenvironmental monitoring presented here focused on themineralogical and chemical characterization of thedifferent waste materials, consisting of dumpings with orefragments, flotation tailings and medieval metallurgicalslags. Leaching experiments with these materials, usingeluents of different compositions and pHs were carried outto assess the conditions governing the mobilization and re-fixation of these species. It was shown, that the carbonatehost rock of the mineralization, the loess blanket coveringthe area and the organically rich municipal sewage sludgesdeposited on top of the tailings, represent potentialbarriers to the dispersion of toxic elements over a muchlarger area. Moreover, particulate emissions from thesteep, unvegetated escarpments of the tailing heapsrepresent a continuous thread to the environment.     

Uptake of heavy metals by some edible vegetables irrigated using wastewater: a preliminary study in Accra, Ghana

The heavy metals (Fe, Zn, Pb, Ni, Cr, Co, and Cd) burden in wastewater, soil, and vegetable samples from a wastewater irrigated farm located at KorleBu, Accra has been investigated. Flame atomic absorption spectrometry after microwave digestion using a combination of HNO₃, HCl, and H₂O₂ (for water), and HNO₃ and HCl (for soil and vegetables). The mean concentrations (in milligrams per kilogram) of heavy metals in the soil samples were in the order of Fe (171?±?5.22)?>?Zn (36.06?±?4.54)?>?Pb (33.35?±?35.62)?>?Ni (6.31?±?8.15)?>?Cr (3.40?±?3.63)?>?Co (1.36?±?0.31)?>?Cd (0.43?±?0.24), while the vegetables were in the order of Fe (183.11?±?161.2)?>?Zn (5.38?±?3.50)?>?Ni (3.52?±?1.27)?>?Pb (2.49?±?1.81)?>?Cr (1.46?±?0.51)?>?Co (0.66?±?0.25)?>?Cd (0.36?±?0.15). The bioconcentration factors suggest environmental monitoring for the heavy metals as follows: Cd (0.828), Cr (0.431), Ni (0.558), Co (0.485), and Fe (1.067). Estimated daily intakes were very low for both children and adults except Fe (0.767 mg/kg/day) in children. The population that consume vegetables from the study area were, however, estimated to be safe based on the results obtained from the health risk index, which were all?<?<1. The sodium absorption ratio according to FAO (1985) classifications indicate that the wastewater in the study area is unsuitable for irrigation purposes.     

Cement for stabilisation of industrial residues containing heavy metals

A method aimed at decreasing the toxicity of heavy metals [namely, Zn(II) and Cr(III)] in real polluted residues by immobilisation has been developed. The residues were processed either with two cement-type stabilisers or lime. The cement-type stabilisers were Portland cement and Depocrete SM/2 at the self-generated pH (ca. 11) which afforded physical as well as chemical potential for the immobilisation of heavy metals. The other stabiliser, lime, reduced organic compounds, thus favouring the decrease of the chemical oxygen demand (COD) and endowing the residue with better mechanical properties for transport. After leaching the stabilised residues using the standard leaching test [Order 13/10/89, Boletín Oficial del Estado (BOE) 270 10/11/89], three ways for establishing the toxicity of the treated residues were used, namely: (1) the ecotoxicity test using Photobacterium phosphoreum (DIN 38 412); (2) determination of the concentration of heavy metals by atomic absorption spectrometry (AAS); (3) determination of the COD or oxygen required for complete chemical oxidation of a water sample. Portland cement (20%) blended with Depocrete SM/2 (3%) acted as an effective stabiliser for residues containing heavy metals as it increased the ecotoxicity index (EC50) by more than five times. Thus the heavy metal concentration in the leaching liquid was lowered to less than 0.1 mg l-1. The addition of 5% of lime afforded a residue easily transportable from the place of treatment to the landfill. The precision of the method was studied in terms of both repeatability and reproducibility. The values found with respect to EC50 and expressed as the relative standard deviation (RSD) were 1.6% and 5.1%, respectively.     

Passive monitoring of atmospheric heavy metals in a historical city of central India by Lepraria lobificans Nyl.

Using an organism living in situ for monitoring is referred as passive monitoring. Lepraria lobificans Nyl., a leprose lichen growing naturally on monuments and buildings in the city Mandav in central India is used for passive monitoring of atmospheric metals. Seven metals (Cd, Cr, Ni, Al, Fe, Cu, and Zn) were analyzed. Samples collected from road site exhibit the maximum concentration of Fe, Cd, Cr, Ni, and Zn. Iron exhibit maximum accumulation both in lichen thallus and the substratum with mean values of 2,195.63 μg g^???1 dry weight. As compared with other growth form of lichens, L. lobificans exhibits the higher accumulation of Fe than foliose and fruticose lichens. On the basis of these results, it can be hypothesized that L. lobificans is an excellent accumulator of different metals. The statistical analysis applied to the element concentration between the metals as well as between the sites by analysis of variance found the difference to be significant at 1% and 5%, respectively. Student–Newman–Keuls test also shows significant difference for iron between the different metals.     

Distribution of heavy metals in polluted creek sediment

Sediments affect the water quality substantially. The metal concentrations in sediments often reflect the degree of pollution of the aquatic environment. The sediments of Thane Creek near Bombay were analyzed for nine metals Cd, Co, Cr, Cu, Fe, Mn, Ni, Pb, and Zn on twenty stations. The average metal concentrations in the sediments were 2.29, 37.25, 39.89, 103.38, 69825.0, 860.15, 105.08, 54.08, and 169.60 g g^-1 dwt respectively. The metal concentrations at certain stations were relatively higher than those of uncontaminated sediments, possibly due to industrial activity in the former area.     

Study of heavy metals in some environmental samples

Fuels like coal and rubber are frequently used for brick burning. However, both coal and rubber contain heavy metals. These heavy metals may elutriate in the wake of fly ash or may adsorb or absorb in the product. The present work deals with the analysis of heavy metals in some samples collected from brick burning industries located in the vicinity of a metropolitan city, Peshawar, Pakistan. Samples from raw clay, product, chimney scale and fossil fuel & rubber were collected and leached with acid mixture. The leachates were concentrated and analyzed by atomic absorption spectrophotometer for the determination of chromium (Cr), lead (Pb), cadmium (Cd) and antimony (Sb). It was observed that heavy metals are present in clay, brick and chimney scale. However, significant amount of these metals was observed in chimney scale. It is inferred that such emanations laden with heavy metals are accompanying the stack gases which are being dumped in to the environment. In order to avoid environmental problems, strict environmental regulations shall be enforced and a constant check on these emanations to the environment must be made to ensure clean air act.     

Performance optimisation of a passive sampler for monitoring hydrophobic organic pollutants in water

The performance of an integrative passive sampler that consists of a C18 Empore disk sorbent receiving phase fitted with low density polyethylene membrane was optimised for the measurement of time-weighted average concentrations of hydrophobic micropollutants in water. A substantial improvement of sampling characteristics including the rate of sampling and the sampling precision was achieved by decreasing the internal sampler resistance to mass transfer of hydrophobic organic chemicals. This was achieved by adding a small volume of n-octanol, a solvent with high permeability (solubility [times] diffusivity) for target analytes, to the interstial space between the receiving sorbent phase and the polyethylene diffusion-limiting membrane.     

Concentrations of cadmium and lead heavy metals in Dardanelles seawater

Cadmium and lead were determined simultaneously in seawater by differential pulse stripping voltammetry (DPSV) preceded by adsoptive collection of complexes with 8-hydroxyquinoline (oxine) on to a hanging mercury drop electrode (HMDE). In preliminary experiments the optimal analytical condition for oxine concentration was found to be 2.10⁻⁵ M, at pH 7.7, the accumulation potential was −1.1 V, and the initial scannig potential was −0.8 V. The peak potentials were found −0.652 V for Cd and −0.463 V for Pb At the 60 s accumalation time. The limit of detection (LOD) and limit of quantitatification (LOQ) were found to be by voltammetry as 0.588 and 1.959 μg l⁻¹ (RSD, 5.50%) for Cd and 0.931 and 3.104 μg l⁻¹ (RSD, 4.10%) for Pb at 60 s stirred accumulation time respectively. In these conditions the most of the seawater samples are amenable for direct voltammetric determination of cadmium and lead using a HMDE. An adsorptive stripping mechanism of the electrode reaction was proposed. For the comparison, seawater samples were also analysed by ICP-atomic emission spectrometry method (ICP-AES). The applied voltammetric technique was validated and good recoveries were obtained.     

Assessment of long-term wastewater irrigation impacts on the soil geochemical properties and the bioaccumulation of heavy metals to the agricultural products

An extensive field survey was employed for assessing the impacts of long-term wastewater irrigation of forage crops and orange orchards in three suburban agricultural areas in Cyprus (areas I, II, and III), as compared to rainfed agriculture, on the soil geochemical properties and the bioaccumulation of heavy metals (Zn, Ni, Mn, Cu, Co) to the agricultural products. Both ryegrass fields and orange orchards in areas I and II were continuously wastewater irrigated for 10 years, whereas clover fields in area III for 0.5, 4, and 8 years. The results revealed that wastewater reuse for irrigation caused a slight increase in soil salinity and Cl^? content in areas I and II, and a remarkable increase, having strong correlation with the period in which wastewater irrigation was practiced, in area III. Soil salinization in area III was due to the high electrical conductivity (EC) of the wastewater applied for irrigation, attributed to the influx of seawater to the sewage collection network in area III. In addition, the wastewater irrigation practice resulted in a slight decrease of the soil pH values in area III, while a subtle impact was identified regarding the CaCO₃, Fe, and heavy metal content in the three areas surveyed. The heavy metal content quantified in the forage plants’ above-ground parts was below the critical levels of phytotoxicity and the maximum acceptable concentration in dairy feed, whereas heavy metals quantified in orange fruit pulp were below the maximum permissible levels (MPLs). Heavy metal phytoavailability was confined due to soil properties (high pH and clay content), as evidenced by the calculated low transfer factor (TF).     

Development of a hybrid pollution index for heavy metals in marine and estuarine sediments

Heavy metal pollution of sediments is a growing concern in most parts of the world, and numerous studies focussed on identifying contaminated sediments by using a range of digestion methods and pollution indices to estimate sediment contamination have been described in the literature. The current work provides a critical review of the more commonly used sediment digestion methods and identifies that weak acid digestion is more likely to provide guidance on elements that are likely to be bioavailable than other traditional methods of digestion. This work also reviews common pollution indices and identifies the Nemerow Pollution Index as the most appropriate method for establishing overall sediment quality. Consequently, a modified Pollution Index that can lead to a more reliable understanding of whole sediment quality is proposed. This modified pollution index is then tested against a number of existing studies and demonstrated to give a reliable and rapid estimate of sediment contamination and quality.     

Multivariate analysis of heavy metals concentrations in river estuary

Multivariate statistical techniques such as multivariate analysis of variance (MANOVA) and discriminant analysis (DA) were applied for analyzing the data obtained from two rivers in the Penang State of Malaysia for the concentration of heavy metal ions (As, Cr, Cd, Zn, Cu, Pb, and Hg) using a flame atomic absorption spectrometry (F-AAS) for Cr, Cd, Zn, Cu, Pb, As and cold vapor atomic absorption spectrometry (CV-AAS) for Hg. The two locations of interest with 20 sampling points of each location were Kuala Juru (Juru River) and Bukit Tambun (Jejawi River). MANOVA showed a strong significant difference between the two rivers in terms of heavy metal concentrations in water samples. DA gave the best result to identify the relative contribution for all parameters in discriminating (distinguishing) the two rivers. It provided an important data reduction as it used four parameters (Zn, Pb, Cd and Cr) affording 100% correct assignations. Results indicated that the two rivers were different in terms of heavy metals concentrations in water, and the major difference was due to the contribution of Zn. A negative correlation was found between discriminate functions (DF) and Cr and As, whereas positive correlation was exhibited with other heavy metals. Therefore, DA allowed a reduction in the dimensionality of the data set, delineating a few indicator parameters responsible for large variations in heavy metal concentrations. Correlation matrix between the parameters exhibited a strong evidence of mutual dependence of these metals.