Multiplex-PCR and PCR-RFLP assays to monitor water quality against pathogenic bacteria

In this work we developed and optimized two molecular-based approaches to monitor rapidly, sensitively and specifically bacterial pathogens from three different genera, Escherichia coli, Pseudomonas aeruginosa, and Salmonella spp., directly in waters. To achieve this aim, firstly a multiplex-PCR assay (M-PCR) was optimized using a primer pair specific for each pathogen. Secondly, as a molecular confirmatory test after isolation of the pathogens by classical microbiological methods, PCR-RFLP of their amplified 16S rDNA genes was performed. It was observed from the results that the developed M-PCR assay has significant impact on the ability to detect sensitively, rapidly and specifically the three pathogens directly in water within a short time (5 h from sampling to obtain final results), therefore it represents a considerable advancement over other known more time-consuming and less-sensitive methods for identification and characterization of these kinds of pathogens.     

Sediment quality in the Douro river estuary based on trace metal contents, macrobenthic community and elutriate sediment toxicity test (ESTT)

The aim of this study was to evaluate the sediment quality in the mesotidal Douro River estuarine environment, in order to identify areas where sediment contamination could cause ecosystem degradation. Samples were obtained in five locations and sediment characterised for grain size, total organic matter, total-recoverable metals (Al, Fe, Cu, Pb, Cr, Ni, Cd, Zn and Mn), as well as acid volatile sulfide (AVS) and simultaneously extracted metals (SEM). In situ effects were evaluated by examining the macrobenthic community structure. An elutriate sediment toxicity test (ESTT) was used to estimate the amount of metals and nutrients that could be exchanged with the water column through resuspension, and its positive or negative effects on the growth of the micro-alga Emiliania huxleyi in a 10 day test. Anthropogenic metal contamination was identified at the north bank of the Douro estuary, with deleterious effects on the macrobenthic community, namely decrease in number of species and diversity. This contamination could possibly also be toxic for water column organisms, in case of resuspension, as shown by the ESTT. Sediments from the salt marsh at the south bank showed an impoverished macrobenthic community and elutriate toxicity, which appeared to be due to anaerobic conditions. This study clearly shows the usefulness of the ESST approach to assess the biological effect of resuspension of estuarine sediments.     

Comparative Metal Distribution in Hair of Pakistani and Libyan Population and Source Identification by Multivariate Analysis

Using nitric acid-perchloric acid wet digestion based FAAS method ten selected metals (Cd, Co, Cr, Fe, K, Mn, Na, Ni, Pb and Zn) were determined in the scalp hair of male Pakistani and Libyan donors (n = 62), between 3–54 years age and residing in typical urban areas. The study was taken up to identify sources of metal distribution in two diverse population segments exposed to different environments. Sodium emerged with the highest mean hair concentration of 511.0 μg/g and 496.9 μg/g for Libyan and Pakistani donors respectively, followed by K, at 266.2 μg/g and 242.4 μg/g for the two donor categories. Cadmium showed the lowest mean hair concentration for both Pakistani (0.380 μg/g) and Libyan (0.530 μg/g) donors. Levels of Na, K, Pb, Cr and Cd were higher in hair of Libyan donors compared with Pakistani counterparts which showed higher comparative levels of Zn, Fe, Co, Ni and Mn. Strong metal-to-metal correlations were found between Na–K (r = 0.638) and Cd–Co (r = 0.574) for Pakistani donors, while for Libyan donors, Na–K (r = 0.680) and Fe–Mn (r = 0.624) correlations emerged as strongly significant. Only K was found to have a strong positive correlation (r = 0.611) with age for Pakistani donors while this correlation was significantly negative (r = −0.500) for Libyan donors, probably arising from individual food habits. Other metals showed no viable relationship with age. Principal Component Analysis (PCA) and Cluster Analysis (CA) of the data evidenced food, industrial emissions and automobile emissions as possible sources of metal distribution. The results of the present study are compared with those reported for subjects from other regions of the world.     

Influence of organic waste type and soil structure on the bacterial filtration rates in unsaturated intact soil columns

Organic wastes are considered to be a source for the potentially pathogenic microorganisms found in surface and sub-surface water resources. Following their release from the organic waste matrix, bacteria often infiltrate into soil and may be transported to significant depths contaminating aquifers. We investigated the influence of soil texture and structure and most importantly the organic waste properties on the transport and filtration coefficients of Escherichia coli and total bacteria in undisturbed soil columns. Intact soil columns (diameter 16 cm and height 25 cm) were collected from two soils: sandy clay loam (SCL) and loamy sand (LS) in Hamadan, western Iran. The cores were amended with cow manure, poultry manure and sewage sludge at a rate of 10 Mg ha(-1) (dry basis). The amended soil cores were leached at a steady-state flux of 4.8 cm h(-1) (i.e. 0.12 of saturated hydraulic conductivity of the SCL) to a total volume of up to 4 times the pore volume of the columns. The influent (C(0)) and effluent (C) were sampled at similar time intervals during the experiments and bacterial concentrations were measured by the plate count method. Cumulative numbers of the leached bacteria, filtration coefficient (lambda(f)), and relative adsorption index (S(R)) were calculated. The preferential pathways and stable structure of the SCL facilitated the rapid transport and early appearance of the bacteria in the effluent. The LS filtered more bacteria when compared with the SCL. The effluent contamination of poultry manure-treated columns was greater than the cow manure- and sewage sludge-treated ones. The difference between cow manure and sewage sludge was negligible. The lambda(f) and S(R) values for E. coli and total bacteria were greater in the LS than in the SCL. This indicates a predominant role for the physical pore-obstruction filtration mechanisms as present in the poorly structured LS vs. the retention at adsorptive sites (chemical filtration) more likely in the better structured SCL. While the results confirmed the significant role of soil structure and preferential (macroporous) pathways, manure type was proven to have a major role in determining the maximum penetration risk of bacteria by governing filtration of bacteria. Thus while the numbers of bacteria in waste may be of significance for shallow aquifers, the type of waste may determine the risk for microbial contamination of deep aquifers.     

Relevance of joint action toxicity evaluations in setting realistic environmental safe limits of heavy metals

The evaluation of types of toxicological interactions existing between heavy metals, which are prominent in effluents of some industrial establishments in Lagos State, Nigeria and the Lagos lagoon sediment was carried out against benthic animals, Tympanotonus fuscatus, Clibanarius africanus and Sesarma huzardi of the Lagos lagoon. In order to determine the type of interactions existing between the metals, acute toxicity tests of the metal compounds when acting singly and in joint action studies, by adopting mixture ratios that depict (i) the proportions of the concentrations of the metal ions in the sediment of the Lagos lagoon and (ii) equitoxic mixtures, i.e. based on the 96 hLC(50) values of the metal compounds under single action studies were carried out in laboratory biotests. The joint action evaluations of the test metal mixtures, prepared on the basis of the proportion of the availability of the metals ions in lagoon sediment and equitoxic ratio against the test animals agreed mainly with the model of antagonism (reduction in toxicity) except for test mixtures prepared on the basis of equitoxic ratio and tested against T. fuscatus, where interactions between the test metals was in conformity with the model of synergism, indicating that the toxicity of the constituent metals in the mixture was enhanced. Furthermore, on the basis of classification of synergistic ratio model, the toxic effect of Pb compound which was the least toxic metal compound in the single action toxicity studies was found to be enhanced (synergised) in the presence of other metals when tested jointly. The significance of the results in setting water quality criteria aimed at protecting aquatic biota was discussed.     

Conservation strategies for effective land management of protected areas using an erosion prediction information system (EPIS)

This research demonstrates the predictive modeling capabilities of a geographic information system (GIS)-based soil erosion potential model to assess the effects of implementing land use change within a tropical watershed. The Revised Universal Soil Loss Equation (RUSLE) was integrated with a GIS to produce an Erosion Prediction Information System (EPIS) and modified to reflect conditions found in the mountainous tropics. Research was conducted in the Zenzontia subcatchment of the Río Ayuquíla, located within the Sierra de Manantlán Biosphere Reserve (SMBR), México. Expanding agricultural activities within this area will accentuate the already high rate of soil erosion and resultant sediment loading occurring in the Río Ayuquíla. Two land-use change scenarios are modeled with the EPIS: (1) implementation of soil conservation practices in erosion prone locations; and (2) selection of sites for agricultural expansion which minimize potential soil loss. Confronted with limited financial resources and the necessity for expedient action, managers of the SMBR can draw upon the predictive capacity of the EPIS to facilitate rapid and informed land-use planning decisions.     

Denitrification in constructed wetlands used for treatment of swine wastewater

Constructed wetland treatment of swine wastewater probably involves substantial denitrification. Our objective was to assess denitrification and denitrification enzyme activity (DEA) in such wetlands in relation to plant communities, N loading, carbon or nitrogen limitations, and water depth. Two wetland cells each 3.6 m wide and 33.5 m long were connected in series. One set of cells was planted with rushes and bulrushes, including soft rush (Juncus effusus L.), softstem bulrush [Schoenoplectus tabernaemontani (K.C. Gmel.) Pallal, American bulrush [Schoenoplectus americanus (Pers.) Volkart ex Schinz & R. Keller], and woolgrass bulrush [Scirpus cyperinus (L.) Kunth]. Another set was planted with bur-reeds and cattails, including American bur-reed (Sparganium americanum Nutt.), broadleaf cattail (Typha latifolia L.), and narrowleaf cattail (Typha angustifolia L.). The sets will be referred to herein as bulrush and cattail wetlands, respectively. Denitrification and DEA were measured via the acetylene inhibition method in intact soil cores and disturbed soil samples that were taken during four years (1994-1997). Although DEA in the disturbed samples was greater than denitrification in the core samples, the measurements were highly correlated (r2 > or = 0.82). The DEA was greater in the bulrush wetlands than the cattail wetlands, 0.516 and 0.210 mg N kg(-1) soil h(-1), respectively; and it increased with the cumulative applied N. The DEA mean was equivalent to 9.55 kg N ha(-1) d(-1) in the bulrush wetlands. We hypothesized and confirmed that DEA was generally limited by nitrate rather than carbon. Moreover, we determined that one of the most influential factors in DEA was wetland water depth. In bulrush wetlands, the slope and r2 values of the control treatment were -0.013 mg N kg(-1) soil h(-1) mm(-1) depth and r2 = 0.89, respectively. Results of this investigation indicate that DEA can be very significant in constructed wetlands used to treat swine wastewater.     

Impact of biochar enriched with dairy manure effluent on carbon and nitrogen dynamics

Amending soils with biochar can have multiple environmental benefits, including improvement in soil physicochemical properties, carbon sequestration, reduction in leaching losses of essential nutrients, and reduction in greenhouse gas (GHG) emissions. This study was conducted to determine the effect of enriched biochar amendments on leaching losses of essential nutrients and GHG emissions from soil. The enriched biochar was prepared by shaking biochar with dairy manure effluent for 24 h, which increased the C and N concentration of biochar by 9.3 and 8.3%, respectively. Incubation and leaching experiments were conducted for 8 wk with three treatments: soil, soil + 1% biochar, and soil + 1% enriched biochar. Amendment with biochar and enriched biochar relative to unamended soil resulted in 68 and 75% reduction in net nitrification, 221 and 229% reduction in net ammonification, 67 and 68% reduction in cumulative CO flux, respectively, and 26% reduction in cumulative NO flux for both biochar treatments. There were no significant differences among treatments in total leaching losses of C, N, and base cations. Our findings suggest that enrichment of biochar with dairy manure effluent can promote C and N storage in soil and provide additional environmental benefits.     

A framework for assessing the impact of land use policy on community exposure to air toxics

Our research focuses on the linkage between land use planning policy and the spatial pattern of exposure to air toxics emissions. Our objective is to develop a modeling framework for assessment of the community health risk implications of land use policy. The modeling framework is not intended to be a regulatory tool for small-scale land use decisions, but a long-range planning tool to assess the community health risk implications of alternative land use scenarios at a regional or subregional scale. This paper describes the development and application of an air toxic source model for generating aggregate emission factors for industrial and commercial zoning districts as a function of permitted uses. To address the uncertainty of estimating air toxics emission rates for planned general land use or zoning districts, the source model uses an emissions probability mass function that weights each incremental permitted land use activity by the likelihood of occurrence. We thus reduce the uncertainty involved in planning for development with no prior knowledge of the specific industries that may locate within the land use district. These air toxics emission factors can then be used to estimate pollutant atmospheric mass flux from land use zoning districts, which can then be input to air dispersion and human health risk assessment models to simulate the spatial pattern of air toxics exposure risk. The model database was constructed using the California Air Toxics Inventory, 1997 US Economic Census, and land assessment records from several California counties. The database contains information on more than 200 air toxics at the 2-digit Standard Industrial Classification (SIC) level. We present a case study to illustrate application of the model. LUAIRTOX, the interactive spreadsheet model that applies our methodology to the California data, is available at http://www2.bren.ucsb.edu/~mwillis/LUAIRTOX.htm.     

Dissolved phosphorus retention and release from a coastal plain in-stream wetland

Dissolved phosphorus (DP) can be released from wetlands as a result of flooding or shifts in water column concentrations. Our objectives were to determine the long-term (1460 d) DP retention and release characteristics of an in-stream wetland, and to evaluate how these characteristics respond to flooding, draining, and changes in DP concentrations. The studied in-stream wetland drains an agriculturally intensive subwatershed in the North Carolina Coastal Plain region. The wetland's DP retention and release characteristics were evaluated by measuring inflow and outflow DP concentrations, DP mass balance, and DP movement across the sediment-water column interface. Phosphorus sorption isotherms were measured to determine the sediment's equilibria P concentration (EPCo), and passive samplers were used to measure sediment pore water DP concentrations. Initially, the in-stream wetland was undersized (0.31 ha) and released 1.5 kg of DP. Increasing the in-stream wetland area to 0.67 ha by flooding resulted in more DP retention (28 kg) and low outflow DP concentrations. Draining the in-stream wetland from 0.67 to 0.33 ha caused the release of stored DP (12.1 kg). Shifts both in sediment pore water DP concentrations and sediment EPCo values corroborate the release of stored DP. Reflooding the wetland from 0.33 to 0.85 ha caused additional release of stored DP into the outflowing stream (10.9 kg). We conclude that for a time period, this in-stream wetland did provide DP retention. During other time periods, DP was released due to changes in wetland area, rainfall, and DP concentrations.