Costantino Masciopinto

Quantitative PCR Monitoring of Antibiotic Resistance Genes and Bacterial Pathogens in Three European Artificial Groundwater Recharge Systems

ABSTRACT Aquifer recharge presents advantages for integrated water management in the anthropic cycle, namely, advanced treatment of reclaimed water and additional dilution of pollutants due to mixing with natural groundwater. Nevertheless, this practice represents a health and environmental hazard because of the presence of pathogenic microorganisms and chemical contaminants. To assess the quality of water extracted from recharged aquifers, the groundwater recharge systems in Torreele, Belgium, Sabadell, Spain, and Nardò, Italy, were investigated for fecal-contamination indicators, bacterial pathogens, and antibiotic resistance genes over the period of 1 year. Real-time quantitative PCR assays for Helicobacter pylori, Yersinia enterocolitica, and Mycobacterium avium subsp. paratuberculosis, human pathogens with long-time survival capacity in water, and for the resistance genes ermB, mecA, blaSHV-5, ampC, tetO, and vanA were adapted or developed for water samples differing in pollutant content. The resistance genes and pathogen concentrations were determined at five or six sampling points for each recharge system. In drinking and irrigation water, none of the pathogens were detected. tetO and ermB were found frequently in reclaimed water from Sabadell and Nardò. mecA was detected only once in reclaimed water from Sabadell. The three aquifer recharge systems demonstrated different capacities for removal of fecal contaminators and antibiotic resistance genes. Ultrafiltration and reverse osmosis in the Torreele plant proved to be very efficient barriers for the elimination of both contaminant types, whereas aquifer passage followed by UV treatment and chlorination at Sabadell and the fractured and permeable aquifer at Nardò posed only partial barriers for bacterial contaminants.

Quantification of pathogenic microorganisms and microbial indicators in three wastewater reclamation and managed aquifer recharge facilities in Europe

Managed Aquifer Recharge (MAR) is becoming an attractive option for water storage in water reuse processes as it provides an additional treatment barrier to improve recharged water quality and buffers seasonal variations of water supply and demand. To achieve a better understanding about the level of pathogenic microorganisms and their relation with microbial indicators in these systems, five waterborne pathogens and four microbial indicators were monitored over one year in three European MAR sites operated with reclaimed wastewater. Giardia and Cryptosporidium (oo)cysts were found in 63.2 and 36.7% of the samples respectively. Salmonella spp. and helminth eggs were more rarely detected (16.3% and 12.5% of the samples respectively) and Campylobacter cells were only found in 2% of samples. At the Belgian site advanced tertiary treatment technology prior to soil aquifer treatment (SAT) produced effluent of drinking water quality, with no presence of the analysed pathogens. At the Spanish and Italian sites amelioration of microbiological water quality was observed between the MAR injectant and the recovered water. In particular Giardia levels decreased from 0.24-6.14 cysts/L to 0-0.01 cysts/L and from 0.4-6.2 cysts/L to 0-0.07 cysts/L in the Spanish and Italian sites respectively. Salmonella gene copies and Giardia cysts were however found in the water for final use and/or the recovered groundwater water at the two sites. Significant positive Spearman correlations (p<0.05, r(s) range: 0.45-0.95) were obtained, in all the three sites, between Giardia cysts and the most resistant microbial markers, Clostridium spores and bacteriophages.

A combined PHREEQC-2/parallel fracture model for the simulation of laminar/non-laminar flow and contaminant transport with reactions.

A combination of a parallel fracture model with the PHREEQC-2 geochemical model was developed to simulate sequential flow and chemical transport with reactions in fractured media where both laminar and turbulent flows occur. The integration of non-laminar flow resistances in one model produced relevant effects on water flow velocities, thus improving model prediction capabilities on contaminant transport. The proposed conceptual model consists of 3D rock-blocks, separated by horizontal bedding plane fractures with variable apertures. Particle tracking solved the transport equations for conservative compounds and provided input for PHREEQC-2. For each cluster of contaminant pathways, PHREEQC-2 determined the concentration for mass-transfer, sorption/desorption, ion exchange, mineral dissolution/precipitation and biodegradation, under kinetically controlled reactive processes of equilibrated chemical species. Field tests have been performed for the code verification. As an example, the combined model has been applied to a contaminated fractured aquifer of southern Italy in order to simulate the phenol transport. The code correctly fitted the field available data and also predicted a possible rapid depletion of phenols as a result of an increased biodegradation rate induced by a simulated artificial injection of nitrates, upgradient to the sources.

Stygofauna Abundance and Distribution in the Fissures and Caves of the Nardò (Southern Italy) Fractured Aquifer Subject to Reclaimed Water Injections

The demographic growth in developing countries and the increasing pressure of anthropological activities in industrialized states around the world, are leading to a gradual contamination of the natural habitats of our planet. Although the extent of these effects is unclear, the results can already seen in the quality of natural resources, which are intensely stressed by climate changes (greenhouse effect, nutrients load, water consumption, etc.) and by direct contamination of toxic wastes. This could progressively destroy the variety of faunal species and, indeed, recent warming has caused changes in species distribution and abundance. This paper presents an investigation into the possible effects of climate change and anthropological pressures on the ground water fauna present at the Nardò site (Salento peninsula, Southern Italy). Three ecological categories were examined: stygoxenes, stygophiles and stygobionts. The latter are anophthalmic, without pigment, measure up to 10–12 mm, and live in water which moves throughout fissures and karstic caves of carbonate aquifers. These stygofauna are very sensitive to changes, due to environmental stresses, such as water temperature, dissolved oxygen, water salinity, pH and chemical constituents, in their hypogeous habitat. The stygofauna categories are active organisms which contribute to the biodegradation of organic compounds in wastewater artificially (or naturally) injected in the fractured subsoil. Weak information has been available until now about Salento stygofauna ability to resist water pollution caused by human activities. At the Nardò site 12000 m3/d of 2 y effluent from municipal treatment plants have been injected since 1991 in a natural sinkhole. Here, the abundance of the stygofauna, recovered in three wells (Colucci, Brusca and Spundurata cave) at progressive distances from sinkhole, and their distribution have been correlated with ground water constituents. Groundwater quality was monitored on each occasion that stygofauna were collected, during the spring-autumn seasons.

Assessment of the impact of sea-level rise due to climate change on coastal groundwater discharge

An assessment of sea intrusion into coastal aquifers as a consequence of local sea-level rise (LSLR) due to climate change was carried out at Murgia and Salento in southern Italy. The interpolation of sea-level measurements at three tide-gauge stations was performed during the period of 2000 to 2014. The best fit of measurements shows an increasing rate of LSLR ranging from 4.4mm/y to 8.8mm/y, which will result in a maximum LSLR of approximately 2m during the 22nd century. The local rate of sea-level rise matches recent 21st and 22nd century projections of mean global sea-level rise determined by other researchers, which include increased melting rates of the Greenland and Antarctic ice sheets, the effect of ocean thermal expansion, the melting of glaciers and ice caps, and changes in the quantity of stored land water. Subsequently, Ghyben-Herzbergs equation for the freshwater/saltwater interface was rewritten in order to determine the decrease in groundwater discharge due to the maximum LSLR. Groundwater flow simulations and ArcGIS elaborations of digital elevation models of the coast provided input data for the Ghyben-Herzberg calculation under the assumption of head-controlled systems. The progression of seawater intrusion due to LSLR suggests an impressive depletion of available groundwater discharge during the 22nd century, perhaps as much as 16.1% of current groundwater pumping for potable water in Salento.

A Suitable Tool for Sustainable Groundwater Management

Artificial recharge is used to increase the availability of groundwater storage and reduce saltwater intrusion in coastal aquifers, where pumping and droughts have severely impaired groundwater quality. The implementation of optimal recharge methods requires knowledge of physical, chemical, and biological phenomena involving water and wastewater filtration in the subsoil, together with engineering aspects related to plant design and maintenance operations. This study uses a novel Decision Support System (DSS), which includes soil aquifer treatment (SAT) evaluation, to design an artificial recharge plant. The DSS helps users make strategic decisions on selecting the most appropriate recharge methods and water treatment technologies at specific sites. This will enable the recovery of safe water using managed aquifer recharge (MAR) practices, and result in reduced recharge costs. The DSS was built using an artificial intelligence technique and knowledge-based technology, related to both quantitative and qualitative aspects of water supply for artificial recharge. The DSS software was implemented using rules based on the cumulative experience of wastewater treatment plant engineers and groundwater modeling. Appropriate model flow simulations were performed in porous and fractured coastal aquifers to evaluate the suitability of this technique for enhancing the integrated water resources management approach. Results obtained from the AQUASTRESS integrated project and DRINKADRIA IPA CBC suggest the most effective strategies for wastewater treatments prior to recharge at specific sites.

Flow and Transport in Fractured Aquifers: New Conceptual Models Based on Field Measurements

We derived new equations of fracture aperture (or tube diameter) as functions of a tortuosity factor that can be used in discrete models and even in continuum equivalent models to simulate fluid flow and pollutant transport in fractured aquifers. MODFLOW/MT3DMS water velocity predictions have been compared with those obtained using a specific software application which solves flow and transport problems in a 3D set of parallel fissures. The results of a pumping/tracer test carried out in a fractured limestone aquifer in Bari (Southern Italy) have been used to calibrate advective/dispersive tracer fluxes given by the applied models. The mean tracer velocity given by a breakthrough curve was greater than values predicted by continuum models. This discrepancy increased when the hydraulic conductivity of the considered fractured medium decreased. Successful simulations of flow and transport in the fractured limestone aquifer are then achieved by accommodating a new tortuosity factor in models. The importance of the proposed tortuosity factor correction lies in the possibility of taking into account the effective tracer velocity during flow and transport simulations in fractures even when using a continuum model.

A New Method to Infer Advancement of Saline Front in Coastal Groundwater Systems by 3D: The Case of Bari (Southern Italy) Fractured Aquifer

A new method to study 3D saline front advancement in coastal fractured aquifers has been presented. Field groundwater salinity was measured in boreholes of the Bari (Southern Italy) coastal aquifer with depth below water table. Then, the Ghyben-Herzberg freshwater/saltwater (50%) sharp interface and saline front position were determined by model simulations of the freshwater flow in groundwater. Afterward, the best-fit procedure between groundwater salinity measurements, at assigned water depth of 1.0 m in boreholes, and distances of each borehole from the modelled freshwater/saltwater saline front was used to convert each position (x, y) in groundwater to the water salinity concentration at depth of 1.0 m. Moreover, a second best-fit procedure was applied to the salinity measurements in boreholes with depth z. These results provided a grid file (x, y, z, salinity) suitable for plotting the actual Bari aquifer salinity by 3D maps. Subsequently, in order to assess effects of pumping on the saltwater-freshwater transition zone in the coastal aquifer, the Navier-Stokes (N-S) equations were applied to study transient density-driven flow and salt mass transport into freshwater of a single fracture. The rate of seawater/freshwater interface advancement given by the N-S solution was used to define the progression of saline front in Bari groundwater, starting from the actual salinity 3D map. The impact of pumping of 335 L·s−1 during the transition period of 112.8 days was easily highlighted on 3D salinity maps of Bari aquifer.

Mass-transfer impact on solute mobility in porous media: A new mobile-immobile model

The theory for modeling non-equilibrium solute transport in porous media is still based on approximations to a model proposed by Lapidus and Amundson in 1952 that has not been updated. This Mobile-Immobile Model (MIM) is based on the definition of a mass-transfer coefficient (α), which has been proven subject to some severe limitations. Measurements at both laboratory and field scales have demonstrated the scale-dependency of α values. This means that the MIM theory fails in real applications, since α is not constant, as defined in the kinetic model theory, but is a time-residence (or distance) dependent coefficient. Multi-rate mass-transfer models have been proposed in recent literature to capture real-world solute transport with a multiple mass transfer. In this study, we propose a novel model, which implements the analytical solution of Ficks second law of diffusion directly in the nonequilibrium advection/dispersion equation of solute transport in porous media. New model solutions properly fitted data collected during tracer tests carried out at the CNR-IRSA Laboratory (Bari, Italy) in a horizontal sandbox, 2 m of length, by using sodium chloride as the conservative tracer. Selected breakthrough curves at specific positions were used to validate the proposed model solution and estimate both conventional and proposed coefficients of mass transfer. Results have shown a decreasing trend of α from 0.09 to 0.04 h-1 after about 1.2 m of filtration for the investigated sand, whereas new solutions provide two scale-invariant tracer coefficients of rate of tracer mass-transfer (0.004 ± 0.005 h-1) and of tracer time delay (1.19 ± 0.01). The proposed model performs very well, since it provides a readily solved analytical solution with respect to the conventional MIM. Results of the proposed MIM are very similar to those provided by the conventional MIM. The new model solution can be implemented in particle tracking or random walk software in order to solve two-dimensional nonequilibrium solute transport in groundwater.