Cryptosporidium spp. and Giardia duodenalis in two areas of Galicia (NW Spain)

The aim of the present study was to investigate the environmental dispersal of Cryptosporidium spp. and Giardia duodenalis in two distinct areas (coastal and inland) in Galicia (NW Spain). Faecal samples were collected from healthy asymptomatic domestic (cows and sheep) and wild animals (deer and wild boars) in the selected areas. In each of the selected areas, samples of untreated water (influent) and of treated water (final effluent) were collected from each of the 12 drinking water treatments plants (DWTPs) and 12 wastewater treatment plants (WTPs) under study. Analysis of a single sample from each of the 635 (coastal) and 851 (inland) domestic and wild animals selected at random revealed that the prevalences of cryptosporidiosis and giardiosis in coastal area were 9.2% and 15.9% respectively, and in inland area, 13.7% and 26.7% respectively. In the coastal area, Cryptosporidium spp. oocysts were detected in influent and effluent samples from 2/12 (16.6%) DWTPs and 8/12 (66.6%) WTPs, while G. duodenalis cysts were detected in influent and effluent samples from 3/12 (25.0%) DWTPs and 12/12 (100%) WTPs. The concentrations were notably higher in WTPs; the mean parasite concentrations in the final treated effluent were 10 oocysts per litre and 137.8 cysts per litre for Cryptosporidium and Giardia, respectively. The mean concentration of G. duodenalis cysts per litre was significantly higher (P < 0.05) than the mean concentration of Cryptosporidium spp. oocysts per litre in both the influent and the effluent samples from all the treatment plants. In the coastal area, C. parvum, C. hominis and G. duodenalis assemblages A (I and II) and E were most repeatedly detected. In the inland area, C. parvum, C. andersoni and G. duodenalis assemblages A (I and II), B and E were most frequently identified.     

Detection and genotyping of Giardia duodenalis in wastewater: relation between assemblages and faecal contamination origin

Among the seven assemblages identified in Giardia duodenalis species, only assemblages A and B infect humans and numerous other mammals as well. On the other hand, assemblage E is considered to be host restricted to livestock. The aim of the present study was to compare the presence of G. duodenalis assemblages A, B and E in wastewater samples from two municipal treatment plants (n=24) and one slaughterhouse (n=12). Thus, PCR assays targeting the tpi gene were developed to detect specifically these three G. duodenalis assemblages. Assemblages A and B were detected in urban wastewater with a predominance of assemblage A, especially for one treatment plant. Concerning slaughterhouse wastewater, assemblage A was found in 58% of the samples, whereas assemblage B was not detected. Assemblage E was not detected in urban wastewater, but was found in 92% of the samples from slaughterhouse. Thus, combination of assemblages A and B seemed to indicate a human contamination origin, while combination of assemblages A and E appeared to correspond to a livestock contamination origin.     

Detection of Cryptosporidium spp. and Giardia duodenalis in surface water: A health risk for humans and animals

The objective of the present study was to determine the degree of contamination by Cryptosporidium spp. and Giardia duodenalis in a river basin in a livestock farming area in Galicia (NW, Spain). Water samples (50 l) were collected at 22 points in the main basin (including 5 recreational areas), and at the source and mouth of the 3 most important rivers and at the mouth of a smaller, secondary river. Faecal samples were collected from dairy cattle selected at random from 18 herds farmed in the area. A total of 139 neonatal calves, 480 heifers and 697 cows were sampled. The prevalence, intensity of infection and the risk associated with the spread of infection by both enteropathogens were determined. Water and faecal samples were collected in spring, summer, autumn and winter of 2007. The species and genotypes of these parasites present in the water samples were identified. In both water and faecal samples, more parasitic stages were collected in spring and summer than in autumn and winter. In spring, Cryptosporidium spp. oocysts were detected in 33 (9.4%) cows from 13 (72.2%) herds, and G. duodenalis cysts were detected in 56 (16.0%) cows from 15 farms (83.3%); the intensity of infection ranged from 5 to 7895 G. duodenalis cysts per gram of faeces. Infective stages of Cryptosporidium spp. and G. duodenalis were also detected in respectively 26 (89.6%) and 27 (93.1%) water samples, in spring. The mean concentrations of parasites ranged from 2 to 1200 Cryptosporidium spp. oocysts per litre and from 2 to 400 G. duodenalis cysts per litre. Cryptosporidium parvum, C. andersoni, C. hominis and assemblages A-I, A-II, E of G. duodenalis were detected. The presence of both protozoans must be monitored in cattle, in sources of water used for recreational purposes and in artificial waterways used by farmers (water channels, animal drinking water and drainage systems).     

Monitoring of Cryptosporidium and Giardia river contamination in Paris area

This study evaluates the protozoan contamination of river waters, which are used for drinking water in Paris and its surrounding area (about 615,000 m(3) per day in total, including 300,000 m(3) for Paris area). Twenty litre samples of Seine and Marne Rivers were collected over 30 months and analyzed for Cryptosporidium oocysts and Giardia cysts detection according to standard national or international methods. Cryptosporidium oocysts and Giardia cysts were found, respectively, in 45.7% and 93.8% of a total of 162 river samples, with occasional high concentration peaks. A significant seasonal pattern was observed, with positive samples for Cryptosporidium more frequent in autumn than spring, summer and winter, and positive samples for Giardia less frequent in summer. Counts of enterococci and rainfalls were significantly associated with Giardia concentration but not Cryptosporidium. Other faecal bacteria were not correlated with monitored protozoan. Marne seems to contribute mainly to the parasitic contamination observed in Seine. Based on seasonal pattern and rainfall correlation, we hypothesize that the origin of contamination is agricultural practices and possible dysfunction of sewage treatment plants during periods of heavy rainfalls. High concentrations of protozoa found at the entry of drinking water plants justify the use of efficient water treatment methods. Treatment performances must be regularly monitored to ensure efficient disinfection according to the French regulations.     

Impact of bathers on levels of Cryptosporidium parvum oocysts and Giardia lamblia cysts in recreational beach waters

Recreational beach water samples collected on weekends and weekdays during 11 consecutive summer weeks were tested for potentially viable Cryptosporidium parvum oocysts and Giardia lamblia cysts using the multiplexed fluorescence in situ hybridization (FISH) method. The levels of oocysts and cysts on weekends were significantly higher than on the weekdays (P<0.01). Concentrations of oocysts in weekend samples (n=27) ranged from 2 to 42 oocysts/L (mean: 13.7 oocysts/L), and cyst concentration ranged from 0 to 33 cysts/L (mean: 9.1 cysts/L). For the samples collected on weekdays (n=33), the highest oocyst concentration was 7 oocysts/L (mean: 1.5 oocysts/L), and the highest cyst concentration was 4 cysts/L (mean: 0.6 cysts/L). The values of water turbidity were significantly higher on weekends than on weekdays, and were correlated with the number of bathers and concentration of C. parvum oocysts and G. lamblia cysts (P<0.04). The study demonstrated positive relationships between number of bathers and levels of waterborne C. parvum oocysts and G. lamblia cysts in recreational beach water. It is essential to test recreational waters for Cryptosporidium and Giardia when numbers of bathers are greatest, or limit the number of bathers in a recreational beach area.     

Applying fluorescence based technology to the recovery and isolation of Cryptosporidium and Giardia from industrial wastewater streams

As increasing water shortages continue, water re-use is posing new challenges with treated wastewater becoming a significant source of non-potable water. Rapid detection strategies that target waterborne pathogens of concern to industry are gaining importance in the assessment of water quality. This study reports on the ability to recover spiked Cryptosporidium and Giardia from a variety of industrial wastewater streams of varied water quality. Incorporation of an internal quality control used commonly in finished water-enabled quantitative assessments of pathogen loads and we describe successful analysis of pre- and part-treated wastewater samples from four industrial sites. The method used combined calcium carbonate flocculation followed by flow cytometry and epifluorescence microscopy. Our focus will now aim at characterising the ambient parasites isolated from industrial wastewater with the objective of developing a suite of highly specific platform detection technologies targeted to industrial needs.     

Contribution of treated wastewater to the microbiological quality of Seine River in Paris

Urban part of Seine River serving as drinking water supply in Paris can be heavily contaminated by Cryptosporidium spp. and Giardia duodenalis. In the absence of agricultural practice in this highly urbanized area, we investigated herein the contribution of treated wastewater to the microbiological quality of this river focusing on these two parasites. Other microorganisms such as faecal bacterial indicators, enteroviruses and oocysts of Toxoplasma gondii were assessed concurrently. Raw wastewaters were heavily contaminated by Cryptosporidium and Giardia (oo)cysts, whereas concentrations of both protozoa in treated wastewater were lower. Treated wastewater, flowed into Seine River, had a parasite concentration closed to the one found along the river, in particular at the entry of a drinking water plant (DWP). Even if faecal bacteria were reliable indicators of a reduction in parasite concentrations during the wastewater treatment, they were not correlated to protozoal contamination of wastewater and river water. Oocysts of T. gondii were not found in both raw and treated wastewater, or in Seine River. Parasitic contamination was shown to be constant in the Seine River up to 40 km upstream Paris. Altogether, these results strongly suggest that treated wastewater does not contribute to the main parasitic contamination of the Seine River usually observed in this urbanized area.     

Removal of Giardia and Cryptosporidium in drinking water treatment: a pilot-scale study

Giardia and Cryptosporidium have emerged as waterborne pathogens of concern for public health. The aim of this study is to examine both parasites in the water samples taken from three pilot-scale plant processes located in southern Taiwan, to upgrade the current facilities. Three processes include: conventional process without prechlorination (Process 1), conventional process plus ozonation and pellet softening (Process 2), and integrated membrane process (MF plus NF) followed conventional process (Process 3). The detection methods of both parasites are modified from USEPA Methods 1622 and 1623. Results indicated that coagulation, sedimentation and filtration removed the most percentage of both protozoan parasites. The pre-ozonation step can destruct both parasites, especially for Giardia cysts. The microfiltration systems can intercept Giardia cysts and Cryptosporidium oocysts completely. A significant correlation between water turbidity and Cryptosporidium oocysts was found in this study. The similar results were also found between three kinds of particles (phi=3-5,5-8 and 8-10 microm) and Cryptosporidium oocysts.     

Settling velocity of Cryptosporidium parvum and Giardia lamblia

Understanding transport behavior of Cryptosporidium parvum oocysts and Giardia lamblia cysts (together referred to as (oo)cysts) in overland flow is important for beneficial uses of receiving water bodies. Like sediment, (oo)cysts are subjected to deposition once they are present in overland flow or low flow environments like reservoirs, wetlands and sedimentation basins. The objectives of this paper are to present the theory and experiment to determine the free settling velocity (v(s)) of (oo)cysts and to compare experimental settling velocities to estimates using Stokes' law. A settling experiment was designed to quantify the v(s) of (oo)cysts in an aqueous column. C. parvum oocysts used were spherical with average diameter (+/-1SD) of 6.6+/-1.1 microm. G. lamblia cysts were oval shaped (average eccentricity = 1.48+/-0.19) with average size of 11.8 +/-1.3 microm. Average densities were 1009 kg m(-3) for C. parvum oocysts and 1013 kg m(-3) for G. lamblia cysts. Observed experimental settling velocities are 0.27 microm s(-1) and 0.67 microm s(-1) for C. parvum and G. lamblia, respectively. Estimated average settling velocities using Stokes' law were 0.36 microm s(-1) for C. parvum and 0.84 microm s(-1) for G. lamblia. R-squared values of the observations from the settling experiments with the Stokes' law estimation are 0.87 and 0.88 for G. lamblia and C. parvum, respectively. Our results suggest that Stokes' law can be used to estimate settling velocities of (oo)cysts. Qualitatively, the low settling velocities indicate that (oo)cysts will very slowly settle out of suspension.     

Detection of enteric viruses, Giardia and Cryptosporidium in two different types of drinking water treatment facilities

In this study, two types of drinking water treatment facilities (two conventional drinking water treatment plants (DWTPs) and two compact units (Cus)) were compared referring to their production capacity. Water samples were collected from three main points: (a) different water treatment steps (b) washings of sand filters and (c) distribution system at different distances from the water treatment plants. Both viruses and protozoa were concentrated from each water sample by adsorption and accumulation on the same nitrocellulose membrane filters (0.45 microm pore size). Enteroviruses were detected by plaque infectivity assay in BGM cells and HAV, HEV and Norovirus were detected by RT-PCR. Giardia and Cryptosporidium were detected by conventional staining methods and PCR. The results revealed that enterovirus load at the intake ranged between 10-15 PFU/L for the two compact units and between 4.5 and 75 PFU/L for the two conventional DWTPs. The virus load in distribution system of the first type DWTPs at 1 km from the plant was the same as that of the intake. Viruses in the other type of treatment plants CUs at 1, 5 and 7 km, were much reduced. Investigation of raw water sediments of the two DWTPs showed enterovirus counts between 12 and 17.5 PFU/L. Virus count was reduced in sand of filters after washing. Giardia cysts were equally detected by microscopy and PCR in only intake samples of EL-Hawamdia CU (33.3%) and Meet Fares DWTP (50%). Cryptosporidium oocysts were equally detected by microscopy and PCR in intake samples of Abo EL-Nomros CU (100%), EL-Hawamdia CU (66.7%) and Fowa DWTP (50%). At Meet Fares DWTP three positive intake samples for Cryptosporidium were detected by PCR, compared with only two positive samples by microscopy. Giardia cysts and Cryptosporidium oocysts were detected in raw water sediment and sand of filters before washing. Only one sample from Meet Fares DWTP sand of filters after washing was positive for both Giardia and Cryptosporidium. It can be concluded that the poor microbial quality of the water may be due to improper operational skills and management of the various water treatment plants (especially at the two high capacity treatment plants).     

Comparing the partitioning behavior of Giardia and Cryptosporidium with that of indicator organisms in stormwater runoff

Microbial association with particles can significantly affect the fate and transport characteristics of microbes in aquatic systems as particle-associated organisms will be less mobile in the environment than their free phase (i.e. unattached) counterparts. As such, similarities or dissimilarities in the partitioning behavior of indicator organisms and pathogens may have an impact on the suitability of a particular indicator to act as a surrogate for a pathogen. This research analyzed the partitioning behavior of two pathogens (Cryptosporidium, Giardia) and several common indicator organisms (fecal coliform, Escherichia coli, Enterococci, Clostridium perfringens spores, and coliphage) in natural waters under both dry and wet weather conditions. Samples were taken from several streams in two distinct sampling phases: (i) single grab samples; and (ii) intrastorm samples obtained throughout the duration of four storms. Partitioning behavior varied by microbial type, with 15-30% of bacterial indicators (fecal coliform, E. coli, and Enterococci) associated with settleable particles compared to 50% for C. perfringens spores. Both pathogens exhibited similar levels of particle association during dry weather (roughly 30%), with increased levels observed during wet weather events (Giardia to 60% and Cryptosporidium to 40%). The settling velocities of particle-associated microbes were also estimated, with those of the bacterial indicators (fecal coliform, E. coli, and Enterococci), as well as C. perfringens spores, being similar to that of the Giardia and Cryptosporidium, suggesting these organisms may exhibit similar transport behavior. With respect to intrastorm analysis, the highest microbial concentrations, in both particle-associated and free phase, occurred during the earlier stages of a storm. The total loadings of both indicators and pathogens were also estimated over the course of individual storms.     

Intensive exploitation of a karst aquifer leads to Cryptosporidium water supply contamination

Groundwater from karst aquifers is an important source of drinking water worldwide. Outbreaks of cryptosporidiosis linked to surface water and treated public water are regularly reported. Cryptosporidium oocysts are resistant to conventional drinking water disinfectants and are a major concern for the water industry. Here, we examined conditions associated with oocyst transport along a karstic hydrosystem, and the impact of intensive exploitation on Cryptosporidium oocyst contamination of the water supply. We studied a well-characterized karstic hydrosystem composed of a sinkhole, a spring and a wellbore. Thirty-six surface water and groundwater samples were analyzed for suspended particulate matter, turbidity, electrical conductivity, and Cryptosporidium and Giardia (oo)cyst concentrations. (Oo)cysts were identified and counted by means of solid-phase cytometry (ChemScan RDI^®), a highly sensitive method. Cryptosporidium oocysts were detected in 78% of both surface water and groundwater samples, while Giardia cysts were found in respectively 22% and 8% of surface water and groundwater samples. Mean Cryptosporidium oocyst concentrations were 29, 13 and 4/100 L at the sinkhole, spring and wellbore, respectively. Cryptosporidium oocysts were transported from the sinkhole to the spring and the wellbore, with respective release rates of 45% and 14%, suggesting that oocysts are subject to storage and remobilization in karst conduits. Principal components analysis showed that Cryptosporidium oocyst concentrations depended on variations in hydrological forcing factors. All water samples collected during intensive exploitation contained oocysts. Control of Cryptosporidium oocyst contamination during intensive exploitation is therefore necessary to ensure drinking water quality.     

Infectivity of Giardia lamblia cysts obtained from wastewater treated with ultraviolet light

Several waterborne outbreaks of giardiasis have been linked to discharge of wastewater effluents into surface water. Little is known about the infectivity of Giardia lamblia cysts present in UV treated wastewater effluents. In this study, the infectivity of G. lamblia cysts, recovered from primary effluent and secondary effluent, both upstream and downstream of operating full-scale UV reactors at four wastewater treatment plants, was assessed using the Mongolian gerbil model. Infectivity of cysts obtained from the primary effluents was scored as either strong or moderate for induction of infection in gerbils at three out of four wastewater treatment plants. G. lamblia recovered from secondary effluent both upstream and downstream of the UV reactors caused weak infections in the gerbils. The probability of weak infections caused by inoculums of 50-1400 cysts per gerbil was, on the average, reduced by approximately 10% at the four wastewater UV installations with coliform reduction equivalent doses ranging from 6 to 18 mJ/cm². The UV systems provided considerably less inactivation of the parasite than expected based on the UV dose response of Giardia reported in the literature.     

The prevalence and characterisation of Cryptosporidium spp. in beef abattoir water supplies

The prevalence of Cryptosporidium spp. in 50 l samples of water used to wash beef carcasses at (a) an abattoir with a borehole water (BH) supply (n = 46) and (b) an abattoir with a river water (RW) supply (n = 48) was determined. In addition, a 100 l water sample and post-wash carcass samples (n = 24) were collected from the RW supply on a single day in July. Cryptosporidium spp. was detected in 0% and 26.1% of samples from the BH and RW supply abattoirs, respectively, with oocyst concentrations ranging from 0.02 to 8.6/l. Cryptosporidium spp. was not isolated from post-wash beef carcasses, while it was detected in water samples from that day at a concentration of 0.06 oocysts/l. The species of 3/5 isolates were identified as C. parvum, and the remaining were C. andersoni. This study has demonstrated that water used to wash beef carcasses can be contaminated with Cryptosporidium of human health importance and is a potential source of carcass contamination.     

Detection of microsporidia in drinking water, wastewater and recreational rivers

Diarrhea is the main health problem caused by human-related microsporidia, and waterborne transmission is one of the main risk factors for intestinal diseases. Recent studies suggest the involvement of water in the epidemiology of human microsporidiosis. However, studies related to the presence of microsporidia in different types of waters from countries where human microsporidiosis has been described are still scarce. Thirty-eight water samples from 8 drinking water treatment plants (DWTPs), 8 wastewater treatment plants (WWTPs) and 6 recreational river areas (RRAs) from Galicia (NW Spain) have been analyzed. One hundred liters of water from DWTPs and 50 L of water from WWTPs and RRAs were filtered to recover parasites, using the IDEXX Filta-Max® system.Microsporidian spores were identified by Weber’s stain and positive samples were analyzed by PCR, using specific primers for Enterocytozoon bieneusi, Encephalitozoon intestinalis, Encephalitozoon cuniculi, and Encephalitozoon hellem. Microsporidia spores were identified by staining protocols in eight samples (21.0%): 2 from DWTPs, 5 from WWTPs, and 1 from an RRA. In the RRA sample, the microsporidia were identified as E. intestinalis.To the best of our knowledge, this is the first report of human-pathogenic microsporidia in water samples from DWTPs, WWTPs and RRAs in Spain. These observations add further evidence to support that new and appropriate control and regulations for drinking, wastewater, and recreational waters should be established to avoid health risks from this pathogen.     

Molecular and phylogenetic approaches for assessing sources of Cryptosporidium contamination in water

The high sequence diversity and heterogeneity observed within species or genotypes of Cryptosporidium requires phylogenetic approaches for the identification of novel sequences obtained from the environment. A long-term study on Cryptosporidium in the agriculturally-intensive South Nation River watershed in Ontario, Canada was undertaken, in which 60 sequence types were detected. Of these sequence types 33 were considered novel with no identical matches in GenBank. Detailed phylogenetic analysis identified that most sequences belonged to 17 previously described species: Cryptosporidium andersoni, Cryptosporidium baileyi, Cryptosporidium hominis, Cryptosporidium parvum, Cryptosporidium ubiquitum, Cryptosporidium meleagridis, muskrat I, muskrat II, deer mouse II, fox, vole, skunk, shrew, W12, W18, W19 and W25 genotypes. In addition, two new genotypes were identified, W27 and W28. C. andersoni and the muskrat II genotype were most frequently detected in the water samples. Species associated with livestock made up 39% of the total molecular detections, while wildlife associated species and genotypes accounted for 55% of the Cryptosporidium identified. The human pathogenic species C. hominis and C. parvum had an overall prevalence of 1.6% in the environment, indicating a small risk to humans from the Cryptosporidium present in the watershed. Phylogenetic analysis and knowledge of host-parasite relationships are fundamental in using Cryptosporidium as a source-tracking or human health risk assessment tool.     

Wastewater treatment contributes to selective increase of antibiotic resistance among Acinetobacter spp.

The occurrence and spread of multi-drug resistant bacteria is a pressing public health problem. The emergence of bacterial resistance to antibiotics is common in areas where antibiotics are heavily used, and antibiotic-resistant bacteria also increasingly occur in aquatic environments. The purpose of the present study was to evaluate the impact of the wastewater treatment process on the prevalence of antibiotic resistance in Acinetobacter spp. in the wastewater and its receiving water. During two different events (high-temperature, high-flow, 31 °C; and low-temperature, low-flow, 8 °C), 366 strains of Acinetobacter spp. were isolated from five different sites, three in a wastewater treatment plant (raw influent, second effluent, and final effluent) and two in the receiving body (upstream and downstream of the treated wastewater discharge point). The antibiotic susceptibility phenotypes were determined by the disc-diffusion method for 8 antibiotics, amoxicillin/clavulanic acid (AMC), chloramphenicol (CHL), ciprofloxacin (CIP), colistin (CL), gentamicin (GM), rifampin (RA), sulfisoxazole (SU), and trimethoprim (TMP). The prevalence of antibiotic resistance in Acinetobacter isolates to AMC, CHL, RA, and multi-drug (three antibiotics or more) significantly increased (p < 0.01) from the raw influent samples (AMC, 8.7%; CHL, 25.2%; RA, 63.1%; multi-drug, 33.0%) to the final effluent samples (AMC, 37.9%; CHL, 69.0%; RA, 84.5%; multi-drug, 72.4%), and was significantly higher (p < 0.05) in the downstream samples (AMC, 25.8%; CHL, 48.4%; RA, 85.5%; multi-drug, 56.5%) than in the upstream samples (AMC, 9.5%; CHL, 27.0%; RA, 65.1%; multi-drug, 28.6%). These results suggest that wastewater treatment process contributes to the selective increase of antibiotic resistant bacteria and the occurrence of multi-drug resistant bacteria in aquatic environments.     

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.     

Bioaccumulation of heavy metals by the aquatic plants Potamogeton pectinatus L. and Potamogeton malaianus Miq. and their potential use for contamination indicators and in wastewater treatment

The concentrations of heavy metals in the leaves of two aquatic plants Potamogeton pectinatus L. and Potamogeton malaianus Miq., and the corresponding water and sediment samples from the Donghe River in Jishou City of Hunan Province, China were studied to investigate metal contamination from the intensive industrial activities in the surrounding area. Results showed that the concentrations of heavy metals in the sediments, especially Cd, Mn and Pb, were much higher than the eco-toxic threshold values developed by the U. S. Environmental Protection Agency. Between the two plant species, P. pectinatus showed the higher capacity in metal accumulation. The highest concentrations of Cd, Pb, Cu, Zn and Mn were found in the leaves of P. pectinatus, reaching 596, 318, 62.4, 6590 and 16,000 mg kg(-1) (DW), respectively. Significantly positive relationships were observed among the concentrations of Zn, Cu and Mn in the leaves of both aquatic plants and those in water, indicating the potential use of the two plants for pollution monitoring of these metals. In addition, a laboratory experiment was conducted to investigate the ability of P. pectinatus and P. malaianus to remove heavy metals from contaminated river water. The average removal efficiencies by P. pectinatus and P. malaianus for Cd, Pb, Mn, Zn and Cu from the spiked Donghe River water were 92%, 79%, 86%, 67% and 70%, respectively. The results indicated that P. pectinatus and P. malaianus had high capabilities to remove heavy metals directly from the contaminated water. The potential use of these plants in wastewater treatment is worth further exploration.     

Development of a Ct equation for the inactivation of Cryptosporidium oocysts with chlorine dioxide

Cryptosporidium parvum, a protozoan parasite, has been implicated in a number of waterborne disease outbreaks and is difficult to inactivate using free chlorine. It appears, however, to be inactivated more easily by other oxidants such as chlorine dioxide or ozone. A major element of the EPA (US EPA) strategy for controlling C. parvum (oocysts) in drinking water is the possible use of Ct (concentration of disinfectant in mg/L times time in minutes) values. To support this strategy a Ct equation, based on first-order kinetics, is proposed to provide guidance to drinking water utilities for the application of chlorine dioxide for controlling C. parvum oocysts. The equation is based on standard statistical techniques using available bench scale data. It can predict mean inactivation levels as well as a statistically conservative upper bound Ct value. This upper bound could be used to insure an appropriate safety factor for the protection of public health.